具有降低的吸附作用和增加的水解作用的熱酶變異體的制作方法

專利名稱：具有降低的吸附作用和增加的水解作用的熱酶變異體的制作方法
技術領域：
本發明涉及新的用于各種洗滌組合物的酶變異體和編碼這種酶變異體的DNA序列。
背景技術：
酶構成了最大種類的天然產生的蛋白質。每一類酶通常催化(加速反應而不被消耗)不同種類的化學反應。稱為蛋白酶的一種酶已知能夠水解(分解化合物成兩個或更多個更簡單的化合物，在斷裂的化學鍵的兩側接受水分子的H和OH部分)其它蛋白質。將天然產生的和蛋白質工程蛋白酶作為添加劑加入洗衣洗滌劑制劑中即利用了水解蛋白質的這種能力。在衣服上的許多污垢是蛋白質的，廣譜的蛋白酶可顯著改善這種污垢的除去。
不幸的是，這些蛋白質在其天然的，細菌的環境中的功效水平通常不能轉移至相對非天然的洗滌環境。尤其是蛋白酶特性，例如熱穩定性、pH穩定性、氧化穩定性和底物特性對于酶的天然環境之外的應用未必達到最佳。
蛋白酶的氨基酸序列決定了蛋白酶的特征。根據氨基酸序列中改變的位點、性質和/或幅度，改變蛋白酶的氨基酸序列可改變酶的特性至各不相同的程度，甚至可失活酶。人們已做了許多嘗試以改變蛋白酶的野生型氨基酸序列，以改善它們的性質，從而增加蛋白酶在洗滌環境中的效率。這些嘗試包括改變氨基酸序列以便在相當不同的條件下提高熱穩定性和改善氧化穩定性。
盡管在現有技術中描述了各種各樣的嘗試，但人們仍不斷需要用于洗滌各種表面的蛋白酶的新有效變異體。
發明目的本發明的目的在于提供與野生型酶相比較具有改善的水解作用的熱酶(Thermitase)變異體。
本發明的另一個目的在于提供含有這些枯草桿菌蛋白酶變異體的洗滌組合物。
發明概述本發明涉及具有野生型熱酶氨基酸序列的修飾氨基酸序列的熱酶變異體，野生型氨基酸序列包括第一環區域、第二環區域、第三環區域、第四環區域和第五環區域；其中，修飾的氨基酸序列在一個或多個環區域中的特定標記的位點上含有與野生型熱酶不同的氨基酸(即取代)，與野生型熱酶相比，熱酶變異體具有對不溶解底物降低的吸附作用和增加的水解作用。本發明還涉及編碼這種熱酶變異體的DNA序列。本發明還涉及含有這種熱酶變異體的用于洗滌各種表面的組合物。
發明詳述I.熱酶變異體本發明涉及枯草桿菌蛋白酶類，尤其是熱酶，它們通過編碼酶的各種核苷酸順序的突變，從而改變酶的氨基酸序列而被修飾。本發明的修飾枯草桿菌蛋白酶(下文稱為“熱酶變異體”)與野生型熱酶相比，具有對不溶解底物降低的吸附作用和增加的水解作用。本發明還涉及編碼這種熱酶變異體的DNA序列。
本發明的枯草桿菌蛋白酶屬于稱為蛋白酶類。蛋白酶是斷裂肽鍵的催化劑。一種類型的蛋白酶是絲氨酸蛋白酶。絲氨酸的特征在于在活性位點上存在必需的絲氨酸殘基。
隨著酶濃度的增加酶對可溶解底物的水解速率增加的現象已經作了充分報導。因而，對于表面結合的底物，例如在許多洗滌應用中遇到的情況，水解速率隨著表面濃度的增加而增加似乎是可能的，這已由事實說明。(Brode，P.F.III和D.S.Rauch，LANGMUIR，“枯草桿菌蛋白酶BPN’“固定化底物上的活性”，卷8，1325-1329頁(1992))。事實上，當酶的表面濃度變化時，對于不溶解底物已經發現了表面濃度與速率的直線關系。(Rubingh，D.N.和M.D.Bauer.“在蛋白質溶液界面上的蛋白酶的催化水解”多聚物溶液混和與界面(，POLYMER SOLUTIONS，BLENDSAND INTERFACES)，I.Noda和D.N.Rubingh，Elsevier，編著第464頁(1992))。出乎意料的是，當試圖將這種原理用于研究得到較好洗滌性能的變異體蛋白酶時，我們不能發現吸附更多的酶得到較好的性能。事實上，我們驚奇地發現與之相反的事實酶與底物的下降的吸附導致增加的底物水解(即，較好的洗滌性能)。
盡管不希望受任何理論束縛，但我們相信，當將變異體相互比較時，改善的性能是如下事實的結果，即吸附較少的酶也不太緊密地結合，因而在欲除去不溶解蛋白質底物的表面更易移動。在可比的酶溶液濃度下，增加的移動能力足以勝過通過向表面提供較高濃度酶而給予的任何優點。
本發明描述的突變用于改變(即降低)酶與表面結合的污垢的吸附。在熱酶中，某些氨基酸在酶分子上形成外環。為了討論，這些環可稱為第一、第二、第三、第四和第五環區域。具體地說，位點66-73形成第一環區域；位點103-115形成第二環區域；位點134-141形成第三環區域；位點162-171形成第四環區域；位點191-195形成第五環區域；和位點204-224形成第六環區域(位點編號類似于野生型枯草桿菌蛋白酶熱酶(SEQ ID NO1)的氨基酸序列)。
我們相信，這些環區域在酶分子與表面結合的肽的吸附中起著重要的作用，在一個或多個環區域中具體的突變對吸附具有明顯的效果。盡管不希望受任何理論束縛，但我們相信出于至少兩個原因，這些環區域對熱酶分子的吸附是重要的。首先，含有環區域的氨基酸能夠密切地與分子暴露的任何表面接觸。其次，熱酶分子的環區域與活性位點的接近和結合囊使得它們在酶與表面結合的底物(肽/蛋白質污垢)的催化產生的吸附中起作用。
本文使用的術語“變異體”是指含有不同于野生型氨基酸序列的酶。
本文使用的術語“突變熱酶DNA”指的是編碼熱酶變異體的DNA序列。
本文使用的術語“野生型熱酶”指的是由SEQ ID NO1表示的酶。該熱酶的氨基酸序列在Meloun，B.，Baudys，M，Kostka，V.，Hausdorf，G，Frommel，C.，和Hohne，W.E.，FEBA LETT.，卷183，第195-200頁(列為參考文獻)中進一步描述。
本文使用的術語“熱酶野生型氨基酸序列”包括SEQ ID NO1以及除位點66、67、68、69、70、72、73、103、104、105、106、107、108、109、110、111、112、113、114、115、134、135、136、137、138、139、140、141、162、163、164、165、166、167、168、169、170、171、191、192、193、194、195、204、205、206、207、208、209、210、211、212、213、214、215、216、217、218、219、220、221、222、223或224之外對氨基酸序列修飾的SEQ ID NO1。
本文使用的術語“更親水的氨基酸”指的是參考下表中的親水性值比目標氨基酸具有更大親水性值的任何其它氨基酸。如下親水性值表(表1)以增加的親水性值的下降順序列出氨基酸(參見Hopp，T.P.，和Woods.K.R.，“由氨基酸序列判斷蛋白質抗原決定簇”，美國國家科學院院刊，卷78，3824-3828頁，1981，列為參考文獻)。
表1氨基酸 親水性值Trp-3.4Phe-2.5Tyr-2.3Leu，Ile -1.8Val-1.5Met-1.3Cys-1.0Ala，His -0.5Thr-0.4Pro，Gly -0.0Gln，Asn 0.2Ser0.3Arg+，Lys+，Glu-，Asp-3.0表l還說明了帶有電荷(該特征基于約8-9的pH)。正電荷的氨基酸是Arg和Lys，負電荷氨基酸是Glu和Asp，其余的氨基酸是中性的。在本發明的優選實施方案中，取代氨基酸是中性的或負電荷的，更優選負電荷(即Glu或Asp)。
因而，例如說明“用相等或更親水的中性或帶有負電荷的氨基酸取代的Gln”指的是用Asn(它與Gln的親水性相等)或Ser、Glu或Asp(它們比Gln更親水)取代的Gln。它們每個均是中性或具有負電荷，與Gln相比較，它們具有較大的親水性值。同樣，說明“用更親水的中性或帶有負電荷的氨基酸取代的Pro”指的是用Gln、Asn、Ser、Glu或Asp取代的Pro。
在本發明的一個實施方案中，熱酶變異體具有熱酶野生型氨基酸序列的修飾氨基酸序列，其中野生型氨基酸序列含有在第一環區域、第二環區域、第三環區域、第四環區域、第五環區域或第六環區域的一個或多個中的一個或多個位點上的取代基；從而熱酶變異體與野生型熱酶相比較，對不溶解底物具有降低的吸附作用和增加的水解作用。
在本發明的優選實施方案中，在一個或多個環區域中一個或多個位點取代氨基酸，參考表1，是中性的或負電荷的，與在野生型氨基酸序列中目標位點的氨基酸相比較，是相等或更親水性的，優選是更親水性的。A.第一環區域中的取代當取代發生在第一環區域中時，取代發生在位點66、67、68、69、70、72或73的一個或多個上。
當取代發生在位點66時，取代氨基酸是Asn、Asp、Glu或Ser。
當取代發生在位點67時，取代氨基酸是Asn、Asp、Glu或Ser。
當取代發生在位點68時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser。
當取代發生在位點69時，取代氨基酸是Asp、Gln、Glu或Ser。
當取代發生在位點70時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser。
當取代發生在位點72時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser。
當取代發生在位點73時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、Pro或Ser。B.第二環區域的取代當取代發生在第二環區域中時，取代發生在位點103、104、105、106、107、108、109、110、111、112、113、114或115的一個或多個上。
當取代發生在位點103時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。
當取代發生在位點104時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Met、Pro、Ser、Thr或Val。
當取代發生在位點105時，取代氨基酸是Glu。
當取代發生在位點106時，取代氨基酸是Asp、Gln、Glu或Ser。
當取代發生在位點107時，取代氨基酸是Asp或Glu。
當取代發生在位點108時，取代氧基酸是Asn、Asp、Gln、Glu、Pro或Ser。
當取代發生在位點109時，取代氨基酸是Asp或Glu。
當取代發生在位點110時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser。
當取代發生在位點111時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、Pro或Ser。
當取代發生在位點112時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Phe、Pro、Ser、Thr、Tyr或Val。
當取代發生在位點113時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、Pro或Ser。
當取代發生在位點114時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
當取代發生在位點115時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。C.第三環區域中的取代當取代發生在第三環區域中時，取代發生位點134、135、136、137、138、139、140或141的一個或多個上。其中當取代發生在位點134時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Met、Pro、Ser、Thr或Val。
當取代發生在位點135時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser。
當取代發生在位點136時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser。
當取代發生在位點137時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、Pro或Ser。
當取代發生在位點138時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr。
當取代發生在位點139時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser。
當取代發生在位點140時，取代氨基酸是Asp、Gln、Glu或Ser。和當取代發生在位點141時，取代氨基酸是Asp或Glu。D.第四環區域中的取代當取代發生在第四環區域中時，取代發生在位點162、163、164、165、166、167、168、169、170或171的一個或多個上。
當取代發生在位點162時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser。
當取代發生在位點163時，取代氨基酸是Asp、Gln、Glu或Ser。
當取代發生在位點164時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
當取代發生在位點165時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser。
當取代發生在位點166時，取代氨基酸是Asp、Gln、Glu或Ser。
當取代發生在位點167時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、Pro或Ser。
當取代發生在位點168時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
當取代發生在位點169時，取代氨基酸是Asn、Asp、Gln、Glu、Gly或Ser。
當取代發生在位點170時，取代氨基酸是Asp、Gln、Glu或Ser；和當取代發生在位點171時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro、Ser、Thr或Val。E.第五環區域中的取代當取代發生在第五環區域中時，取代發生在位點191、192、193、194或195一個或多個上。
當取代發生在位點191時，取代氨基酸是Asp或Glu。
當取代發生在位點192時，取代氨基酸是Asp或Glu。
當取代發生在位點193時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro、Ser、Thr、Tyr或Val。
當取代發生在位點194時，取代氨基酸是Asp或Glu。
當取代發生在位點195時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、Pro或Ser。F第六環區域中的取代當取代發生在第六環區域中時，取代發生在位點204、205、206、207、208、209、210、211、212、213、214、215、216、217、218、219、220、221、222、223或224的一個或多個上。
當取代發生在位點204時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
當取代發生在位點205時，取代氨基酸是Asn、Asp、Gln、Glu、Gly或Ser。
當取代發生在位點206時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser。
當取代發生在位點207時，取代氨基酸是Asp或Glu。
當取代發生在位點208時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Phe、Pro、Ser、Thr、Tyr或Val。
當取代發生在位點209時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Leu、Met、Pro、Ser、Thr或Val。
當取代發生在位點210時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro、Ser、Thr或Val。
當取代發生在位點211時，取代氨基酸是Asp或Glu。
當取代發生在位點212時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、Pro或Ser。
當取代發生在位點213時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Phe、Pro、Ser、Thr或Val。
當取代發生在位點214時，取代氨基酸是Asn、Asp、Gln、Glu、Gly或Ser。
當取代發生在位點215時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、Pro或Ser。
當取代發生在位點216時，取代氨基酸是Asp或Glu。
當取代發生在位點217時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、Pro或Ser。
當取代發生在位點218時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Leu、Met、Pro、Ser、Thr或Val。
當取代發生在位點219時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr。
當取代發生在位點220時，取代氨基酸是Asp或Glu。
當取代發生在位點221時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、 Gly、His、Ile、Met、Pro、Ser、Thr或Val。
當取代發生在位點222時，取代氨基酸是Asp或Glu。
當取代發生在位點223時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser。
當取代發生在位點224時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、Pro或Ser。G.酶變異體的制備實施例1突變體熱酶DNA構建了含有野生型熱酶基因的噬菌粒(“TP”)。將質粒pUC119的2.8Kbp Pvu II限制酶片段(Vieira，J.和Messing，J，“單鏈的質粒DNA的制備”，153酶學方法3-11(1989))克隆至質粒pUB110(Bacillus GeneticStock Center，Columbus，OH1E9)的PvuII位點。pUC 119-pUB110雜交質粒被稱為pJMA601。在pJMA601的Bam H1限制位點克隆入具有TP的聚合酶鏈反應擴增的熱酶基因。噬菌粒TP被轉化入大腸桿菌Ung-菌株CJ236，用VCSM13輔助噬菌體(Kunkel，T.A，J.D.Roberts和R.A.Zakour，“無需表型選擇的快速和有效的定點突變”，酶學方法，卷154，367-382頁(1987)；由Yuckenberg，P.D.，F.Witney，J.Geisselsoder和J.McClary改進，“使用含尿嘧啶的DNA和噬菌粒載體的體外定點突變”，定點突變 操作指南M.J.McPherson編，第27-48頁(1991)；均列為本文參考文獻)制備單鏈的含尿嘧啶DNA模板。使用Zoller和Smith方法的單引物定點誘變修飾方法(Zoller，M.J.和M Smith，“使用M13衍生的載體的寡核苷酸定向誘變一種在DNA任何片段中制備點突變的有效和常用方法”，核酯研究，卷10，6487-6500頁(1982)，列為本文參考文獻)制備所有突變體(基本上如上述Yuckenberg等人，1991中說明)。寡核苷酸使用Applied Biosystem Inc.380B DNA合成儀制備。突變反應產物轉移至大腸桿菌菌株MM294(美國典型培養物保藏中心大腸桿菌33625)中。所有突變體由DNA測序證實，分離的DNA轉化入至枯草芽孢桿菌表達菌株BG2036(Yang，M.Y.，E.Ferrari和D.J.Henner，(1984)，“枯草芽孢桿菌的中性蛋白酶基因的克隆和克隆的基因用于產生體外得到的缺失突變”，細菌學雜志，卷160，15-21頁)。對于某些突變體，用在相應環中移碼終止的密碼子突變的修飾TP制備尿嘧啶模板。寡核苷酸用來恢復合適的閱讀框和編碼在位點66、67、68、69、70、72、73、103、104、105、106、107、108、109、110、111、112、113、114，115、134、135、136、137、138、139、140、141、162、163、164、165、166、167、168、169、170、171、191、192、193、194、195、204、205、206、207、208、209、210、211、212、213、214、215、216、217、218、219、220、221、222、223或224的隨機取代(在這些密碼子中對于所有三種堿基所有四種寡核苷酸的等摩爾和/或可變混合物)。校正移碼終止和制備功能酶的突變由它們消化酪蛋白的能力說明。隨機取代用DNA測序確定。
實施例2發酵將含有感興趣的枯草芽孢桿菌突變體的枯草芽孢桿菌細胞(BG2036)在1升LB-葡萄糖肉湯培養基中的培養至對照中期，在總體積10升的Biostat ED發酵罐(B.Braun Biotech，Inc.，Allentown，Pennsylvania)中培養。發酵培養基含有酵母提取物、淀粉、消泡劑、緩沖劑和痕量礦物質(參見發酵，操作指南，B.McNeil和L.M.Harvey編，1990)。在發酵試驗過程中，肉湯保持恒定pH7.0。加入氯霉素以進行誘變質粒的抗生素選擇。細胞在37℃生長過夜至A600約60，并收獲。
實施例3提純將發酵肉湯進行如下步驟以得到純酶。通過離心除去肉湯中的枯草芽孢桿菌細胞，通過用100K截斷膜除去細的粒子而澄清。然后用10K截斷膜上濃縮，進行流體透析以降低離子強度，用0.025M MES緩沖劑(2-(N-嗎啉代)乙磺酸)調節pH至5.5。通過將酶加入陽離子交換層析柱或親合吸附層析柱，由柱中用氯化鈉或丙二醇梯度洗脫而進一步純化(參見，Scopes，R.K.，蛋白質純化理論和實踐，Springer-Verlag，NewYork(1984)，列為本文參考文獻)。
使用pNA試驗(DelMar，E.G.，C.Largman，J.W.Brodrick和M.C.Geokas，生物化學年鑒，卷99，316-320頁(1979)，列為本文參考文獻)測定在梯度洗脫過程中收集的級分的活性酶濃度。該試驗測量當酶水解溶解的合成底物，琥珀酰-丙氨酰-丙氨酰-脯氨酰-苯基丙氨酰-對-硝基苯胺(sAAPF-pNA)時釋放對-硝基苯胺的速率。用分光光度計在410nm測量水解反應產生黃色的速率，與活性酶濃度成比例。此外，在280nm的吸附測量用來測定總蛋白質濃度。活性酶/總蛋白質比率得到酶的純度，并用于鑒定用作存貯溶液的收集的級分。
為避免在貯存過程中酶的自溶，在由層析柱得到的收集的級分中加入相等重量的丙二醇。在提純過程完成后，存貯酶溶液用SDS-PAGE(十二烷基磺酸鈉聚丙烯酰胺凝膠電泳)校驗，用胰蛋白酶抑制劑型II-T由Sigma Chemical Company(St.Louis，Missouri)購買的火雞卵白經活性位滴定法測定絕對酶濃度。測量的轉化系數將說明在酶分子中不同位點發生的改變使得酶變異體與野生型相比較對溶解底物pNA具有增加的活性。
在使用的制劑中，酶存貯溶液通過Sephadex-G25(Pharmacia，Piscataway，New Jersey)大小排除柱以除去丙二醇和交換緩沖劑。用含有0.01M氯化鈣的0.1M Tris緩沖劑(三(羥基甲基-氨基)甲烷)代替在酶存貯溶液中的MES緩沖劑，用鹽酸調節pH至8.6。在恒溫25℃下在Tris緩沖劑中在pH8.6下進行所有試驗。H.酶變異體的特性實施例4模型表面制備由CPG Inc.(Fairfield，New Jersey)購買的氨基丙基控制的孔玻璃(CPG)用作共價連接由Bachem，Inc.(Torence，California)購買的sAAPD-pNA底物的載體。反應在二甲基亞砜中進行，用(1-乙基-3-[3-(二甲基氨基)丙基]碳化二亞胺鹽酸鹽)(EDC)作偶聯劑。在反應完成后(用pNA試驗監測)，除去過量的溶劑，用二甲基亞砜(DMSO)和雙蒸餾水漂洗CPGsAAPF-pNA。然后在烘箱中氮氣吹掃在約70℃下干燥。反應方案和固定的底物的制備方法如Brode，P.F.III，和D.S.Rauch，“枯草桿菌蛋白酶BPN’在固定底物上的活性”，LANGMUIR，卷8，1325-1329頁(1992)(列為本文參考文獻)中描述進行。
CPG表面含有62000±7000pNA分子/μm2。從CPG Inc.報導的CPG的50.0m2/g的值來看，表面積保持未變。這說明用于在CPG中加入sAAPf-pNA的方法未破壞孔結構(平均直徑486A)。
實施例5表面水解試驗使用CPGsAAPf-pNA，酶變異體的吸附和CPG-結合的肽的水解可在同一試驗中測量。在含有Tris緩沖劑和已脫氣的CPGsAAPF-pNA的燒瓶中加入少量的酶變異體存貯溶液。燒瓶在活塞作用的振蕩器中振蕩90分鐘，在此期間，以不同的時間間隔停止振蕩器(例如，在吸附水解作用的最初階段，例如最初20分鐘為每2分鐘-在實驗結束時為每10分鐘)。使CPGsAAPF-pNA靜置，取樣溶液。如上述Brode等人，1991中所述進行吸附和水解的實驗過程和計算。
測定所有的酶的自溶穩定性，在實驗的時間內，應未顯示明顯的自溶損失。因此，酶吸附可通過測量溶液消耗測定。最初酶變異體濃度與在每個間隔的時間點測量的濃度之間的差值得到吸附的酶變異體的量。由表面水解的pNA的量由在410nm時試樣的吸附讀數測定。水解的pNA的總量由取樣的量加上殘余在燒瓶中的量計算。該值通過減去當不存在酶時在pH8.6下由Tris緩沖劑水解的pNA的量而校準。根據酶的效率，基準水解范圍為總水解的7-29％。
實施例6可溶解底物的動力學分析可溶解底物sAAPF-pNA的水解速率通過用DU-70分光光度計在410nm下測量作為時間函數的吸附增量而監測。酶濃度保持恒定，并制備成6-10納摩爾的范圍，而對于每次動力學測定，底物濃度在90-700μMsAAPF-pNA之間變化。在900秒的時間內，每秒收集吸附數據點，數據傳遞至LOTUSTM記錄紙(Lotus Development Corporation，Cambridge，Massachu setts)。用標準Linewweaver Burk分析法進行動力學參數的分析，其中在試驗最初(通常為最初幾分鐘)的數據符合于線性回歸曲線以得到vO。以標準倒數方式標繪的vO和sO數據得到KM和Kcat。I.熱酶變異體的實例降低表面結合的底物的吸附作用和增加的水解作用的本發明熱酶變異體在如下表2-36中說明。在描述的具體突變中，首先給出野生型中出現的原始氨基酸，其次是位點編號，第三是取代氨基酸。
表2環1-單突變變異體Gln66AsnGln66AspGln66GluGln66SerAsn67AspAsn67GlnAsn67GluAsn67SerGly68AsnGly68AspGly68GlnGly68GluGly68ProGly68SerAsn69AspAsn69GlnAsn69GluAsn69SerGly70AsnGly70AspGly70GlnGly70GluGly70ProGly70SerGly72AsnGly72AspGly72GlnGly72GluGly72ProGly72SerThr73AsnThr73AspThr73GlnThr73GluThr73GlyThr73ProThr73Ser
表3環1-雙突變變異體Asn68Asp + Gly72ProGly69Asn + Thr73ProAsn66Glu + Gly69SerAsn66Glu + Gly67SerGly69Gln + Thr73ProGly67Ser + Gly72SerGly67Asp + Thr73GlyGly69Pro + Ser70AspGly67Pro + Gly69ProAsn66Glu + Thr73GlyGly69Gln + Gly72AspAsn66Gln + Thr73GlyAsn68Asp + Thr73SerAsn66Gln + Gly69AsnGly67Glu + Thr73SerGly72Glu + Thr73ProAsn66Glu + Thr73ProGly72Asp + Thr73GlnAsn68Ser + Gly69GluGly67Pro + Gly69SerGly67Pro + Gly72AspGly69Ser + Gly72GluSer70Asp + Thr73GlnGly67Ser + Asn68GluGly69Ser + Thr73SerAsn68Asp + Gly69SerAsn66Ser + Gly72AspGly69Asp + Gly72SerAsn68Ser + Thr73ProSer70Asp + Thr73SerSer70Glu + Gly72AsnGly69Asp + Thr73ProGly67Glu + Thr73GlyGly72Asp + Thr73SerSer70Asp + Thr73AsnAsn66Ser + Gly69GluGly72Ser + Thr73GlnSer70Glu + Thr73SerAsn66Gln + Gly69Ser表4環1-三突變變異體Ser70Asp + Gly72Gln + Thr73GlyGly67Ser + Asn68Gln + Ser70AspAsn66Ser + Gly67Asn + Thr73GlnAsn66Gln + Ser70Asp + Gly72AsnSer70Glu + Gly72Gln + Thr73SerGly67Pro + Asn68Glu + Thr73SerAsn66Ser + Gly69Gln + Gly72GlnAsn66Ser + Ser70Asp + Thr73GlnGly67Gln + Gly69Asp + Thr73GlnGly69Pro + Gly72Glu + Thr73SerGly67Gln + Gly69Asn + Thr73AsnAsn66Ser + Gly69Asp + Thr73AsnGly69Asp + Gly72Ser + Thr73SerAsn68Asp + Gly72Gln + Thr73SerAsn66Gln + Asn68Ser + Thr73AsnAsn68Ser + Gly69Ser + Ser70AspAsn68Gln + Gly69Ser + Thr73GlnGly67Ser + Gly69Pro + Gly72GluGly67Asn + Gly69Ser + Thr73AsnGly67Gln + Gly72Asn + Thr73GlnGly67Pro + Asn68Gln + Gly69ProGly69Asn + Ser70Glu + Thr73SerAsn66Ser + Ser70Glu + Gly72AsnGly69Gln + Ser70Asp + Gly72AsnGly67Ser + Ser70Glu + Thr73GlnAsn66Glu + Gly67Gln + Thr73ProGly67Ser + Gly72Glu + Thr73SerGly67Pro + Gly69Gln + Gly72AspAsn66Ser + Gly67Glu + Thr73ProAsn66Ser + Gly67Glu + Asn68SerGly67Asn + Asn68Glu + Gly69AspAsn68Glu + Gly69Glu + Gly72ProAsn68Glu + Gly69Glu + Thr73AsnGly67Glu + Asn68Asp + Gly69GlnGly67Glu + Asn68Asp + Thr73GlnAsn66Asp + Gly67Asp + Gly72SerAsn66Glu + Gly67Glu + Thr73Pro表5環1-四突變變異體Asn66Gln + Gly67Ser + Gly69Asp + Gly72ProAsn68Ser + Ser70Glu + Gly72Gln + Thr73ProAsn66Gln + Gly67Gln + Ser70Asp + Thr73AsnGly67Ser + Gly69Asp + Gly72Gln + Thr73GlyAsn66Glu + Gly67Ser + Asn68Ser + Thr73AsnGly67Ser + Asn68Asp + Gly72Asn + Thr73GlyAsn66Ser + Gly67Glu + Gly69Gln + Thr73GlyAsn68Glu + Gly69Asp + Gly72Asn + Thr73AsnGly67Glu + Asn68Glu + Gly72Asn + Thr73AsnAsn66Glu + Gly67Glu + Asn68Gln + Gly72ProAsn66Glu + Gly67Glu + Asn68Ser + Gly72AsnAsn66Glu + Gly67Glu + Gly72Pro + Thr73AsnGly67Ser + Gly69Asp + Ser70Glu + Thr73GlnAsn66Ser + Gly69Glu + Ser70Glu + Thr73GlyGly67Pro + Asn68Glu + Gly69Glu + Ser70GluGly67Ser + Asn68Glu + Gly69Asp + Ser70AspGly67Asn + Asn68Asp + Gly69Asp + Ser70GluGly67Asp + Asn68Gln + Gly69Glu + Ser70AspAsn66Gln + Gly67Asp + Gly69Asp + Ser70AspGly67Asp + Asn68Asp + Ser70Asp + Thr73AsnGly67Asp + Asn68Asp + Gly69Pro + Ser70GluGly67Asp + Asn68Gln + Ser70Glu + Gly72ProAsn66Gln + Gly67Glu + Ser70Asp + Gly72GlnGly67Glu + Ser70Glu + Gly72Gln + Thr73SerGly67Asp + Asn68Gln + Ser70Asp + Gly72SerGly67Glu + Ser70Asp + Gly72Ser + Thr73Ser表6環2-單突變變異體Val103AlaVal103AsnVal103AspVsl103GlnVal103GlnVal103GluVal103GlyVal103HisVal103MetVal103ProVal103SerVal103ThrLeu104AlaLeu104AsnLeu104AspLeu104CysLeu104GlnLeu104GluLeu104GlyLeu104HisLeu104IleLeu104MetLeu104ProLeu104SerLeu104ThrLeu104ValAsp105GluAsn106AspAsn106GlnAsn106GluAsn106SerSer107AspSer107GluGly108AsnGly108AspGly108GlnGly108GluGly108Pro
Gly108SerSer109AspSer109GluGly110AsnGly110AspGly110GlnGly110GluGly110ProGly110SerThr111AsnThr111AspThr111GlnThr111GluThr111GlyThr111ProThr111SerTrp112AlaTrp112AsnTrp112AspTrp112CysTrp112GlnTrp112GluTrp112GlyTrp112HisTrp112IleTrp112LeuTrp112MetTrp112PheTrp112ProTrp112SerTrp112ThrTrp112TyrTrp112ValThr113AsnThr113AspThr113GlnThr113GluThr113GlyThr113ProThr113SerAla114AsnAla114AspAla114GlnAla114GluAla114GlyAla114HisAla114ProAla114SerAla114ThrVal115AlaVal115AsnVal115AspVal115Cys
Val115GlnVal115GluVal115GlyVal115HisVal115MetVal115ProVal115SerVal115Thr表7環2-雙突變變異體Val103Gln + Ser109GluAsp105Glu + Gly108GlnAsp105Glu + Gly110AsnAsp105Glu + Gly110ProVal103Asp + Thr111GlnLeu104Gln + Gly108AspLeu104Gly + Ser107GluThr111Pro + Ala114SerAsn106Gln + Ser109AspAsp105Glu + Gly108ProThr113Asp + Val115GlyVal103Met + Leu104GlnGly108Gln + Thr113AsnGly110Glu + Ala114GlyThr111Gln + Val115GlnGly110Glu + Thr111GlnGly108Pro + Ala114GluGly108Ser + Val115GlyAsp105Glu + Thr113GlnLeu104His + Ser107AspAsp105Glu + Ala114GlnGly110Gln + Thr113SerVal103His + Ala114GlnAsp105Glu + Trp112IleThr111Ser + Thr113AspLeu104His + Thr111GluAsn106Gln + Trp112TyrLeu104Gly + Thr113GlnVal103Thr + Val115AsnVal103Thr + Asn106SerVal103Pro + Thr111ProSer109Glu + Val115MetVal103Ala + Asn106GlnVal103Cys + Thr111GlnTrp112Gly + Thr113GlyVal103Gly + Asp105GluGly110Asn + Thr113GlyThr111Asn + Thr113GluVal103Glu + Trp112MetLeu104Pro + Ser109AspGly108Gln + Ala114GlyLeu104Met + Val115GluLeu104Gly + Ala114SerThr113Ser + Val115AlaAsp105Glu + Asn106GlnVal103Thr + Asp105GluGly110Glu + Val115MetVal103Gly + Gly110GluSer109Asp + Gly110GlnThr111Asp + Ala114SerAsn106Glu + Ala114ProSer109Glu + Thr113GlyGly108Asn + Trp112GluVal103Pro + Asn106GluAsp105Glu + Trp112LeuLeu104Cys + Asp105GluLeu104Cys + Thr111SerLeu104Thr + Ser109AspVal103Asn + Val115GluVal103Ser + Gly108Glu表8環2-三突變變異體Val103Gln + Leu104Thr + Ser109GluLeu104Ser + Asp105Glu + Trp112PheAsn106Gln + Gly108Ser + Gly110GlnSer109Asp + Thr111Asn + Trp112ThrAsn106Ser + Gly108Gln + Ser109AspAsp105Glu + Gly108Pro + Val115GlyAsn106Ser + Ser107Asp + Val115GlyAsn106Ser + Trp112Gln + Val115GluLeu104Pro + Ser109Asp + Thr113GlyLeu104Asp + Gly110Gln + Val115ProGly108Asn + Thr111Ser + Ala114ProLeu104Ser + Asn106Ser + Thr111GluVal103His + Leu104Gln + Asn106AspAsn106Gln + Trp112Asn + Ala114AspVal103Met + Gly110Pro + Thr111ProLeu104Thr + Gly108Glu + Gly110AsnLeu104Ala + Ser109Asp + Trp112CysGly108Glu + Gly110Pro + Val115GlyLeu104Pro + Thr113Asn + Ala114AspVal103Asn + Leu104Met + Asn106GluVal103Cys + Ser109Glu + Val115CysVal103Gly + Leu104Val + Ser107AspGly108Pro + Ser109Glu + Trp112GlnLeu104Met + Trp112Cys + Thr113AsnAsn106Asp + Trp112Ile + Ala114SerVal103Gly + Ser109Glu + Ala114ProLeu104Ile + Trp112Leu + Thr113GlyAsn106Gln + Gly108Asn + Ser109GluVal103Asn + Trp112Pro + Ala114AspGly108Asn + Gly110Ser + Trp112GlyGly110Asp + Trp112Leu + Val115HisVal103Gly + Leu104Gly + Gly108AspLeu104Ala + Thr111Ser + Val115AlaThr111Ser + Thr113Asn + Val115HisAsp105Glu + Gly108Gln + Val115GlyLeu104Gly + Ala114Ser + Val115AlaVal103His + Leu104Gly + Asn106GluVal103Ser + Gly110Glu + Thr113ProVal103Pro + Gly110Asn + Ala114GluLeu104Met + Gly110Asp + Val115ProLeu104Asp + Thr111Gln + Trp112AlaSer109Asp + Gly110Pro + Trp112AsnLeu104Asp + Asp105Glu + Trp112CysGly108Glu + Ser109Glu + Ala114ThrGly108Asp + Ser109Asp + Thr111ProGly108Glu + Ser109Glu + Ala114AsnAsn106Ser + Ser109Glu + Gly110AspAsn106Gln + Ser109Asp + Gly110AspVal103Asp + Leu104Asp + Gly108ProLeu104Thr + Asp105Glu + Asn106GluAsp105Glu + Asn106Asp + Thr113AsnGly108Gln + Thr113Glu + Ala114GluLeu104Ser + Thr111Glu + Trp112GluAsn106Asp + Ser107Asp + Gly108AspAsp105Glu + Ser109Asp + Gly110GluTrp112Ala + Thr113Asp + Val115AspVal103Gln + Asp105Glu + Gly108AspLeu104Gln + Asp105Glu + Ser107GluSer107Asp + Ser109Asp + Thr111GlyVal103Cys + Ser107Asp + Ser109Glu表9環2-四突變變異體Asp105Glu + Gly110Ser + Trp112Phe + Thr113SerVal103Ala + Ser107Asp + Gly110Pro + Ala114AsnVal103Gln + Gly108Pro + Thr111Ser + Ala114GluVal103Thr + Ser107Asp + Thr113Gln + Val115CysLeu104Asn + Gly110Ser + Thr113Glu + Val115GlnVal103Pro + Gly108Glu + Trp112His + Val115MetVal103Ala + Asn106Ser + Gly108Asp + Thr111AsnVal103Ala + Leu104Asp + Gly110Ser + Val115AlaSer109Glu + Trp112Pro + Ala114Asn + Val115MetVal103Thr + Asn106Asp + Gly108Ser + Gly110GlnVal103Pro + Leu104Pro + Asn106Ser + Gly110AsnSer109Glu + Gly110Pro + Thr111Ser + Thr113SerAsn106Gln + Gly108Ser + Trp112Tyr + Ala114ThrLeu104Cys + Gly110Gln + Thr111Asp + Thr113GlyLeu104Ser + Asp105Glu + Gly108Gln + Val115AsnVal103Asp + Gly108Asn + Thr113Pro + Ala114ProVal103Asp + Leu104Asn + Thr113Asn + Val115AlaVal103Asn + Gly110Asp + Thr111Asn + Val115CysVal103Ser + Asn106Glu + Gly110Gln + Val115MetLeu104Val + Thr111Gln + Trp112His + Ala114SerLeu104Ser + Asp105Glu + Gly110Gln + Thr111AsnVal103Ala + Asp105Glu + Asn106Ser + Gly110AsnVal103Glu + Gly110Ser + Trp112Phe + Val115GlyAsn106Glu + Thr113Asn + Ala114Pro + Val115AsnLeu104Glu + Asp105Glu + Gly108Ser + Gly110AsnGly108Glu + Ser109Glu + Thr111Asn + Ala114ProGly108Glu + Ser109Glu + Thr111Gln + Ala114ThrLeu104Ile + Gly108Pro + Ser109Glu + Gly110AspVal103Glu + Leu104Asp + Trp112Cys + Thr113SerAsp105Glu + Asn106Glu + Gly110Pro + Thr113GlyGly110Gln + Thr111Glu + Trp112Glu + Thr113GlySer107Asp + Gly108Asp + Ser109Asp + Trp112ValAsn106Asp + Ser107Asp + Gly108Glu + Val115ProAsp105Glu + Ser107Glu + Gly108Asp + Gly110SerAsp105Glu + Gly108Asp + Ser109Glu + Thr111GlnAsp105Glu + Gly108Glu + Ser109Glu + Thr111ProLeu104Asp + Ser109Glu + Trp112Asn + Ala114GlyVal103Ser + Leu104Glu + Gly108Asn + Ser109GluAsp105Glu + Ser109Glu + Gly110Glu + Trp112GlnVal103Ser + Asp105Glu + Gly108Glu + Val115ProAsp105Glu + Ser107Asp + Trp112Leu + Thr113GlyLeu104His + Asp105Glu + Asn106Ser + Ser107AspAsp105Glu + Ser107Glu + Ser109Asp + Val115GlnAsp105Glu + Ser107Glu + Ser109Glu + Thr113GlySer107Glu + Ser109Glu + Thr111Gln + Thr113GlnSer107Glu + Ser109Glu + Trp112Val + Val115ProSer107Glu + Ser109Glu + Trp112Ile + Val115GlnAsp105Glu + Ser109Glu + Gly110Gln + Val115ProLeu104Glu + Asp105Glu + Ser107Glu + Ala114GlyAsp105Glu + Asn106Glu + Gly108Pro + Ser109AspLeu104Asn + Asp105Glu + Gly110Asp + Ala114GlySer107Glu + Gly108Gln + Ser109Asp + Gly110AspGly108Gln + Gly110Glu + Trp112Asp + Val115GluVal103Asp + Leu104Ser + Asp105Glu + Val115GlnLeu104Glu + Asn106Glu + Gly110Glu + Val115AsnLeu104Asp + Gly110Glu + Thr111Ser + Trp112AspSer109Asp + Gly110Glu + Thr113Asn + Val115GluVal103Ser + Gly108Asp + Gly110Glu + Thr111GluLeu104Gly + Asn106Glu + Gly108Asn + Ser109GluVal103Asp + Leu104Gly + Thr111Glu + Val115Met表10環3-單突變變異體Leu134AlaLeu134AsnLeu134AspLeu134Cys
Leu134GlnLeu134GluLeu134GlyLeu134HisLeu134IleLeu134MetLeu134ProLeu134SerLeu134ThrLeu134ValGly135AsnGly135AspGly135GlnGly135GluGly135ProGly135SerGly136AsnGly136AspGly136GlnGly136GluGly136ProGly136SerThr137AsnThr137AspThr137GlnThr137GluThr137GlyThr137ProThr137SerVal138AlaVal138AsnVal138AspVal138CysVal138GlnVal138GluVal138GlyVal138HisVal138MetVal138ProVal138SerVal138ThrGly139AsnGly139AspGly139GlnGly139GluGly139ProGly139SerAsn140AspAsn140GlnAsn140GluAsn140SerSer141AspSer141Glu
表11環3-雙突變變異體Leu134Val + Ser141AspLeu134Val + Thr137GluGly135Asn + Ser141AspGly135Ser + Thr137SerLeu134Met + Gly139AspLeu134Thr + Gly139GlnGly139Gln + Asn140GluGly139Pro + Ser141AspLeu134Glu + Val138AsnLeu134Asn + Val138AlaLeu134Ser + Asn140GluVal138Asn + Ser141AspGly135Glu + Val138SerLeu134Ile + Gly139GluLeu134Ile + Gly135AsnLeu134Met + Asn140AspGly136Ser + Gly139AspGly136Pro + Val138AsnLeu134Gly + Gly135ProLeu134Gly + Ser141GluGly135Pro + Gly139ProLeu134Thr + Gly139ProGly135Glu + Asn140SerThr137Gln + Gly139AspGly135Asp + Asn140SerLeu134Ser + Asn140AspGly136Gln + Gly139AspLeu134Pro + Ser141GluLeu134His + Asn140GlnLeu134Pro + Gly135GluGly135Pro + Gly136AspVal138Thr + Ser141AspVal138Pro + Ser141AspLeu134Gln + Thr137GluVal138Ser + Ser141AspVal138Thr + Asn140GlnLeu134Glu + Val138GlnGly139Asn + Ser141GluGly139Pro + Ser141GluGly135Gln + Gly139GluThr137Pro + Val138GlyGly135Asp + Gly136GlnGly135Asn + Val138ThrThr137Asn + Ser141GluGly135Pro + Val138GluGly135Glu + Thr137SerGly135Asp + Gly139ProLeu134Ala + Gly135GlnVal138Gln + Ser141AspGly135Ser + Gly139AspThr137Ser + Val138MetLeu134Ala + Gly136GlnLeu134Glu + Asn140GlnGly136Asn + Asn140GlnGly136Gln + Ser141AspVal138Ser + Ser141GluLeu134Thr + Thr137GluGly135Ser + Gly139ProVal138Cys + Gly139AspThr137Gly + Gly139Asp表12環3-三突變變異體Gly135Ser + Val138Cys + Ser141AspGly135Gln + Thr137Glu + Val138CysLeu134Ser + Gly135Pro + Gly136GluLeu134Pro + Val138Cys + Asn140SerLeu134Val + Gly135Asp + Val138MetLeu134Pro + Gly135Pro + Gly139GluGly135Gln + Val138Asp + Gly139SerGly136Pro + Val138His + Ser141AspLeu134Val + Gly136Ser + Asn140AspThr137Asp + Val138Met + Gly139GlnLeu134Val + Gly136Pro + Thr137GlyGly135Gln + Val138Gln + Asn140AspLeu134His + Gly136Glu + Asn140GlnGly135Pro + Gly139Gln + Ser141AspGly136Asn + Val138Asn + Ser141AspGly135Asp + Val138Thr + Gly139GlnGly136Pro + Thr137Asp + Gly139SerLeu134Gly + Gly136Gln + Gly139SerLeu134Val + Gly135Glu + Thr137GlyLeu134Thr + Val138Gln + Gly139SerGly136Asp + Thr137Asn + Val138GlyGly135Asp + Val138Ser + Asn140GlnGly136Asn + Val138Glu + Asn140GlnLeu134Cys + Thr137Glu + Val138GlyLeu134Asn + Gly135Glu + Gly139SerGly135Ser + Val138His + Asn140GluLeu134Ala + Gly139Ser + Asn140GlnGly135Asp + Thr137Pro + Val138CysLeu134Cys + Val138Gly + Asn140AspLeu134Thr + Val138Ser + Gly139AspLeu134Cys + Thr137Gln + Ser141GluLeu134Val + Gly136Pro + Gly139GluLeu134Ala + Gly135Asn + Ser141GluGly136Gln + Thr137Gln + Val138ProLeu134Asp + Gly135Pro + Gly139SerGly135Ser + Val138Gly + Ser141AspLeu134Val + Thr137Asp + Gly139GlnLeu134His + Gly135Pro + Gly139GlnThr137Pro + Val138Gln + Asn140AspThr137Gln + Val138Cys + Gly139GluThr137Ser + Gly139Glu + Asn140GlnGly135Gln + Thr137Gln + Asn140AspGly135Gln + Thr137Gln + Val138AspLeu134His + Gly139Gln + Asn140AspLeu134Ala + Thr137Asn + Gly139GluVal138Pro + Gly139Pro + Asn140GlnGly135Asn + Asn140Ser + Ser141GluLeu134Ile + Gly135Asp + Val138CysThr137Gln + Gly139Ser + Asn140SerLeu134Ala + Gly136Pro + Asn140AspGly135Ser + Gly139Asp + Asn140SerLeu134Thr + Gly135Asn + Ser141GluLeu134Gly + Thr137Ser + Val138AlaLeu134Thr + Gly135Gln + Val138ProGly135Gln + Gly136Asp + Thr137SerGly135Ser + Val138Thr + Asn140AspLeu134Cys + Thr137Ser + Val138GluGly136Ser + Thr137Asn + Ser141AspLeu134Thr + Gly135Glu + Gly139GlnLeu134Thr + Gly135Asp + Thr137Ser表13環3-四突變變異體Leu134Met + Thr137Pro + Asn140Ser + Ser141GluLeu134Asn + Gly136Pro + Thr137Ser + Val138GlnLeu134Gln + Gly136Gln + Val138Ser + Gly139SerGly135Glu + Gly136Asn + Thr137Pro + Asn140GlnLeu134Ser + Gly136Asn + Gly139Gln + Ser141GluLeu134Gln + Val138Pro + Gly139Ser + Ser141AspLeu134Pro + Gly135Asn + Thr137Glu + Asn140GlnGly135Asp + Thr137Gly + Val138Gln + Gly139SerLeu134Val + Gly135Asp + Thr137Asn + Asn140SerLeu134Thr + Gly135Glu + Gly136Ser + Thr137SerGly136Pro + Thr137Gln + Gly139Gln + Asn140GluLeu134His + Gly135Glu + Val138Asn + Asn140GlnLeu134Gln + Gly135Ser + Thr137Asn + Ser141GluGly135Gln + Gly139Asn + Asn140Gln + Ser141AspThr137Gln + Val138His + Gly139Asn + Ser141GluLeu134Thr + Gly135Gln + Val138Thr + Asn140AspLeu134Met + Gly135Glu + Thr137Ser + Val138SerVal138Thr + Gly139Ser + Asn140Ser + Ser141GluLeu134Met + Gly136Pro + Val138Cys + Gly139ProLeu134His + Gly136Pro + Thr137Gln + Gly139GluGly135Asn + Gly136Gln + Val138Cys + Ser141GluLeu134Asn + Thr137Ser + Val138Gln + Ser141AspGly135Gln + Gly136Pro + Thr137Asn + Asn140SerGly135Asn + Gly139Ser + Asn140Gln + Ser141AspLeu134Met + Gly136Pro + Val138Met + Ser141GluLeu134Cys + Thr137Gln + Val138Pro + Ser141AspGly136Asn + Thr137Gln + Gly139Asn + Asn140SerLeu134Asn + Thr137Ser + Gly139Ser + Asn140AspGly135Asp + Gly136Asp + Val138Met + Asn140SerGly135Glu + Gly136Asp + Val138His + Gly139GlnLeu134Ser + Gly136Glu + Thr137Glu + Val138ThrLeu134Thr + Val138Gln + Asn140Glu + Ser141GluGly135Gln + Thr137Asn + Asn140Glu + Ser141GluGly135Gln + Thr137Gly + Val138Asp + Gly139AspLeu134Thr + Gly135Glu + Gly136Asp + Thr137AspVal138Asn + Gly139Asp + Asn140Glu + Ser141AspLeu134Ile + Gly139Glu + Asn140Asp + Ser141AspGly136Pro + Gly139Glu + Asn140Glu + Ser141AspLeu134Gln + Gly139Asp + Asn140Asp + Ser141GluLeu134Val + Gly136Glu + Val138Asp + Asn140GlnGly136Asp + Thr137Gln + Val138Asp + Gly139AsnGly136Glu + Thr137Glu + Val138His + Gly139AspLeu134His + Thr137Gln + Val138Glu + Asn140GluLeu134Gly + Gly135Ser + Thr137Glu + Gly139GluLeu134Gln + Gly135Pro + Thr137Asp + Gly139AspLeu134His + Gly135Asn + Thr137Asp + Gly139GluLeu134Val + Gly135Glu + Thr137Asp + Gly139GlnGly135Asn + Thr137Ser + Gly139Asp + Ser141AspLeu134Cys + Thr137Asn + Gly139Glu + Ser141AspLeu134Ile + Thr137Pro + Gly139Asp + Ser141AspLeu134Thr + Val138Asn + Gly139Glu + Ser141AspGly135Pro + Thr137Gly + Gly139Glu + Ser141AspGly136Ser + Val138Ala + Gly139Glu + Ser141GluLeu134Thr + Gly139Asp + Asn140Ser + Ser141AspGly135Ser + Thr137Gly + Gly139Glu + Ser141AspLeu134Met + Gly135Asn + Gly139Asp + Ser141GluThr137Pro + Val138Gly + Gly139Asp + Ser141AspGly136Glu + Thr137Pro + Val138Gln + Gly139AspThr137Asn + Val138Glu + Gly139Asp + Ser141AspLeu134Thr + Val138Glu + Gly139Glu + Ser141Glu表14環4-單突變變異體Gly162AsnGly162AspGly162GlnGly162GluGly162ProGly162SerAsn163AspAsn163GlnAsn163GluAsn163SerAla164AsnAla164Asp
Ala164GlnAla164GluAla164GlyAla164HisAla164ProAla164SerAla164ThrGly165AsnGly165AspGly165GlnGly165GluGly165ProGly165SerAsn166AspAsn166GlnAsn166GluAsn166SerThr167AsnThr167AspThr167GlnThr167GluThr167GlyThr167ProThr167SerAla168AsnAla168AspAla168GlnAla168GluAla168GlyAla168HisAla168ProAla168SerAla168ThrPro169AsnPro169AspPro169GlnPro169GluPro169GlyPro169SerAsn170AspAsn170GlnAsn170GluAsn170SerTyr171AlaTyr171AsnTyr171AspTyr171CysTyr171GlnTyr171GluTyr171GlyTyr171HisTyr171IleTyr171Leu
Tyr171MetTyr171ProTyr171SerTyr171ThrTyr171Val表15環4-雙突變變異體Ala164Gln + Pro169GluAsn163Asp + Thr167SerAla164Pro + Ala168AspAla164Gln + Asn166SerGly165Asp + Thr167ProAla164Gln + Ala168SerGly165Gln + Asn166GlnGly162Gln + Asn163GluGly165Gln + Thr167GluGly165Ser + Ala168GlnAla164Gln + Ala168GluAla164His + Tyr171MetGly165Pro + Pro169GlnGly165Gln + Ala168ProGly162Pro + Ala164GlyThr167Pro + Asn170SerAsn163Gln + Tyr171HisAsn163Ser + Ala164GlnAla164Asp + Asn166GlnGly165Asn + Tyr171IleAla168Asp + Asn170GlnAsn166Asp + Ala168GlyAsn163Glu + Thr167SerAsn166Glu + Ala168SerGly162Gln + Ala168GluGly162Asn + Asn170SerAsn163Gln + Ala168HisThr167Pro + Ala168ThrGly162Asn + Gly165GluAsn166Glu + Tyr171ProAla168Asn + Asn170GlnGly162Glu + Thr167SerThr167Asp + Tyr171ValGly162Asp + Ala168HisGly165Ser + Ala168GluThr167Asp + Tyr171MetAla168Thr + Pro169GlyGly165Ser + Pro169GluGly162Glu + Ala164ThrAla168Ser + Tyr171AspAsn163Ser + Gly165ProAla168Asn + Tyr171SerAla164Gln + Gly165GlnThr167Asp + Asn170SerGly165Ser + Asn170GlnThr167Gly + Tyr171GluGly162Asn + Tyr171GlnGly165Asn + Asn166GluAla168Thr + Tyr171GluGly165Ser + Asn166GluThr167Gly + Ala168ThrAsn166Asp + Ala168AsnPro169Glu + Tyr171ValGly162Glu + Asn170SerGly162Ser + Pro169GlyAla164Asn + Asn166GluThr167Gly + Asn170AspThr167Glu + Asn170SerAsn163Asp + Ala164GlnGly165Gln + Pro169Glu表16環4-三突變變異體Ala164Gly + Gly165Glu + Thr167SerGly162Asn + Ala164Thr + Asn166AspAla164Asn + Gly165Asp + Pro169GlnGly165Pro + Thr167Asn + Pro169GluAla168Gly + Pro169Gly + Asn170AspGly162Asp + Gly165Pro + Ala168SerGly165Asp + Thr167Asn + Tyr171GlyThr167Ser + Pro169Asn + Tyr171ThrAsn166Ser + Ala168Gly + Tyr171IleGly165Gln + Thr167Pro + Ala168GluGly162Ser + Asn163Asp + Gly165GlnGly165Glu + Thr167Gly + Ala168SerThr167Gly + Ala168Asp + Tyr171ValAsn163Ser + Gly165Gln + Ala168AspAsn163Asp + Ala164Gly + Gly165AsnAla164Gln + Pro169Ser + Asn170GlnAla164His + Thr167Gln + Pro169GlyThr167Ser + Ala168Asn + Tyr171AsnGly162Asp + Gly165Ser + Ala168AsnGly162Pro + Asn163Gln + Tyr171ValAsn166Glu + Ala168Ser + Asn170GlnThr167Pro + Pro169Ser + Tyr171HisGly162Gln + Asn163Gln + Gly165AspAla164Ser + Thr167Ser + Ala168GluThr167Gln + Pro169Glu + Asn170GlnAsn163Gln + Pro169Asn + Asn170GlnGly162Glu + Ala168Gln + Asn170GlnAla164Gly + Gly165Ser + Asn166AspAla164Ser + Ala168Gly + Asn170GlnAsn163Glu + Ala164Asn + Ala168SerThr167Ser + Pro169Glu + Tyr171SerAla168Glu + Pro169Gln + Tyr171IleGly162Pro + Ala164Pro + Tyr171ProGly162Pro + Asn166Ser + Pro169GluAla164Gly + Ala168Asn + Tyr171SerAla164Thr + Asn170Asp + Tyr171LeuAsn163Gln + Thr167Glu + Pro169SerAsn166Asp + Ala168Gln + Tyr171ProGly162Asn + Ala164Glu + Tyr171AlaAsn166Glu + Thr167Gln + Ala168ProAla164Pro + Ala168Gln + Asn170AspAla164Thr + Gly165Gln + Ala168AspAla164Ser + Ala168Gly + Asn170GluAla164His + Gly165Glu + Tyr171MetAla164Gln + Gly165Asp + Asn166GlnGly162Gln + Thr167Asn + Ala168AsnAla168His + Pro169Gln + Tyr171GluAla164Gln + Asn166Gln + Tyr171ValGly162Pro + Ala164Ser + Asn170GlnThr167Glu + Ala168Gly + Asn170SerThr167Ser + Pro169Gly + Asn170SerAsn163Glu + Asn166Ser + Ala168SerAsn163Gln + Ala168Glu + Tyr171AlaThr167Ser + Ala168Ser + Asn170AspGly165Asp + Asn166Glu + Tyr171ProAsn163Asp + Ala164Asp + Thr167SerAla164Asp + Gly165Glu + Asn166GlnGly162Ser + Ala164Glu + Gly165AspGly165Asn + Asn170Glu + Tyr171GluGly165Pro + Asn170Glu + Tyr171Glu表17環4-四突變變異體Ala164Asn + Asn166Asp + Thr167Asn + Ala168GlnGly162Asn + Asn163Gln + Ala164Asn + Asn170GluAla164Thr + Asn166Ser + Ala168Asn + Pro169GluAla164Gln + Pro169Asn + Asn170Glu + Tyr171ThrAsn163Ser + Ala164Thr + Thr167Asp + Tyr171ThrThr167Ser + Pro169Ser + Asn170Asp + Tyr171SerGly162Asn + Thr167Gly + Pro169Gln + Tyr171SerGly162Pro + Asn163Ser + Ala168Glu + Tyr171ValAsn163Gln + Ala168Asp + Pro169Gln + Asn170GlnAla164Asn + Gly165Ser + Pro169Gln + Tyr171IleGly162Glu + Thr167Gln + Ala168Ser + Tyr171SerGly165Ser + Ala168Gly + Asn170Asp + Tyr171LeuAla164His + Gly165Asp + Asn166Gln + Tyr171ThrAsn163Gln + Ala164Ser + Asn166Ser + Ala168GluAsn166Asp + Thr167Asn + Asn170Ser + Tyr171ThrGly162Asp + Gly165Gln + Asn170Ser + Tyr171GlnGly162Pro + Ala168Asn + Pro169Glu + Tyr171GlyAsn166Gln + Thr167Gln + Ala168Thr + Asn170AspAsn163Gln + Thr167Asp + Ala168Ser + Tyr171ProAla164Pro + Thr167Glu + Pro169Ser + Tyr171ValGly165Asn + Asn166Gln + Pro169Gly + Asn170AspAsn166Ser + Thr167Pro + Ala168Asn + Asn170GluAla164Gly + Ala168Asn + Pro169Asp + Tyr171AlaAla164Gln + Gly165Gln + Thr167Ser + Pro169GluAsn163Glu + Gly165Ser + Thr167Gln + Ala168ThrGly162Pro + Ala164Thr + Thr167Ser + Pro169GlnGly162Gln + Asn163Glu + Gly165Gln + Thr167ProAsn163Ser + Pro169Gly + Asn170Glu + Tyr171SerAsn163Gln + Gly165Ser + Asn170Asp + Tyr171LeuAla164His + Gly165Pro + Pro169Ser + Asn170GlnGly162Pro + Asn166Gln + Ala168Asn + Asn170GluAsn163Gln + Gly165Pro + Thr167Ser + Tyr171GlyAsn163Gln + Gly165Asn + Asn166Ser + Tyr171CysAsn163Asp + Gly165Pro + Asn166Ser + Tyr171GlyAla164Pro + Thr167Pro + Pro169Glu + Asn170SerAsn163Asp + Ala164Glu + Pro169Gln + Tyr171HisAsn163Glu + Ala164Glu + Ala168Gly + Tyr171HisThr167Asp + Ala168Glu + Pro169Gln + Asn170SerAla164Asp + Gly165Asp + Ala168Asn + Pro169GlnGly162Ser + Ala164Gly + Asn170Asp + Tyr171GluAsn163Gln + Thr167Asn + Ala168Asp + Pro169GluAsn163Ser + Gly165Pro + Ala168Asp + Pro169GluGly165Ser + Ala168Asp + Pro169Glu + Asn170SerAsn166Gln + Ala168Gln + Pro169Asp + Asn170AspAla164Gly + Thr167Gly + Pro169Glu + Asn170AspGly162Pro + Ala164Gly + Pro169Asp + Asn170GluAla164Pro + Thr167Glu + Ala168Asp + Pro169AspGly162Asp + Asn163Gln + Ala168His + Asn170AspGly162Glu + Ala164Ser + Asn166Ser + Asn170GluGly162Glu + Ala164Ser + Asn166Gln + Asn170GluThr167Asp + Ala168His + Pro169Glu + Tyr171ProAsn163Glu + Ala164His + Gly165Asp + Asn166GlnAsn163Asp + Ala164Gln + Gly165Asp + Asn166SerAsn163Glu + Ala164Asp + Gly165Gln + Ala168AspGly162Glu + Ala164Glu + Asn166Ser + Pro169AspAla164Glu + Asn166Asp + Asn170Ser + Tyr171GlyAla164Glu + Gly165Gln + Asn166Glu + Tyr171CysAla164Asp + Asn166Glu + Thr167Asn + Pro169AsnThr167Asp + Pro169Glu + Asn170Glu + Tyr171SerAsn166Asp + Thr167Ser + Ala168Asp + Tyr171His表18環5-單突變變異體Ser191AspSer191GluSer192AspSer192GluPhe193AlaPhe193AsnPhe193Asp
Phe193CysPhe193GlnPhe193GluPhe193GlyPhe193HisPhe193IlePhe193LeuPhe193MetPhe193ProPhe193SerPhe193ThrPhe193TyrPhe193ValSer194AspSer194GluThr195AsnThr195AspThr195GlnThr195GluThr195GlyThr195ProThr195Ser表19環5-雙突變變異體Ser192Asp + Phe193SerSer192Asp + Phe193MetSer191Glu + Phe193AsnSer191Asp + Phe193IleSer191Glu + Phe193GlnSer191Asp + Thr195SerPhe193Gly + Thr195AspSer192Asp + Phe193CysSer191Glu + Thr195ProSer191Glu + Phe193MetPhe193Thr + Thr195GluSer192Asp + Thr195GlnPhe193Asp + Thr195AsnSer191Asp + Phe193GlnPhe193Gly + Thr195ProSer192Asp + Thr195SerSer191Glu + Phe193IlePhe193Gln + Thr195AsnSer191Glu + Thr195GlySer191Glu + Phe193HisSer191Asp + Phe193LeuSer192Glu + Phe193ProPhe193His + Thr195GluSer192Asp + Thr195GlySer192Glu + Phe193AlaSer192Glu + Phe193ValSer192Asp + Phe193LeuSer192Glu + Phe193AsnPhe193Tyr + Thr195SerPhe193His + Thr195AsnSer192Glu + Thr195SerPhe193Ile + Thr195GluPhe193Val + Thr195AsnPhe193Met + Thr195GlnSer191Asp + Thr195ProSer192Glu + Phe193TyrPhe193Ser + Thr195AsnSer192Glu + Phe193GlnSer191Asp + Phe193SerSer192Asp + Phe193IleSer192Asp + Phe193HisSer192Asp + Phe193GlySer191Glu + Phe193SerSer192Glu + Thr195GlnSer191Glu + Thr195SerSer191Asp + Thr195AsnSer191Glu + Phe193ValPhe193Tyr + Thr195AspPhe193His + Thr195SerSer192Glu + Phe193IlePhe193Met + Thr195GlySer191Glu + Phe193TyrSer191Asp + Thr195GlnSer192Asp + Phe193AsnSer192Asp + Phe193ThrSer192Glu + Phe193CysPhe193Asp + Thr195GlnPhe193Val + Thr195GlnSer192Glu + Phe193ThrPhe193Leu + Thr195Glu表20環5-三突變變異體Ser192Glu + Phe193Thr + Thr195GlySer191Glu + Phe193His + Thr195SerSer192Asp + Phe193Leu + Thr195AsnSer191Asp + Phe193Ala + Thr195SerSer192Asp + Phe193Val + Thr195GlnSer191Asp + Phe193Ser + Thr195ProSer191Asp + Phe193Ile + Thr195GlySer192Glu + Phe193His + Thr195GlySer192Glu + Phe193Leu + Thr195AsnSer192Asp + Phe193Gly + Thr195GlnSer192Asp + Phe193Ile + Thr195ProSer191Glu + Phe193Asn + Thr195SerSer192Asp + Phe193Met + Thr195AsnSer191Glu + Phe193Gly + Thr195AsnSer192Glu + Phe193His + Thr195AsnSer192Asp + Phe193Met + Thr195ProSer191Glu + Phe193Leu + Thr195AsnSer191Glu + Phe193Met + Thr195AsnSer192Glu + Phe193Ala + Thr195GlnSer191Glu + Phe193Tyr + Thr195GlySer192Asp + Phe193Ala + Thr195GlySer191Glu + Phe193Met + Thr195GlySer192Glu + Phe193Thr + Thr195GlnSer191Asp + Phe193Asn + Thr195GlySer192Glu + Phe193Asn + Thr195SerSer192Asp + Phe193Ser + Thr195GlySer192Glu + Phe193Leu + Thr195ProSer191Asp + Phe193Cys + Thr195GlySer191Asp + Ser192Asp + Phe193TyrSer191Asp + Ser192Asp + Phe193GlySer191Asp + Ser192Glu + Phe193MetSer191Asp + Ser192Glu + Thr195GlnSer191Glu + Ser192Asp + Phe193ValSer191Glu + Ser192Glu + Phe193ThrSer191Glu + Ser192Asp + Phe193ThrSer191Asp + Ser192Asp + Thr195GlySer191Glu + Ser192Glu + Phe193AsnSer191Asp + Ser192Glu + Phe193AlaSer191Asp + Ser192Asp + Phe193AsnSer191Asp + Ser192Asp + Phe193ProSer191Asp + Ser192Glu + Phe193ThrSer191Asp + Ser192Asp + Thr195AsnSer191Asp + Ser192Asp + Thr195GlnSer191Glu + Ser192Asp + Phe193AsnSer191Glu + Ser192Glu + Phe193GlnSer191Glu + Ser192Asp + Phe193LeuSer191Asp + Ser192Asp + Thr195SerSer191Asp + Ser192Glu + Phe193SerSer191Glu + Ser192Asp + Phe193TyrSer191Asp + Ser192Asp + Phe193ThrSer191Asp + Ser192Glu + Phe193ProSer191Glu + Ser192Asp + Phe193IleSer191Asp + Ser192Glu + Phe193TyrSer191Glu + Ser192Glu + Phe193TyrSer191Asp + Ser192Asp + Phe193SerSer191Glu + Ser192Glu + Thr195ProSer191Asp + Ser192Glu + Thr195SerSer191Glu + Ser192Asp + Phe193MetSer191Asp + Ser192Glu + Thr195ProSer191Glu + Ser192Asp + Phe193His表21環5-四突變變異體Ser191Asp + Ser192Glu + Phe193Gly + Thr195GlnSer191Glu + Ser192Asp + Phe193His + Thr195GlySer191Glu + Ser192Glu + Phe193Cys + Thr195ProSer191Asp + Ser192Glu + Phe193Cys + Thr195GlnSer191Asp + Ser192Glu + Phe193Thr + Thr195ProSer191Glu + Ser192Glu + Phe193Val + Thr195AsnSer191Asp + Ser192Glu + Phe193Val + Thr195AsnSer191Glu + Ser192Glu + Phe193Leu + Thr195SerSer191Glu + Ser192Asp + Phe193Cys + Thr195GlnSer191Glu + Ser192Asp + Phe193Pro + Thr195ProSer191Glu + Ser192Asp + Phe193Ser + Thr195GlySer191Glu + Ser192Asp + Phe193Pro + Thr195AsnSer191Glu + Ser192Glu + Phe193Ser + Thr195ProSer191Glu + Ser192Asp + Phe193Ala + Thr195GlySer191Asp + Ser192Glu + Phe193His + Thr195AsnSer191Asp + Ser192Glu + Phe193Asn + Thr195AsnSer191Glu + Ser192Glu + Phe193Gln + Thr195SerSer191Glu + Ser192Glu + Phe193Thr + Thr195GlnSer191Glu + Ser192Glu + Phe193Asn + Thr195GlnSer191Glu + Ser192Glu + Phe193Tyr + Thr195SerSer191Asp + Ser192Glu + Phe193Ile + Thr195ProSer191Glu + Ser192Asp + Phe193Val + Thr195GlnSer191Glu + Ser192Asp + Phe193Val + Thr195AsnSer191Asp + Ser192Glu + Phe193Tyr + Thr195SerSer191Asp + Ser192Glu + Phe193Ala + Thr195SerSer191Glu + Ser192Glu + Phe193His + Thr195AsnSer191Asp + Ser192Asp + Phe193Pro + Thr195AsnSer191Asp + Ser192Asp + Phe193Gln + Thr195SerSer191Asp + Ser192Asp + Phe193Leu + Thr195AsnSer191Asp + Ser192Glu + Phe193Val + Thr195GlnSer191Glu + Ser192Glu + Phe193Gly + Thr195GlySer191Glu + Ser192Glu + Phe193Thr + Thr195ProSer191Glu + Ser192Glu + Phe193Thr + Thr195GlySer191Glu + Ser192Asp + Phe193Ile + Thr195SerSer191Glu + Ser192Glu + Phe193Ile + Thr195GlySer191Asp + Ser192Asp + Phe193Pro + Thr195ProSer191Asp + Ser192Asp + Phe193Ile + Thr195GlySer191Glu + Ser192Asp + Phe193Thr + Thr195GlySer191Asp + Ser192Glu + Phe193Ser + Thr195GlySer191Glu + Ser192Asp + Phe193Tyr + Thr195SerSer191Asp + Ser192Asp + Phe193Thr + Thr195GlnSer191Asp + Ser192Glu + Phe193Ser + Thr195GlnSer191Glu + Ser192Glu + Phe193His + Thr195SerSer191Glu + Ser192Asp + Phe193Leu + Thr195AsnSer191Asp + Ser192Glu + Phe193Leu + Thr195ProSer191Asp + Ser192Asp + Phe193Ile + Thr195ProSer191Asp + Ser192Asp + Phe193Tyr + Thr195SerSer191Asp + Ser192Glu + Phe193Ile + Thr195GlnSer191Asp + Ser192Glu + Phe193Pro + Thr195SerSer191Asp + Ser192Glu + Phe193Leu + Thr195GlnSer191Glu + Ser192Asp + Phe193Asn + Thr195SerSer191Glu + Ser192Glu + Phe193Ser + Thr195AsnSer191Asp + Ser192Asp + Phe193Glu + Thr195GlnSer191Asp + Ser192Glu + Phe193Glu + Thr195ProSer191Glu + Ser192Asp + Phe193Glu + Thr195GlnSerl91Glu + Ser192Glu + Phe193Glu + Thr195GlySer191Glu + Ser192Glu + Phe193Asp + Thr195GlySer191Asp + Ser192Asp + Phe193Glu + Thr195SerSer191Glu + Ser192Asp + Phe193Asp + Thr195ProSer191Glu + Ser192Glu + Phe193Asp + Thr195Gln表22環6-單突變變異體Ala204AsnAla204AspAla204GlnAla204GluAla204GlyAla204HisAla204ProAla204SerAla204ThrPro205AsnPro205AspPro205GlnPro205GluPro205GlyPro205SerGly206AsnGly206AspGly206GlnGly206GluGly206ProGly206SerSer207AspSer207GluTrp208AlaTrp208AsnTrp208AspTrp208CysTrp208GlnTrp208GluTrp208GlyTrp208HisTrp208IleTrp208LeuTrp208MetTrp208PheTrp208ProTrp208SerTrp208ThrTrp208TyrTrp208ValIle209AlaIle209AsnIle209Asp
Ile209CysIle209GlnIle209GluIle209GlyIle209HisIle209LeuIle209MetIle209ProIle209SerIle209ThrIle209ValTyr210AlaTyr210AsnTyr210AspTyr210CysTyr210GlnTyr210GluTyr210GlyTyr210HisTyr210IleTyr210LeuTyr210MetTyr210ProTyr210SerTyr210ThrTyr210ValSer211AspSer211GluThr212AsnThr212AspThr212GlnThr212GluThr212GlyThr212ProThr212SerTyr213AlaTyr213AsnTyr213AspTyr213CysTyr213GlnTyr213GluTyr213GlyTyr213HisTyr213IleTyr213LeuTyr213MetTyr213ProTyr213SerTyr213ThrTyr213ValPro214AsnPro214AspPro214Gln
Pro214GluPro214GlyPro214SerThr215AsnThr215AspThr215GlnThr215GluThr215GlyThr215ProThr215SerSer216AspSer216GluThr217AsnThr217AspThr217GlnThr217GluThr217GlyThr217ProThr217SerTyr218AlaTyr218AsnTyr218AspTyr218CysTyr218GlnTyr218GluTyr218GlyTyr218HisTyr218IleTyr218LeuTyr218MetTyr218ProTyr218SerTyr218ThrTyr218ValAla219AsnAla219AspAla219GlnAla219GluAla219GlyAla219HisAla219ProAla219SerAla219ThrSer220AspSer220GluLeu221AlaLeu221AsnLeu221AspLeu221CysLeu221GlnLeu221GluLeu221GlyLeu221His
Leu221IleLeu221MetLeu221ProLeu221SerLeu221ThrLeu221ValSer222AspSer222GluGly223AsnGly223AspGly223GlnGly223GluGly223ProGly223SerThr224AsnThr224AspThr224GlnThr224GluThr224GlyThr224ProThr224Ser表23環6-雙突變變異體Ser207Glu + Thr215GlnPro205Gly + Ala219GlyThr217Asn + Thr224GlyGly206Gln + Pro214AspAla204Asn + Thr212SerPro205Gly + Ser220AspTyr213Gln + Tyr218GluPro205Gln + Leu221GluThr212Gln + Ser220AspTyr210Gly + Thr224GlyTyr213Gly + Ser216GluSer207Glu + Gly223AsnTyr210Ser + Tyr218LeuSer216Glu + Thr217SerSer216Glu + Thr224AsnPro205Asn + Ala219AsnThr217Gly + Leu221ThrThr215Glu + Ala219GlnThr217Asn + Tyr218GluAla204Asn + Gly206ProThr212Ser + Thr224GlnThr217Gly + Ser222GluTrp208Glu + Ile209AsnPro205Ser + Ala219GlyTyr213Ser + Ala219SerAla204Ser + Gly206SerAla204Gly + Thr215Glu
42Thr212Gln + Ala219GluAla204Gly + Trp208GlnAla204Thr + Ser216AspThr212Gln + Tyr218GlnSer207Glu + Leu221MetThr215Pro + Gly223GlnAla204Gly + Ser216AspTyr210Asp + Ala219HisIle209Gln + Tyr218AlaIle209Ala + Ala219ProAla204Asn + Ser222GluTyr210Asp + Tyr213AlaTyr218Leu + Thr224GluTyr218Asn + Ser220GluTrp208Met + Ala219ProPro205Gly + Ser222AspLeu221Val + Thr224GlnTrp208Ile + Pro214SerTrp208Asp + Thr212ProAla204Thr + Gly223GlnThr217Gln + Tyr218IleTyr213His + Ser216AspThr212Ser + Thr215AspThr217Asn + Leu221AsnPro214Asp + Thr217ProSer207Glu + Ala219HisThr217Asp + Tyr218HisTrp208Ala + Thr217SerPro205Gln + Ser220AspIle209Gly + Ser216GluTyr218Ser + Ser220GluThr215Asn + Ser216GluThr215Gly + Tyr218GlnThr217Ser + Leu221AlaAla219Pro + Leu221HisPro205Gln + Leu221MetTyr213Met + Leu221IleTrp208Asn + Thr212SerSer207Glu + Thr224GlyPro205Ser + Leu221CysAla204Gly + Tyr210ValTrp208Phe + Ser222AspIle209Thr + Ser222GluLeu221Ile + Ser222GluTyr213Pro + Thr217GluIle209Thr + Ser216GluTyr210Cys + Ser222AspLeu221Glu + Thr224SerTyr213Ala + Thr217ProTrp208Cys + Thr215GluAla204Ser + Thr217SerIle209His + Gly223SerAla204Asn + Trp208Tyr
43Pro205Gln + Ser207GluAla204Asn + Trp208MetTyr210Pro + Leu221AspTyr210Gln + Thr215SerAla204Pro + Ser216GluAla204Gly + Ser222AspSer216Asp + Leu221MetTyr210Ile + Ala219AsnPro214Glu + Thr224SerPro205Gln + Thr215SerAla204Gly + Pro205AsnTyr210Asp + Tyr218ValTrp208Ser + Thr215GlySer222Glu + Thr224SerTrp208Gln + Ser216AspSer207Asp + Gly223SerPro205Gly + Tyr210CysThr212Gly + Thr224AspSer220Glu + Gly223ProGly206Pro + Tyr210MetTyr210Ser + Ser222AspSer207Glu + Pro214GlnPro205Gly + Gly223AsnTyr213Ser + Pro214AspPro214Gln + Ser220AspAla204Thr + Ser222GluAla219Asn + Leu221AspGly206Ser + Thr215ProThr212Asn + Tyr218ValThr217Pro + Gly223AspGly206Pro + Tyr218AsnGly206Gln + Thr217AspTyr210Ala + Ser220AspPro214Asp + Tyr218ValLeu221His + Gly223AspPro214Gln + Gly223AspIle209Pro + Ser216AspIle209Thr + Gly223AspPro214Asn + Ala219AspTyr218Cys + Ser220GluSer207Glu + Tyr218AsnTyr218Gly + Leu221GlnIle209Thr + Tyr218ValPro214Glu + Gly223SerTyr213Met + Ala219ThrSer222Glu + Thr224AsnTrp208Asp + Ile209MetThr212Gln + Pro214GlnSer207Asp + Tyr213LeuThr212Gln + Gly223ProTyr210Leu + Ser216GluSer216Asp + Ala219ThrTyr213Gln + Thr217AspGly206Pro + Ser222GluAla204Thr + Ser220AspAla219Thr + Thr224ProTrp208Ile + Ala219GluIle209Gly + Thr215SerGly206Gln + Tyr210GluPro214Gln + Ser220GluSer207Glu + Thr217AsnAla204Pro + Gly223AspIle209Cys + Ser222AspPro214Gly + Gly223ProIle209Met + Thr224ProPro214Glu + Thr217SerPro205Ser + Ser207AspTyr210Thr + Thr212GlnGly206Gln + Tyr218ProTyr218Val + Ser222GluLeu221Asn + Gly223GluPro205Gln + Tyr218ValPro205Gln + Thr217AsnTrp208His + Ile209AlaPro205Ser + Gly223GlnPro214Ser + Thr217GlyTyr218His + Ser222AspThr212Gln + Gly223SerThr215Glu + Ala219ThrTyr213Gly + Tyr218AspThr212Gln + Tyr213GlnTyr210Ile + Thr215AspAla204Asn + Gly223ProSer207Glu + Ile209AsnThr215Gln + Ala219AspSer220Asp + Gly223ProPro205Ser + Tyr213AlaTrp208Gly + Ser222GluThr215Glu + Tyr218AlaAla204Gln + Ala219GlnPro205Asn + Tyr213ThrTyr218Thr + Thr224GlnSer222Asp + Thr224SerAla204Pro + Tyr213CysGly206Gln + Leu221GlySer216Glu + Thr224SerSer207Asp + Ala219AsnTrp208Leu + Ile209HisPro205Ser + Tyr213ValThr212Gly + Thr215Asp表24環6-三突變變異體Gly206Ser+Thr212Ser+Ala219GluAla204Asn + Thr212Ser + Ser216GluPro205Gln + Leu221Glu + Gly223AsnTrp208Thr + Ile209Cys + Thr224AspIle209Met + Thr212Gln + Ser220AspTyr210Gly + Tyr213Gly + Thr224GlySer207Glu + Tyr210Ser + Gly223AsnTyr213Gly + Ser216Asp + Thr217SerPro205Asn + Ser216Glu + Thr224AsnThr215Glu + Ala219Gln + Leu221ThrAla204Asn + Thr217Asn + Tyr218GluThr212Ser + Thr217Gly + Thr224GlnAla204Ser + Tyr218His + Ala219GlnPro205Ser + Tyr213Ser + Leu221AsnAla204Ser + Gly206Ser + Ala219SerAla204Gly + Trp208Gln + Ser216AspSer207Glu + Tyr218Gln + Leu221MetAla204Gly + Thr215Pro + Gly223GlnIle209Gln + Tyr218Ala + Ala219ProAla204Asn + Trp208Met + Ser222GluTrp208Met + Ala219Pro + Ser222AspPro205Gly + Leu221Val + Thr224GlnAla204Thr + Thr212Pro + Gly223GlnGly206Gln + Ile209Gln + Thr212GlnGly206Pro + Ser220Glu + Leu221SerGly206Gln + Tyr210Gly + Ser222AspAla204Gly + Tyr213Thr + Pro214GlnAla204His + Thr212Gln + Thr224GlnGly206Gln + Pro214Asp + Tyr218SerThr212Asn + Ser216Asp + Thr217SerSer207Asp + Tyr213Ile + Thr224ProPro205Gln + Tyr213Met + Leu221HisTrp208Asn + Thr212Ser + Gly223AspSer207Glu + Leu221Gly + Thr224GlyPro205Ser + Tyr210Val + Leu221CysPro205Gly + Ile209His + Tyr213ThrIle209Pro + Tyr210Asn + Ser220AspPro205Gly + Thr212Pro + Thr224GlyTrp208Cys + Thr217Asp + Ala219AsnPro205Gly + Leu221Cys + Gly223ProAla204His + Ser216Glu + Leu221SerThr217Asp + Gly223Ser + Thr224GlyPro205Gln + Trp208Gln + Ile209AlaThr212Gly + Thr217Gly + Leu221HisGly206Asn + Thr212Gly + Tyr213GlyGly206Asn + Ser207Glu + Thr212SerThr212Gln + Leu221Glu + Gly223SerTyr210His + Thr215Asp + Thr224ProPro214Gln + Thr215Pro + Ser216GluPro205Gly + Tyr213Val + Gly223AspTrp208Phe + Thr215Asn + Thr224GlyTyr213Met + Ala219Pro + Ser220GluTrp208Phe + Ile209Ser + Tyr213GlnGly206Ser + Ser216Glu + Tyr218ProAla204Gln + Tyr210His + Thr224GluPro205Ser + Thr212Pro + Ala219SerPro205Gln + Tyr210Met + Thr217ProAla204Gly + Ile209His + Ser216GluThr212Asn + Thr217Ser + Thr224AsnGly206Pro + Tyr210Gly + Gly223GlnPro205Gln + Thr217Asp + Ala219ThrPro205Ser + Ile209Met + Ser216AspThr215Asn + Ser216Glu + Thr224SerPro205Gln + Leu22lThr + Ser222GluTyr210Cys + Ala219Asn + Leu221ThrAla204Ser + Gly206Ser + Ala219GlnIle209Pro + Ser216Asp + Leu221CysTrp208Thr + Tyr210Val + Thr212SerThr212Gly + Pro214Asp + Gly223GlnPro214Gln + Thr215Ser + Thr224GluThr212Ser + Pro214Gln + Leu221HisTrp208Gly + Ser220Glu + Gly223SerPro205Ser + Ser220Asp + Leu221ThrGly206Asn + Thr215Pro+ Leu221AlaTyr213Leu + Leu221His + Gly223AspSer207Asp + Thr212Gly + Tyr218CysGly206Gln + Ile209Val + Ala219ThrTrp208Ile + Tyr218Ile + Gly223AsnAla204His + Ile209Gly + Ser216AspTrp208Leu + Thr212Gly + Ala219SerThr212Gln + Pro214Glu + Ala219ThrPro205Gln + Ser207Asp + Thr212ProPro205Ser + Tyr210Pro + Tyr218IleLeu221Ala + Gly223Asn + Thr224GluAla204Thr + Gly206Gln + Ser207AspSer216Glu + Ala219Gln + Gly223GlnPro205Gly + Trp208Leu + Leu221GlnThr217Gln + Gly223Gln + Thr224GluThr212Gly + Thr217Ser + Tyr218AlaGly206Asn + Gly223Asp + Thr224AsnAla204Gly + Tyr210Cys + Tyr213ThrGly206Ser + Thr212Asn + Thr215GlyAla204Asn + Thr212Asn + Ser216GluGly206Gln + Tyr218Cys + Gly223GluAla204Ser + Pro205Ser + Gly206AsnIle209Gly + Leu221Val + Thr224ProGly206Asn + Trp208His + Thr217GlyGly206Ser + Trp208Leu + Ser222AspPro205Gly + Thr212Gln + Ser222AspAla204Thr + Tyr213Gln + Thr224ProThr212Ser + Thr217Ser + Gly223AspPro205Ser + Tyr213Ile + Ser222GluIle209Pro + Tyr210Asp + Ala219ProGly206Gln + Thr215Pro + Ser220GluTrp208Cys + Tyr210Leu + Thr215GlyTrp208Tyr + Thr212Gln + Ser222GluGly206Asn + Ser222Asp + Gly223AsnPro205Gly + Gly206Asn + Leu221CysPro205Gln + Tyr213Ile + Ser216GluAla204His + Tyr210His + Ser220AspAla204Thr + Thr212Asn + Ala219AspSer207Glu + Tyr210Ala + Thr212GlyTyr210His + Thr217Gly + Leu221MetAla204Thr + Ile209Met + Ser222GluTrp208Pro + Ala219Pro + Ser222AspAla204His + Ile209Ser + Ser220GluPro214Ser + Leu221Glu + Gly223GlnGly206Pro + Leu221Met + Gly223AspAla204His + Tyr213Thr + Ser222AspTyr213Ala + Leu221Met + Ser222AspTrp208Leu + Thr212Ser + Pro214AspTyr213Ile + Ser216Glu + Thr217ProPro205Gln + Pro214Gln + Thr224GlyTyr213Cys + Thr217Gly + Ser220GluGly206Asn + Thr212Gly + Tyr218ProGly206Pro + Ser216Glu + Gly223ProThr212Gln + Thr215Gly + Ser222GluAla204Pro + Thr217Asp + Thr224SerPro205Ser + Thr212Pro + Ala219AspIle209Cys + Pro214Ser + Thr215AspIle209Cys + Ser220Asp + Gly223GlnThr217Gly + Ser220Asp + Leu221GlnAla204His + Pro205Asn + Trp208GluAla204Gln + Thr212Ser + Thr215AspPro214Asn + Ser220Glu + Leu221GlyIle209Gly +Thr215Asn + Ala219ProSer207Glu + Thr217Gln + Thr224SerAla204Gly + Thr215Asn + Gly223ProIle209Ser + Tyr213Ile + Leu221MetTyr210Thr + Pro214Ser + Ser222AspTyr210Leu + Thr212Ser + Leu221AsnGly206Asn + Pro214Asp + Tyr218MetGly206Asn + Pro214Gln + Ala219AsnSer207Glu + Pro214Ser + Ala219HisAla204Asn + Trp208Ala + Ile209GlnThr217Glu + Tyr218Leu + Gly223AsnTrp208Pro + Pro214Gly + Leu221AlaGly206Pro + Thr215Ser + Leu221CysAla204Ser + Ile209Val + Thr224AspTrp208Ile + Tyr210Pro + Thr224AspThr212Gln + Thr215Asn + Thr217AspTyr210Ser + Tyr213Cys + Thr217AsnThr212Gln + Ser222Asp + Gly223GlnTyr210Asn + Thr217Glu + Tyr218MetPro214Asn + Ala219Gln + Ser220GluPro205Ser + Trp208Met + Leu221AspAla204Asn + Tyr210Cys + Tyr213AsnTyr218Leu + Leu221His + Ser222GluAla204His + Tyr218Ser + Ser222AspGly206Pro + Tyr213Asn + Pro214AsnGly206Pro + Thr215Gln + Tyr218SerAla204Pro + Ile209His + Leu221ThrGly206Gln + Thr212Gly + Gly223GluThr215Gly + Tyr218Ala + Thr224ProPro205Gln + Tyr213Gly + Gly223GluAla204Gly + Tyr213Asn + Leu221ThrTrp208Gly + Ile209His + Thr215SerAla204His + Ser207Glu + Tyr213CysTrp208His + Tyr210His + Ser222AspPro205Gly + Tyr210Asp + Leu221GlnPro205Asn + Tyr210Ala + Pro214SerThr212Pro + Thr215Gln + Thr224AspAla204Thr + Tyr213Val + Pro214GluAla204Gly + Tyr210Pro + Pro214GluPro205Gly + Trp208His + Tyr218GlyGly206Ser + Ala219Gln + Gly223SerTyr213Thr + Tyr218Cys + Ser220GluThr215Pro + Ser216Glu + Leu221AsnPro205Gly + Thr212Asn + Thr224AspPro205Gly + Ser207Asp + Thr212Ser表25環6-四突變變異體Gly206Asn + Thr215Gln + Tyr218Ile + Leu221IleTyr210Gly + Thr212Gln + Tyr213Gly + Thr224GlyPro205Gly + Gly206Pro + Trp208Asn + Thr224SerIle209Thr + Pro214Asp + Thr217Asn + Tyr218CysAla204Ser + Tyr218His + Ala219Gln + Leu221AsnAla204Ser + Pro205Ser + Tyr213Ser + Ala219SerAla204Gly + Thr212Gln + Ser216Asp + Gly223SerSer207Glu + Thr215Pro + Tyr218Gln + Leu221MetAla204Gly + Ser216Asp + Ala219His + Gly223GlnIle209Gln + Tyr210Asp + Tyr218Ala + Ala219ProTrp208Met + Tyr218Leu + Ala219Pro + Ser222GluPro205Gly + Leu221Val + Ser222Asp + Thr224GlnAla204Thr + Thr212Pro + Ser222Glu + Gly223GlnGly206Gln + Ile209Gln + Thr212Gln + Leu221HisAla204Ser + Gly206Pro + Ser220Asp + Leu221SerAla204Gly + Thr212Gln + Tyr213Thr + Thr224GlnTyr210Ala + Pro214Ser + Tyr218Val + Ser222AspAla204Gln + Pro205Ser + Gly206Ser + Ser222GluTrp208Cys + Pro214Gln + Ala219Thr + Thr224AspAla204Gly + Gly206Ser + Tyr210Asp + Leu221GlyAla204Gln + Gly206Asn + Tyr210Gln + Ala219AspPro205Ser + Gly206Pro + Ile209Val + Tyr213ProGly206Pro + Pro214Glu + Thr215Pro + Leu221MetSer207Asp + Trp208Met + Tyr218Val + Leu221ProTyr210Gly + Thr212Pro + Pro214Gly + Ser222AspGly206Pro + Thr212Gln + Thr217Ser + Ser222AspAla204Pro + Trp208Met + Ile209Val + Ser216GluTrp208Ser + Tyr218His + Ala219His + Gly223Pro
49Ala204Gln + Trp208Ala + Tyr210Asn + Ser222GluGly206Gln + Ser220Asp + Leu221Ala + Thr224SerIle209Gln + Thr212Ser + Ala219Gln + Ser220GluPro205Asn + Thr212Gly + Tyr218Ala + Ser222GluPro205Gln + Trp208Cys + Tyr213Cys + Pro214AspPro205Gly + Thr215Gln + Thr217Pro + Gly223SerAla204His + Gly206Pro + Ser216Glu + Leu221SerSer207Asp + Leu221Gly + Gly223Gln + Thr224AsnTyr210His + Thr212Ser + Leu221Glu + Gly223SerPro205Gly + Thr215Asn + Gly223Asp + Thr224GlyTrp208Phe + Tyr213Met + Thr215Pro + Ser220GluGly206Ser + Trp208Phe + Ile209Ser + Tyr213GlnAla204Gln + Pro205Ser + Thr212Pro + Ala219SerAla204Gly + Tyr210Met + Ser216Glu + Thr217ProGly206Pro + Thr212Asn + Thr217Gln + Thr224AsnPro205Ser + Ile209Met + Thr215Asn + Thr224SerTyr210Thr + Thr212Pro + Tyr218Asn + Ser220GluTrp208Thr + Tyr210Val + Thr212Ser + Leu221CysThr212Gly + Pro214Asp + Thr215Ser + Gly223GlnSer207Glu + Thr212Ser + Pro214Gln + Leu221ProPro205Asn + Gly206Gln + Ser207Asp + Leu221ThrPro205Ser + Thr215Pro + Ser220Asp + Leu221CysGly206Asn + Tyr213Leu + Leu221Ala + Gly223AspSer207Asp + Thr212Gly + Pro214Asn + Thr224AsnAla204Pro + Ser207Glu + Ile209Val + Ala219ThrGly206Gln + Trp208Ile + Tyr218Ile + Gly223AsnAla204His + Trp208Leu + Ile209Gly + Ser216AspAla204Gln + Thr212Gly + Pro214Glu + Ala219SerAla204Asn + Pro205Gln + Ser207Asp + Thr212ProPro205Gly + Trp208Leu + Ser216Glu + Ala219GlnTrp208Tyr + Thr217Gln + Leu221Gln + Thr224GluThr212Gly + Thr217Ser + Tyr218Ala + Gly223AspAla204Gly + Gly206Asn + Thr215Asn + Gly223AspGly206Ser + Tyr210Cys + Tyr213Thr + Thr215GlyAla204Gln + Pro205Ser + Gly206Asn + Ala219AspAla204Ser + Ile209Gly + Leu221Val + Thr224ProGly206Asn + Ser207Glu + Trp208His + Thr217GlyGly206Ser + Trp208Pro + Ala219Thr + Ser222AspAla204Thr + Tyr213Gln + Thr217Ser + Thr224ProPro205Ser + Thr212Ser + Tyr213Ile + Gly223AspGly206Gln + Ile209Asn + Thr215Pro + Ser220GluTrp208Cys + Thr212Gln + Thr215Gly + Ser222GluGly206Asn + Trp208Tyr + Ser222Glu + Gly223AsnPro205Gly + Gly206Asn + Leu221Cys + Ser222AspPro205Gln + Tyr210His + Tyr213Ile + Ser220AspAla204His + Trp208Asn + Thr212Asn + Ala219AspAla204His + Gly206Pro + Leu221Met + Gly223AspAla204Ser + Tyr213Thr + Leu221Met + Ser222AspTyr213Ile + Pro214Gln + Ser216Glu + Thr217ProPro205Gln + Tyr213Cys + Ser220Glu + Thr224GlyGly206Asn + Tyr213Asn + Tyr218Pro + Ser220GluPro205Ser + Gly206Pro + Tyr218Asp + Gly223ProAla204Pro + Pro205Ser + Thr217Asp + Thr224SerPro205Asn + Trp208Ile + Thr212Gln + Thr217GlyIle209Cys + Pro214Ser + Ser220Asp + Gly223GlnAla204His + Pro205Asn + Trp208Glu + Tyr210HisIle209Gly + Thr215Asn + Thr217Asn + Ala219ProPro205Gly + Pro214Asn + Tyr218Ile + Ser222AspAla204Ser + Pro205Ser + Thr212Asn + Tyr213IlePro205Gln + Ser207Glu + Tyr210Pro + Thr215SerPro205Gln + Ser207Glu + Thr212Gln + Tyr213SerAla204Ser + Ile209Asn + Tyr213Cys + Ser216AspTrp208Ala + Ile209Gln + Thr217Glu + Gly223AsnTrp208Pro + Pro214Gly + Tyr218Leu + Leu221AlaGly206Pro + Thr215Ser + Ser216Glu + Leu221CysAla204Ser + Trp208Ile + Ile209Val + Thr224AspTyr210Pro + Thr212Gln + Thr215Asn + Thr217AspTyr210Ser + Tyr213Cys + Pro214Asn + Thr217AsnThr212Gln + Tyr218Met + Ser222Asp + Gly223GlnPro205Ser + Trp208Met + Ala219Gln + Leu221AspAla204Asn + Tyr210Cys + Tyr213Asn + Tyr218LeuAla204His + Pro205Gly + Tyr218Ser + Ser222AspGly206Pro + Tyr213Asn + Pro214Asn + Tyr218SerAla204Pro + Ile209His + Thr215Gln + Leu221ThrAla204Asn + Gly206Gln + Thr212Gly + Gly223GluThr215Gly + Tyr218Ala + Gly223Glu + Thr224ProPro205Gln + Tyr213Gly + Ser220Asp + Leu221ThrPro205Gly + Trp208His + Tyr210His + Leu221GlnTyr210Leu + Thr212Pro + Tyr218Gln + Ala219GluGly206Gln + Thr212Gln + Tyr218His + Gly223AspTrp208Ile + Tyr213His + Thr215Ser + Ser222GluTrp208Val + Pro214Asn + Thr215Gly + Leu221AspPro205Asn + Tyr213Met + Pro214Ser + Thr224SerPro205Gly + Trp208Met + Ile209Leu + Tyr213MetPro205Gly + Thr217Glu + Tyr218Leu + Ala219HisTrp208Val + Tyr213Gly + Ser216Asp + Leu221ThrTyr210Gln + Thr212Pro + Tyr218Gly + Gly223GluPro205Gly + Trp208Asp + Ile209His + Tyr210AlaTrp208Ala + Ile209Leu + Tyr210Glu + Tyr213CysGly206Asn + Trp208Leu + Tyr213Val + Ser216AspAla204Pro + Pro205Gly + Thr217Glu + Leu221AlaTrp208Asn + Thr212Asn + Tyr213His + Ser216GluAla204His + Thr212Gly + Ala219Gln + Leu221ThrGly206Gln + Thr212Pro + Tyr213Met + Thr224ProTrp208Thr + Ile209Gln + Tyr213Ser + Ser222AspAla204Asn + Ile209Asn + Tyr213Pro + Leu221ProPro205Gly + Tyr210His + Tyr218Ile + Leu221AsnAla204Pro + Tyr210Cys + Pro214Gln + Thr215ProPro205Ser + Tyr213Ser + Ser216Asp + Thr217GlyPro205Asn + Trp208Ser + Ala219Glu + Ser220AspTyr210Ala + Tyr218Met + Ala219Glu + Ser220AspAla204Gly + Pro205Gln + Pro214Asp + Thr215AspTyr210Ser + Thr215Asn + Ser222Glu + Gly223GluGly206Pro + Tyr213Val + Ser222Glu + Gly223AspPro214Ser + Leu221Glu + Ser222Asp + Gly223GlnPro205Gln + Ser216Asp + Thr217Glu + Ala219ThrTrp208Leu + Thr212Gln + Ser220Glu + Leu221GluGly206Gln + Ser220Asp + Leu221Asp + Gly223AsnThr217Asp + Tyr218Asp + Gly223Ser + Thr224GlyPro205Gln + Ile209Ala + Thr217Glu + Tyr218GluPro214Gln + Thr215Asp + Ser216Glu + Thr224ProAla204His + Ser207Glu + Tyr213Cys + Ser222AspPro205Gly + Ser207Glu + Thr212Gln + Ser222AspSer207Glu + Tyr210Ser + Tyr218Leu + Ser222GluSer207Glu + Tyr218Cys + Ser222Glu + Thr224ProPro205Ser + Trp208Glu + Leu221Glu + Ser222GluTrp208Glu + Ile209Asn + Thr217Gly + Ser222GluSer216Glu + Thr217Asp + Tyr218Glu + Thr224GlyTrp208Asp + Tyr218Leu + Ala219Thr + Leu221GluAla204His + Thr212Gly + Thr215Glu + Thr217GluPro214Asp + Ser216Asp + Ala219His + Gly223AsnTyr210Pro + Pro214Asp + Ser216Glu + Tyr218IlePro205Gln + Tyr210Leu + Pro214Glu + Thr217AspAla204Ser + Ser207Glu + Pro214Ser + Gly223AspSer207Asp + Thr212Pro + Thr217Gly + Gly223AspSer207Glu + Pro214Asn + Ser220Glu + Ser222AspPro205Ser + Thr217Pro + Ala219Glu + Leu221GluTrp208Gln + Tyr210Glu + Thr212Asn + Leu221GluTrp208Gln + Thr215Pro + Ser220Glu + Ser222GluGly206Pro + Thr217Gly + Ser220Asp + Ser222GluGly206Gln + Tyr210Gly + Ser220Glu + Ser222AspSer207Glu + Thr212Asn + Tyr218Pro + Leu221AspAla204Asn + Ser207Glu + Trp208His + Leu221AspTyr213Ile + Tyr218Leu + Ser222Asp + Thr224GluGly206Gln + Tyr210Met + Ser222Asp + Thr224GluIle209Ser + Tyr210His + Ser222Glu + Thr224AspAla204Ser + Tyr218Asp + Ala219Pro + Ser220GluGly206Gln + Thr217Ser + Tyr218Glu + Ser220GluPro214Asp + Tyr218Asp + Gly223Ser + Thr224GlyGly206Gln + Tyr210Gly + Pro214Glu + Tyr218GluTrp208Asp + Thr217Gln + Ser220Asp + Gly223AspSer207Glu + Tyr210Ala + Thr215Asn + Ser220AspSer207Glu + Thr217Gly + Ser220Asp + Leu221GlnSer207Glu + Thr217Gly + Tyr218Asn + Ser220AspPro205Asn + Ser207Asp + Thr212Gly + Ser220AspSer207Glu + Thr212Asn + Ala219Gly + Ser220AspGly206Pro + Ser207Glu + Trp208Asp + Thr224AspSer207Asp + Trp208Thr + Tyr218Gln + Thr224GluAla204Thr + Ser207Asp + Leu221Ala + Thr224GluSer207Asp + Ala219Glu + Ser220Asp + Gly223SerSer207Glu + Tyr210Glu + Thr215Gln + Thr224AsnAla204Gln + Tyr210Glu + Thr212Asn + Ser222Asp表26環6-五取代變異體Gly206Asn + Tyr210Gly + Thr212Gln + Tyr218Ile +Thr224GlyAla204Asn + Pro205Gly + Trp208Glu + Thr215Asn +Gly223ProPro205Gln + Gly206Pro + Trp208Asn + Thr212Gln +Thr224SerAla204Ser + Pro205Ser + Tyr213Ser + Thr215Ser +Ala219GlyAla204Gly + Gly206Gln + Thr212Pro + Ser220Asp +Leu221SerPro214Ser + Thr215Gln + Tyr218Val + Ser222Asp +Thr224AsnAla204Pro + Ser207Asp + Tyr210Cys + Thr212Pro +Thr215ProGly206Asn + Pro214Gln + Ala219Gly + Gly223Asn +Thr224AspAla204Gln + Pro205Ser + Gly206Ser + Tyr210Gln +Ala219AspGly206Pro + Ser207Asp + Trp208Met + Thr215Pro +Leu221ProAla204Asn + Pro205Gln + Tyr210Cys + Thr212Asn +Leu221SerAla204Asn + Gly206Gln + Trp208Thr + Ile209Ser +Ser222AspPro205Gln + Tyr213Leu + Thr217Glu + Tyr218Met +Leu221GlnAla204Pro + Trp208Met + Ile209Val + Tyr210Leu +Ser216GluTrp208Ser + Ile209Ser + Tyr218His + Ala219His +Gly223ProTrp208Pro + Ile209Thr + Thr212Gly + Thr215Gly +Thr224GluIle209Val + Pro214Ser + Ser216Asp + Tyr218Met +Leu221MetGly206Gln + Ile209Gln + Ser220Asp + Leu221Gly +Thr224SerPro205Gln + Trp208Cys + Thr212Asn + Tyr213Cys +Pro214AspPro205Gln + Trp208Gln + Thr217Asp + Gly223Ser +Thr224GlySer207Glu + Thr212Ser + Thr217Gly + Tyr218Gln +Leu221HisTyr210His + Thr212Ser + Leu221Glu + Gly223Ser +Thr224ProTrp208Phe + Tyr213Met + Thr215Pro + Ser220Glu +Thr224GlyGly206Ser + Trp208Phe + Ile209Ser + Tyr213Cys +Ser216GluAla204Gln + Pro205Ser + Tyr210His + Tyr218Pro +Thr224GluPro205Gln + Gly206Gln + Leu221His + Ser222Asp +Gly223GlnAla204Gly + Ile209His + Tyr210Met + Ser216Glu +Thr217ProGly206Pro + Ile209Ala + Tyr210Gly + Leu221Asp +
Gly223GlnGly206Asn + Tyr210Met + Thr215Glu + Gly223Asn +Thr224GlyGly206Gln + Ile209Pro + Ser216Asp + Leu221Cys +Gly223ProTrp208Thr + Tyr210Val + Thr212Ser + Pro214Asp +Gly223GlnThr212Gly + Pro214Gln + Thr215Ser + Leu221Gly +Thr224GluAla204Thr + Thr212Ser + Thr217Ser + Gly223Asp +Thr224ProPro205Ser + Ile209Pro + Tyr213Ile + Ala219Pro +Ser222GluTrp208Gly + Thr212Gln + Tyr218Gly + Ser222Glu +Gly223SerGly206Pro + Ile209Pro + Tyr213Ser + Thr217Gly +Tyr218ValPro205Gln + Tyr213Cys + Pro214Gln + Ser220Glu +Thr224GlyGly206Asn + Thr212Gly + Tyr213Asn + Tyr218Pro +Ser220GluAla204Pro + Pro205Ser + Thr212Gln + Thr217Asp +Thr224SerPro205Asn + Trp208Ile + Thr212Gln + Pro214Ser +Thr217GlyIle209Cys + Thr217Gly + Ser220Asp + Leu221Gln +Gly223GlnIle209Thr + Thr215Gly + Ser216Glu + Tyr218Leu +Ala219ProIle209Met + Pro214Gly + Thr217Gly + Tyr218Cys +Leu221MetPro205Gly + Pro214Asn + Tyr218Ile + Ala219Gly +Ser222AspTyr210Thr + Thr212Ser + Pro214Ser + Leu221Asn +Ser222AspGly206Asn + Tyr210Ile + Pro214Gln + Ala219Asn +Leu221ValPro205Asn + Thr215Ser + Ala219Ser + Leu221Ala +Thr224AspGly206Pro + Pro214Asn + Thr215Gln + Tyr218Ser +Leu221ThrAla204Pro + Gly206Gln + Ile209His + Thr212Gly +Gly223GluPro205Gln + Tyr213Gly + Tyr218Ala + Gly223Glu +Thr224ProPro205Ser + Pro214Gly + Thr217Pro + Ala219Thr +Leu221GluAla204Pro + Pro205Asn + Ser207Asp + Tyr210Ala +Pro214SerGly206Ser + Tyr213Thr + Ala219Gln + Ser220Glu +Gly223SerPro205Gly + Thr212Asn + Thr215Pro + Ser216Glu +Leu221AsnSer207Glu + Ile209Ala + Thr212Asn + Thr215Gln +Tyr218MetTyr213His + Thr215Gln + Thr217Asn + Ala219Glu +Leu221SerGly206Gln + Tyr213Asn + Thr215Ser + Tyr218Ala +Leu221AlaTrp208Met + Tyr210Met + Tyr218Gln + Ser220Glu +Thr224SerAla204Thr + Pro205Ser + Ile209Leu + Thr217Asn +Thr224ProPro205Gly + Ile209Leu + Thr215Asn + Ser216Glu +Gly223SerIle209Ser + Tyr213Leu + Thr217Asp + Tyr218Asn +Ala219HisAla204Ser + Pro205Ser + Ser207Glu + Leu221Val +G1y223ProAla204His + Thr212Gly + Ala219Gln + Leu221Thr +Gly223GluPro205Ser + Gly206Gln + Thr212Pro + Gly223Pro +Thr224ProAla204Asn + Trp208Thr + Tyr213Pro + Leu221Pro +Ser222AspPro205Gly + Ile209Met + Tyr218Met + Leu221Val +Gly223AsnThr212Gly + Tyr218Ile + Ala219Asn + Ser220Glu +Thr224ProAla204Gln + Pro205Gln + Gly206Gln + Ile209Met +Ser216AspAla204Thr + Thr212Gly + Ala219Asn + Gly223Asp +Thr224AsnAla204Pro + Trp208His + Tyr213Leu + Thr217Ser +Ser222GluPro205Ser + Ser207Asp + Ile209Asn + Tyr213Met +Thr217GlnGly206Pro + Trp208Ser + Thr212Gly + Tyr213Cys +Gly223GluAla204Asn + Gly206Pro + Trp208Val + Pro214Asn +Tyr218GluIle209Gly + Tyr210Ala + Tyr218Pro + Gly223Ser +Thr224SerPro205Gln + Tyr213His + Tyr218Met + Ser220Glu +Leu221GlyAla204Asn + Tyr210Cys + Thr212Gln + Tyr218Leu +Ser222AspGly206Ser + Ile209Leu + Thr212Gly + Pro214Gln +Thr215GluSer207Asp + Pro214Ser + Thr215Asn + Thr217Asn +Leu221SerPro205Gly + Trp208Thr + Thr217Gly + Ser220Glu +Gly223SerGly206Ser + Trp208Glu + Tyr210Gln + Thr217Gln +Leu221ProGly206Gln + Ser207Glu + Thr212Asn + Pro214Ser +
Leu221AlaAla204Ser + Trp208Leu + Ile209Asn + Thr215Asp +Thr224AsnGly206Ser + Ile209Pro + Thr217Ser + Ser220Glu +Gly223GlnPro205Gln + Trp208Pro + Thr212Pro + Pro214Glu +Ala219SerGly206Pro + Ser207Glu + Tyr210Met + Thr212Gly +Tyr213LeuAla204His + Tyr210Pro + Tyr213Ala + Thr217Glu +Thr224SerTrp208Cys + Ile209Met + Tyr210Ala + Tyr213Met +Ala219AsnTrp208Thr + Tyr210Ala + Tyr218Met + Ala219Glu +Ser220AspTrp208Asn + Ile209Pro + Ala219Asp + Ser220Asp +Gly223SerPro205Ser + Tyr213Thr + Pro214Asp + Thr215Asp +Tyr218ValIle209Ser + Tyr213Pro + Leu221Met + Ser222Asp +Gly223AspAla204Thr + Thr217Gln + Tyr218Ile + Ser222Glu +Gly223AspPro205Ser + Tyr210Ala + Thr215Asn + Ser222Glu +Gly223GluTyr210Pro + Thr212Pro + Leu221Asn + Ser222Asp +Gly223AspTrp208Pro + Ile209Pro + Tyr213Ile + Ser222Asp +Gly223AspGly206Gln + Tyr213Gly + Thr217Gly + Ser222Glu +Gly223GluTrp208Ser + Ile209Pro + Thr212Pro + Ser216Glu +Thr217AspGly206Asn + Tyr218Met + Ala219His + Ser220Glu +Leu221AspIle209Ala + Thr215Glu + Ser216Glu + Gly223Gln +Thr224AsnIle209Gly + Pro214Ser + Thr215Glu + Ser216Asp +Tyr218IleAla204Ser + Trp208Val + Ile209Ser + Thr215Glu +Ser216AspThr212Pro + Thr215Asp + Ser216Glu + Thr217Glu +Thr224SerAla204His + Ser207Glu + Trp208His + Tyr210His +Ser222AspSer207Glu + Tyr213Asn + Leu221Thr + Ser222Asp +Thr224GlyGly206Ser + Ser207Glu + Trp208Pro + Ala219Thr +Ser222AspPro205Gly + Ser207Glu + Thr212Gln + Tyr213Gln +Ser222AspPro205Ser + Ser207Glu + Tyr213His + Thr217Pro +Ser222GluGly206Pro + Ser207Asp + Tyr213Ile + Tyr218Asn +Ser222GluGly206Pro + Trp208Glu + Tyr210Cys + Pro214Gln +Ser220AspTyr210Glu + Tyr213Gly + Tyr218Leu + Ala219Asp +Thr224SerGly206Pro + Ser207Asp + Leu221Asp + Ser222Glu +Thr224GlnTyr213Ser + Pro214Ser + Thr215Glu + Thr217Asp +Tyr218AlaAla204Asn + Tyr210Gln + Thr212Asn + Pro214Glu +Ser216AspPro205Gln + Gly206Pro + Pro214Glu + Ser216Asp +Thr224SerSer207Asp + Thr212Pro + Thr215Gly + Thr217Gly +Gly223AspSer207Glu + Tyr210Val + Pro214Asn + Leu221Thr +Gly223GluSer207Asp + Ile209Thr + Tyr210Met + Thr212Pro +Gly223AspSer207Glu + Tyr210Ser + Tyr218Leu + Ser220Asp +Ser222GluSer207Asp + Thr217Gln + Ser220Asp + Ser222Asp +Thr224AsnTrp208Thr + Thr217Glu + Tyr218Ser + Ala219Glu +Leu221CysPro205Gly + Thr212Ser + Ala219Gln + Ser220Glu +Ser222GluGly206Pro + Trp208His + Tyr210Gly + Ser220Glu +Ser222AspAla204Gln + Gly206Ser + Trp208Gln + Ser220Glu +Ser222GluAla204Ser + Tyr213Thr + Ser220Asp + Leu221Met +Ser222AspThr212Gly + Thr215Asp + Ser216Asp + Thr217Gly +Tyr218AspAla204Asn + Tyr210Glu + Tyr213Ala + Ser220Glu +Ser222GluAla204His + Gly206Pro + Ser216Glu + Tyr218Asp +Leu221SerGly206Pro + Ile209Ser + Tyr210His + Ser222Glu +Thr224AspTrp208Met + Tyr218Leu + Ala219Pro + Ser222Glu +Thr224GluTrp208Val + Tyr210Leu + Thr212Asn + Tyr218Asp +Ser220GluAla204Ser + Tyr213Gln + Tyr218Asp + Ala219Pro +Ser220GluThr212Asn + Tyr213Gln + Thr217Ser + Tyr218Glu +Ser220GluPro214Ser + Leu221Glu + Ser222Asp + Gly223Ser +Thr224AspPro205Gly + Tyr210Cys + Ala219Asp + Ser220Glu +
Ser222AspGly206Gln + Tyr210Gly + Pro214Glu + Thr217Pro +Tyr218GluTrp208Asp + Pro214Ser + Thr217Gln + Ser220Asp +Gly223AspSer207Glu + Tyr210Ala + Thr215Asn + Ser220Asp +Leu221AlaSer207Asp + Trp208Cys + Ile209Ala + Thr215Asn +Ser220GluSer207Asp + Tyr210Met + Tyr218Asn + Ser220Glu +Leu221CysSer207Glu + Tyr210Pro + Thr215Gln + Tyr218Cys +Ser220GluSer207Glu + Thr212Ser + Pro214Gln + Ser220Asp +Leu221ProPro205Asn + Ser207Asp + Thr215Pro + Ser220Asp +Leu221ThrTrp208Glu + Ile209Asn + Thr217Gly + Ala219Asp +Ser222GluPro205Asn + Ser207Glu + Tyr213Leu + Ser220Asp +Gly223GluAla204Ser + Pro205Gly + Ser207Asp + Thr212Ser +Thr224AspAla204Pro + Gly206Gln + Ser207Glu + Tyr218Asn +Thr224GluGly206Ser + Ser207Glu + Tyr210Met + Tyr213Cys +Thr224AspPro205Asn + Ser207Glu + Ile209Val + Tyr213Pro +Thr224GluSer207Asp + Ile209Val + Tyr213Ala + Gly223Gln +Thr224GluPro205Asn + Ser207Glu + Ala219Glu + Ser220Glu +Gly223GlnGly206Asn + Ser207Glu + Trp208Asp + Ala219Asp +Thr224ProAla204Gly + Gly206Ser + Ser207Asp + Tyr210Asp +Leu221GlyAla204Pro + Ser207Glu + Ile209Thr + Tyr210Glu +Tyr218LeuIle209Thr + Tyr213Gly + Ala219Asp + Ser222Glu +Gly223AspPro205Gln + Tyr213Gln + Tyr218Glu + Leu221Glu +Ser222AspGly206Gln + Ser207Glu + Thr212Asn + Ser220Glu +Thr224GluSer207Glu + Thr217Gly + Tyr218Asn + Ser220Asp +Thr224GluAla204Thr + Tyr210Cys + Ser220Glu + Leu221Ser +Gly223GluIle209Pro + Ser216Glu + Ala219Glu + Ser220Asp +Gly223AsnGly206Gln + Tyr210Met + Thr212Pro + Ala219Glu +Ser222AspPro205Asn + Tyr210Glu + Thr217Asp + Tyr218Ala +Thr224ProAla204Gly + Trp208Gln + Thr212Gln + Ser216Asp +Ala219GluAla204Asn + Trp208Ala + Ser216Asp + Ala219Glu +Thr224GlyAla204Asn + Tyr210Asp + Thr215Glu + Thr217Glu +Ala219GlyTrp208Leu + Ile209Gly + Thr215Asp + Thr217Glu +Ser220AspThr212Pro + Tyr213Leu + Ser216Glu + Tyr218Glu +Leu221GluThr217Asp + Ala219Glu + Leu221Asn + Ser222Glu +Thr224SerPro205Ser + Trp208Glu + Tyr218Glu + Leu221Gly +Ser222GluSer207Glu + Thr215Pro + Thr217Asp + Ser220Glu +Gly223ProThr217Asp + Ala219Asn + Leu221Glu + Ser222Glu +Gly223AsnTrp208Tyr + Ile209Asn + Thr217Glu + Leu221Glu +Ser222AspAla204Gly + Tyr210Glu + Pro214Glu + Ser216Glu +Thr217AsnPro214Asp + Thr215Gln + Ser216Glu + Ser220Glu +Leu221MetAla204Gln + Tyr210Glu + Thr212Asn + Thr217Glu +Ser222AspTrp208Leu + Ile209Val + Thr215Asp + Thr217Glu +Leu221GluGly206Pro + Ile209Val + Tyr213Pro + Thr215Asp +Ala219GluPro205Asn + Ser207Glu + Tyr218Asp + Ser222Asp +Thr224GlyGly206Ser + Tyr210Asp + Pro214Asp + Thr215Pro +Leu221AspTyr210Ser + Pro214Asp + Tyr218Cys + Ser220Glu +Ser222GluSer207Asp + Ile209Ser + Thr212Ser + Pro214Asp +Thr215Asp表27環6-六取代變異體Gly206Asn + Tyr210Gly + Thr212Gln + Tyr213Gly +Tyr218Ile + Thr224GlyPro205Asn + Gly206Ser + Tyr213Gly + Ser216Asp +Thr217Ser + Thr224AsnAla204Ser + Pro205Ser + Tyr213Ser + Tyr218His +Ala219Gln + Leu221AsnSer207Glu + Thr212Gln + Thr215Pro + Tyr218Gln +Leu221Met + Gly223GlnPro205Ser + Ser207Asp + Tyr210Val + Tyr213Leu +Leu221Gln + Thr224GlyAla204Ser + Trp208Cys + Ile209His + Thr215Glu +Thr217Pro + Gly223SerAla204Gly + Ile209Asn + Tyr210Ile + Ser216Asp +Ala219Asn + Leu221MetAla204Gly + Pro205Gln + Pro214Glu + Thr215Ser +Tyr218Asn + Thr224SerGly206Asn + Trp208Cys + Thr212Gly + Tyr213Gly +Ser216Asp + Leu221HisTrp208Pro + Ile209Gly + Thr215Ser + Ala219Thr +Leu221Thr + Thr224AspPro205Gly + Gly206Gln + Ile209Gly + Thr212Asn +Pro214Asn + Ser220GluPro205Gly + Trp208Leu + Thr217Gln + Leu221Gln +Gly223Gln + Thr224GluAla204Gly + Gly206Asn + Tyr210Cys + Tyr213Thr +Gly223Asp + Thr224AsnGly206Asn + Trp208His + Ile209Gly + Thr217Gly +Leu221Val + Thr224ProAla204Thr + Thr212Gln + Tyr213Gln + Thr217Ser +Ser222Asp + Thr224ProTrp208Gly + Thr212Gln + Tyr213Leu + Tyr218Gly +Ser222Glu + Gly223SerPro205Ser + Trp208Ser + Tyr213Asn + Tyr218Gln +Ser222Asp + Thr224GlnAla204His + Pro205Gly + Ile209Asn + Tyr213Gln +Thr215Ser + Thr217SerAla204Thr + Gly206Asn + Trp208Leu + Thr215Ser +Ser216Asp + Ala219HisGly206Ser + Thr212Gln + Tyr213Val + Tyr218Cys +Ser220Glu + Thr224GlnGly206Asn + Trp208Ile + Ile209Ser + Tyr218Gln +Gly223Glu + Thr224GlyGly206Asn + Ser207Glu + Ile209Cys + Thr212Ser +Thr217Pro + Ala219HisAla204Asn + Tyr210Cys + Tyr213Asn + Tyr218Leu +Leu221His + Ser222GluGly206Pro + Tyr213Asn + Pro214Asn + Thr215Gln +Tyr218Ser + Leu221ThrAla204Pro + Pro205Asn + Gly206Gln + Ile209His +Thr212Gly + Gly223GluPro205Gln + Tyr213Gly + Thr215Gly + Tyr218Ala +Gly223Glu + Thr224ProAla204His +Ser207Glu + Trp208Gly + Ile209His +Tyr213Cys + Thr215SerGly206Ser + Trp208His + Tyr213Thr + Ala219Gln +Ser220Glu + Gly223SerTyr213His + Thr215Gln + Thr217Asn + Tyr218Ala +Ala219Glu + Leu221SerAla204Asn + Pro205Ser + Gly206Asn + Ile209Leu +Thr215Asp + Thr224ProPro205Ser + Ile209Gly + Tyr213Leu + Thr217Asp +Tyr218Asn + Ala219HisAla204Asn + Pro205Gln + Ser207Asp + Tyr210Val +Thr212Gly + Leu221ThrAla204Asn + Trp208Thr + Ile209Gln + Tyr213Pro +Leu221Pro + Ser222AspAla204Ser + Gly206Ser + Tyr213Gly + Pro214Gln +Tyr218Val + Ser222AspAla204Pro + Tyr210Cys + Tyr213Ser + Pro214Gln +Thr215Pro + Thr217GlyThr212Ser + Pro214Asp + Thr217Gly + Ala219Thr +Leu221Pro + Thr224GlnPro205Gln + Gly206Gln + Ser207Glu + Tyr210Gly +Thr212Pro + Tyr218GlyPro205Gln + Gly206Pro + Trp208Asn + Ile209Met +Tyr213Ser + Ala219AspAla204Asn + Ser207Glu + Trp208Ala + Tyr213Asn +Leu221Thr + Thr224GlyAla204Pro + Trp208His + Ile209Asn + Tyr213Leu +Thr217Ser + Ser222GluGly206Pro + Trp208Ser + Tyr210Val + Thr212Gly +Tyr213Cys + Ala219AspPro205Gly + Gly206Gln + Ile209Cys + Tyr210His +Thr215Gln + Ala219ThrTrp208Gly + Ile209Ser + Thr212Gln + Tyr218Met +Leu221Ser + Gly223SerPro205Asn + Ile209Asn + Thr212Asn + Thr217Glu +Gly223Pro + Thr224SerPro205Gly + Trp208Thr + Thr212Ser + Ser216Glu +Thr217Gly + Gly223SerGly206Ser + Trp208Glu + Tyr210Gln + Thr215Gly +Thr217Gln + Leu221ProGly206Ser + Ser207Glu + Trp208Leu + Thr212Asn +Leu221Ala + Thr224AsnAla204Asn + Pro205Gln + Gly206Gln + Trp208Ile +Ser216Asp + Tyr218CysAla204Thr + Tyr210Cys + Tyr213Ser + Thr215Ser +Ser216Asp + Ala219HisTrp208Leu + Ile209Asn + Thr212Gly + Thr215Ser +Tyr218Pro + Leu221GlyPro205Gly + Gly206Gln + Tyr218Gly + Ala219Asn +Leu221Gln + Thr224SerPro205Gln + Trp208Val + Tyr210Pro + Thr215Gly +Ala219Ser + Thr224GluPro205Gly + Trp208Met + Ile209Ser + Thr212Asn +Tyr218Ala + Ala219ThrThr212Gly + Tyr213Asn + Pro214Ser + Ser216Glu +Thr217Pro + Leu221ThrAla204Gly + Trp208Pro + Ile209Pro + Tyr213Gln +Tyr218Ser + Ser220GluAla204Gly + Gly206Asn + Ile209Met + Thr212Pro +Ala219Glu + Leu221MetPro205Asn + Gly206Pro + Trp208Leu + Tyr210Pro +Thr217Gln + Tyr218GluPro205Asn + Gly206Gln + Tyr213Val + Ser220Asp +Gly223Ser + Thr224AsnAla204Thr + Pro205Gly + Gly206Gln + Pro214Gln +Leu221Val + Gly223GlnPro205Ser + Ile209Gln + Thr212Pro + Ala219Gly +Ser220Glu + Gly223GlnAla204Gly + Gly206Gln + Ile209Leu + Thr212Asn +Pro214Gln + Thr215SerGly206Asn + Trp208Ser + Thr212Ser + Pro214Asn +Ser220Glu + Gly223SerAla204His + Tyr213Leu + Pro214Asn + Thr217Pro +Ser220Glu + Gly223GlnGly206Pro + Ile209Cys + Pro214Gln + Thr217Ser +Tyr218Cys + Ala219GlnAla204Thr + Tyr210Ala + Thr215Gly + Thr217Gln +Ala219Pro + Ser220AspSer207Asp + Ile209Thr + Thr212Gly + Thr217Pro +Ala219Gly + Thr224ProAla204Gly + Ser207Glu + Thr212Asn+ Pro214Ser +Tyr218Ser + Ala219SerPro205Gln + Gly206Gln + Thr212Ser + Tyr213Ala +Thr215Ser + Gly223GluPro205Ser + Tyr210Ala + Pro214Ser + Thr215Asn +Ser222Asp + Gly223GluIle209Pro + Tyr210Pro + Thr212Pro + Leu221Asn +Ser222Asp + Gly223AspPro205Ser + Thr212Ser + Tyr213Ile + Ala219Pro +Ser222Glu + Gly223AspAla204Asn + Ile209Cys + Tyr213Cys + Tyr218Ala +Leu221Asp + Ser222GluIle209Val + Thr212Asn + Tyr213Met + Thr217Asn +Gly223Asp + Thr224AspPro205Gly + Ile209Asn + Thr212Asn + Ser216Glu +Thr217Glu + Tyr218GlnAla204His + Trp208Ser + Thr212Pro + Tyr213Pro +Ser216Glu + Thr217AspPro205Ser + Ile209Ala + Thr212Gly + Ser216Asp +Thr217Glu + Ala219ThrPro205Ser + Trp208Met + Pro214Asn + Ala219Gln +Ser220Glu + Leu221AspAla204Pro + Gly206Ser + Tyr213Ile + Ser220Glu +Leu221Asp + Gly223SerGly206Pro + Tyr210Gly + Tyr213His + Thr217Glu +Tyr218Asp + Leu221AsnThr212Pro + Tyr213Ser + Thr215Asp + Ser216Asp +Tyr218Pro + Gly223GlnSer207Glu + Ile209Asn + Thr212Ser + Thr217Gly +Ser222Glu + Thr224GlnPro205Gly + Ser207Glu + Tyr210Ser + Pro214Gln +Ser222Asp + Gly223AsnGly206Pro + Ser207Asp + Trp208Ser + Pro214Ser +Leu221Asn + Ser222AspGly206Gln + Ser207Asp + Ile209Cys + Tyr210Met +Thr217Asn + Ser222GluGly206Pro + Ser207Glu + Ile209Pro + Tyr210Ile +Leu221Asn + Ser222GluPro205Ser + Ser207Glu + Trp208His + Thr215Pro +Leu221Asn + Ser222GluPro205Asn + Ser207Asp + Thr215Gln + Tyr218Ile +Leu221Gly + Ser222GluAla204Pro + Ser207Asp + Tyr210Cys + Thr212Pro +Thr215Pro + Ser222GluPro205Gly + Pro214Asp + Thr215Glu + Ser216Asp +Thr217Pro + Gly223GlnIle209Thr + Thr217Gly + Ala219Glu + Ser220Asp +Leu221Glu + Gly223SerAla204Gln + Ser207Asp + Ile209Thr + Tyr218Asn +Ser222Glu + Gly223AspPro205Ser + Ser207Glu + Pro214Gly + Thr215Asn +Ser222Asp + Gly223GluPro205Asn + Gly206Pro + Trp208Asp + Thr212Gln +Ser222Asp + Gly223SerGly206Gln + Pro214Asp + Thr215Asp + Thr217Asp +Gly223Ser + Thr224GlyAla204His + Ile209Thr + Ser216Asp + Thr217Asp +Tyr218Asp + Leu221SerThr212Asn + Pro214Gln + Tyr218Met + Ser222Glu +Gly223Asp + Thr224GluAla204Gln + Gly206Pro + Tyr210Glu + Pro214Gly +Ala219Asp + Leu221GlnAla204Gly + Pro205Gln + Trp208Asp + Ile209Gln +Pro214Gly + Leu221GluAla204Asn + Gly206Gln + Ser207Glu + Trp208His +Leu221Asp + Ser222AspTrp208Gly + Tyr213Ser + Pro214Ser + Thr215Glu +Thr217Asp + Tyr218AlaAla204Asn + Pro205Ser + Tyr210Pro + Pro214Asp +Ser216Glu + Tyr218IleGly206Gln + Ile209Leu + Thr212Gly + Pro214Asp +Ser216Glu + Leu221HisPro205Gln + Trp208Glu + Tyr210Asp + Thr212Gln +Tyr213Asn + Thr215SerSer207Asp + Thr212Pro + Thr215Gly + Thr217Gln +Leu221Met + Gly223AspAla204Asn + Pro205Gln + Ser207Glu + Trp208Met +Ala219Pro + Gly223AspAla204Thr + Ser207Asp + Thr212Gly + Ala219Asn +Gly223Asp + Thr224AsnSer207Asp + Pro214Asn + Thr215Gly + Tyr218Ala +Leu221Ser + Gly223AspSer207Glu + Thr212Asn + Tyr213Gln + Tyr218Pro +Ser220Glu + Leu221AspAla204Asn + Gly206Ser + Ser207Asp + Tyr210Thr +Ser220Asp + Leu221AspGly206Ser + Thr212Asn + Thr215Pro + Tyr218Val +Leu221Asp + Gly223AspTrp208Met + Tyr210Thr + Thr212Asn + Ala219Gln +Leu221Asp + Gly223AspGly206Ser + Ser207Glu + Pro214Ser + Ala219Asn +Ser220Glu + Ser222GluAla204His + Ser207Glu + Ala219Ser + Ser220Glu +Leu221His + Ser222AspPro205Ser + Ser207Glu + Tyr213His + Thr217Pro +Ser220Glu + Ser222GluSer207Glu + Ile209Met + Tyr210Gly + Tyr213Leu +Ser220Glu + Ser222AspSer207Asp + Tyr213Pro + Pro214Gln + Ser220Glu +Ser222Glu + Gly223ProGly206Ser + Trp208Gln + Tyr210Glu + Thr212Asn +Thr215Gly + Leu221GluPro205Gly + Thr212Ser + Tyr218Leu + Ala219Gln +Ser220Glu + Ser222GluTrp208Phe + Thr212Asn + Pro214Ser + Ser220Glu +Ser222Asp + Gly223GlnAla204Gly + Gly206Pro + Tyr210Ser + Tyr218Cys +Ser220Glu + Ser222AspPro205Ser + Trp208Leu + Tyr213Ala + Thr217Ser +Ser220Asp + Ser222AspPro205Gly + Trp208His + Tyr210His + Ser220Glu +Leu221Gln + Ser222AspPro205Gly + Thr212Gln + Ala219Thr + Ser220Glu +Ser222Asp + Gly223ProAla204Ser + Gly206Pro + Tyr210Gly + Ser220Glu +Leu221Ser + Ser222AspIle209Ser + Tyr210Ser + Thr212Asn + Tyr213Cys +Ser220Asp + Ser222GluAla204Asn + Pro205Gly + Ser207Asp + Ile209Ser +Tyr213Val + Leu221GluAla204Asn + Gly206Pro + Thr215Glu + Ser216Glu +Thr217Asn + Tyr218GluGly206Pro + Ile209Ser + Tyr210His + Thr217Gly +Ser222Glu + Thr224AspGly206Gln + Thr217Gly + Ser220Asp + Ser222Glu +Gly223Glu + Thr224SerGly206Gln + Trp208Gin + Ile209Gln + Ser216Asp +Tyr218Asp + Ala219GluGly206Asn + Thr212Pro + Pro214Ser + Thr217Glu +Tyr218Glu + Ser220GluGly206Gln + Trp208Ser + Tyr210Asp + Pro214Gln +Thr217Glu + Ala219AspGly206Ser + Ser207Glu + Trp208Pro + Tyr210Asp +Ala219Thr + Ser222AspTrp208Gly + Tyr210His + Thr212Ser + Pro214Glu +Tyr218Asp + Leu221IleAla204Gln + Gly206Gln + Tyr210Gly + Pro214Glu +Thr217Pro + Tyr218GluAla204His + Pro214Glu + Thr215Gln + Thr217Gln +Tyr218Glu + Leu221ProSer207Asp + Ile209Cys + Thr215Pro + Tyr218His +Ala219Thr + Ser220GluPro205Asn + Ser207Glu + Trp208Tyr + Ile209Asn +Thr217Gln + Ser220GluSer207Asp + Pro214Ser + Thr215Asn + Thr217Asn +Ser220Glu + Leu221SerPro205Gly + Ser207Asp + Tyr210Ser + Ser220Asp +Gly223Pro + Thr224SerGly206Gln + Ser207Glu + Ile209Gln + Ser220Asp +Leu221Ala + Thr224SerAla204Ser + Pro205Gly + Ser207Asp + Thr212Ser +Thr215Gln + Ser220GluPro205Ser + Ser207Asp + Thr217Gln + Ala219His +Ser220Glu + Gly223GluSer207Glu + Thr215Gln + Tyr218Val + Ser220Asp +Gly223Glu + Thr224AsnPro205Gly + Ile209Pro + Tyr210Asp + Ala219Asp +Ser222Asp + Gly223SerGly206Asn + Tyr210Gln + Pro214Asn + Thr215Glu +Tyr218Glu + Leu221AlaIle209Asn + Tyr210Asp + Thr212Gln + Ser222Glu +Gly223Pro + Thr224AsnAla204Gly + Gly206Gln + Ser220Asp + Leu221Asp +Gly223Asn + Thr224GluGly206Pro + Thr212Pro + Thr217Pro + Tyr218Asn +Leu221Glu + Thr224GluIle209His + Tyr210Leu + Tyr213Leu + Pro214Asp +Thr217Asp + Ser220GluIle209Asn + Tyr210Ile + Thr212Asn + Thr215Gly +Ser220Asp + Gly223GluGly206Gln + Tyr210Ala + Thr215Asn + Ser216Glu +Thr217Asp + Ser220AspTrp208Thr + Tyr213Pro + Tyr218His + Ala219Glu +Leu221Cys + Ser222AspAla204Gly + Trp208Gln + Thr212Gln + Ser216Asp +Ala219Glu + Gly223SerGly206Ser + Pro214Gly + Ser216Asp + Tyr218Pro +Ala219Glu + Leu221ProPro205Asn + Tyr210Asn + Thr217Glu + Ala219Gln +Ser220Glu + Gly223GlnAla204Thr + Tyr213Ala + Pro214Gln + Thr215Gln +Thr217Asp + Ser220GluTrp208Gln + Tyr210His + Thr217Asp + Tyr218Met +Ser220Asp + Thr224SerAla204Asn + Gly206Pro + Tyr210Asp + Pro214Glu +Ser220Glu + Thr224AsnTyr213Ile + Ser216Glu + Tyr218His + Ala219Glu +Leu221Asp + Thr224SerPro205Ser + Trp208Glu + Tyr218Glu + Leu221Gly +Ser222Glu + Thr224GlySer207Asp + Tyr213Met + Pro214Gly + Thr217Gln +Tyr218Asp + Ala219GluTrp208Phe + Ala219Asp + Leu221Ala + Ser222Asp +Gly223Asn + Thr224GluGly206Pro + Ile209His + Pro214Glu + Ser220Asp +Leu221Glu + Thr224ProTrp208Tyr + Ile209Asn + Thr212Asn + Thr217Glu +Leu221Glu + Ser222AspPro205Gly + Ser216Glu + Thr217Glu + Ala219Gln +Leu221Glu + Gly223GlnPro214Asn + Thr217Pro + Ala219Asp + Ser220Asp +Leu221Gln + Thr224AspAla204His + Gly206Asn + Ile209His + Pro214Glu +Ser216Asp + Ser220GluSer207Asp + Tyr210Glu + Thr217Asp + Tyr218Ala +Leu221Ile + Thr224ProThr212Gly + Tyr213Ala + Pro214Glu + Leu221Glu +Ser222Asp + Thr224SerPro205Gln + Gly206Asn + Thr212Gln + Tyr218Glu +Ser222Asp + Gly223AspPro205Gln + Thr212Gly + Tyr213Ala + Thr217Glu +Tyr218Glu + Ser222GluIle209Gln + Tyr213Ala + Pro214Glu + Ala219Asp +Ser222Asp + Thr224ProAla204Gly + Tyr210Pro + Pro214Glu + Ala219Glu +Leu221Asn + Ser222AspGly206Pro + Thr212Asn + Tyr213Thr + Pro214Asp +Ser220Asp + Ser222GluAla204Thr + Tyr213Ser + Pro214Asp + Ala219Asn +Ser220Asp + Ser222GluAla204His + Gly206Asn + Tyr210Pro + Pro214Glu +Ser220Glu + Ser222GluSer207Asp + Trp208Met + Pro214Glu + Thr215Asp +Thr217Ser + Gly223ProTrp208Cys + Ile209Ser + Pro214Glu + Ser222Asp +Gly223Asp + Thr224GlyPro205Asn + Thr212Asn + Tyr213Leu + Thr215Asp +Ser222Asp + Gly223Glu表28環6-七取代突變變異體Gly206Asn + Tyr210Gly + Thr212Gln + Thr215Gln +Tyr218Ile + Leu221Ile + Thr224GlyAla204Ser + Pro205Ser + Ile209Met + Tyr213Ser +Tyr218His + Ala219Gln + Leu221AsnAla204Gly + Trp208Gln + Thr212Gln + Ser216Asp +Tyr218Gln + Leu221Met + Gly223SerGly206Gln + Ile209Leu + Tyr210Ala + Thr212Gly +Tyr213Gly + Ser216Asp + Gly223GlnAla204Gln + Pro205Asn + Gly206Asn + Trp208Ile +Tyr210Gln + Thr215Gln + Ala219AspAla204Asn + Pro205Gln + Ile209Ala + Tyr213Leu +Thr217Glu + Tyr218Met + Leu221GlnPro205Gly + Ile209Gln + Thr212Ser + Ala219Gln +Ser220Glu + Leu221Cys + Thr224SerGly206Asn + Trp208Cys + Thr212Gly + Tyr213Gly +Ser216Asp + Thr217Gly + Leu221HisAla204Gln + Pro205Ser + Tyr210His + Thr212Pro +Tyr218Pro + Ala219Ser + Thr224GluAla204Gly + Gly206Asn + Tyr210Cys + Tyr213Thr +Thr215Gly + Gly223Asp + Thr224AsnAla204Gln + Pro205Ser + Gly206Gln + Thr212Asn +Tyr218Cys + Ala219His + Gly223GluAla204Ser + Gly206Asn + Ile209Gly + Pro214Gly +Thr217Gly + Leu221Val + Thr224ProPro205Gly + Ile209Gln + Pro214Asn + Tyr218Ile +Ala219Gly + Leu221Met + Ser222AspAla204Asn + Trp208Ala + Ile209Gln + Pro214Gly +Thr217Glu + Tyr218Leu + Gly223AsnGly206Asn + Tyr210Glu + Tyr213Gly + Thr217Gln +Tyr218Leu + Ala219Thr + Thr224SerGly206Pro + Ile209His + Tyr213Asn + Pro214Asn +Thr215Gln + Tyr218Ser + Leu221ThrPro205Gly + Gly206Ser + Trp208His + Tyr213Thr +Ala219Gln + Ser220Glu + Gly223SerPro205Gln + Gly206Ser + Trp208Val + Tyr210Leu +Thr212Asn + Tyr218Met + Ser220GluAla204His + Gly206Asn + Tyr210Ser + Pro214Ser +Thr217Gln + Tyr218His + Ser222AspGly206Gln + Tyr213Asn + Thr215Gly + Ser216Glu +Thr217Gly + Tyr218His + Gly223AsnGly206Gln + Ser207Glu + Tyr210Gly + Thr212Pro +Tyr213Met + Gly223Pro + Thr224ProAla204Asn + Trp208Thr + Ile209Gln + Tyr213Pro +Thr217Ser + Leu221Pro + Ser222AspAla204Ser + Gly206Gln + Trp208Asn + Thr212Ser +Ala219Gln + Ser220Glu + Leu221AsnPro205Gly + Gly206Gln + Ile209Asn + Tyr213Val +Thr215Gln + Thr217Ser + Ala219ThrAla204Ser + Pro205Gln + Ile209Asn + Thr212Asn +Thr217Glu + Gly223Pro + Thr224SerThr212Gly + Tyr213Ile + Pro214Gly + Tyr218Gly +Ala219Pro + Leu221Asn + Thr224GluAla204Asn + Tyr210Asp + Thr212Pro + Tyr213Asn +Thr217Asn + Tyr218Cys + Leu221HisAla204Gln + Ile209Cys + Tyr210Gln + Thr212Gly +Tyr213Gln + Gly223Pro + Thr224AspPro205Gln + Gly206Gln + Trp208Thr + Tyr210Leu +Thr215Gly + Thr217Gly + Tyr218LeuTyr210Ala + Tyr213Val + Pro214Gln + Ala219Ser +Leu221Cys + Ser222Glu + Thr224AsnAla204Asn + Gly206Pro + Ser207Asp + Trp208His +Ile209His + Pro214Gln + Leu221GlnAla204Asn + Gly206Asn + Trp208Ile + Ile209Asn +Ser216Glu + Ala219Pro + Gly223AsnAla204Ser + Gly206Ser + Ile209Ala + Thr212Ser +Pro214Gln + Tyr218Asp + Leu221HisGly206Asn + Ile209Pro + Tyr213Cys + Thr215Gln +Ser216Glu + Thr217Pro + Ala219HisAla204Gln + Trp208His + Tyr210Gly + Thr212Gln +Tyr218Met + Ala219Asn + Ser220GluAla204Pro + Ser207Asp + Tyr213Val + Pro214Gly +Thr215Gly + Tyr218Ala + Leu221GlnPro205Asn + Gly206Pro + Trp208Leu + Tyr210Pro +Pro214Asn + Thr217Gln + Tyr218GluPro205Asn + Gly206Gln + Tyr213Val + Ala219Gln +Ser220Asp + Gly223Ser + Thr224AsnGly206Pro + Ile209Cys + Tyr210Ser + Thr212Asn +Tyr213Cys + Pro214Gln + Tyr218CysAla204His + Trp208Asn + Thr212Pro + Pro214Gln +Thr215Asn + Thr217Gly + Thr224AspPro205Ser + Gly206Ser + Tyr210Ser + Thr215Pro +Ala219Asn + Leu221Met + Ser222AspAla204Ser + Gly206Gln + Trp208Pro + Thr212Asn +Thr215Glu + Tyr218Ile + Gly223AsnAla204Ser + Pro205Asn + Trp208Tyr + Thr215Gly +Tyr218Pro + Ser220Glu + Thr224GlyAla204Asn + Tyr210His + Pro214Asn + Tyr218Pro +Ala219Gln + Ser220Glu + Leu221IleAla204Gly + Gly206Asn + Ser207Asp + Thr212Asn +Tyr213Leu + Thr215Pro + Gly223ProAla204Asn + Tyr210Met + Thr212Gln + Pro214Gly +Ala219Gly + Ser222Asp + Gly223GlnTyr210Pro + Thr212Gly + Tyr213Leu + Thr215Gly +Thr217Gly + Ala219Glu + Ser220AspGly206Ser + Trp208Asn + Ile209Pro + Ala219Asp +Ser220Asp + Gly223Ser + Thr224GlyPro205Ser + Tyr210Cys + Tyr213Thr + Pro214Asp +Thr215Asp + Tyr218Val + Leu221SerTrp208His + Ile209Val + Thr212Pro + Tyr213Leu +Thr217Ser + Ser222Glu + Gly223GluPro205Gly + Gly206Gln + Tyr210Asn + Tyr213Gly +Thr217Gly + Ser222Glu + Gly223GluAla204Thr + Thr212Gln + Tyr213Gln + Thr217Ser +Ser222Asp + Gly223Asp + Thr224ProPro205Gly + Ile209Cys + Tyr213Val + Pro214Gln +Thr215Pro + Ser222Glu + Gly223AspTrp208Ser + Ile209Ser + Ser216Glu + Thr217Glu +Tyr218Gly + Ala219His + Gly223ProGly206Pro + Tyr210Gln + Thr212Pro + Tyr213Leu +Ser216Glu + Thr217Asp + Leu221ProTrp208Met + Ile209Gln + Tyr210Met + Tyr218Gln +Ser220Glu + Leu221Glu + Thr224SerIle209Gln + Thr212Ser + Tyr213Val + Thr215Asp +Ser216Glu + Tyr218Met + Ala219ProTyr213Pro + Pro214Gln + Thr215Asp + Ser216Glu +Thr217Asp + Tyr218Met + Leu221AlaAla204His + Pro205Gly + Ser207Glu + Trp208His +Tyr210His + Tyr213Cys + Ser222AspGly206Pro + Ser207Asp + Trp208Ser + Pro214Ser +Leu221Asn + Ser222Asp + Gly223ProSer207Glu + Trp208Tyr + Ile209Gly + Tyr213Thr +Pro214Ser + Tyr218Met + Ser222GluPro205Ser + Ser207Asp + Trp208Ser + Thr212Gly +Tyr213Asn + Tyr218Gln + Ser222AspAla204Thr + Gly206Asn + Trp208Thr + Thr212Gln +Pro214Glu + Thr215Asp + Ser216AspAla204Thr + Gly206Asn + Ser207Glu + Tyr210Ile +Leu221Thr + Ser222Glu + Gly223GluAla204His + Ser207Glu + Thr215Gln + Thr217Gln +Tyr218Asn + Ser222Glu + Gly223GluAla204Gln + Trp208Asp + Tyr210Thr + Thr212Ser +Thr217Asn + Ser222Asp + Gly223AsnIle209Thr + Ser216Asp + Thr217Asp + Tyr218Asp +Leu221Ser + Gly223Ser + Thr224GlyAla204Asn + Gly206Gln + Ser207Glu + Trp208His +Thr212Ser + Leu221Asp + Ser222AspGly206Ser + Ser207Asp + Tyr210Val + Thr212Gln +Tyr218Ile + Leu221Asp + Ser222AspAla204Ser + Gly206Pro + Pro214Gly + Thr215Asp +Thr217Asp + Tyr218Pro + Gly223ProGly206Gln + Ile209His + Pro214Asp + Thr215Pro +Ser216Asp + Leu221Gly + Thr224SerGly206Asn + Ser207Glu + Trp208Asp + Ile209Gly +Tyr213Gly + Ser220Glu + Ser222GluPro205Ser + Ser207Asp + Ile209Asn + Tyr213Met +Thr217Gln + Ala219His + Gly223GluPro205Gly + Gly206Gln + Ser207Glu + Ala219Pro +Ser220Asp + Leu221Glu + Thr224SerTrp208Met + Tyr210Thr + Thr212Asn + Ala219Gln +Leu221Asp + Gly223Asp + Thr224GlnPro205Ser + Ser207Glu + Tyr213His + Thr217Pro +Ala219Ser + Ser220Glu + Ser222GluSer207Asp + Tyr210Met + Pro214Ser + Tyr218Asn +Ser220Glu + Leu221Cys + Ser222GluAla204Pro + Ser207Glu + Pro214Gln + Ser220Asp +Leu221His + Ser222Asp + Thr224SerSer207Glu + Ile209Met + Tyr210Gly + Tyr213Leu +Thr215Gln + Ser220Glu + Ser222AspGly206Pro + Ser207Asp + Ile209Ser + Tyr210His +Thr217Gly + Ser222Glu + Thr224AspPro205Asn + Tyr210Cys + Tyr213Gly + Thr217Asn +Tyr218Met + Ser220Glu + Ser222AspIle209Gly + Thr212Pro + Thr215Asn + Thr217Gly +Ala219His + Ser220Glu + Ser222GluAla204Gln + Gly206Gln + Thr212Pro + Ser220Glu +Leu221Val + Ser222Asp + Thr224GlnAla204Gln + Pro205Ser + Gly206Ser + Trp208Gln +Thr215Pro + Ser220Glu + Ser222GluSer207Glu + Trp208Leu + Tyr210Ile + Thr212Gly +Tyr218Cys + Leu221Asp + Gly223AsnPro205Gly + Gly206Pro + Ser207Asp + Ile209Met +Leu221Asp + Ser222Glu + Thr224AspPro205Gly + Thr215Asn + Thr217Pro + Tyr218Val +Leu221Thr + Ser222Glu + Thr224AspTrp208Phe + Tyr213Met + Thr215Asn + Tyr218Asp +Ala219Pro + Ser220Glu + Thr224GlyAla204Pro + Pro205Gly + Trp208Leu + Thr217Gln +Leu221Asp + Ser222Glu + Thr224AspIle209Ala + Thr212Gln + Thr217Gln + Tyr218Thr +Ser220Glu + Leu221Asp + Gly223GluAla204Asn + Ile209Cys + Tyr213Cys + Tyr218Ala +Ala219Glu + Leu221Asp + Ser222GluGly206Ser + Ser207Glu + Trp208His + Tyr210Asp +Tyr218Val + Ala219Thr + Ser222AspPro205Gly + Ser207Asp + Trp208Thr + Thr212Ser +Thr217Gly + Ser220Glu + Gly223SerSer207Glu + Trp208Asn + Tyr210Pro + Pro214Asn +Tyr218Ser + Ser220Asp + Gly223SerAla204Gln + Ser207Asp + Trp208Gly + Ile209Ala +Thr212Asn + Ser220Glu + Gly223ProPro205Gln + Ser207Asp + Ile209Gln + Pro214Asn +Thr215Gln + Ser220Asp + Thr224GlyPro205Asn + Ser207Glu + Ile209Val + Thr212Gly +Tyr213Pro + Ala219Asn + Thr224GluAla204Gly + Ser207Glu + Ile209Ala + Thr212Pro +Tyr213Cys + Ala219Glu + Ser220AspPro205Ser + Trp208Glu + Thr212Asn + Tyr218Glu +Ala219His + Leu221Glu + Ser222GluPro205Ser + Ile209Pro + Tyr210Asp + Thr212Ser +Tyr213Ile + Ala219Pro + Ser222GluGly206Ser + Trp208Thr + Tyr210Ala + Pro214Asp +Tyr218Met + Ala219Glu + Ser220AspIle209Leu + Pro214Glu + Thr215Pro + Ala219Glu +Ser220Glu + Gly223Pro + Thr224GlyTrp208Tyr + Thr212Asn + Thr217Glu + Ala219Gly +Ser220Asp + Leu221Glu + Ser222AspAla204Thr + Tyr210Pro + Tyr213Asn + Pro214Glu +Tyr218Asp + Ser220Asp + Thr224GlyGly206Pro + Thr212Pro + Pro214Gly + Thr215Pro +Ser220Asp + Leu221Asp + Thr224GluThr212Asn + Pro214Gln + Tyr218Met + Ala219Asp +Ser222Glu + Gly223Asp + Thr224GluTrp208Gly + Tyr210Pro + Thr212Gly + Ser220Glu +Leu221Met + Gly223Asp + Thr224GluGly206Gln + Tyr213Asn + Pro214Asn + Thr217Asp +Ala219His + Ser220Glu + Leu221AspPro205Gln + Ile209Pro + Tyr210Asn + Tyr218Thr +Ala219Glu + Ser220Asp + Gly223AspAla204Thr + Ser207Asp + Tyr210Asp + Tyr213Met +Ala219Asn + Ser220Glu + Thr224AspPro205Ser + Trp208Pro + Ile209Pro + Tyr213Ile +Ala219Asp + Ser222Asp + Gly223AspAla204Asn + Thr212Pro + Thr217Glu + Tyr218Asp +Ser220Glu + Ser222Asp + Thr224GlnGly206Pro + Ser207Asp + Trp208Val + Tyr210Thr +Tyr213Ala + Ser220Asp + Thr224AspPro205Gly + Ser207Glu + Tyr210Ile + Pro214Ser +Thr217Gly + Ser220Asp + Thr224GluAla204Thr + Tyr210Cys + Thr212Pro + Tyr218Ile +Ser220Glu + Leu221Ser + Gly223GluAla204Gln + Trp208Pro + Pro214Asn + Ala219Thr +Ser220Glu + Gly223Asp + Thr224GlnPro205Gln + Tyr213Gly + Thr215Gly + Tyr218Ala +Ser220Asp + Gly223Glu + Thr224ProPro205Gly + Gly206Ser + Thr217Pro + Ala219His +Ser220Asp + Leu221Cys + Gly223AspPro205Ser + Tyr210Thr + Thr212Asn + Tyr213Cys +Pro214Ser + Ala219Asp + Ser222AspTrp208Asn + Thr217Glu + Tyr218Cys + Ala219Glu +Ser220Asp + Gly223Glu + Thr224SerPro205Ser + Gly206Pro + Trp208Pro + Ile209Ala +Thr215Asp + Tyr218Asp + Ser220AspTrp208Thr + Thr212Gly + Tyr213Ile + Thr215Asp +Leu221Asp + Ser222Glu + Gly223GluAla204Gly + Pro205Gly + Tyr210Glu + Thr217Glu +Tyr218Cys + Ala219Gly + Leu221ValSer207Asp + Thr215Pro + Thr217Asp + Tyr218Ile +Leu221Ala + Ser222Glu + Gly223AspGly206Pro + Trp208Glu + Tyr210Cys + Pro214Gln +Thr217Asp + Ser220Asp + Gly223GlnTrp208Thr + Tyr213Pro + Pro214Ser + Ser216Asp +Tyr218Met + Ala219Asp + Thr224ProAla204Asn + Trp208Ala + Tyr213Asn + Ser216Asp +Ala219Glu + Leu221Cys + Thr224GlyGly206Ser + Tyr210Ser + Pro214Gly + Ser216Asp +Tyr218Cys + Ala219Glu + Leu221ProAla204Thr + Tyr210Gln + Thr212Gln + Tyr213Ala +Pro214Gln + Thr217Asp + Ser220GluPro205Gln + Gly206Ser + Ser207Glu + Tyr210Ser +Ser216Asp + Thr217Glu + Tyr218GluTyr210Met + Tyr213Met + Ser216Asp + Ala219Glu +Ser220Glu + Leu221Cys + Ser222GluAla204Gln + Ile209His + Ser216Glu + Thr217Ser +Ala219Glu + Ser220Asp + Ser222GluIle209Ser + Tyr210Leu + Tyr213Leu + Pro214Asp +Thr217Asp + Ser220Glu + Leu221GluAla204His + Pro205Gln + Pro214Asn + Tyr218Gln +Ala219Asp + Leu221Glu + Thr224AspAla204Gly + Ser207Glu + Pro214Glu + Tyr218Pro +Ser220Glu + Ser222Asp + Thr224AsnAla204Pro + Pro205Gly + Trp208Glu + Ile209Ser +Tyr210Asp + Ala219Thr + Thr224GluAla204Gly + Ser207Glu + Trp208Glu + Tyr210Asp +Ser216Glu + Thr217Gln + Thr224GlyTrp208Ala + Tyr210Cys + Thr212Ser + Thr217Glu +Ala219Gln + Ser220Asp + Ser222GluGly206Asn + Ser207Glu + Trp208Glu + Thr212Ser +Ser216Glu + Leu221Met + Gly223GluAla204Thr + Ser207Asp + Ile209Cys + Thr215Asp +Tyr218Ile + Ser220Asp + Leu221AspSer207Asp + Thr212Pro + Pro214Gln + Ser216Glu +Tyr218Val + Ser220Asp + Ser222AspIle209Pro + Thr212Gly + Tyr213Ser + Pro214Glu +Thr215Glu + Ala219Gln + Ser220GluPro205Asn + Ser207Glu + Ile209Met + Pro214Ser +Ser216Asp + Ala219Glu + Leu221AspAla204Thr + Ser207Glu + Tyr210Leu + Ser216Glu +Ala219Asp + Leu221Asp + Gly223AsnGly206Asn + Ser207Glu + Pro214Gly + Thr215Glu +Thr217Glu + Tyr218Thr + Ala219GluGly206Gln + Ser207Glu + Tyr210Ala + Thr215Asn +Thr217Asp + Ser220Asp + Leu221AlaSer207Glu + Trp208Leu + Tyr213Val + Pro214Asp +Ala219Glu + Ser222Glu + Thr224ProAla204Thr + Ser207Asp + Trp208Leu + Tyr210Asp +Thr212Pro + Thr215Asp + Leu221GluPro205Gln + Ser207Asp + Ile209Ala + Thr217Asn +Tyr218Glu + Ala219Asp + Gly223AspSer207Glu + Ile209Thr + Tyr213Gly + Ser216Asp +Ala219Asp + Ser222Glu + Gly223ProGly206Ser + Ser207Asp + Ile209Ala + Thr212Gln +Thr217Glu + Ala219Glu + Leu221AsnPro205Asn + Tyr210Glu + Thr217Asp + Tyr218Ala +Leu221Ile + Ser222Glu + Thr224ProSer216Asp + Thr217Asp + Tyr218Cys + Ala219His +Leu221Glu + Ser222Glu + Gly223AsnAla204Gln + Trp208Asp + Pro214Gln + Ser216Glu +Thr217Gln + Ser220Asp + Gly223AspPro205Gln + Tyr210Leu + Tyr213Thr + Ser216Glu +Tyr218Glu + Ser220Glu + Gly223GluPro214Gly + Thr217Asp + Tyr218Gly + Ala219Glu +Leu221Asn + Ser222Glu + Thr224GluGly206Pro + Ser207Asp + Trp208Asp + Thr212Gln +Thr217Asp + Gly223Ser + Thr224GluAla204Pro + Pro205Ser + Gly206Pro + Ile209Val +Tyr213Pro + Thr215Asp + Ala219GluAla204Pro + Gly206Pro + Tyr210Val + Pro214Glu +Ser220Glu + Ser222Glu + Thr224ProAla204Gly + Gly206Pro + Thr212Gln + Pro214Glu +Ser220Glu + Leu221Val + Ser222AspAla204Asn + Gly206Gln + Ile209Ser + Tyr210Ala +Pro214Glu + Ser220Glu + Ser222AspPro205Gln + Trp208Gln + Pro214Glu + Thr217Pro +Leu221Met + Ser222Asp + Gly223GluAla204Thr + Pro205Asn + Pro214Asp + Thr215Pro +Leu221Cys + Ser222Asp + Gly223AspIle209Pro + Tyr210Pro + Thr212Pro + Thr217Asp +Leu221Asn + Ser222Asp + Gly223AspGly206Gln + Trp208Thr + Ile209Ser + Tyr213Met +Thr215Glu + Ser222Asp + Gly223GluPro205Asn + Trp208Ser + Thr212Gly + Tyr213Asn +Thr215Asp + Ser222Asp + Gly223GluTyr210Gly + Thr212Pro + Pro214Gly + Ser216Asp +Ala219His + Leu221Glu + Ser222AspPro205Gly + Gly206Pro + Ile209His + Thr215Asp +Ala219Gly + Leu221Glu + Ser222GluPro205Gln + Tyr210Gly + Thr212Asn + Ser216Asp +Ala219Pro + Ser220Glu + Leu221AspGly206Ser + Trp208Glu + Tyr210Gln + Thr215Gly +Ser216Glu + Thr217Glu + Leu221ProPro205Gln + Gly206Gln + Ser216Glu + Thr217Asp +Leu221Ala + Ser222Glu + Thr224ProGly206Ser + Ser207Glu + Trp208Asp + Ile209Leu +Thr215Asp + Ser216Glu + Tyr218GlnTyr210His + Thr212Gln + Thr215Asp + Ser216Glu +Leu221Glu + Gly223Ser + Thr224ProAla204Ser + Pro205Ser + Gly206Asn + Thr215Asp +Ser216Glu + Tyr218Val + Gly223AspAla204Gln + Gly206Asn + Ser207Glu + Trp208Pro +Pro214Asp + Thr215Asp + Ser222AspSer207Asp + Tyr210Gln + Thr212Gln + Thr215Asp +Ser216Asp + Leu221Gly + Ser222AspIle209Met + Tyr210Leu + Thr217Glu + Tyr218Thr +Ser220Glu + Gly223Glu + Thr224AsnPro205Ser + Ser207Glu + Trp208His + Thr215Pro +Thr217Asp + Leu221Asn + Ser222GluAla204Thr + Ser207Glu + Thr212Gly + Tyr213Leu +Pro214Asn + Ser216Glu + Ser222GluAla204Pro + Ser207Asp + Tyr210Leu + Thr212Gln +Tyr213Thr + Thr215Glu + Ser222Glu表29多環雙突變變異體Leu104Gly + Tyr210ProAsn 66Gln + Thr217GluGly 67Ser + Gly 69SerVal103Gln + Gly223AspTrp112Cys + Thr217GlyAla164Glu + Trp208GlnLeu134Ile + Trp208GlnAla219Gly + Ser220GluAsn 66Asp + Leu104GlyAla168Thr + Ser216GluLeul04Ala + Ser109GluThr111Gln + Val115AlaSer109Asp + Tyr210ProGly 69Ser + Serl91AspVal103Asp + Gly206GlnGly 67Glu + Pro214AsnTrp208Ile + Thr215AspSer107Glu + Thr113GlyGly108Ser + Leu221GluSer 70Glu + Pro169AsnGly165Gln + Leu221AspGly108Glu + Ala164GlyLeu104His + Thr167ProAla164His + Ser220GluThr195Gln + Ser222AspTrp208His + Ala219ProGly 69Glu + Asn106GlnTrp112Cys + Thr113ProGly 67Gln + Trp208AspAla114Glu + Leu134IleSer109Glu + Gly136SerGly110Ser + Ser207GluVal103Ala + Thr212SerAla114Glu + Thr167ProThr195Glu + Ala204ThrGly 67Ser + Thr217GluTrp208Tyr + Ser220GluAla168His + Thr217AspAla168Glu + Tyr213AsnGly110Pro + Ser220AspTyr171Asn + Leu221MetThr 73Asp + Leu134SerThr137Asn + Thr217ProSer 70Asp + Ala168ThrThr167Glu + Tyr210ProThr195Glu + Tyr210GlyTyr210Gln + Thr217GluGly 67Glu + Val115CysSer 70Glu + Ala219ThrGly136Pro + Leu221AlaGly 67Ser + Thr195SerPhe193His + Ser222AspAsn 68Glu + Tyr210SerGly165Asp + Pro169SerTrp208Pro + Thr215GlyThr 73Gly + Leu104GlyThr113Asn + Ser216GluGly 72Gln + Gly139AspTyr210Ile + Leu221GlyThr 73Pro + Tyr171AspLeu104Gln + Thr137GlyGly 69Pro + Tyr210AlaThr217Gly + Gly223GluThr195Asp + Tyr213SerGly 72Asn + Ala219AsnVal138Pro + Tyr210AlaTyr218Met + Gly223AsnSer220Glu + Leu221AsnTrp112Ala + Gly139AsnGly135Glu + Leu221GlyThr113Asn + Thr217SerThr195Asn + Thr217AspTyr210Val + Ser220GluVal138Gly + Ala219AspAsn166Glu + Leu221HisGly165Gln + Leu221HisThr113Gln + Ser192AspAsn 66Asp + Thr 73SerThr167Gln + Thr217GlyVal115Pro + Gly135SerAla219Gln + Gly223AspTrp208His + Thr217AsnTrp208Leu + Tyr210ValGly108Asn + Ser191GluSer107Glu + Tyr210AlaAsn 68Gln + Ser220GluTrp112Cys + Leu221AsnGly108Gln + Ser109AspVal103Met + Ala168HisAla164Pro + Ser192GluPro214Ser + Ser220GluThr215Asn + Thr217GlnSer220Glu + Leu221ProGly136Ser + Trp208PheThr195Asp + Tyr210GlnLeu104Gln + Ala219AspGly 69Pro + Leu221ProGly 69Asn + Ala204GlyGly135Glu + Ala219AsnGly136Asn + Ser220GluGly136Ser + Trp208ValGly139Asp + Asn140GlnTrp208Cys + Tyr210AsnGly165Ser + Ser220AspGly110Ser + Trp208ThrAla114Asp + Gly162AsnSer192Asp + Leu221CysVal103Ser + Gly135GluAsn140Glu + Trp208AlaThr113Asp + Leu221SerSer109Asp + Gly110SerSer109Glu + Thr113GlnThr 73Gly + Gly136AspAla164His + Leu221AsnGly 67Pro + Va1103HisAla114Asn + Thr212GlnAsn140Asp + Gly223ProGly139Gln + Pro214AsnAsp105Glu + Thr195GlyGly135Pro + Ser207GluLeu104Glu + Ala114ProAsn 66Glu + Gly 69ProVal115Cys + Ser207GluAsn 68Glu + Leu221AlaAsn 66Ser + Gly 69AspSer109Glu + Phe193ProTrp208Glu + Thr212ProGly136Gln + Trp208CysSer 70Asp + Leu221HisSer 70Glu + Phe193SerThr217Asn + Ser222GluTyr213Thr + Tyr218GlyTrp208Ser + Leu221CysAsn163Asp + Leu221SerAsn 66Gln + Gly223ProGly139Asn + Ser220AspThr 73Ser + Pro214AspSer 70Asp + Leu221IleGly165Gln + Ser220AspAsn166Asp + Thr217AsnAsp105Glu + Thr111ProAsn140Asp + Trp208PheAsn166Glu + Pro214AsnTrp208Ala + Thr217SerGly108Ser + Gly110GluVal138Asn + Asn170AspGly110Glu + Thr224GlnTrp208Met + Ser220AspAsn 68Glu + Val115SerAsn170Glu + Tyr210GlyGly110Asn + Ser192GluTrp208Gln + Ser220GluAsn140Gln + Pro214GlnThr217Ser + Ser220GluSer141Glu + Ala168ThrGly162Glu + Tyr210HisLeu104Gly + Thr224AspTrp112Pro + Ser191AspTrp208Thr + Tyr210GluGly108Gln + Thr111AspLeu134Val + Thr217ProAsn170Asp + Tyr210HisVal115Ser + Ser220GluGly 69Pro + Ser 70AspThr 73Gly + Tyr210GluGly 67Asp + Gly108GlnGly 69Ser + Gly110AspTyr210Thr + Ser220GluTrp208Phe + Leu221MetGly136Gln + Leu221AspThr113Ser + Trp208ValTyr171Leu + Tyr210ThrLeu134Asn + Thr167GlnAsn140Ser + Tyr210AspAsn 66Gln + Gly 67GluLeu134Cys + Pro214AsnGly136Ser + Leu221AlaThr111Pro + Trp208ValThr113Asp + Tyr171GlyTrp208Ile + Ser220GluPro214Ser + Thr224GluGly165Pro + Ser220GluThr137Gln + Tyr171GluTrP208Glu + Pro214GlySer216Asp + Leu221MetGly136Ser + Thr217GlnSer141Glu + Tyr210LeuTrp208Thr + Gly223SerAsn 66Ser + Asn170GlnAsn 66Gln + Asn170GluPro169Glu + Tyr210ThrTyr171Leu + Leu221AspGly139Glu + Leu221ProThr 73Pro + Leu134GlyTyr210Glu + Leu221AlaGly139Ser + Tyr210GlyAsn163Asp + Ala164SerAsn163Gln + Thr217AspAsn140Asp + Tyr218SerGly108Ser + Ala164GlnVal138Ser + Trp208LeuThr217Ser + Thr224AspSer109Asp + Tyr218AlaAsn 66Glu + Phe193ValAsn140Asp + Ala204GlyVal138Ser + Ser220AspGly108Asn + Tyr210ThrGly139Asn + Tyr171ProThr113Pro + Trp208ThrSer141Glu + Leu221GlyAsn106Glu + Thr167GlyTrp112Ser + Ala219SerAsp105Glu + Asn166GlnAla114Pro + Tyr171AsnThr217Gln + Ser220AspThr113Gly + Tyr210AspTyr171Cys + Tyr210GluAsn166Asp + Leu221ValAla114His + Ser191AspGly 67Glu + Thr217AsnAsn 66Glu + Thr215GlnThr111Asn + Ser220GluTrp208Gln + Leu221SerGly 69Gln + Ala114ThrAsn163Asp + Leu221AlaPro214Gly + Leu221IleThr217Gly + Ser220AspAla219Thr + Ser220AspTyr210Cys + Ser220GluVal138Asp + Gly223ProThr195Asn + Leu221GlyLeu104Thr + Thr215SerGly 69Gln + Leu134ThrGly165Pro + Leu221GlyThr167Ser + Thr217GluThr212Pro + Thr217GluAsn166Ser + Tyr218AspGly108Glu + Tyr171GlnGly 67Asn + Tyr210IleVal115Cys + Pro214GlyThr137Asp + Tyr210ValLeu104Met + Trp208GlyLeu134Ala + Ser192AspTrp112Gln + Ala219GlnGly108Glu + Val115HisThr113Asn + Gly139ProTrp208Gln + Gly223AsnSer207Glu + Trp208GlnAsn 66Ser + Tyr210AsnSer207Glu + Tyr210ProVal103Ala + Thr215GluVal103Asn + Tyr210AlaAsn 66Ser + Trp208PheThr215Asn + Ala219ThrGly136Asn + Gly139AsnTyr171Thr + Ser216GluLeu104Glu + Tyr210AsnSer109Glu + Thr217AsnLeu134Ile + Asn166SerTyr171Gly + Thr217GlnVal115Cys + Thr224AspPro169Ser + Thr215AspGly 72Gln + Gly135AsnTyr210Thr + Ser220AspAsn 68Glu + Thr113SerAsn106Gln + Thr113AspSer109Glu + Trp208AsnLeu134Gln + Thr137GluAla114Thr + Asn170GluAla219Pro + Ser220GluTyr171Val + Trp208ProGly139Ser + Tyr213SerSer107Asp + Thr113AsnThr167Gly + Leu221MetTyr171Gly + Ser207GluGly 67Ser + Thr217SerVal115Gly + Thr217SerAsn 66Ser + Trp208IleSer191Glu + Trp208CysThr137Asp + Trp208SerLeu221Gly + Ser222GluSer216Glu + Leu221ThrGly162Pro + Ser207GluThr217Asp + Ala219ThrLeu104Cys + Ser207GluGly 69Gln + Ser220GluGly 69Gln + Val138AspPro169Gly + Tyr210LeuThr 73Asp + Gly110SerThr215Gly + Thr217SerGly 69Ser + Gly 72AsnSer191Glu + Thr217AsnGly135Ser + Tyr171SerThr111Asn + Phe193LeuTrp112Leu + Ser207AspLeu104Glu + Trp208AlaLeu221Ile + Thr224AspGly136Glu + Gly165AsnTyr210Glu + Thr217ProTrp112Phe + Pro214AspGly110Gln + Trp112MetAsn 68Ser + Trp208AspGly 69Ser + Leu104CysThr137Glu + Thr215ProAsn166Gln + Thr217GluTyr210Thr + Thr212SerLeu134His + Trp208AspSer141Glu + Tyr210ValSer191Glu + Tyr210LeuGly139Pro + Ser207AspGly162Ser + Tyr171MetThr 73Gln + Val103AlaTyr171Val + Phe193AspLeu221Thr + Ser222GluGly 69Asp + Ala114GlnGly108Glu + Trp208MetGly 72Ser + Tyr210GlnThr137Asn + Trp208AspAsn140Asp + Phe193ThrSer192Glu + Trp208AlaAsn170Glu + Pro205SerTyr210Ala + Tyr213SerTyr171Gln + Tyr218IleAsn 66Asp + Leu221CysAsn163Gln + Gly223AsnVal103Thr + Leu134SerThr167Ser + Asn170AspSer 70Glu + Gly162SerTyr171Gln + Ser192GluTyr210Val + Gly223SerThr137Glu + Leu221ProGly 69Asn + Thr217AspPro205Ser + Tyr218GluTyr210Met + Leu221AsnThr215Asp + Thr217ProLeu134Val + Gly136GluThr 73Pro + Leu221GluTyr213Val + Ser220GluThr 73Ser + Ser107GluTrp208Ser + Leu221ProThr113Asp + Leu221HisAla114Thr + Thr217AspAla168Ser + Tyr218GluTrp208Asp + Leu221AsnThr137Asp + Trp208IleGly135Gln + Ser191GluVal103Glu + Trp208GlyAsn140Asp + Leu221HisSer192Asp + Tyr210CysGly108Asn + Tyr210HisAsp105Glu + Trp208AsnVal103Pro + Tyr210ProThr 73Gln + Trp112TyrLeu134Cys + Asn170SerAsn166Glu + Tyr210CysAsn106Ser + Leu221MetSer109Glu + Leu221CysSer107Asp + Leu221AlaAla114Ser + Leu221AsnGly162Asn + Leu221Val表33多環三突變變異體Leu104Gly + Tyr210Pro + Thr217GluAsn 66Gln + Gly 67Ser + Gly 69SerVal103Gln + Trp112Cys + Gly223AspAla164Glu + Trp208Gln + Thr217GlyLeu134Ile + Trp208Gln + Ser220GluAsn 66Asp + Leu104Gly + Ala219GlyLeu104Ala + Thr111Gln + Val115AlaGly 69Ser + Ser109Asp + Tyr210ProGly139Asn + Thr167Asn + Thr217SerGly108Glu + Ala164Gly + Gly165GlnLeu104His + Thr167Pro + Ser220AspGly 69Glu + Asn106Gln + Trp208HisTrp112Cys + Thr113Pro + Trp208AspGly110Ser + Ser207Glu + Thr212SerAla168His + Trp208Tyr + Ser220GluTyr171Asn + Thr217Asp + Leu221MetSer 70Asp + Thr137Asn + Thr217ProThr167Glu + Ala168Thr + Tyr210ProSer 70Glu + Gly136Pro + Leu221AlaGly 67Ser + Thr195Ser + Ser222AspPro169Ser + Trp208Pro + Thr215GlyThr 73Gly + Leu104Gly + Tyr171GluGly 72Gln + Tyr210Ile + Leu221GlyLeu104Gln + Thr137Gly + Tyr210AlaTyr213Ser + Thr217Gly + Gly223GluGly 72Asn + Thr195Asp + Ala219AsnThr137Asp + Val138Pro + Tyr210AlaSer109Glu + Val115Met + Trp208CysGly139Asn + Ser220Glu + Leu221AsnTrp112Ala + Gly135Glu + Leu221GlyThr113Asn + Thr195Asn + Thr217SerAla164His + Ser220Asp + Leu221HisAsn 66Asp + Thr 73Ser + Thr113GlnGly135Ser + Thr167Gln + Thr217GlyTyr171Asp + Trp208His + Thr217AsnGly108Asn + Trp208Leu + Tyr210ValTrp112Cys + Ser220Glu + Leu221AsnVal103Met + Ala168His + Ser191AspPro214Ser + Thr217Gln + Ser220GluThr215Asn + Ser220Glu + Leu221ProGly136Ser + Trp208Phe + Tyr210GlnGly 69Pro + Ala219Asp + Leu221ProGly 69Asn + Ala204Gly + Ser220AspGly136Ser + Gly139Asp + Trp208ValAsn140Gln + Trp208Cys + Tyr210AsnGly110Ser + Gly165Ser + Trp208ThrVal103Ser + Gly135Glu + Leu221CysAsn140Glu + Trp208Ala + Leu221SerGly 67Pro + Val103His + Ala164HisAsn140Asp + Thr212Gln + Gly223ProGly139Gln + Thr195Gly + Pro214AsnGly 69Pro + Leu104Glu + Ala114ProPhe193Pro + Trp208Glu + Thr212ProGly136Gln + Trp208Cys + Leu221HisSer 70Asp + Thr 73Gly + Phe193SerTyr213Thr + Tyr218Gly + Gly223AspThr195Glu + Trp208Ser + Leu221CysAsn163Asp + Leu221Ser + Gly223ProAsn 66Gln + Gly139Asn + Ser220AspThr 73Ser + Pro214Asp + Leu221IleThr111Pro + Asn140Asp + Trp208PheAsn166Glu + Pro214Asn + Thr217SerGly108Ser + Gly110Glu + Trp208AlaLeu134Asp + Gly165Asn + Tyr210ThrPhe193Thr + Thr217Gln + Ser220AspGly 67Pro + Trp112Gln + Ser192AspTyr210Ile + Thr217Gln + Ser222AspVal103Ser + Leu104Ile + Ala164AsnAla114Asp + Thr167Gly + Thr217AsnThr111Asp + Thr195Gly + Tyr210LeuTyr210Thr + Ser220Glu + Leu221MetGly136Gln + Trp208Phe + Leu221AspThr113Ser + Trp208Val + Tyr210ThrLeu134Asn + Asn140Ser + Thr167GlnGly136Ser + Trp208Val + Leu221AlaGly165Pro + Pro214Ser + Ser220GluGly136Ser + Tyr210Leu + Thr217GlnSer141Glu + Trp208Thr + Gly223SerAsn 66Ser + Thr113Pro + Asn170GluThr 73Pro + Tyr210Thr + Ser216GluThr 73Pro + Tyr210Glu + Leu221AlaGly139Ser + Ala164Ser + Tyr210GlyGly108Ser + Val138Ser + Ala164GlnTrp208Leu + Thr217Ser + Thr224AspAsn 66Glu + Phe193Val + Tyr218AlaGly108Asn + Tyr171Pro + Tyr210ThrThr113Pro + Gly139Asn + Trp208ThrAsp105Glu + Asn166Gln + Tyr171AsnAla114Pro + Thr217Gln + Ser220AspGly 67Glu + Ala114His + Thr217AsnSer141Glu + Trp208Gln + Leu221SerGly 69Gln + Ala114Thr + Asn170AspAsn163Asp + Ala219Thr + Leu221AlaSer 70Asp + Thr217Gly + Ala219ThrThr195Asn + Leu221Gly + Gly223ProGly 69Gln + Leu104Thr + Leu134ThrGly139Asp + Gly165Pro + Leu221GlyGly 67Asn + Gly108Glu + Tyr210IleVal115Cys + Tyr210Val + Pro214GlyLeu104Met + Thr137Asp + Trp208GlyTrp112Gln + Leu134Ala + Ser192AspGly108Glu + Val115His + Ala219GlnThr113Asn + Gly139Pro + Thr167AspThr137Asp + Trp208Gln + Gly223AsnSer207Glu + Trp208Gln + Tyr210AsnAsn 66Ser + Ser207Glu + Tyr210ProVal103Asn + Tyr210Ala + Thr215GluAsn 66Ser + Trp208Phe + Ala219ThrGly136Asn + Gly139Asn + Thr215AsnSer109Glu + Asn166Ser + Thr217AsnLeu134Ile + Tyr171Gly + Thr217GlnGly 72Gln + Gly110Asp + Gly135AsnAsn 68Glu + Asn106Gln + Thr113SerAla114Thr + Leu134Gln + Thr137GluSer107Asp + Gly139Ser + Tyr213SerThr113Asn + Thr167Gly + Leu221MetGly 67Ser + Ser207Glu + Thr217SerAsn 66Ser + Trp208Ile + Thr217SerTrp208Ser + Leu221Gly + Ser222GluGly162Pro + Ser216Glu + Leu221ThrLeu104Cys + Asn106Gln + Ser207GluGly 69Gln + Pro169Gly + Tyr210LeuThr 73Asp + Gly110Ser + Thr217SerGly 69Ser + Gly 72Asn + Thr215GlyGly135Ser + Tyr171Ser + Ser191GluThr111Asn + Phe193Leu + Thr217GluTrp208Ala + Leu221Ile + Thr224AspTrp112Phe + Pro214Asp + Thr217ProAsn 68Ser + Gly110Gln + Trp112MetGly 69Ser + Leu104Cys + Trp208AspThr137Glu + Asn166Gln + Thr215ProGly108Glu + Tyr210Thr + Thr212SerThr 73Gln + Gly162Ser + Tyr171MetTyr171Val + Phe193Asp + Leu221ThrGly 72Ser + Gly108Glu + Trp208MetThr137Asn + Trp208Asp + Tyr210GlnAsn140Asp + Phe193Thr + Trp208AlaGly 69Glu + Tyr210Ala + Tyr213SerAsn 66Asp + Tyr218Ile + Leu221CysVal103Thr + Leu134Ser + Asn163GlnTyr171Gln + Ser192Glu + Gly223SerGly 69Asn + Thr137Glu + Leu221ProPro205Ser + Tyr218Glu + Leu221AsnTyr210Met + Thr215Asp + Thr217ProThr 73Pro + Leu134Val + Gly136GluThr 73Ser + Ser107Glu + Trp208SerGly162Gln + Ser220Asp + Leu221ProVal103Glu + Gly135Gln + Trp208GlyAsn140Asp + Tyr210Cys + Leu221HisGly108Asn + Ser192Asp + Tyr210HisAsp105Glu + Trp208Asn + Tyr210ProThr 73Gln + Val103Pro + Trp112TyrLeu134Cys + Asn170Ser + Tyr210CysAsn106Ser + Ser109Glu + Leu221CysSer107Asp + Ala114Ser + Leu221AlaGly 69Asn + Gly162Asn + Leu221AsnGly 67Ser + Gly135Asp + Gly162AsnVal115Gly + Leu134Ser + Ala164GlnAsn 66Glu + Leu104Asn + Ala168GlyGly 69Ser + Asn166Glu + Tyr218AsnThr113Asp + Ala204Ser + Tyr218SerAsn166Gln + Trp208Thr + Thr217AspGly108Glu + Thr217Ser + Tyr218GlyAsn106Asp + Trp208Met + Thr217GlnGly 69Glu + Tyr171Val + Trp208HisGly139Asn + Pro169Gly + Trp208AsnVal115Ala + Thr215Gln + Leu221GluThr111Pro + Pro169Gln + Ala219GluThr 73Glu + Thr137Asn + Trp208HisAsn 66Asp + Asn106Ser + Leu221ThrGly 67Pro + Gly 69Ser + Thr224SerVal115His + Ser191Asp + Tyr218ValThr137Glu + Thr195Asn + Trp208GlyLeu134Glu + Trp208Ser + Leu221SerVal103Ser + Trp208Glu + Thr217AsnGly139Asn + Phe193Pro + Leu221CysAsn106Gln + Ala164Pro + Asn166AspAsp105Glu + Val138Ser + Thr217ProGly135Pro + Tyr171Thr + Ser222GluSer216Asp + Thr217Asn + Thr224GlyAla114Gly + Asn140Gln + Tyr218AspSer107Asp + Pro214Gly + Leu221SerAsp105Glu + Asn170Ser + Thr195ProSer 70Glu + Tyr171Leu + Pro214SerGly 67Asp + Leu104Thr + Gly223AsnGly108Asp + Ala114Asn + Tyr210CysGly110Asn + Pro214Gly + Ser220AspGly 69Asn + Pro169Gln + Ser220GluGly162Glu + Trp208Ala + Thr217AsnTrp112Tyr + Val115Pro + Ser220GluTrp208Gly + Tyr210Ile + Pro214AspLeu134Asn + Pro169Ser + Leu221AspThr113Ser + Trp208Leu + Leu221GluAsn166Ser + Tyr218Ile + Ser220GluTrp112Pro + Phe193His + Pro214AspLeu104Glu + Val115Ser + Ala164SerGly 69Pro + Pro169Asp + Leu221HisGly108Gln + Val138Ser + Ser220GluGly110Asn + Thr137Gln + Ser220GluAla164Thr + Asn166Ser + Tyr210AspTrp112Pro + Val115Pro + Pro214GluVal115Glu + Gly139Asn + Tyr210GlnGly 69Ser + Thr 73Ser + Tyr210ProAla114Ser + Ser207Asp + Thr224ProThr 73Pro + Asn166Glu + Thr212GlyGly 72Gln + Gly110Asp + Leu221HisTyr210Cys + Pro214Asn + Tyr218AspGly 69Asn + Gly136Pro + Asn166SerThr 73Gly + Gly108Ser + Ser220GluThr 73Gly + Pro205Ser + Thr224SerVal103Met + Ala164Asp + Leu221MetThr111Asn + Thr167Glu + Phe193MetThr 73Asp + Tyr210Asn + Thr217GlnAsn 66Glu + Thr 73Gly + Leu221GlnThr113Asp + Asn166Ser + Thr167ProAsn 66Gln + Tyr171Gly + Tyr210ProAsn 68Ser + Gly 69Pro + Asn170GlnAsn 68Ser + Val115Glu + Tyr210AsnThr113Gln + Ser191Asp + Leu221HisThr167Pro + Trp208Val + Tyr210LeuGly162Glu + Trp208Ala + Thr217ProLeu104Met + Ser207Asp + Thr212GlyThr137Gln + Asn163Asp + Pro205GlnVal115Thr + Thr217Gln + Ser220AspSer 70Asp + Val138Ser + Tyr210ValPro214Gly + Leu221Ile + Thr224AspLeu134Val + Gly165Gln + Trp208ThrSer109Asp + Ala114Asn + Tyr210SerAsn 66Ser + Val115Asn + Trp208GlnThr 73Pro + Phe193Glu + Gly206GlnLeu134Ser + Leu221Pro + Ser222AspVal115Cys + Ala164Glu + Asn170GlnAla164Asn + Trp208Cys + Thr217GluAsn 66Glu + Ala168Pro + Thr215ProGly135Ser + Ser207Asp + Thr217GlnAsn140Gln + Thr167Asp + Trp208MetGly165Gln + Trp208Thr + Leu221AsnThr 73Pro + Asn106Glu + Leu221CysThr113Ser + Leu221Ile + Gly223AspAsn 66Asp + Gly136Gln + Gly162GlnSer191Glu + Thr217Asn + Thr224ProLeu104Gln + Gly162Glu + Tyr210GlnTrp112Gln + Asn163Asp + Thr217GlyLeu134Pro + Thr217Asn + Leu221GluGly165Pro + Tyr210Met + Thr217GluAla164Gly + Ser191Glu + Trp208CysVal115Cys + Gly139Asn + Pro205AsnGly135Pro + Trp208Ala + Tyr210ProPro169Asn + Thr195Glu + Tyr210HisSer107Asp + Trp208Met + Tyr210ProThr 73Gly + Ala164Gly + Tyr210GluLeu104Val + Pro205Gln + Leu221AspThr111Gln + Ser141Asp + Leu221AsnTrp112Cys + Thr167Gln + Thr217ProVal138Thr + Asn170Glu + Leu221ProSer 70Asp + Leu134Ser + Thr224AsnThr137Asn + Ser207Asp + Tyr218ThrGly135Glu + Ala164Asn + Ala203SerLeu134Gly + Trp208Glu + Thr212SerAsn163Asp + Trp208Asn + Tyr210ThrVal115Thr + Ser192Glu + Thr217SerGly 72Pro + Ser109Asp + Tyr210GlnSer109Glu + Asn163Gln + Phe193ProGly136Pro + Pro169Glu + Leu221SerGly110Asn + Ser220Glu + Leu221MetTrp112Gly + Tyr210Asp + Thr217ProTrp208Ser + Tyr210Ser + Leu221ProTrp112Met + Ser192Glu + Leu221AlaThr137Gly + Thr167Ser + Thr217GluSer109Asp + Gly162Pro + Tyr210AlaAla114Thr + Gly162Asn + Ser191GluThr195Asn + Tyr218Pro + Leu221GluThr 73Glu + Ala164Gln + Pro214SerAsn 68Glu + Trp208Tyr + Thr217AsnGly165Pro + Thr212Ser + Ser220GluGly 67Asn + Gly139Gln + Ala168AspSer141Glu + Pro214Gln + Leu221GlnGly206Ser + Trp208Tyr + Thr215GlnLeu134Pro + Gly162Asn + Phe193IleTrp112Ala + Thr217Asn + Ser220AspThr111Gly + Gly139Glu + Thr212AsnSer107Glu + Tyr213Ala + Thr217ProGly165Ser + Thr217Glu + Gly223ProTyr210His + Thr217Asp + Leu221AlaTrp208Cys + Thr217Asp + Tyr218AsnGly 69Gln + Val103Ala + Ser220GluAsn106Ser + Tyr210Gly + Gly223SerTrp208Cys + Tyr210Asn + Thr217AspAsn 66Gln + Gly139Gln + Thr217GlyThr 73Glu + Leu104Ile + Tyr210IleThr113Ser + Pro169Asn + Thr217SerTyr210Gly + Pro214Gly + Leu221GluLeu134Ile + Ala219Glu + Ser220AspAsn140Asp + Ser141Glu + Tyr218SerTyr213Val + Ser220Glu + Leu221GluTyr210Val + Ser220Glu + Leu221AspAla164His + Ser220Glu + Leu221GluTyr171His + Ser220Asp + Leu221GluTyr210Ile + Ser220Glu + Leu221AspTrp112Leu + Ser220Asp + Leu221GluGly136Pro + Ser191Asp + Ser192GluAla168Ser + Thr217Asp + Tyr218GluAsn166Ser + Thr217Glu + Tyr218AspGly162Asp + Asn163Glu + Leu221MetTrp208Ser + Tyr210Glu + Ser220AspAla114Thr + Gly165Asp + Phe193GluSer207Glu + Thr217Asn + Ser222GluAsn140Ser + Trp208Glu + Ser220AspTrp208Asp + Leu221Asn + Ser222GluGly139Gln + Trp208Glu + Leu221AspAsn 68Glu + Asp105Glu + Asn106AspThr137Gln + Gly139Asp + Tyr171GluTrp208Asp + Tyr210Glu + Leu221AlaGly 69Pro + Asn166Glu + Ala168GluTyr171Cys + Tyr210Glu + Leu221AspThr137Glu + Pro169Asp + Asn170SerGly162Glu + Tyr171Asp + Thr217SerLeu134Ile + Ser191Glu + Gly223GluVal115Pro + Ser207Glu + Ser220AspGly139Pro + Ser207Asp + Ser220GluSer 70Glu + Val103Pro + Asn106AspGly 69Asp + Thr 73Asp + Tyr218IleAsn 66Asp + Trp208Gly + Pro214GluAsn 66Asp + Ser 70Glu + Gly136SerAsn163Asp + Ser191Asp + Ser207GluAsn 66Ser + Gly 69Asp + Ser109GluGly 67Glu + Trp208Ile + Thr215AspSer 70Glu + Tyr210Gln + Thr217GluSer 70Glu + Gly135Pro + Thr217GluVal138Glu + Ser141Asp + Tyr210GlnVal138Glu + Ser141Glu + Pro214AsnSer107Asp + Gly110Asp + Ala114GluSer109Glu + Ala114Glu + Gly136SerGly110Glu + Gly136Glu + Tyr210GlnGly135Ser + Tyr210Asp + Thr217GluSer 70Glu + Pro169Asn + Leu221AspVal115Gly + Gly162Asp + Thr167AspGly110Pro + Thr217Asp + Ser220AspAla164His + Thr217Asp + Ser220AspTyr210Cys + Thr217Asp + Ser220AspVal103Ser + Thr217Asp + Ser220AspThr217Glu + Tyr218Val + Ser220GluGly 69Ser + Thr217Glu + Ser220GluTyr210Val + Thr217Asp + Ser220GluGly 67Glu + Ser107Glu + Thr111AsnAsn166Asp + Ser191Asp + Leu221ValLeu134Asp + Phe193Glu + Gly223AspAsn 66Asp + Ser216Asp + Thr217GluVal103Glu + Asn106Glu + Thr113GluSer 70Asp + Gly139Ser + Ser220AspLeu104Asp + Asp105Glu + Ser191GluAla114Glu + Ser216Asp + Thr217GluThr195Glu + Ser216Asp + Thr217GluVal115Glu + Ser220Asp + Leu221GluSer109Asp + Ser220Asp + Leu221GluThr215Glu + Ser216Asp + Ser220AspSer192Asp + Phe193Glu + Thr217AspGly165Asp + Tyr210Glu + Ser220AspSer 70Glu + Asp105Glu + Tyr218AspGly 69Glu + Ala168Glu + Thr195Glu表34多環四突變變異體Asn 66Gln + Leu104Gly + Tyr210Pro + Thr217GluGly 67Ser + Gly 69Ser + Val103Gln + Gly223AspTrp112Cys + Ala164Glu + Trp208Gln + Thr217GlyLeu134Ile + Trp208Gln + Ala219Gly + Ser220GluLeu104Ala + Ser109Glu + Thr111Gln + Val115AlaGly108Glu + Ala164Gly + Gly165Gln + Thr167ProGly 69Glu + Asn106Gln + Trp112Cys + Thr113ProThr 73Asp + Leu134Ser + Thr137Asn + Thr217ProGly 67Ser + Gly136Pro + Thr195Ser + Leu221AlaThr 73Gly + Leu104Gly + Trp208Pro + Thr215GlyGly110Glu + Gly136Pro + Phe193Met + Thr217ProAla164Gln + Tyr171Ile + Phe193Ile + Thr224GlyGly136Pro + Ala164Thr + Ser207Asp + Trp208LeuGly108Gln + Trp112Pro + Trp208Thr + Tyr210GluThr111Asp + Leu134Val + Tyr210His + Thr217ProGly136Gln + Trp208Phe + Ala219Asp + Leu221MetThr113Ser + Tyr171Leu + Trp208Val + Tyr210ThrGly136Ser + Ser141Glu + Trp208Thr + Tyr210LeuAsn 66Ser + Thr 73Pro + Thr113Pro + Asn170GluThr 73Pro + Gly139Ser + Tyr210Glu + Leu221AlaGly108Ser + Val138Ser + Trp208Leu + Thr217SerAsn 68Ser + Gly110Gln + Trp112Met + Ser192AspThr 73Gln + Val103Ala + Gly162Ser + Ala164GluThr137Asn + Asn140Asp + Phe193Thr + Trp208AlaLeu134Ser + Asn163Gln + Ser220Glu + Gly223AsnAsn140Glu + Tyr171Gln + Tyr210Val + Gly223SerThr 73Pro + Leu134Val + Gly136Glu + Thr217ProSer107Glu + Gly162Gln + Trp208Ser + Leu221ProGly108Asn + Gly162Asn + Ser192Asp + Tyr210CysVal103Pro + Ser109Glu + Trp208Asn + Tyr210ProThr 73Gln + Trp112Tyr + Leu134Cys + Asn170SerGly 69Asn + Gly162Asn + Asn166Ser + Leu221AsnGly 67Ser + Leu134Ser + Gly162Asn + Ala164GlnLeu134Ile + Gly135Pro + Ala168Thr + Trp208MetTrp112Ser + Ala168His + Thr217Ser + Leu221AlaGly110Ser + Thr111Gln + Thr215Ser + Ser222AspThr111Pro + Thr137Asn + Pro169Gln + Ala219GluThr 73Glu + Asn106Ser + Trp208His + Leu221ThrAsn 66Asp + Gly 67Pro + Gly 69Ser + Thr224SerLeu104Cys + Val115His + Ser191Asp + Tyr218ValLeu134Glu + Thr195Asn + Trp208Ser + Leu221SerVal103Ser + Gly139Asn + Thr217Asn + Leu221CysAsn106Gln + Ala164Pro + Asn166Asp + Phe193ProAsp105Glu + Gly135Pro + Val138Ser + Thr217ProGly 67Asp + Thr113Asn + Ala114Asn + Tyr210CysAsn166Asp + Thr195Gly + Tyr210Gly + Thr224GlnTrp112Pro + Asn166Ser + Phe193His + Tyr218IleGly 69Pro + Leu104Glu + Val115Ser + Ala164SerTrp112Pro + Val115Glu + Gly139Asn + Tyr210GlnGly 69Ser + Thr 73Ser + Tyr210Pro + Thr224ProGly 72Gln + Thr 73Pro + Gly110Asp + Leu221HisThr111Asn + Gly136Pro + Tyr210Thr + Tyr218AspGly108Asp + Gly136Asn + Asn166Ser + Leu221ThrVal103Met + Pro205Ser + Leu221Met + Thr224SerGly 69Asn + Thr 73Asp + Trp208Gln + Tyr210LeuThr 73Gly + Asn166Ser + Thr167Pro + Leu221GlnAsn 66Gln + Thr113Asp + Tyr171Gly + Tyr210ProGly162Glu + Thr167Pro + Trp208Val + Tyr210LeuSer207Asp + Trp208Ala + Thr212Gly + Thr217ProLeu104Met + Thr137Gln + Asn163Asp + Pro205GlnSer109Asp + Leu134Val + Gly165Gln + Trp208ThrAla114Asn + Val115Asn + Trp208Gln + Tyr210SerAsn 66Glu + Ala164Asn + Trp208Cys + Thr215ProGly135Ser + Ala168Pro + Ser207Asp + Thr217GlnThr167Asp + Trp208Met + Thr217Gln + Leu221AsnThr 73Pro + Asn106Glu + Thr113Ser + Leu221CysGly162Gln + Ser191Glu + Thr217Asn + Thr224ProGly 69Gln + Leu104Gln + Gly162Glu + Tyr210GlnLeu104Gln + Tyr171Pro + Thr217Gly + Leu221GluGly110Glu + Phe193Asn + Trp208Cys + Gly223ProAla164Gly + Ser191Glu + Pro205Asn + Trp208CysGly135Pro + Ser207Glu + Trp208Ala + Tyr210CysThr 73Glu + Leu104Val + Pro205Gln + Leu221CysTrp112Cys + Val138Thr + Thr167Gln + Thr217ProSer109Asp + Ala164His + Gly223Pro + Thr224AsnAsn 68Gln + Tyr210Pro + Thr212Ser + Ser220AspLeu134Gly + Trp208Glu + Tyr210Met + Thr212SerAsn163Asp + Asn170Gln + Trp208Asn + Tyr210ThrTrp112Gly + Trp208Ser + Tyr210Asp + Thr217ProTrp112Met + Ser192Glu + Trp208Ser + Tyr210ValTrp112Leu + Ser220Asp + Leu221Ala + Thr224AsnLeu104Met + Thr111Asp + Ala164Gln + Pro214SerGly139Gln + Gly165Pro + Thr212Ser + Ser220GluThr113Asp + Leu134His + Trp208Ile + Leu221AlaAsp105Glu + Pro214Gln + Thr215Gln + Leu221GlnLeu134Pro + Gly162Asn + Gly206Ser + Trp208TyrTrp112Ala + Phe193Ile + Thr217Asn + Ser220AspSer107Glu + Thr212Asn + Tyr213Ala + Thr217ProGly 67Gln + Gly 69Gln + Thr113Asn + Ser220GluAsn 66Gln + Trp208Cys + Tyr210Asn + Thr217AspThr 73Glu + Leu104Ile + Gly139Gln + Thr217GlyThr113Ser + Pro169Asn + Tyr210Ile + Thr217SerLeu134Pro + Thr167Gln + Tyr210Met + Ser220AspThr 73Glu + Asn166Gln + Tyr171His + Leu221AlaVal138Met + Asn170Gln + Thr217Asn + Ala219HisGly136Gln + Ala219Ser + Ser220Asp + Thr224GlnGly 69Asn + Asn166Gln + Tyr210Glu + Thr217GlyThr 73Ser + Gly165Gln + Thr212Pro + Ser222GluThr137Pro + Asn140Gln + Tyr171Gly + Ser220GluTyr210Pro + Tyr213Ile + Ser220Asp + Leu221ProVal103Asp + Asn106Ser + Phe193His + Thr217GlyAsn 68Asp + Thr113Asn + Val138Ala + Trp208HisAsp105Glu + Leu134Met + Gly165Ser + Leu221GlnAsn 68Gln + Trp208Thr + Thr217Ser + Ser220AspTyr171Asn + Thr195Asn + Tyr210Asn + Tyr218AsnAsn 66Ser + Pro169Ser + Tyr210Cys + Gly223GluAla114Gln + Phe193Glu + Tyr210Gly + Thr224ProAsn 66Ser + Gly136Ser + Ala164Asp + Ala168GlnLeu104Gly + Gly162Pro + Ser192Asp + Leu221AsnGly108Asp + Gly162Pro + Trp208Thr + Gly223AsnAsn 66Glu + Thr113Gly + Ala164Pro + Gly223ProGly110Pro + Trp112Ala + Ser216Glu + Leu221HisVal115Met + Pro169Ser + Trp208His + Tyr210AspLeu104Ser + Ser109Asp + Gly110Pro + Thr195GlyLeu104Asn + Thr137Glu + Trp208Ser + Leu221ValAsn106Gln + Ala114Asp + Tyr171Thr + Leu221ProPro205Asn + Tyr210Asn + Thr217Gly + Tyr218LeuLeu104Pro + Asn106Ser + Thr137Asp + Thr217ProGly110Ser + Thr137Gln + Asn170Glu + Thr217AsnThr113Gly + Phe193Met + Trp208His + Tyr213IleAsn166Gln + Ser207Asp + Thr217Ser + Tyr218MetGly162Asp + Phe193Pro + Trp208Tyr + Thr217GlnGly 69Ser + Ser107Glu + Trp208Met + Tyr210LeuVal103Asp + Thr167Pro + Ile209Val + Tyr210GlnThr111Asn + Asn170Ser + Trp208Tyr + Thr212GlySer107Glu + Thr137Pro + Gly162Ser + Trp208CysVal103Thr + Ala164Thr + Ala168Glu + Thr217SerGly139Pro + Tyr210Asn + Leu221Pro + Thr224ProLeu134Ser + Ser141Glu + Tyr210Ala + Thr217GlnVal103Asn + Leu134Ala + Tyr218Met + Thr224AsnAsn 66Asp + Gly135Gln + Asn163Ser + Leu221ValGly 69Glu + Val115Pro + Thr137Pro + Trp208IleGly 69Pro + Gly165Pro + Tyr210Ser + Leu221IleAsn 66Gln + Gly110Glu + Trp112Gly + Trp208ProTrp112Ala + Ala114Pro + Gly135Ser + Asn163AspGly 67Asn + Ser207Glu + Tyr210Thr + Thr217ProAsn166Asp + Thr167Pro + Ile209His + Tyr210ValThr 73Ser + Asn106Gln + Tyr171Glu + Thr217AsnVal115Glu + Tyr210Ser + Thr217Gln + Ala219GlyLeu104Pro + Thr195Gln + Trp208Tyr + Ser222AspSer141Asp + Ala204Gln + Trp208Val + Tyr210LeuGly 67Ser + Gly 69Glu + Val103Gly + Tyr210GlnTyr171Met + Phe193Leu + Thr215Asp + Thr217GlnAsn 68Gln + Asp105Glu + Trp208Phe + Leu221AlaLeu104Asn + Val115Gln + Leu134Gln + Pro214GluGly165Ser + Ser191Asp + Pro214Gly + Leu221ThrGly 67Ser + Asn 68Gln + Tyr210Val + Ser220AspSer109Asp + Val138Gly + Trp208Ser + Thr217ProThr111Gly + Gly136Pro + Ser141Asp + Leu221IleTrp112Asn + Gly162Ser + Ile209Cys + Thr224GlnVal115Ala + Pro169Glu + Tyr210Val + Leu221CysGly135Ser + Tyr210Leu + Thr217Glu + Leu221ProAsn 66Gln + Ser109Asp + Gly110Gln + Tyr218ThrThr 73Gly + Trp208Ile + Thr217Glu + Leu221AsnLeu134Ser + Trp208His + Tyr210Leu + Ser220GluGly135Pro + Thr137Gln + Ala219Ser + Leu221GlnAsn 68Gln + Asn106Glu + Pro169Gln + Thr215GlnVal138Gln + Gly165Glu + Tyr218Leu + Leu221CysPhe193Gln + Ser207Asp + Trp208Cys + Thr217GlnGly 67Asn + Gly165Asn + Phe193Tyr + Pro205SerAsn 66Ser + Ala168Ser + Tyr210Cys + Thr217AspPro214Gly + Ser216Asp + Thr217Gln + Leu221HisAsn 66Gln + Val115Gln + Asn166Asp + Tyr218MetThr111Glu + Gly165Ser + Ala219His + Leu221ThrSer141Asp + Asn163Ser + Phe193Tyr + Ala204SerGly108Pro + Asn163Ser + Gly165Asp + Tyr213SerTrp208His + Tyr210Thr + Tyr218Gln + Ser220AspGly 69Glu + Ala164Pro + Trp208Val + Thr224SerSer109Asp + Ala203Pro + Trp208His + Thr217SerThr 73Asp + Tyr171Cys + Tyr210Pro + Leu221ProGly162Pro + Tyr218Val + Ala219Thr + Ser220GluAsn170Gln + Ser192Glu + Trp208Pro + Tyr210GlyThr113Ser + Thr137Gln + Phe193Ser + Pro205SerTrp112Pro + Thr137Gly + Ser220Asp + Leu221AsnGly162Glu + Ala164Ser + Ala168His + Pro214AsnAla114Pro + Leu134Glu + Asn140Ser + Phe193IleAla164Gln + Ala168Glu + Trp208Asn + Tyr210LeuVal103Pro + Gly139Pro + Asn163Asp + Gly165GlnGly162Ser + Trp208Glu + Tyr210Ser + Leu221GlnSerl41Asp + Tyr171His + Trp208Ala + Leu221MetAsn 66Ser + Gly139Gln + Asn163Gln + Tyr210ThrLeu134Asp + Gly136Ser + Asn163Gln + Trp208ValSer191Asp + Phe193Leu + Pro214Gln + Thr217SerGly 67Asn + Gly108Asp + Thr195Asn + Thr215ProAla114Thr + Asn140GIu + Asn166Ser + Ala203ProSer191Glu + Ala203Gly + Trp208Asn + Tyr210AlaAsn140Asp + Ser141Glu + Ala164Gln + Tyr218SerThr 73Ser + Tyr213Val + Ser220Glu + Leu221GluPhe193Met + Thr215Gln + Ser220Glu + Leu221GluThr113Ser + Trp208Leu + Ser220Glu + Leu221GluThr 73Pro + Val115Ala + Ser192Asp + Phe193GluAla114Asn + Trp208Pro + Tyr210Glu + Ser220GluTrp208Cys + Tyr210Glu + Ala219Asn + Ser220AspAla114Pro + Gly139Pro + Tyr210Asp + Ser220GluTyr210Glu + Thr217Gln + Tyr218Gln + Ser220GluAsn140Ser + Trp208Glu + Ser220Asp + Leu221AspVal103Pro + Trp208Glu + Tyr210Ser + Ser220GluThr 73Gln + Ser107Asp + Ser109Glu + Asn163GlnGly 67Gln + Ser107Asp + Ser109Glu + Leu221CysAsp105Glu + Asn106Ser + Ser109Glu + Thr217ProLeu104Glu + Asp105Glu + Ser107Asp + Leu221SerGly206Pro + Pro214Asp + Thr217Glu + Leu221AsnSer207Asp + Trp208Glu + Ser220Asp + Leu221SerGly135Asp + Gly162Glu + Tyr171Asp + Thr217SerGly 69Glu + Leu104Asp + Asp105Glu + Tyr210AlaAla168Asn + Ser220Asp + Leu221Pro + Ser222AspPhe193Gln + Ser207Glu + Thr217Ser + Leu221GluGly110Gln + Gly165Gln + Ser222Asp + Thr224AspAsn106Glu + Ser109Asp + Phe193Ile + Trp208LeuGly110Asn + Thr137Glu + Pro169Asp + Asn170SerGly136Glu + Gly162Glu + Trp208Ala + Thr217AsnLeu134Ile + Ser191Glu + Leu221His + Gly223GluGly139Pro + Tyr171Met + Ser207Asp + Ser220GluVal103Pro + Thr111Gly + Ser207Asp + Ser220AspGly108Asn + Thr195Gly + Ser207Asp + Ser220AspSer 70Glu + Val103Pro + Asn106Asp + Ala168ProThr113Gly + Gly162Ser + Asn163Asp + Asn170GluLeu104Ile + Ser107Glu + Gly110Asp + Asn170GlnThr 73Gly + Gly135Asp + Pro169Asp + Trp208ThrTyr210Met + Thr215Asp + Tyr218Glu + Leu221AsnLeu104Thr + Gly136Glu + Pro169Asp + Leu221ValAsn 66Asp + Thr 73Glu + Tyr218Ile + Leu221CysAsp105Glu + Thr111Asp + Ala114Pro + Val138MetAsn 68Glu + Thr 73Glu + Trp208Tyr + Thr217AsnSer207Glu + Tyr213Ile + Ala219Asp + Ser222AspAsn163Asp + Ala164Ser + Ser191Asp + Ser207GluThr 73Glu + Ala168Ser + Tyr210Glu + Tyr218GluGly 67Glu + Trp208Ile + Pro214Asn + Thr215AspAsp105Glu + Thr111Glu + Thr113Glu + Asn163GlnSer 70Glu + Tyr210His + Thr217Asp + Leu221AlaGly 67Glu + Gly108Asp + Thr137Gln + Thr167ProSer107Asp + Gly110Asp + Ala114Glu + Tyr218AsnSer 70Asp + Tyr210Glu + Thr217Asn + Ser220GluThr113Gly + Trp208Glu + Ser220Asp + Thr224AspVal138Met + Ala219Glu + Ser222Asp + Thr224GlnSer 70Glu + Trp112Tyr + Tyr218Cys + Leu221GluGly 67Ser + Ser 70Glu + Pro169Asn + Leu221AspVal115Cys + Gly139Asn + Thr217Asp + Ser220AspGly 69Ser + Val115Thr + Thr217Glu + Ser220GluAla168His + Trp208Tyr + Thr217Asp + Ser220GluThr111Gln + Gly135Pro + Thr217Asp + Ser220GluAla164Thr + Ser191Glu + Trp208Glu + Thr215AsnAsn 68Glu + Val103Glu + Ser109Glu + Asn166GlnSer 70Asp + Gly139Ser + Thr217Glu + Ser220AspLeu134Asp + Phe193Glu + Ala203Ser + Gly223AspAsn 66Asp + Asn 68Ser + Ser216Asp + Thr217GluAla164Glu + Trp208Cys + Leu221Glu + Ser222AspSer 70Glu + Ser207Glu + Ala219Ser + Ser220GluGly 67Asp + Ser 70Asp + Thr 73Gly + Tyr210GluTrp208Ile + Thr217Glu + Tyr218Asp + Ser222AspLeu104Ile + Thr111Glu + Gly135Asp + Asn163GlnAla114Glu + Leu134Ile + Ala219Glu + Ser220AspAsn166Glu + Thr167Gly + Ser222Asp + Gly223AspGly139Glu + Tyr210Ala + Ser222Asp + Gly223GluVal103Gly + Gly110Glu + Thr111Glu + Ser222GluAla114Glu + Asn170Ser + Ser216Asp + Thr217GluGly108Asp + Ser109Asp + Tyr210Gln + Thr217GluAsn 68Ser + Ser109Asp + Gly110Glu + Ser220GluAsn 68Glu + Asn163Ser + Ser207Asp + Trp208GluSer207Asp + Trp208Glu + Thr217Glu + Leu221CysGly 69Pro + Asp105Glu + Asn106Asp + Asn166AspSer109Asp + Tyr210Gln + Ser220Glu + Leu221AspTyr171Gly + Ser192Glu + Ser220Asp + Leu221GluAsn170Glu + Ser191Glu + Ser192Glu + Pro205SerAsn 66Asp + Tyr210Gln + Thr217Glu + Tyr218GluAsn106Asp + Ser107Glu + Thr137Asp + Tyr210GlnSer192Asp + Phe193Glu + Pro205Ser + Thr217AspSer 70Asp + Gly162Asp + Asn163Glu + Leu221SerGly 69Asn + Asn106Glu + Tyr210Asp + Ser220GluVal103Ser + Asn140Asp + Tyr210Glu + Ser220AspGly 69Glu + Phe193His + Ser207Glu + Ser222AspGly162Glu + Asn163Gln + Trp208Asp + Ser220GluVal115Cys + Asn170Asp + Trp208Asp + Ser220AspGly110Glu + Trp208Asp + Leu221Asn + Ser222GluPro169Asp + Tyr171Glu + Pro214Asn + Gly223GluAsn163Glu + Ser207Glu + Trp208Met + Tyr210ThrGly 67Glu + Ser 70Glu + Val115Cys + Gly162GluGly162Glu + Ala164Glu + Ser220Glu + Leu221GlyAsn 66Glu + Val103Asp + Gly108Asn + Leu221MetAsp105Glu + Ser109Glu + Tyr218Asp + Leu221AlaGly 67Asp + Ala164Asp + Ala168Glu + Tyr213AsnGly 69Pro + Ser141Asp + Pro214Asp + Thr217AspThr 73Glu + Trp112Ile + Trp208Gly + Ser220AspGly 72Ser + Ser107Asp + Thr111Asp + Asn166GlnGly165Glu + Ser192Asp + Thr217Asp + Leu221IleGly135Glu + Asn140Glu + Ala204His + Pro214SerAsp105Glu + Gly110Asp + Asn170Asp + Ala219GlyAsn 66Asp + Asn 68Asp + Leu134Gly + Asn163GluLeu104Thr + Ala168Asp + Asn170Asp + Ser220GluThr 73Asp + Ser109Glu + Pro214Glu + Thr217AsnThr 73Asp + Val115Cys + Pro214Asp + Thr224GluSer109Asp + Thr113Asp + Leu134Asn + Phe193ProGly 69Ser + Gly108Asp + Gly110Asp + Ser220GluGly139Asp + Ser141Asp + Ala164Asp + Leu221ValAsn 66Asp + Trp208Met + Ser220Asp + Ser222GluTrp112Asp + Tyr210Thr + Ser220Glu + Ser222GluSer109Asp + Ser207Glu + Tyr210Gly + Leu221GluGly 69Asp + Asp105Glu + Thr137Asp + Trp208AsnSer109Asp + Thr111Glu + Ser191Glu + Tyr210LeuGly139Gln + Pro169Asp + Ser191Asp + Ser207GluGly 67Ser + Asn140Asp + Ser222Asp + Thr224GluVal138Asp + Asn170Glu + Trp208His + Thr215GluAsn 68Asp + Thr111Gly + Val138Asp + Asn170AspLeu104Glu + Ser107Asp + Gly135Gln + Tyr171AspLeu104Pro + Trp112Asp + Asn140Glu + Ser222GluGly139Glu + Asn170Asp + Gly206Pro + Ser220GluAla164Asp + Asn170Asp + Ser216Asp + Thr217ProThr113Glu + Ala114His + Ser207Glu + Ser220GluGly110Glu + Val115Gln + Ser207Asp + Thr224GluAsn106Glu + Asn163Glu + Asn170Glu + Leu221HisGly 69Asp + Thr 73Glu + Thr113Asn + Ser141GluGly 69Asp + Thr 73Asp + Thr111Gly + Gly139GluSer107Asp + Asn163Glu + Ser192Asp + Trp208MetAsn 68Asp + Gly108Asp + Ser207Asp + Ile209AlaGly135Asp + Gly139Pro + Pro169Asp + Trp208GluAsn 66Glu + Thr167Gly + Trp208Glu + Pro214AspAsn 66Asp + Ser109Asp + Trp208Gly + Pro214GluGly 69Glu + Asn140Ser + Ser192Glu + Pro214AspLeu104Gly + Gly139Glu + Ser191Asp + Thr224AspAsn106Gln + Thr113Asp + Asn163Asp + Asn166GluLeu134Glu + Ser207Asp + Trp208Pro + Tyr210GluSer191Glu + Thr195Asn + Tyr218Pro + Leu221GluAsn170Asp + Phe193Ser + Ser220Glu + Thr224AspAsp105Glu + Ala114Glu + Ser192Glu + Thr224AsnGly 67Asp + Gly108Asp + Ser207Asp + Tyr210CysAsn 68Asp + Gly110Glu + Thr113Gly + Leu221AlaLeu104Ile + Ala114Glu + Ser220Glu + Gly223GluSer107Asp + Ser220Glu + Leu221Pro + Gly223AspVa1103Asp + Val138Glu + Ser141Glu + Gly206GlnVal138Glu + Ser141Asp + Tyr210Gln + Ser220AspAsn 68Asp + Val115Gly + Thr217Asp + Ser220GluAsn 66Asp + Leu104Gly + Ala168Thr + Ser216GluVal103Glu + Asn106Glu + Thr113Pro + Ser220AspAsp105Glu + Thr113Asp + Gly139Asn + Ser141GluSer 70Glu + Asp105Glu + Gly139Asp + Tyr210AlaAsn 66Glu + Tyr210Asp + Thr217Glu + Leu221AlaGly108Glu + Gly135Ser + Tyr210Asp + Thr217GluGly 69Asp + Gly108Gln + Trp112Asp + Ser141GluSer107Glu + Gly110Pro + Thr217Asp + Ser220AspGly135Glu + Thr217Glu + Tyr218Val + Ser220GluAsn163Asp + Tyr171Thr + Thr217Asp + Ser220AspAla114Thr + Thr137Glu + Thr217Asp + Ser220AspAsn166Glu + Thr212Pro + Thr217Asp + Ser220AspGly 67Asp + Ser107Asp + Tyr210Asn + Ser222GluAsn140Ser + Trp208Gly + Thr217Glu + Leu221AspAsn166Gln + Thr215Pro + Thr217Glu + Leu221AspVal115Gln + Ala168Gln + Phe193Asp + Leu221AspTrp208Ile + Pro214Ser + Ser220Glu + Thr224GluGly108Glu + Thr137Glu + Asn163Asp + Trp208SerGly139Pro + Ala164Glu + Ser191Asp + Pro214GluThr 73Asp + Thr111Pro + Asn166Asp + Ser191AspSer 70Glu + Ala168His + Thr195Glu + Ser216AspSer109Glu + Thr137Asp + Gly139Ser + Ala164GlnAsn170Gln + Tyr210Asp + Pro214Glu + Gly223SerSer 70Glu + Gly139Asn + Ala164Glu + Ser220AspAsn 66Asp + Thr113Glu + Ser141Asp + Trp208GlyAsn 66Asp + Gly108Glu + Thr215Gly + Leu221SerAsn 68Ser + Thr 73Glu + Asn166Glu + Leu221AspThr 73Asp + Ser192Asp + Ser220Asp + Leu221MetSer107Asp + Thr111Asp + Ser192Glu + Thr217Ser表35多環五突變變異體Asn 66Gln + Gly 67Ser + Leu104Gly + Tyr210Pro +Thr217GluGly 69Ser + Val103Gln + Trp112Cys + Thr217Gly +Gly223AspVal115Pro + Leu134Ile + Ala164Glu + Phe193Asn +Trp208GlnLeu104Gly + Thr113Ser + Ala168Glu + Asn170Ser +Ala219GlyGly 69Ser + Ser109Asp + Thr111Gln + Val115Ala +Tyr210ProVal103Thr + Gly108Glu + Val115Pro + Tyr171Ser +Trp208AsnLeu134Ala + Val138Cys + Asn163Gln + Tyr171Ser +Ser220AspAsn 66Ser + Val115Gly + Ser191Glu + Trp208Ile +Thr217SerGly 72Asn + Trp112Phe + Val115Ala + Asn166Gln +Trp208GlyLeu134His + Gly165Gln + Trp208Pro + Tyr210Val +Thr217GluLeu104Pro + Tyr171Leu + Trp208Leu + Ala219Glu +Leu221SerAsn166Glu + Thr167Pro + Tyr210Pro + Tyr213Ile +Pro214AsnGly139Gln + Asn170Glu + Trp208Cys + Thr215Ser +Leu221AsnAla114Glu + Gly135Ser + Ala168Thr + Ala203Thr +Trp208MetAla164His + Ala168His + Phe193Ala + Thr217Ser +Ser220AspGly 67Pro + Gly 69Ser + Ser191Asp + Tyr218Val +Thr224SerGly139Asn + Ala164Pro + Phe193Pro + Thr217Asn +Leu221CysLeu104Met + Ser207Asp + Trp208Ala + Thr212Gly +Thr217ProLeu134Val + Trp208Thr + Pro214Gly + Leu221Ile +Thr224AspSerl09Asp + Ala114Asn + Val115Asn + Gly165Gln +Tyr210SerLeu104Ser + Gly110Pro + Gly136Glu + Ala168Thr +Ala219GlnGly110Pro + Leu134Pro + Asn166Asp + Asn170Ser +Trp208AsnThr113Gln + Pro169Gly + Asn170Gln + Tyr218Gln +Leu221SerThr137Ser + Thr195Gln + Ser207Glu + Trp208Gln +Thr217ProGly 69Gln + Val103Gly + Trp112Met + Trp208Asn +Pro214GlyGly136Pro + Gly162Ser + Trp208Ile + Tyr210Gly +Pro214GlnThr137Asp + Gly165Gln + Thr195Ser + Trp208Pro +Thr212SerAsn 66Ser + Pro169Ser + Tyr210Asn + Tyr218Asn +Gly223GluSer 70Glu + Asn106Gln + Val138Ala + Ala168Gln +Leu221ValGly 69Ser + Thr113Asp + Ala114Gln + Trp208Thr +Ala219SerThr113Gln + Ala168Asp + Pro169Ser + Asn170Ser +Ala219ThrGly139Ser + Asn170Gln + Tyr171Leu + Trp208Cys +Thr215SerGly 67Ser + Ala114Asn + Gly139Ser + Tyr210Thr +Tyr218SerTrp112Glu + Ala164Pro + Tyr171Pro + Thr195Asn +Thr217SerGly 67Glu + Val115Asn + Gly162Gln + Gly165Gln +Phe193ValGly135Ser + Tyr171Cys + Trp208His + Tyr210Met +Leu221ValGly 69Asn + Asn106Ser + Asn166Glu + Tyr210Cys +Leu221GlyGly135Ser + Tyr171Asn + Trp208His + Ser216Glu +Leu221GlyGly 69Asn + Ser109Asp + Gly165Asn + Phe193Asn +Trp208HisPro169Ser + Ser207Glu + Pro214Asn + Thr215Asn +Thr217GlySer 70Asp + Asn163Ser + Tyr210Val + Tyr213Asn +Leu221AlaGly108Gln + Gly110Ser + Gly135Gln + Thr137Asn +Trp208GluAsn 68Ser + Ala164Gln + Asn170Glu + Thr217Ser +Gly223SerGly 69Ser + Trp112Cys + Ala164Gly + Leu221Ser +Gly223GluGly 69Glu + Gly 72Gln + Thr137Pro + Trp208Ile +Tyr218ProVal103Gln + Val138His + Gly162Ser + Asn166Asp +
Trp208HisVal115Gln + Leu134Cys + Ala168Ser + Pro169Asp +Tyr210SerGly 67Gln + Trp112Leu + Phe193His + Thr195Glu +Thr217SerGly 67Gln + Leu104Asn + Ala164Glu + Ala204Ser +Pro205SerGly135Ser + Val138Ser + Asn140Glu + Tyr210Leu +Thr217GlyGly 67Asn + Thr 73Ser + Tyr171Leu + Phe193Tyr +Tyr210ProGly135Pro + Trp208His + Tyr210Leu + Ala219Ser +Leu221GlnAsn 68Gln + Asn106Glu + Thr137Gln + Pro169Gln +Thr215GlnThr137Asp + Val138Gln + Trp208Ile + Tyr218Leu +Leu221CysGly 67Asn + Gly165Asn + Phe193Tyr + Pro205Ser +Trp208CysAsn 66Ser + Thr 73Ser + Ser107Asp + Ala168Ser +Tyr210CysThr137Gly + Tyr210Pro + Thr215Pro + Ser216Asp +Thr217SerThr 73Pro + Trp112Asp + Gly139Pro + Thr215Pro +Leu221HisLeu104Cys + Thr111Pro + Trp208Pro + Leu221Cys +Ser222AspThr113Gln + Ala204His + Trp208Phe + Thr217Asp +Leu221CysThr111Gly + Gly139Gln + Tyr171Leu + Tyr210Met +Thr217GluGly110Asn + Val115Gln + Pro169Gln + Thr195Gly +Ser220AspTyr171Asp + Gly206Ser + Thr217Gln + Ala219Gly +Leu221ThrSer109Glu + Thr111Ser + Val115Cys + Val138Cys +Tyr171SerLeu104His + Asn163Asp + Trp208Thr + Tyr210Cys +Thr217AsnGly136Asn + Gly139Pro + Ser141Glu + Pro169Gln +Ala219HisGly 69Ser + Gly136Ser + Thr137Asn + Thr195Pro +Ala219GluAsn106Asp + Gly162Ser + Asn163Gln + Pro169Gly +Leu221ProAla168Gly + Phe193Met + Pro205Asn + Tyr210Asn +Leu221AsnGly 69Pro + Gly139Asp + Ala164Gly + Tyr213Ser +Thr217SerGly 72Ser + Leu104Gln + Asn166Glu + Pro169Asn +Thr215GlyLeu134Cys + Trp208Asn + Tyr210Ala + Tyr218Gly +Thr224GlyLeu104Gln + Thr195Asn + Trp208Val + Tyr210Pro +Leu221CysGly 67Pro + Thr 73Asn + Trp208Pro + Ala219Asn +Ser220AspThr 73Gln + Gly108Pro + Gly139Pro + Ser192Glu +Gly223GlnAsn 68Glu + Thr 73Gln + Pro169Gln + Tyr210Pro +Leu221MetLeu104His + Thr113Gln + Asn140Gln + Thr167Asn +Phe193AspGly 67Pro + Asn 68Ser + Gly139Asp + Thr215Gly +Thr217ProGly136Ser + Val138Met + Thr167Gln + Tyr218Asn +Ser220AspAsp105Glu + Gly110Ser + Trp112His + Tyr210Gln +Ala219ThrAsn 68Asp + Asn106Gln + Ala164Ser + Thr195Gln +Trp208CysLeu104Val + Leu134Ala + Thr137Asn + Asn170Asp +Leu221IleGly 67Ser + Leu134Met + Trp208Ala + Tyr210Ser +Thr217ProAla114Gly + Gly135Asp + Gly139Ser + Asn170Ser +Thr217SerThr111Asn + Phe193Ser + Pro205Gln + Trp208Thr +Leu221GluSer109Asp + Val138Asn + Phe193Ala + Trp208Cys +Thr212GlnSer 70Asp + Asn166Ser + Ile209Pro + Tyr210Met +Leu221SerAsn106Ser + Leu134Cys + Gly136Ser + Ala168Thr +Tyr210GluGly 67Gln + Leu134Gln + Gly135Asp + Phe193His +Thr224GlyAsn 68Ser + Leu104Ala + Gly162Asp + Tyr171Met +Thr217GlyAsn163Glu + Thr212Gln + Thr215Asn + Thr217Asn +Leu221MetGly 69Gln + Asn106Gln + Ser207Glu + Trp208Thr +Thr217GlyGly139Glu + Ala168His + Thr195Gly + Leu221His +Thr224AsnGly 72Ser + Ala114Asn + Leu134His + Pro169Glu +Trp208TyrVal138Ser + Ala164Asn + Tyr210Cys + Tyr218Leu +Ser220AspLeu104Gln + Tyr171Glu + Trp208Gly + Thr217Gln +Ala219HisAsn106Ser + Tyr171Gly + Phe193Gln + Thr215Gln +Ala219GlnGly 72Ser + Gly108Asn + Trp112Ile + Asn166Glu +Leu221CysAsn 68Ser + Leu104Thr + Thr215Ser + Ala219Thr +
Thr224AspVal138Ser + Asn163Asp + Phe193Cys + Pro205Asn +Thr217SerGly 67Ser + Tyr210Pro + Tyr218Thr + Ser220Asp +Leu221SerGly135Asn + Gly136Gln + Gly162Ser + Tyr210Asn +Pro214AsnThr195Ser + Trp208Pro + Tyr210Gly + Ala219Ser +Leu221IleAsn 68Glu + Thr111Pro + Asn163Ser + Phe193Tyr +Leu221ValGly 67Ser + Thr 73Pro + Asn170Glu + Tyr210Leu +Leu221ValGly135Asn + Gly139Ser + Phe193Asp + Thr217Gly +Leu221GlyGly136Gln + Gly139Gln + Gly165Ser + Tyr210Asp +Thr217ProSer141Glu + Gly162Pro + Thr167Gly + Tyr210Ile +Thr217SerLeu104Pro + Gly110Pro + Gly136Pro + Pro205Asn +Thr217GluVal115Ala + Ala164Thr + Ser192Asp + Trp208Leu +Tyr210MetThr 73Asn + Val103Asn + Thr215Asn + Thr217Gln +Ser222AspVal103Pro + Trp208Tyr + Ile209Cys + Tyr210Thr +Thr217GlnTrp112Phe + Ala114Ser + Pro169Gln + Ser220Glu +Leu221HisGly108Asn + Leu134Met + Tyr210Ser + Tyr213Ile +Pro214GluAla168Pro + Trp208His + Tyr210Ala + Ser220Glu +Leu221GlnGly139Pro + Asn140Glu + Ser141Asp + Asn166Ser +Trp208ThrVal103Asn + Thr111Gly + Tyr210Gln + Ser216Asp +Thr217GluVal138His + Gly139Asn + Ser207Asp + Trp208Glu +Leu221GlnAla168Asn + Pro169Gly + Thr217Pro + Ser220Asp +Leu221AspTyr210Leu + Ala219Gly + Ser220Asp + Leu221Asp +Gly223SerVal103Ala + Pro169Asn + Thr217Gln + Ser220Asp +Leu221AspThr 73Asn + Asn170Asp + Tyr171Glu + Tyr210Ile +Leu221SerGly136Ser + Ser191Asp + Ser192Glu + Tyr218Cys +Leu221AsnThr 73Pro + Val115Ala + Ser192Asp + Phe193Glu +Trp208GlnThr137Gln + Gly162Asp + Asn163Asp + Ala203His +Pro205GlnAsn163Gln + Tyr210Asp + Thr215Pro + Ser220Glu +Gly223ProAsp105Glu + Asn106Glu + Ser107Asp + Trp208Phe +Tyr210SerTrp112Gly + Phe193Gln + Ser207Asp + Trp208His +Ser222AspGly108Asn + Ser207Glu + Tyr213Thr + Thr215Pro +Ser222AspLeu134Met + Ser207Asp + Trp208Val + Tyr210His +Ser222AspGly 69Glu + Asn106Asp + Ala114His + Ala168Ser +Thr217GlnLeu104Pro + Trp208Glu + Thr212Asn + Thr215Ser +Ser222AspThr111Pro + Thr137Asn + Pro169Gln + Tyr210Glu +Ala219GluThr 73Gly + Val115Gly + Trp208Glu + Tyr210Asp +Ser220GluGly135Gln + Gly136Glu + Asn170Ser + Tyr171Glu +Trp208GlnThr 73Ser + Ser107Asp + Ser109Asp + Gly136Pro +Tyr210GlyLeu134Pro + Pro169Gln + Pro214Glu + Ser216Glu +Thr217GlyAsp105Glu + Ser109Glu + Pro169Gly + Tyr171Gln +Thr212SerGly135Pro + Val138Ala + Ser207Asp + Tyr210Thr +Gly223GluGly135Pro + Ser207Glu + Trp208Ala + Tyr210Pro +Gly223GluPro169Gln + Phe193Val + Tyr210Glu + Thr215Asn +Leu221AspAla114Gln + Gly136Ser + Ala219Asn + Ser220Asp +Ser222AspLeu134Ser + Thr167Gly + Ser220Asp + Leu221Pro +Ser222AspGly108Gln + Trp112Leu + Tyr218Glu + Ser220Asp +Leu221GluGly108Pro + Ala114Asn + Ser207Asp + Tyr210Glu +Ser220GluThr137Glu + Gly162Glu + Pro169Asn + Asn170Ser +Tyr171AspAsn 68Gln + Gly162Glu + Ala164Thr + Gly165Asp +Tyr171AlaThr137Pro + Gly139Asp + Asn170Glu + Trp208Met +Tyr210ThrAsn 68Gln + Val138Ser + Gly162Pro + Ala164Glu +Asn170AspAsn 68Gln + Ser207Glu + Trp208Thr + Thr217Ser +Ser220AspVal138Ala + Phe193Thr + Ser207Asp + Tyr210Met +Ser220AspPhe193Ala + Trp208Glu + Ile209Ser + Leu221Cys +
Gly223GluThr111Gly + Phe193Gln + Ser207Asp + Ser220Asp +Gly223GluSer 70Asp + Asp105Glu + Gly108Glu + Thr111Ser +Tyr210ValGly136Glu + Gly162Asp + Gly165Ser + Ala168Pro +Thr195GluLeu104Gly + Gly165Glu + Pro169Glu + Thr217Asn +Leu221AlaThr195Asn + Tyr210Asp + Thr217Glu + Ser220Glu +Gly223ProTrp208Phe + Tyr210Asp + Tyr213Cys + Thr217Glu +Ser220GluSer 70Asp + Gly108Glu + Tyr171Gly + Thr217Gln +Leu221ProThr195Gly + Ser207Glu + Tyr210Glu + Tyr213Cys +Ala219SerAsn 68Glu + Asp105Glu + Gly110Asp + Thr111Pro +Asn140SerSer 70Asp + Val103Gln + Pro214Gln + Ser220Asp +Leu221GluAsn 66Asp + Thr 73Glu + Asn106Ser + Trp208His +Leu221ThrAla164Asp + Thr167Asp + Tyr171Pro + Ala219Gln +Leu221ThrAla114His + Ala219Gly + Ser220Glu + Ser222Glu +Thr224GluAsn106Asp + Ser109Asp + Thr111Asp + Thr137Ser +Pro205AsnLeu104Cys + Asn163Glu + Ser192Asp + Ala219Pro +Ser222GluGly162Asp + Ala164Gln + Ser207Asp + Trp208Tyr +Gly223AspSer 70Asp + Ser107Glu + Ser109Glu + Val115Cys +Gly135SerAla114Gly + Ala219Asp + Ser220Glu + Leu221Asn +Gly223GluAsn 66Asp + Asp105Glu + Val115Gln + Thr137Ser +Trp208HisGly 69Glu + Thr 73Glu + Val103His + Ser107Asp +Leu221AlaGly108Pro + Trp112Gln + Gly162Asn + Ser220Glu +Gly223AspAsn 68Asp + Asn170Gln + Tyr171Pro + Thr215Asp +Ser216AspSer 70Asp + Tyr171Ser + Thr195Gly + Tyr210Asp +Ser220GluThr113Gly + Gly162Ser + Trp208Glu + Ser220Asp +Thr224AspSer 70Glu + Asn140Ser + Ala203Gln + Ala219Glu +Gly223ProTrp112Pro + Asn170Ser + Ser216Asp + Thr217Asp +Ser220AspThr 73Asn + Val138Asp + Pro169Glu + Asn170Ser +Thr217GlySer 70Asp + Asp105Glu + Gly162Pro + Trp208Val +Thr217SerGly 67Asn + Asn140Ser + Ala219Glu + Leu221Met +Ser222GluGly 72Ser + Gly110Pro + Trp112Ala + Tyr210Asp +Thr217GluGly108Pro + Trp112Leu + Leu134Cys + Gly135Asp +Ala164AspAsn 66Gln + Trp208Ala + Thr217Glu + Ser220Glu +Leu221ThrThr 73Gly + Leu134Ser + Thr217Glu + Ser220Glu +Leu221AsnGly 67Ser + Ala168Gly + Pro214Gln + Thr217Glu +Ser220AspVal115Cys + Glyl39Asn + Tyr210Cys + Thr217Asp +Ser220AspLeu134Glu + Thr137Glu + Thr195Asn + Trp208Ser +Leu221SerAla164Gln + Tyr210Val + Thr215Asp + Thr217Glu +Ser220AspGly 67Asp + Ser 70Asp + Thr217Gly + Tyr218Asp +Leu221IleSer 70Asp + Thr111Asn + Trp208Cys + Thr217Glu +Ser220GluLeu134Glu + Gly162Ser + Asn166Gln + Ser207Asp +Gly223AspGly162Pro + Thr167Asn + Pro169Glu + Ser191Glu +Phe193AspSer 70Glu + Trp112Tyr + Ser207Glu + Tyr218Cys +Leu221GluAsn 66Glu + Leu104Glu + Ser107Asp + Trp208Ser +Thr215ProLeu104Ala + Gly135Asp + Thr195Asp + Tyr213Ile +Thr217GlyThr167Gln + Ser192Glu + Ser207Glu + Trp208Cys +Ser220AspGly165Pro + Ser191Asp + Trp208Gly + Ser220Asp +Leu221GluGly135Glu + Asn163Asp + Tyr213Met + Leu221His +Ser222AspThr111Asp + Gly136Glu + Gly162Glu + Ala164Asn +Pro214SerThr113Gln + Tyr213Thr + Ser216Glu + Thr217Glu +Leu221GluSer 70Glu + Glyl08Glu + Ala164Gly + Gly165Gln +Leu221AspSer 70Glu + Leu104Gln + Ser220Asp + Leu221His +Gly223ProGly165Asp + Pro169Ser + Ser207Glu + Trp208Gly +Ser222AspAla168Pro + Pro169Asn + Thr215Asp + Thr217Asp +
Leu221GluGly110Pro + Thr113Ser + Gly165Glu + Trp208Asp +Ser222AspGly110Glu + Thr113Asp + Gly139Glu + Trp208Tyr +Tyr210AlaAsn 68Glu + Asn106Glu + Tyr171His + Tyr210Gln +Ser216GluAla164Glu + Ala168His + Ala219Glu + Ser220Asp +Leu221CysAsn 68Asp + Tyr210Gln + Ala219Glu + Ser220Glu +Leu221SerThr 73Asp + Asn163Ser + Ala219His + Ser222Glu +Gly223GluGly 67Asp +Asn 68Glu + Ala168Gln + Pro169Glu +Phe193AsnGly110Asp + Gly139Gln + Asn140Glu + Ser141Glu +Tyr218AsnSer109Glu + Ser216Glu + Thr217Asp + Tyr218Asn +Leu221HisGly135Pro + Gly136Ser + Phe193Asp + Ser216Asp +Thr217GluGly165Gln + Ser192Glu + Ala203Ser + Ser216Asp +Thr217AspGly 67Gln + Leu134His + Gly136Asp + Ser207Asp +Trp208GluVal103Ser + Ser207Asp + Trp208Glu + Thr217Glu +Leu221CysGly 69Pro + Asp105Glu + Asn106Asp + Asn166Asp +Ile209ThrTrp112Tyr + Asn170Glu + Trp208Gln + Ser220Glu +Leu221GluThr137Gly + Gly139Gln + Gly162Asp + Ser220Glu +Leu221AspThr113Asp + Ala164Gly + Ser191Glu + Ser192Glu +Thr195GlySer107Glu + Thr113Glu + Ala114Asp + Tyr210Pro +Leu221AlaAsn 66Asp + Gly 67Asp + Leu134Pro + Tyr210Asn +Ser222AspAsn 68Gln + Leu104Ala + Pro169Asp + Asn170Glu +Leu221AspAsn106Glu + Ser107Asp + Gly139Asp + Ala203Asn +Thr217SerThr 73Gly + Thr137Glu + Tyr210Asp + Thr217Gln +Ser220GluTrp112Ile + Gly136Glu + Phe193Thr + Tyr210Glu +Ser220GluTrp112Ala + Asn163Ser + Ser192Glu + Tyr210Glu +Ser220GluSer 70Glu + Thr 73Ser + Val103Asp + Pro214Glu +Tyr218GlyAsn106Glu + Ser109Glu + Thr195Gln + Trp208Pro +Pro214GluAsn 68Gln + Val103Asp + Thr137Gln + Ser207Asp +Ser222AspThr137Asp + Ser207Glu + Thr217Gly + Leu221Pro +Ser222AspGly 69Asp + Thr 73Glu + Thr111Pro + Gly139Asn +Ser220AspAsn 66Glu + Ser 70Asp + Pro214Gly + Ser220Glu +Leu221SerSer 70Glu + Leu104Cys + Tyr210Ser + Thr215Glu +Ser220GluThr167Pro + Ser216Glu + Ala219Glu + Leu221Met +Ser222AspVal138Asp + Asn166Glu + Thr167Asp + Ala168Thr +Leu221AlaLeu134Pro + Serl41Glu + Trp208Glu + Thr217Ser +Ser220GluGly135Glu + Tyr171Asn + Pro205Gln + Trp208Glu +Ser220AspTrp112Asn + Thr167Glu + Ala168Gly + Trp208Asp +Ser220GluAsn140Asp + Pro169Gln + Trp208Glu + Thr215Gln +Ser220GluAsn106Gln + Gly136Asp + Trp208Glu + Thr217Ser +Ser220GluAsn106Asp + Thr111Asn + Thr167Gly + Tyr210Asp +Ala219AspAsn 66Gln + Pro169Asp + Tyr171Glu + Pro214Asn +Gly223GluTrp112Gly + Asn163Asp + Thr167Glu + Ser191Glu +Gly223SerTrp112Asp + Thr113Ser + Ala114Glu + Thr217Glu +Gly223GlnAsn 66Asp + Thr 73Gly + Val103Asp + Thr111Pro +Thr195AsnAsn 68Asp + Trp112Met + Trp208Asn + Pro214Asp +Ser220AspAsp105Glu + Ser109Glu + Asn163Glu + Asn166Ser +Tyr218GlyGly110Gln + Trp112Asp + Leu134Pro + Pro214Asp +Thr217AspGly 67Pro + Thr113Glu + Gly135Asp + Gly162Asp +Thr215GlnSer 70Glu + Thr195Ser + Pro214Ser + Ser220Glu +Gly223GluGly110Pro + Val138Glu + Trp208Glu + Tyr210Glu +Thr217GlyGly108Asn + Asn163Glu + Ala219Asp + Leu221Val +Thr224AspThr 73Asp + Gly108Asn + Ser109Glu + Pro214Glu +Thr217AsnThr137Asp + Gly139Glu + Asn170Ser + Trp208Cys +Tyr210AspTrp112Phe + Gly139Glu + Tyr213Thr + Ala219Glu +
Leu221GluLeul04Pro + Pro169Glu + Tyr210Glu + Leu221Asp +Thr224GlyAsp105Glu + Gly165Glu + Thr167Gln + Ala168Glu +Tyr210ProThr137Asp + Trp208Asn + Tyr218Ser + Ser220Asp +Ser222AspGly 69Glu + Thrll3Gln + Thr217Ser + Ser220Glu +Ser222GluLeu104His + Thr113Asp + Ser207Asp + Thr217Ser +Leu221GluAla114Thr + Gly135Asp + Ser207Asp + Thr217Ser +Leu221AspGly 69Ser + Leu104Asp + Asn106Glu + Ser191Glu +Thr217GlyVal103Ala + Tyr171Val + Ser207Asp + Ser216Asp +Tyr218AspGly 69Ser + Leu134Ile + Gly162Asp + Ser207Glu +Ser220GluGly 72Pro + Thr113Glu + Asn166Asp + Thr167Asp +Thr217GlnAsn166Glu + Thr167Asp + Ser207Glu + Thr217Gly +Leu221ValThr111Asn + Trp112Glu + Val138Gly + Asn163Asp +Tyr171GluGly 67Pro + Asn106Glu + Leu134Pro + Thr137Asp +Pro169GluThr195Glu + Ser207Asp + Ser220Asp + Leu221Val +Thr224GlyAsn 68Asp + Gly 69Gln + Gly162Asp + Pro169Ser +Gly223AspThr 73Ser + Gly165Asn + Ser207Asp + Tyr210Asp +Ser216GluVal103Glu + Ala164Asp + Asn170Glu + Trp208Ala +Tyr210IleAla114Thr + Ala164Asp + Asn170Glu + Tyr210Ala +Thr215AspAsn 68Glu + Gly139Pro + Ala164Pro + Ser191Asp +Pro214GluAsn 68Glu + Ala164Glu + Gly165Ser + Tyr210Gly +Pro214AspGly 67Ser + Thr167Gly + Ser207Glu + Ser216Asp +Ser220AspAsp105Glu + Gly110Asn + Ser207Asp + Ala219Gly +Ser220GluVal103Gly + Asn140Asp + Ser207Asp + Tyr213Asn +Ser220AspSer109Glu + Gly165Asn + Ser207Glu + Tyr210Cys +Ser220AspGly110Asn + Asn140Glu + Ser207Glu + Thr217Ser +Ser220GluGly110Asp + Gly136Pro + Gly206Gln + Ser207Glu +Ser220GluThr113Gln + Leu134Ile + Ser192Glu + Thr195Asp +Ser220AspGly136Glu + Trp208Glu + Thr217Gly + Leu221Asn +Gly223AspSer107Asp + Gly110Asp + Leu134Ser + Asn140Glu +Tyr210ThrThr167Glu + Trp208Asp + Thr217Asn + Ala219Asp +Gly223AsnGly 69Glu + Ala114Asp + Thr195Ser + Trp208Ser +Pro214GluVal103Asp + Thr113Glu + Phe193Tyr + Trp208Asp +Ala219ProAsn106Asp + Gly136Ser + Val138Met + Ser191Asp +Ser222AspGly135Asp + Gly139Glu + Tyr210Ala + Thr217Ser +Ser222AspThr113Asp + Asn140Ser + Ala203Gln + Ser207Glu +Tyr210GluGly108Glu + Thr113Gln + Ser207Glu + Tyr210Asp +Pro214AsnGly135Glu + Asn163Asp + Tyr210Met + Tyr218Pro +Ser220AspAsn163Asp + Ala164Thr + Ser191Glu + Thr217Asp +Thr224GlnAsn 68Glu + Leu104Asp + Val115Thr + Ala164Asp +Pro214GlnThr 73Gly + Ser107Glu + Gly108Pro + Leu221Glu +Thr224AspGly 67Ser + Gly162Asp + Thr167Asp + Ser207Glu +Thr217SerGly 67Gln + Ser109Glu + Val138Ser + Pro214Glu +Ala219GluSer107Glu + Gly135Glu + Trp208His + Thr217Ser +Thr224GluAsn 66Glu + Asp105Glu + Val138His + Ser192Glu +Tyr210IleAsn 66Glu + Asp105Glu + Ala219Gln + Ser220Glu +Leu221AsnGly 69Ser + Ser 70Glu + Gly110Glu + Thr217Asp +Leu221AlaTyr171Glu + Ser192Asp + Ser207Glu + Trp208Pro +Tyr210MetGly 67Gln + Asp105Glu + Ala164Asp + Pro169Asn +Gly223AspThr 73Asp + Ser141Asp + Trp208Leu + Ser216Glu +Thr217ProAsn 66Asp + Trp208Gln + Ile209Thr + Tyr210Val +Ser216GluAsn 66Ser + Val103Glu + Asn106Asp + Ser191Glu +Tyr210AlaAsn 68Gln + Leu134Glu + Ser192Glu + Leu221Gly +Ser222GluSer 70Asp + Asp105Glu + Gly108Ser + Gly139Glu +
Thr217GlyGly 67Ser + Asn106Asp + Gly165Gln + Ala219Asp +Ser222GluAsn106Gln + Leu134Met + Asn140Asp + Ala219Asp +Ser222AspGly 69Glu + Val103Gln + Leu134Asp + Thr167Gln +Trp208GlyThr137Glu + Gly165Glu + Tyr210Ser + Thr217Ser +Leu221ProSer107Asp + Gly162Asp + Thr167Asp + Tyr171Cys +Trp208IleAsn106Glu + Trp112Asp + Leu134Gly + Ser141Glu +Phe193SerAla114Thr + Asn140Asp + Phe193Thr + Thr217Glu +Ser220GluThr113Ser + Val115Asp + Leu134Val + Thr217Asp +Ser220GluGly110Asp + Asn170Gln + Thr217Glu + Tyr218Ala +Ser220AspThr167Asp + Phe193Thr + Thr217Asp + Ala219Gln +Ser220AspGly108Glu + Pro214Asn + Thr217Glu + Ser220Asp +Leu221ValAsn106Gln + Gly135Glu + Val138His + Thr217Glu +Ser220GluGly110Glu + Tyr210Met + Thr217Glu + Ser220Asp +Leu221HisGly 67Glu + Ser107Glu + Gly110Ser + Thr113Glu +Thr215SerGly 67Glu + Ser107Glu + Thr111Pro + Ala168His +Ser220GluThr111Asp + Gly136Asp + Trp208Glu + Tyr210Met +Thr217GlnAsp105Glu + Gly162Ser + Asn166Asp + Ser191Asp +Trp208PheAsp105Glu + Asn166Glu + Ser191Glu + Trp208Ala +Leu221HisGly 67Glu + Val103Glu + Ala164Asn + Ser207Asp +Tyr218AlaGly 72Asn + Leu104Asp + Asn106Gln + Thr113Asp +Thr217AspSer 70Asp + Gly135Glu + Ala168Glu + Tyr210Ile +Ala219GlyAsn106Gln + Ser109Glu + Thr113Asp + Thr137Glu +Trp208AsnAla114Ser + Val138Thr + Pro169Asn + Phe193Asp +Trp208GluAsn 68Asp + Val138Asp + Ala164Gly + Ala168Glu +Thr217GlyGly 69Asn + Val138Asp + Ala168Glu + Ser207Asp +Thr217ProGly110Ser + Thr111Gln + Ala164Glu + Thr215Ser +Ser222AspLeu104Ser + Gly108Glu + Val115Glu + Val138His +Ser141GluSer 70Glu + Ala114Ser + Thr137Ser + Ser207Asp +Ser216AspGly 69Asp + Gly110Asp + Thr137Glu + Pro169Ser +Tyr171HisSer 70Asp + Thr137Glu + Ala164His + Trp208Ile +Ser220GluSer 70Glu + Thr137Glu + Thr217Asn + Ser220Glu +Gly223AsnGly110Gln + Trp112Cys + Val115Glu + Asn140Asp +Ser216GluAsn 68Asp + Ser109Glu + Asn166Gln + Trp208Pro +Leu221AspThr113Pro + Thr167Glu + Tyr171Glu + Thr217Gln +Ser220GluTrp112Ile + Leu134His + Gly139Asp + Gly162Glu +Trp208ValGly110Asp + Gly162Gln + Trp208Ile + Tyr210Glu +Gly223AspGly 67Asp + Thr111Asp + Trp208Val + Tyr210Ser +Ser216GluGly 69Asn + Thr 73Glu + Asn106Asp + Ser141Glu +Gly162AsnGly 69Glu + Gly135Asn + Pro169Asn + Trp208Ala +Thr217GluAla203Gly + Trp208Ala + Pro214Glu + Ala219Gln +Leu221AspThr 73Gly + Gly110Glu + Pro214Asp + Ser220Glu +Leu221AsnVal103Ala + Thr167Gly + Ser192Glu + Pro214Glu +Ser220AspAsp105Glu + Tyr171Gln + Pro214Asp + Thr217Ser +Ser220AspTyr171Val + Ser191Asp + Trp208Pro + Pro214Glu +Ser220GluGly 67Asp + Gly162Ser + Gly206Ser + Ser207Glu +Thr217GluGly 67Glu + Trp112Phe + Ser207Asp + Thr217Asp +Ala219GlnAsn 68Asp + Asn163Glu + Phe193Met + Gly206Ser +Leu221GluAsn 66Asp + Leu104Ala + Ser109Glu + Ala168Thr +Ser216GluVal138Met + Gly139Asp + Gly162Ser + Ala168Glu +Ser220Asp表36多環六突變變異體Asn 66Gln + Gly 67Ser + Gly 69Ser + Leu104Gly +Tyr210Pro + Thr217GluSer 70Asp + Gly110Pro + Gly136Gln + Phe193Ser +Trp208Cys + Leu221HisThr113Pro + Gly139Asn + Ser141Glu + Tyr171Pro +Trp208Thr + Leu221GlyGly 69Gln + Leu104Thr + Ser107Asp + Leu134Thr +Thr215Ser + Leu221GlyLeu104Met + Leu134Ala + Thr137Asp + Trp208Gly +Tyr210Val + Pro214GlyGly 69Ser + Thr 73Asp + Gly110Ser + Tyr210Leu +Thr215Gly + Thr217SerAsn 66Ser + Gly 72Gln + Thr 73Pro + Gly110Asp +Trp208Val + Leu221HisThr111Asn + Ala164His + Thr167Glu + Phe193Met +Tyr210Asn + Thr217GlnAsn 66Gln + Thr 73Gly + Thr113Asp + Asn166Ser +Thr167Pro + Tyr210ProLeu104Met + Pro205Gln + Ser207Asp + Trp208Ala +Thr212Gly + Thr217ProLeu134Val + Gly165Gln + Trp208Thr + Pro214Gly +Leu221Ile + Thr224AspAsn 66Ser + Ser109Asp + Ala114Asn + Val115Asn +Trp208Gln + Tyr210SerAsn106Gln + Ala164Pro + Gly165Ser + Tyr171Ala +Ser220Glu + Thr224SerGly110Ser + Gly136Pro + Ile209His + Thr217Gln +Leu221Ile + Thr224AspAsn 66Gln + Val115Pro + Trp208Asp + Thr217Ser +Leu221Ile + Thr224ProGly 67Glu + Thr 73Asn + Asn170Ser + Tyr210Ser +Thr212Gln + Gly223ProVal103Gly + Ala164Thr + Tyr210His + Ala219His +Ser220Glu + Leu221GlyThr 73Pro + Asn140Gln + Thr167Asp + Trp208Met +Thr217Gln + Leu221AsnSer109Asp + Ala164His + Tyr210Pro + Thr212Ser +Gly223Pro + Thr224AsnGly110Gln + Tyr210Pro + Tyr213Thr + Tyr218Met +Ser220Asp + Leu221HisVal138Pro + Ala168Gly + Pro169Gln + Tyr210Ser +Thr217Gln + Ser220AspAla114Thr + Thr137Asn + Gly139Ser + Trp208Pro +Pro214Gln + Ser220GluVal103Met + Thr137Ser + Ala203Ser + Tyr210Thr +Leu221Met + Gly223SerLeu104Glu + Thr111Ser + Val115His + Gly165Asn +Tyr210Met + Ala219HisGly110Pro + Leu134Pro + Asn166Asp + Asn170Ser +Trp208Asn + Thr217AsnThr167Glu + Pro169Gly + Asn170Gln + Trp208Gly +Thr215Ser + Leu221SerThr111Ser + Thr137Ser + Thr195Gln + Ser207Glu +Tyr210Val + Thr217ProAsn106Asp + Gly136Pro + Gly162Ser + Trp208Ile +Tyr210Gly + Pro214GlnSer 70Asp + Gly165Gln + Phe193Asn + Thr195Ser +Trp208Pro + Thr212SerGly 72Ser + Ala114Gln + Asn166Gln + Phe193Glu +Tyr210Gly + Thr224ProAsn 66Ser + Gly136Ser + Gly162Pro + Asn163Ser +Ala164Asp + Ala168GlnGly 67Pro + Asn106Gln + Ser191Asp + Trp208His +Tyr218His + Leu221AsnTrp112Leu + Ala168Pro + Trp208His + Tyr210Ser +Thr217Gly + Ser220AspAsn 66Ser + Thr111Gly + Val115Gly + Ala168Ser +Asn170Gln + Pro214GlyGly110Ser + Trp112Val + Gly162Asn + Thr167Pro +Pro169Gln + Ser216AspGly 69Asn + Asn106Ser + Gly139Glu + Asn170Ser +Tyr210Cys + Leu221GlyGly 69Asn + Ser141Asp + Phe193Asn + Tyr210Ile +Ala219Asn + Leu221SerSer109Asp + Pro169Ser + Trp208His + Pro214Asn +Thr215Asn + Thr217GlyAsn 68Asp + Gly162Gln + Ala164His + Trp208Pro +Tyr210Leu + Leu221ValVal103Thr + Gly139Pro + Ala164Thr + Thr217Ser +Leu221Pro + Thr224ProGly 69Glu + Gly 72Gln + Thr137Pro + Trp208Ile +Tyr218Pro + Gly223GlnGly 67Pro + Leu104Cys + Val138Met + Gly139Ser +Ala203Gln + Ala219AsnTrp112Ser + Phe193Ala + Thr217Gly + Tyr218Glu +Leu221Met + Thr224AsnAsn163Gln + Trp208Leu + Tyr210Ser + Thr217Glu +Ala219Gly + Gly223ProVal103His + Gly110Gln + Gly139Ser + Ser192Asp +Phe193Asn + Thr217GlnGly108Gln + Pro169Asn + Thr195Ser + Trp208Ala +Thr215Pro + Thr217GluSer109Asp + Thr111Asn + Trp112Gln + Thr167Gln +Thr195Gly + Tyr210AsnGly 72Gln + Trp112Leu + Gly136Asp + Tyr171His +Tyr210Thr + Leu221SerGly 69Glu + Ala164Pro + Trp208Val + Tyr210Thr +Tyr218Gln + Thr224SerAla114Pro + Leu134Glu + Asn140Ser + Ala164Gln +Phe193Ile + Trp208AsnAsn 66Ser + Val138Met + Gly139Asn + Asn163Gln +Gly165Gln + Tyr210ThrAsn106Gln + Ser191Asp + Phe193Leu + Trp208Val +Pro214Gln + Thr217SerAla114Thr + Asn166Ser + Tyr171Gly + Ser191Glu +Ala203Pro + Tyr210AlaAsn 68Gln + Tyr171Asp + Phe193Tyr + Gly206Ser +Tyr210Val + Leu221ThrLeu104Ala + Alal14Ser + Asn163Asp + Thr195Gly +Gly206Gln + Leu221AlaGly110Asn + Val115Asn + Trp208Cys + Tyr210His +Ala219Thr + Gly223AspAsn 66Asp + Gly 69Asn + Val115Ala + Ala164Ser +Trp208Gln + Thr217GlnVal115Met + Thr137Ser + Gly162Asn + Asn166Glu +Tyr171His + Pro214SerAsn106Gln + Gly110Gln + Trp208His + Thr217Pro +Ala219Asp + Ser220AspGly 67Asn + Gly 72Asn + Ala114Gly + Asn140Gln +Ser216Asp + Thr217AspGly 69Gln + Thr 73Ser + Pro169Gln + Trp208Ile +Ser220Glu + Leu221AspAsn170Glu + Tyr171Asp + Phe193Ser + Trp208His +Leu221Thr + Thr224GlyGly 67Ser + Gly 69Glu + Ser 70Glu + Val103Gly +Tyr210Gln + Ala219AsnTrp112Gly + Asn166Ser + Trp208Met + Tyr210Glu +Ser220Glu + Thr224AsnLeu134Pro + Thr195Gly + Trp208Glu + Tyr210Asp +Thr217Ser + Ser220GluLeu104Gln + Trp112Gln + Tyr171Pro + Trp208Glu +Thr217Gly + Leu221GluAsp105Glu + Gly108Asp + Gly136Gln + Thr167Gln +Tyr171Gln + Leu221ValSer107Glu + Ser109Glu + Val138Met + Ala168His +Pro169Gln + Phe193SerGly 67Glu + Ser 70Glu + Trp112His + Tyr171Ile +Trp208His + Tyr210AlaVal103Pro + Trp208Glu + Tyr210Ser + Ala219Glu +Ser220Glu + Ser222AspSer191Asp + Ser207Glu + Tyr210Ala + Thr215Gly +Thr217Gln + Gly223SerAsn106Glu + Ser109Asp + Ala168Thr + Tyr171Met +Phe193Ile + Trp208LeuAsn 68Ser + Thr137Gln + Ser191Asp + Ser207Asp +Trp208Asp + Leu221ThrVal103Gln + Val138His + Asn166Asp + Ser192Glu +Trp208His + Leu221GlyVal103Pro + Gly139Pro + Asn163Asp + Gly165Gln +Ala168Glu + Tyr210LeuAsn 68Ser + Thrl13Gln + Leu134Ile + Ser191Glu +Leu221His + Gly223GluGly110Gln + Ser207Glu + Trp208Gln + Thr217Pro +Tyr218Gly + Ser220AspGly162Ser + Phe193Ala + Trp208Glu + Ile209Ser +Leu221Cys + Gly223GluSer 70Asp + Asp105Glu + Gly108Glu + Asn140Gln +Trp208Leu + Thr217GlyGlyl10Asn + Thr137Glu + Pro169Asp + Asn170Ser +Thr195Glu + Tyr210HisAsn 68Gln + Gly162Glu + Ala164Thr + Gly165Asp +Tyr171Ala + Ser192AspVal138Met + Gly139Asn + Asn170Gln + Tyr210Glu +Thr217Asp + Ser220GluGly139Ser + Asn163Asp + Ala164Ser + Ser191Asp +Tyr210Gly + Leu221AlaSer 70Asp + Thr 73Asp + Thr167Ser + Trp208Ser +Thr217Asp + Ala219AspAsn 66Glu + Leu104Glu + Asp105Glu + Ser107Asp +Leu221Ser + Gly223ProAsn 66Glu + Thr 73Asp + Gly110Pro + Ala168Gly +Tyr218Val + Leu221IleGly 67Asn + Asp105Glu + Asn106Glu + Thr111Glu +Gly165Asn + Thr217GlyAsn 66Asp + Tyr210Ser + Pro214Glu + Thr215Asp +Thr217Gln + Tyr218AspThr 73Asn + Ser192Glu + Ala203Thr + Ser207Asp +Trp208Glu + Leu221GluVal138Gly + Pro169Glu + Ser192Glu + Thr195Glu +Trp208Pro + Thr212GlnThr 73Asp + Gly108Asn + Leu134Ser + Tyr210Ala +Thr217Gln + Tyr218GluThr 73Asn + Ala114Thr + Val138Ser + Gly139Asp +Ser141Glu + Asn170GluGly108Gln + Gly135Pro + Gly162Asn + Ser220Glu +Leu221Ala + Gly223GluVal103Gln + Trp112Cys + Ala164Glu + Trp208Gln +Thr217Gly + Gly223AspThr111Gln + Leu134His + Ser191Asp + Phe193Asp +Thr195Gln + Trp208AspVal103Ser + Leu104Ile + Gly110Glu + Gly136Glu +Ala164Asn + Tyr210GlnLeu134Ser + Thr167Gly + Ser191Glu + Ser220Asp +Leu221Pro + Ser222AspGly 69Gln + Trp208Asp + Pro214Glu + Ser220Asp +Ser222Glu + Gly223AsnSer107Asp + Ser109Glu + Trp112Pro + Gly135Asp +Ala168Thr + Thr212GlyAsn140Ser + Thr167Asp + Trp208Glu + Thr217Asn +Ser220Asp + Leu221AspGly135Asp + Gly162Glu + Tyr171Asp + Tyr210Ala +Thr217Ser + Ser222AspVal115Asp + Tyr210Gln + Ser220Glu + Leu221Asp +Ser222Asp + Gly223SerAla114His + Asn166Asp + Tyr171Cys + Tyr210Glu +Ser220Asp + Leu221AspVal115Cys + Gly139Asn + Ala164Gly + Pro205Asn +Thr217Asp + Ser220AspThr167Gln + Tyr210Ile + Thr215Gly + Thr217Asp +Tyr218Asn + Ser220AspThr137Ser + Trp208Leu + Tyr210Leu + Thr217Glu +Ser220Asp + Leu221GlyThr111Asn + Gly135Ser + Trp208Asp + Tyr210Glu +Thr217Glu + Leu221AlaThr113Asn + Ala114Gly + Thr137Asp + Thr167Asp +Trp208Gln + Gly223AsnSer 70Asp + Thr 73Glu + Asn163Asp + Thr167Pro +Pro214Glu + Thr217GlyGly135Glu + Tyr171Asn + Pro205Gln + Ser207Glu +Trp208Glu + Ser220AspGly 69Asp + Asp105Glu + Gly108Asp + Ser207Asp +Trp208Thr + Gly223AsnThr113Gly + Ala114Pro + Tyr210Asp + Pro214Asp +Thr217Gln + Ser220AspAsn 68Glu + Ser 70Asp + Asp105Glu + Ala114Thr +Gly135Ser + Ser220AspThr111Ser + Thr137Glu + Ser207Glu + Trp208Thr +Ser220Glu + Leu221AspGly 69Asp + Leu104Asp + Asp105Glu + Asn140Glu +Asn166Ser + Leu221HisThr137Asp + Gly139Glu + Asn170Glu + Trp208Cys +Tyr210Asp + Leu221ThrGly 67Pro + Trp112Gln + Ser192Asp + Tyr210Ile +Ser220Asp + Ser222AspVal103Gln + Gly108Asp + Gly135Glu + Thr137Glu +Ala168Gln + Tyr210AsnVal115Asp + Leu134Glu + Asn166Gln + Asn170Asp +Tyr210Val + Thr212ProAsn 66Asp +Asn 68Ser + Thr167Gln + Ile209Leu +Ser216Asp + Thr217GluVal103G1u + Gly108Asp + Gly110Glu + Gly165Pro +Thr195Glu + Thr217GlnAsn 66Asp + Ser 70Glu + Thr 73Asp + Ser109Asp +Gly136Pro + Tyr210IleGly 69Asp + Ser 70Glu + Asp105Glu + Asn163Glu +Tyr210Met + Thr217GlnAsn 68Gln + Leu134Glu + Ser207Asp + Trp208Pro +Tyr210Asp + Leu221AspAsn 66Ser + Val115Gly + Gly162Asp + Ser191Glu +Trp208Ile + Thr217SerSer 70Glu + Tyr213Thr + Thr217Asn + Tyr218Gly +Ser222Glu + Gly223AspTrp112Met + Gly165Asp + Pro169Ser + Ser207Glu +Trp208Gly + Ser222AspGly 69Ser + Ser109Asp + Gly110Glu + Ala114Gly +Gly136Asp + Phe193AspAla164Gln + Gly165Pro + Ser192Asp + Phe193Asp +Ser207Glu + Thr217AspAsn 68Glu + Thr 73Asn + Gly162Gln + Trp208Gln +Ala219Glu + Ser220AspSer 70Glu + Leu134Ser + Gly165Asp + Asn166Asp +Ala168Gly + Ile209GlyLeu104Asp + Asp105Glu + Trp208Gly + Tyr210Glu +Tyr218Ala + Gly223ProAsn 68Asp + Gly 69Glu + Thr111Asn + Trp112His +Gly136Asn + Pro169GluGly108Ser + Asn140Asp + Ser141Glu + Ala164Gln +Ser207Glu + Tyr218SerLeu104Cys + Gly162Pro + Ser207Glu + Ser216Glu +Thr217Asp + Ala219ThrAla164Asn + Asn166Asp + Gly206Gln + Tyr210Thr +Ser216Asp + Thr217AspGly108Asp + Ser109Asp + Asn166Ser + Thr217Gln +Ser220Asp + Leu221ThrSer109Asp + Gly110Asp + Val138Ala + Tyr171His +Ala204His + Ser222GluAsn 68Gln + Thr137Gly + Pro169Asp + Ser207Glu +Trp208Glu + Tyr218GlnTrp112Asp + Thr113Glu + Val138Gly + Ser191Glu +Trp208Pro + Leu221SerLeu104Gly + Asp105Glu + Asnl06Glu + Gly206Ser +Trp208His + Tyr218GluGly108Asp + Leu134Asn + Gly136Gln + Trp208Phe +Ser220Asp + Leu221AspGly108Gln + Trp112Tyr + Asn170Glu + Trp208Gln +Ser220Glu + Leu221GluGly 72Asn + Asn166Asp + Phe193Ser + Trp208Tyr +Ser220Glu + Leu221GluAsn 66Glu + Gly 67Glu + Thr111Asn + Thr215Gln +Thr217Asn + Ser220GluAsn106Asp + Ser107Asp + Gly165Asp + Phe193Val +Trp208Val + Leu221ThrLeu104Val + Gly165Gln + Ser192Asp + Phe193Glu +Tyr210Glu + Leu221CysGly136Gln + Pro169Ser + Ser192Asp + Phe193Asp +Trp208Leu + Ser220GluSer107Asp + Val115Thr + Thr137Gln + Gly162Asp +Asn163Asp + Ala203HisVal103Met + Ser107Glu + Trp208Val + Thr215Glu +Ser216Glu + Gly223SerVal103Ser + Asn140Asp + Ser141Glu + Tyr210Glu +Ser220Asp + Gly223AsnAsn163Asp + Asn170Glu + Thr195Asn + Ser207Asp +Tyr210Met + Tyr218ProAsn 66Asp + Gly139Gln + Tyr210Glu + Thr217Ser +Ser220Asp + Thr224AsnGly 69Asp + Asn106Asp + Gly110Asp + Gly165Asn +Pro169Ser + Thr217AspPhe193Ile + Thr215Ser + Thr217Glu + Ser220Asp +Leu221Pro + Gly223GluVal103Gly + Asp105Glu + Thr195Asn + Ser207Asp +Ser222Asp + Gly223AsnGly 69Asp + Asn140Gln + Ala164His + Ala168Ser +Ser207Asp + Ser222AspAsn 68Asp + Thr111Asn + Ala114Gly + Gly135Gln +Ser207Glu + Ser222GluAsn106Asp + Gly139Asn + Gly162Glu + Pro169Ser +Ser191Asp + Thr224AspThr113Asp + Ser192Asp + Ser207Glu + Tyr213Thr +Thr215Pro + Ser222AspGly 69Pro + Asn163Asp + Thr195Glu + Trp208Ser +Tyr210Glu + Leu221CysGly 69Pro + Ser 70Asp + Gly110Ser + Gly135Glu +Asn170Glu + Leu221IleGly 69Glu + Ala164His + Thr195Gln + Ser220Glu +Leu221Glu + Thr224GluGly 69Gln + Gly136Asp + Val138Asp + Pro169Gly +Asn170Gln + Ser220GluGly136Glu + Val138Asp + Thr195Asn + Ser220Glu +Leu221Gly + Gly223ProAsn 68Gln + Ser107Glu + Ser109Glu + Gly139Gln +Ala164Gln + Ser207AspSer107Glu + Ser109Glu + Pro169Gln + Tyr210Met +Thr217Pro + Ser222AspGly 67Asp + Ser 70Glu + Val103Ala + Thr111Glu +Tyr210Asn + Pro214SerAsn 66Glu + Gly 67Asp + Leu134His + Ser191Asp +Ser207Glu + Trp208ThrAsn 66Glu + Val103Asp + Gly108Asn + Asn170Ser +Thr217Asn + Leu221MetThr 73Glu + Trp112Pro + Asn166Ser + Phe193His +Tyr218Ile + Ser220GluVal115His + Ser207Glu + Trp208Tyr + Tyr210Leu +Pro214Glu + Leu221GluAsn 66Gln + Ser107Glu + Ser109Glu + Ser207Glu +Gly223Asp + Thr224GlyAsn 66Glu +Asn 68Asp + Thr 73Gln + Thr195Ser +Ser207Asp + Tyr218ProSer 70Asp + Leu104Ile + Asn163Asp + Thr217Gln +Leu221Val + Thr224GluGly 67Asp + Val103Glu + Asn106Asp + Ala168Asn +Thr195Glu + Trp208LeuThr137Gly + Gly162Glu + Phe193Asp + Trp208Asp +Thr217Asn + Tyr218ValGly135Pro + Gly139Glu + Ser207Glu + Trp208Ala +Tyr210Cys + Gly223GluAsn106Ser + Thr137Asp + Gly139Glu + Ser141Asp +Ala204Gln + Trp208GluTrp112Ala + Asn163Glu + Ser192Glu + Trp208Val +Tyr210Glu + Ser220GluGly110Asp + Trp112Asp + Tyr210Thr + Ser220Glu +Leu221Met + Ser222GluGly 67Asp + Gly 69Asp + Thr 73Pro + Ser109Asp +Trp208Gln + Ser222AspAsn 66Ser + Val103Glu + Asp105Glu + Trp112His +Ser191Glu + Tyr210AlaGly 67Glu + Gly 69Asn + Gly162Glu + Ser207Glu +Thr217Asn + Gly223GluSer 70Asp + Gly165Asp + Ala168Asp + Trp208Pro +Thr217Gly + Leu221IleGly 67Glu + Val103Asn + Gly136Gln + Trp208Pro +Ser216Glu + Leu221MetGly 67Asn + Gly139Glu + Ser141Glu + Tyr213Gly +Ser220Asp + Leu221AsnGly 67Glu + Leu104Ser + Tyr210Ser + Ser220Asp +Ser222Glu + Gly223GlnGly 67Asp +Asn 68Glu + Gly136Asp + Ala168Gln +Pro169Glu + Phe193AsnAsn 66Glu + Gly 69Asp + Asn140Gln + Ser192Asp +Phe193Asp + Tyr210LeuLeu104Asp + Asn106Glu + Asn170Glu + Tyr210Ile +Thr215Ser + Thr224ProLeu104Asp + Ser107Glu + Gly135Asp + Gly139Asn +Ala164Glu + Thr217GlyTrp112Glu + Gly139Glu + Ala164Glu + Thr195Glu +Thr212Ser + Tyr213SerAsn 68Asp + Thr111Gly + Thr113Ser + Val138Asp +Asn170Asp + Phe193SerAsn163Gln + Ala168Asp + Asn170Asp + Ser207Glu +Thr217Gly + Ser220AspGly 67Asp + Asn106Glu + Thr113Ser + Gly136Gln +Asn166Glu + Leu221CysAsn 66Gln + Gly110Asp + Ala114His + Gly136Glu +Thr137Asp + Ser222GluGly 67Asp +Asn 68Ser + Gly 72Pro + Gly139Ser +Thr167Glu + Thr195AspAsn 66Asp + Asn 68Gln + Ser191Glu + Ser207Asp +Trp208Val + Ser220AspSer109Glu + Ala114Glu + Leu134Ile + Gly136Ser +Ala219Glu + Ser220AspVal115Glu + Ala168His + Pro169Asn + Tyr210Asp +Ser216Glu + Ala219AspAsn 66Glu + Asn106Asp + Asnl70Ser + Trp208Thr +Ser216Asp + Tyr218GlyGly162Asn + Asn170Asp + Phe193Glu + Tyr210Gln +Ser220Glu + Thr224AspGly108Ser + Leu134Pro + Thr137Asp + Pro169Glu +Ser207Glu + Thr217GlyThr137Asp + Val138Glu + Asn163Glu + Ala164Asn +Ala168Ser + Thr217GluGly162Glu + Trp208Glu + Tyr210Val + Thr217Glu +Ser220Glu + Gly223ProGly135Asp + Val138Thr + Ser207Asp + Thr217Pro +Tyr218Ile + Ser220GluLeu104Gly + Ser192Asp + Thr195Asp + Thr217Pro +Ser220Glu + Leu221AsnAsn 66Ser + Gly136Glu + Val138Glu + Trp208Leu +Thr217Asp + Ser220GluGly 69Ser + Val103Asp + Ser109Asp + Ser191Asp +Gly206Gln + Tyr210ProAla114Gly + Gly162Asp + Pro214Gly + Thr215Asp +Thr217Glu + Ser220GluThr137Gln + Gly139Asp + Gly165Pro + Tyr171Glu +Thr217Glu + Ser220GluGly 67Glu + Asp105Glu + Asn140Glu + Phe193Gly +Trp208Leu + Leu221CysLeu104Glu + Trp112Leu + Phe193Leu + Ser207Asp +Trp208Ala + Thr224AspSer 70Glu + Thr113Gly + Gly162Ser + Asn163Asp +Ala164Glu + Ser220AspGly 69Glu + Ser 70Asp + Thr111Glu + Ser220Asp +Leu221Pro + Gly223SerAsn 66Gln + Leu104Pro + Gly136Glu + Asn163Asp +Tyr171Ile + Leu221GluSer 70Asp + Leu104Glu + Ala168Asn + Pro169Glu +Thr195Asp + Trp208PheSer 70Glu + Gly162Glu + Trp208Glu + Thr217Gly +Leu221Thr + Ser222AspGly 69Asn + Thr137Glu + Gly139Pro + Asn166Asp +Pro169Asp + Trp208AspGly 69Glu + Leu104Glu + Val115Ser + Asn140Ser +Ser192Glu + Pro214AspLeu104His + Asp105Glu + Val138His + Asn163Asp +Ser192Glu + Thr217AsnAla114Asn + Asn163Glu + Asn170Ser + Ser192Asp +Ala204Asn + Thr217AspAsn 68Gln + Thr 73Asp + Ser109Glu + Trp208Phe +Pro214Glu + Ser220AspVal115Cys + Leu134Asp + Gly135Pro + Ala164Thr +Asn170Asp + Trp208AspAsn 66Glu + Gly 69Glu + Trp112Gln + Gly135Asn +Gly139Asp + Asn170GluThr 73Gly + Ser107Glu + Gly135Asp + Pro169Asp +Trp208Thr + Leu221AsnGly 67Ser + Ser 70Glu + Gly136Asp + Pro169Asn +Leu221Asp + Gly223AspSer 70Asp + Gly110Glu + Val138Ser + Tyr210Val +Leu221Asp + Gly223AspAsn 66Asp + Leu104Glu + Ser109Glu + Tyr210Asn +Thr217Asn + Ser220AspAsn 66Glu + Gly 67Asp + Asn163Glu + Ser207Glu +Thr215Gly + Leu221ThrGly 69Glu + Tyr210Glu + Thr215Asp + Thr217Gln +Tyr218Gln + Ser220GluSer 70Asp + Thr137Glu + Ala164His + Thr167Gly +Trp208Asp + Ser220GluVal138Met + Gly139Gln + Asn140Glu + Ala168Pro +Tyr171Glu + Thr215GluVal103Gly + Asn106Ser + Ser107Asp + Thr111Asp +Ser222Glu + Gly223GluGly 72Gln + Asn106Glu + Gly110Asp + Thr113Glu +Gly135Asn + Ala219GluSer109Glu + Thr113Asp + Trp208Ile + Ser220Glu +Leu221Asp + Thr224SerGly 67Asn + Ser 70Asp + Gly108Asp + Asn170Glu +Thr215Pro + Leu221ThrGly 67Asp + Ser107Asp + Thr167Gly + Tyr210Asn +Ser220Asp + Ser222GluGly 69Asp + Thr 73Glu + Asn106Gln + Thr111Asn +Val138Glu + Thr217AspThr111Asn + Ser191Asp + Thr195Asp + Ala204Thr +Ala219Ser + Ser220GluSer109Asp + Ser191Glu + Tyr210Val + Thr215Asn +Leu221Gly + Ser222GluThr 73Glu + Leu104Asp + Asn106Gln + Thr195Ser +Ser207Glu + Ser220AspAsn 66Asp + Thr 73Asp + Leu104Glu + Gly136Gln +Gly162Asp + Leu221GlyGly 72Asn + Leu104Asp + Asn163Asp + Tyr171Ser +Ser191Glu + Thr217AsnVal115Asp + Gly136Gln + Tyr210Glu + Tyr218Cys +Leu221Asn + Ser222AspThr 73Asp + Asn140Ser + Asn163Glu + Thr195Asp +Ser220Asp + Leu221ProAsn 68Glu + Leu104Asp + Pro169Asn + Trp208Val +Thr217Asn + Gly223GluGly110Glu + Thr167Gln + Ser192Glu + Ser207Glu +Trp208Cys + Ser220AspVal103Gly + Gly136Glu + Asn140Asp + Ser207Asp +Tyr213Asn + Ser220AspThr113Asp + Gly136Asp + Thr137Gln + Asn163Glu +Asn166Glu + Trp208PheAsn 66Glu + Gly 69Glu + Ser109Asp + Ser192Asp +Phe193Val + Tyr218AlaSer 70Glu + Ser141Asp + Asn163Ser + Trp208Met +Tyr210Pro + Thr215AspAsp105Glu + Gly110Asn + Thr111Glu + Leu134Ala +Ser141Glu + Phe193TyrAsn 66Glu + Gly 69Ser + Gly108Asp + Thr111Asp +Thr195Gly + Tyr210LeuGly165Glu + Tyr171Asn + Ser207Glu + Thr215Glu +Thr217Gly + Thr224GluAsn 66Gln + Thr 73Ser + Gly139Asn + Pro214Asp +Ser220Asp + Leu221IleAsn166Glu + Ser192Asp + Trp208Thr + Tyr210Gly +Thr215Asp + Ser222GluAsn 66Asp + Ser191Asp + Thr215Asn + Ala219Asp +Ser222Asp + Gly223GlnAsp105Glu + Gly108Glu + Thr195Asn + Ile209Leu +Pro214Asp + Ser220AspThr111Glu + Val138Asp + Asn163Ser + Tyr218Met +Leu221Glu + Gly223AspGly 67Glu + Val138Glu + Ser141Glu + Trp208Ile +Pro214Asn + Thr215AspTrpl12Leu + Thr113Gln + Gly165Glu + Thr167Asp +Thr217Glu + Ser220AspGly 67Gln + Asn 68Asp + Gly 69Asp + Phe193Ala +Ser216Glu + Ser222GluSer 70Asp + Val115Glu + Gly162Ser + Ala168Thr +Tyr210Gly + Thr217AspSer 70Asp + Gly136Asn + Gly165Glu + Tyr210Asn +Thr217Asp + Leu221HisThr 73Ser + Val103Asn + Ser109Asp + Ser192Asp +Ser207Asp + Trp208IleAsn106Asp + Ser192Glu + Phe193Pro + Ser207Glu +Thr217Gln + Tyr218IleSer141Asp + Thr167Asp + Ala168Asp + Ser191Asp +Ala219Asn + Leu221ThrAsn 66Gln + Ser141Asp + Asn170Ser + Thr217Gln +Ser220Asp + Gly223AspThr 73Gly + Asp105Glu + Ala114His + Ser220Glu +Leu221Asn + Gly223GluAsn106Asp + Asn140Gln + Trp208Ser + Tyr210Cys +Ser220Asp + Gly223GluGly 67Gln + Asp105Glu + Ala164Asp + Pro169Asn +Tyr218Leu + Gly223AspTrp112Val + Asn166Asp + Phe193Gly + Pro214Glu +Ser220Asp + Ser222GluGly108Asp + Thr111Asn + Ser192Glu + Thr217Asn +Ala219Glu + Ser220AspVal103Ser + Val115Asp + Gly139Glu + Asn166Ser +Ser207Glu + Ser220GluLeu104Asp + Asp105Glu + Asn140Glu + Thr167Asp +Pro214Gly + Ala219GlyGly 67Asp + Asn 68Asp + Thr111Asp + Ala164Gln +Phe193Ser + Ser207AspSer109Glu + Trp208Asp + Ser216Glu + Thr217Asp +Tyr218Asn + Leu221HisSer141Asp + Ala168Glu + Tyr210Met + Ser216Glu +Thr217Asp + Tyr218AlaSer109Asp + Gly110Asp + Asn166Asp + Asn170Ser +Thr215Ser + Leu221AspGly110Glu + Thr167Glu + Ala168Asp + Trp208Met +Ser220Asp + Thr224GlnAsp105Glu + Asn106Asp + Gly165Asp + Thr195Ser +Tyr218Ile + Ser220AspAsn106Glu + Ser141Asp + Tyr218Ser + Ala219Thr +Ser220Asp + Leu221GluThr 73Gln + Thr111Glu + Asn166Asp + Trp208Ser +Ser220Asp + Leu221AspSer107Glu + Ala168Asn + Pro169Asn + Ser191Asp +Ser192Asp + Thr217AspAla114Glu + Asn166Ser + Thr212Pro + Thr217Glu +Tyr218Asp + Thr224AspAsn106Glu + Ser107Asp + Gly139Asp + Ala203Asn +Thr217Ser + Ser220AspThr113Asn + Gly139Ser + Ser141Glu + Thr215Glu +Ser216Asp + Ser220AspGly 69Gln + Asn140Glu + Gly162Glu + Asn163Asp +Tyr210Gln + Ser220AspAsn 68Asp + Ala114Pro + Gly135Glu + Trp208Asp +Leu221Ile + Gly223AspAsn 68Asp + Ala164Glu + Trp208Cys + Tyr210Glu +Ala219Thr + Ser220AspThr111Asp + Asn166Asp + Asn170Gln + Tyr210Asp +Thr212Ser + Ser220GluSer141Glu + Gly165Asp + Trp208Ile + Tyr210Asp +Thr217Gln + Ser220AspAsn 66Gln + Val103Asp + Gly108Pro + Thr137Glu +Tyr210Glu + Ser220GluGly108Glu + Thr137Gln + Asn163Glu + Tyr210Glu +Ser220Asp + Leu221CysGly 69Gln + Ser 70Glu + Gly135Glu + Asn163Asp +Thr217Gly + Ser220AspAsn106Glu + Asn140Glu + Asn163Glu + Asn170Glu +Thr195Asn + Leu221HisAsn106Gln + Asn166Asp + Ala168Asn + Asn170Glu +Ser207Glu + Ser220AspAsn 66Asp + Ser 70Asp + Gly110Gln + Gly136Asp +Pro214Gln + Ser220AspAsn 68Ser + Pro169Asp + Ser192Glu + Ala219Asp +Gly223Ser + Thr224AspThr 73Pro + Gly135Ser + Phe193Glu + Gly206Gln +Tyr210Asp + Thr217GluTrp112Glu + Val115Gln + Asn170Gln + Thr217Asp +Tyr218Met + Ser220AspGly110Asn + Val115Glu + Thr137Gln + Ile209Gly +Thr217Asp + Ser220GluGly108Glu + Thr113Gln + Pro214Asn + Thr217Glu +Tyr218Gln + Ser220AspGly110Ser + Thr111Gly + Thr113Glu + Phe193Pro +Thr217Glu + Ser220AspAsn166Glu + Trp208Ile + Tyr210Met + Thr217Glu +Tyr218Val + Ser220AspSer 70Glu + Val103Glu + Val115Gln + Pro169Ser +Tyr210Ala + Ser220AspAsn 66Asp + Ser107Asp + Trp208Met + Tyr210Pro +Ser220Asp + Ser222GluGly110Ser + Thr111Asp + Gly135Gln + Gly136Asp +Thr137Asn + Trp208GluThr167Asp + Ala168Ser + Pro169Glu + Trp208Thr +Thr215Asp + Leu221AspVal115Gln + Ala168Gln + Pro169Gln + Phe193Asp +Tyr210Ser + Leu221AspAsn 66Glu + Gly136Pro + Asn140Gln + Ser191Asp +Trp208Asp + Leu221HisGly136Glu + Gly139Glu + Ser192Asp + Ala203Asn +Trp208Asp + Leu221HisGly 67Asn + Thr113Gln + Thr137Glu + Thr167Asp +Ala219Gln + Leu221GluGly 69Asn + Thr 73Asp + Thr111Pro + Asn166Asp +Ser191Asp + Leu221ValVal138Pro + Asn166Glu + Ser191Asp + Tyr218Asn +Ser220Asp + Leu221GlnAsp105Glu + Gly139Asn + Gly165Glu + Trp208Met +Tyr218Thr + Gly223AspGly110Asn + Thr111Pro + Ala114Asp + Asn170Asp +Trp208Asp + Tyr210AspGly 67Gln + Gly 69Glu + Ser107Asp + Thr167Glu +Ala168Asn + Ser207AspGly 69Glu + Gly165Glu + Trp208Cys + Tyr210Cys +Pro214Asp + Ser220AspSer 70Glu + Gly 72Gln + Val103Thr + Ser109Glu +Thr113Glu + Ser220AspGly 69Pro + Val115Glu + Tyr210Asn + Ser216Asp +Ser220Glu + Ser222GluGly136Glu + Trp208Ser + Ser216Asp + Ser220Asp +Leu221Asn + Ser222AspLeu104Glu + Val115Pro + Gly162Ser + Asn166Asp +Ser220Asp + Ser222AspGly 67Glu + Val138Asp + Thr217Ser + Ser220Asp +Leu221His + Ser222AspAsn 68Asp + Asn170Glu + Gly206Asn + Ser207Asp +Trp208Ile + Leu221AsPAsn106Gln + Ser109Glu + Thr113Asp + Leu134Gln +Thr137Glu + Trp208AsnAsn106Glu + Gly139Gln + Thr167Glu + Pro169Gln +Ser191Asp + Ser207GluSer107Glu + Val115Asp + Gly165Asn + Trp208Thr +Tyr218Asp + Ser220GluGly72Asn + Asn106Gln + Gly139Asp + Pro169Glu +Trp208Pro + Ser220GluVal138Asp + Ser141Glu + Pro169Ser + Ser207Glu +Ala219Glu + Leu221ThrAsn106Gln + Ala114Asp + Gly135Asp + Thr195Asn +Ser207Glu + Ser220AspSer 70Asp + Asn166Glu + Asn170Gln + Ser216Glu +Thr217Gln + Thr224GlyGly 69Asp + Asn140Ser + Gly162Pro + Trp208Gly +Thr217Glu + Leu221AspAsn 68Asp + Ser107Asp + Ala114Gln + Gly162Asp +Thr217Pro + Ser220GluSer 70Asp + Thr111Asp + Ile209Met + Thr217Gly +Ser220Glu + Leu221AsnSer 70Asp + Leu104His + Thr111Gln + Tyr171Ser +Phe193Asp + Ser220GluThr113Gly + Gly139Pro + Ser141Glu + Tyr171Glu +Ser192Asp + Ile209SerTrp112Ala + Thr113Glu + Val138Gln + Ser141Asp +Ser216Asp + Leu221SerGly 69Asn + Asn106Asp + Trp112Gly + Gly162Asp +Phe193Glu + Ser220GluAsn 68Ser + Val103Glu + Asn106Asp + Ala168Pro +Pro169Asp + Pro214GluLeu104Glu + Ala114Ser + Asn140Ser + Gly165Asp +Ser207Asp + Ser220GluAsn 68Asp + Ser109Glu + Gly139Glu + Ala204Ser +Leu221His + Gly223AsnThr111Gly + Val138Pro + Gly139Asp + Ser207Glu +Ser216Glu + Ser220AspAsn106Asp + Asn170Ser + Ser192Glu + Thr195Asp +Trp208His + Ser220GluAsn106Asp + Gly162Asp + Asn166Gln + Thr167Asp +Pro214Asp + Ala219ThrVal115Glu + Gly136Glu + Ser207Glu + Tyr218Gly +Ala219Pro + Leu221ProGly136Ser + Gly162Asp + Asn163Gln + Ala164His +Ser216Asp + Ser222GluAsn 68Ser + Thr 73Glu + Asn166Glu + Asn170Gln +Tyr171Gly + Leu221AspThr113Glu + Val115Cys + Thr137Glu + Ser191Glu +Thr217Ser + Thr224GluGly 67Asp + Ser107Asp + Gly139Asp + Gly165Pro +Thr167Ser + Thr217GluThr73Gly + Gly108Asp + Thr137Glu + Gly162Glu +Tyr210Gln + Ser220AspLeu104Asp + Gly139Pro + Tyr171Asp + Ser191Asp +Ser222Glu + Thr224SerThr113Asp + Pro169Glu + Tyr210Gln + Pro214Ser +Ser220Glu + Gly223AspGly108Asp + Gly136Glu + Ser141Asp + Thr195Asp +Thr215Ser + Thr217GlnSer107Asp + Asn163Glu + Pro169Gly + Ser207Asp +Trp208Asn + Gly223AsnGly110Asp + Asn163Asp + Ser191Asp + Thr195Gln +Tyr210Glu + Thr217ProVal103Glu + Gly108Asp + Ala203Gly + Trp208Met +Ala219Asp + Thr224GluThr73Gln + Thr111Asp + Leu134Asp + Gly136Ser +Asn163Gln + Ser207AspII.洗滌組合物在本發明的另一實施方案中，有效量的一種或多種酶變異體包括在用于洗滌各種需要除去蛋白質污垢的表面的組合物中。這種洗滌組合物包括用于洗滌硬表面的非限制形式洗滌組合物(例如，液體或顆粒)；用于洗滌織物的非限制形式的洗滌組合物(顆粒、液體或條狀制劑)；餐具洗滌組合物(非限制形式)；非限制形式的口腔用洗滌組合物(例如，牙粉、牙膏和漱口劑制劑)；非限制形式的假牙洗滌組合物(例如，液體、片劑)；和非限制形式的接觸透鏡洗滌組合物(例如，液體、片劑)。
除了本文上述的熱酶變異體之外，洗滌組合物還包括與蛋白酶相容的一種或多種洗滌組合物材料。本文使用的術語“洗滌組合物材料”指的是選擇用于所需的洗滌組合物的具體類型和產物形式(例如，液體、顆粒、條狀、噴霧劑、棒狀、膏狀、凝膠)的任何液體、固體或氣態材料，該材料還應與組合物中使用的熱酶變異體相容。通過考慮所洗滌的表面材料、在使用過程中用于洗滌條件下所需的組合物的形式(例如通過洗滌劑使用)很容易地具體選擇洗滌組合物材料。本文中使用的術語“相容”指的是洗滌組合物材料不降低熱酶變異體的蛋白水解活性至在正常使用場合根據需要蛋白酶失效的程度。具體的洗滌組合物材料在下文詳細舉例說明。
本文中使用的“有效量的酶變異體”指的是為獲得在具體洗滌組合物中所需的酶活性而必需的酶變異體的量。這種有效量易于被本領域的技術人員確定，其基于許多因素，例如所用的具體的酶變異體、洗滌應用、洗滌組合物的具體組成，和是否需要液體或無水(例如顆粒、條狀)組合物等等。洗滌組合物優選含有約0.0001％-約10％一種或多種本發明的酶變異體，更優選約0.001％-約1％，還優選約0.01％-約0.1％。其中使用酶變異體的各種洗滌組合物的若干實例在下文中進一步地討論。本文使用的所有份數、百分數和比率均為按重量計，除非另有說明。
本文中使用的“非織物洗滌組合物”包括硬表面洗滌組合物、餐具洗滌組合物、口腔洗滌組合物、假牙洗滌組合物和接觸透鏡洗滌組合物。A.用于硬表面、餐具和織物的洗滌組合物本發明的酶變異體可用于其中需要高泡和高效不溶底物去除的各種洗滌組合物中。因此，酶變異體可與各種常規成分一起使用以提供全配制的硬表面洗滌劑，餐具洗滌組合物、織物洗滌組合物等等。這種組合物可以是液體、顆粒、條狀形式等等。這種組合物可配制成含有多達按表面活性劑重量計30％-60％的現代“濃縮”洗滌劑。
本發明的洗滌組合物可選擇性地，并優選地含有各種陰離子、非離子、兩性離子等等的表面活性劑。這些表面活性劑通常以組合物的約5％-約35％的含量存在。
用于本發明的表面活性劑的非限制性實例包括常規的C11-C18烷基苯磺酸鹽和伯和無規烷基硫酸鹽、式為CH3(CH2)x(CHOSO3-M+)CH3和CH3(CH2)y(CHOSO3-M+)CH2CH3的C10-C18仲(2，3)烷基硫酸鹽，其中x和(y+1)是至少約7，優選至少約9的整數，M是水溶性陽離子，尤其是鈉、C10-C18烷基烷氧基硫酸鹽(尤其是EO 1-5乙氧基硫酸鹽)、C10-C18烷基烷氧基羧酸鹽(尤其是EO 1-5乙氧基羧酸鹽)、C10-C18烷基聚糖苷和它們相應的硫酸化聚糖苷，和C12-C18α-磺化脂肪酸酯、C12-C18烷基和烷基苯酚烷氧基化物(尤其是乙氧基化物和混合的乙氧基化物/丙氧基化物)、C12-C18甜菜堿和磺基甜菜堿(“sultaines”)、C10-C18胺氧化物等等。本發明優選烷基烷氧基硫酸鹽(AES)和烷基烷氧基羧酸鹽(AEC)。(根據配制者的需要，使用這種表面活性劑與上述的胺氧化物和/或甜菜堿或磺基甜菜堿表面活性劑的混合物也是優選的)。其它常規使用的表面活性劑在標準文獻中列出。尤其有用的表面活性劑包括在1993年3月16日頒布的Connor等人的US5194639中的C10-C18N-甲基葡糖酰胺(glucamides)，列為本文參考文獻。
用于洗滌組合物中的各種各樣的其它成分可包括在本發明的組合物中，其包括其它活性成分、載體、增溶劑、加工助劑、染料或顏料、用于液體配方的溶劑等等。如果需要附加地增加泡沫，在組合物中，通常以約1％-約10％的含量加入發泡劑，例如C10-C16烷醇酰胺。C10-C14單乙醇和二乙醇酰胺說明了這種發泡劑的典型種類。與上述的高泡輔助表面活性劑，例如胺氧化物、甜菜堿和磺基甜菜堿一起使用這類發泡劑也是有利的。如果需要，溶解的鎂鹽，例如氯化鎂、硫酸鎂等等可以通常約0.1％-約2％的含量加入以提供附加的起泡。
本發明的液體洗滌組合物可含有作為載體的水和其它溶劑。低分子量的伯或仲醇，例如甲醇、乙醇、丙醇和異丙醇是適宜的。單羥基醇對于表面活性劑的增溶是優選的，但多元醇，例如含有約2-約6個碳原子和約2-約6個羥基的多元醇(例如，1，3-丙二醇、乙二醇、甘油和1，2-丙二醇)也可以使用。組合物可含有約5％-約90％，典型地約10％-約50％的這種載體。
本發明的洗滌組合物將優選配制成在使用過程以含水洗滌操作，洗滌水將具有約6.8-約11.0的pH，最終的產物因此通常在該范圍內配制.控制pH在所需要的范圍內的技術包括使用緩沖劑、堿、酸等等，是本領域技術人員已知的。
在配制本發明的硬表面洗滌組合物和織物洗滌組合物時，配制者可以按重量計約5％-約50％的含量使用各種助劑。典型的助劑包括1-10微米沸石、多羧酸鹽，例如檸檬酸鹽和氧聯二琥珀酸鹽、層狀硅酸鹽、磷酸鹽等等。其它常規的助劑在標準配方中列出。
同樣，配制者可在組合物中，通常以按重量計約0.001％-1％的含量使用各種其它的酶，例如纖維素酶、脂酶、淀粉酶和蛋白酶。在洗衣洗滌劑領域中已知各種洗滌和織物護理酶。
各種漂白化合物，例如過碳酸鹽、過硼酸鹽等等可通常以按重量計約1％-約15％的含量用于該組合物中。如果需要，該組合物還可以含有漂白活性劑，例如四乙酰基乙二胺、壬酰氧基苯磺酸鹽等等，它們也是現有技術中已知的。使用含量通常為按重量計約1％-約10％。
各種污垢解脫劑，尤其是陰離子低聚酯類型、各種螯合劑，尤其是氨基膦酸鹽和乙二胺二琥珀酸鹽、各種粘土去污劑，尤其是乙氧基化四亞乙五胺、各種分散劑，尤其是聚丙烯酸酯和聚天冬氨酸酯、各種增白劑，尤其是陰離子增白劑、各種泡沫抑制劑，尤其硅烷和仲醇、各種織物柔軟劑，尤其是綠土等等都可以按重量計約1％-約35％的含量用于組合物。在標準配方和公開的專利中有此類常規物質的詳細描述。
在洗滌組合物中也可使用酶穩定劑。酶穩定劑包括丙二醇(優選約1％-約10％)、甲酸鈉(優選約0.1％-約1％)和甲酸鈣(優選約0.1％-約1％)。l.硬表面洗滌組合物本文使用的“硬表面洗滌組合物”指的是用于洗滌硬表面，例如地板、墻壁、浴室瓷磚等等的液體和顆粒洗滌組合物。本發明的硬表面洗滌組合物含有有效量的一種或多種本發明的酶變異體，按組合物活性酶重量計優選為約0.001％-約10％，更優選約0.01％-約5％，還優選約0.05％-約1％。除了含有一種或多種酶變異體之外，硬表面洗滌組合物通常含有表面活性劑和水溶性螯合劑。然而，在某些特殊的產物，例如噴霧玻璃清潔劑中，有時是不使用表面活性劑的，因為它們會產生殘留在玻璃表面的膜狀/條紋殘余物。
當存在時，表面活性劑組分可含有少至組合物的0.1％，但通常組合物將含有約0.25％-約10％，更優選約1％-約5％的表面活性劑。
組合物通常含有約0.5％-約50％的洗滌劑助劑，優選為約1％-約10％。
pH值優選在約8-12的范圍內。如果需要進行調節，可使用常規的pH調節劑，例如氫氧化鈉、碳酸鈉或鹽酸。
在組合物中可包括溶劑。有用的溶劑包括，但不限制于，乙二醇醚，例如二乙二醇單己基醚、二乙二醇單丁基醚、乙二醇單丁基醚、乙二醇單己基醚、丙二醇單丁基醚、二丙二醇單丁基醚，和二醇，例如2，2.4-三甲基-1，3-戊二醇和2-乙基-1，3-己二醇。當使用時，溶劑通常以約0.5％-約15％，優選約3％-約11％的含量存在。
此外，當組合物以“完全強度”應用于表面后表面不進行漂洗時在本發明的組合物中可使用高揮發性溶劑，例如異丙醇或乙醇，以有助于組合物從表面快速蒸發。當使用時，揮發性溶劑通常以約2％-約12％的含量存在于組合物中。
本發明的硬表面洗滌組合物的實施方案通過如下實施例說明。
實施例7-12液體硬表面洗滌組合物實施例組分7 8 9 101112Gln66Asn0.05 0.50 0.02 0.03 0.10 0.03Gly206Asn - - - - 0.20 0.02Na2DIDA*EDTA**- - 2.90 2.90 - -檸檬酸鈉- - - - 2.90 2.90C12烷基苯磺酸鈉1.95 - 1.95 - 1.95 -C12烷基硫酸鈉 - 2.20 - 2.20 - 2.20C12(乙氧基)***硫酸鈉 - 2.20 - 2.20 - 2.20C12二甲基胺氧化物 - 0.50 - 0.50 - 0.50枯烯磺酸鈉 1.30 - 1.30 - 1.30 -己基卡必醇***6.30 6.30 6.30 6.30 6.30 6.30水****平衡至100％*N-二乙二醇-N，N-亞氨基二乙酸二鈉**乙二胺二乙酸四鈉***二乙二醇單己基醚****所有配方調節到pH7在實施例7-10中，用表2-36中所列的熱酶變異體代替Gln66Asn，得到基本上類似的結果。
在實施例11-12中，用表2-36中所列的熱酶變異體的任何組合代替Gln66Asn和Gly206Asn，得到基本上類似的結果。
實施例13-18用于清潔硬表面和除去家庭霉變的噴霧組合物實施例組分 131415161718Thr111Asn 0.50 0.05 0.60 0.30 0.20 0.30Thr217Gly+Ser222Glu- - - - 0.30 0.10辛基硫酸鈉 2.00 2.00 2.00 2.00 2.00 2.00十二烷基硫酸鈉 4.00 4.00 4.00 4.00 4.00 4.00氫氧化鈉 0.80 0.80 0.80 0.80 0.80 0.80硅酸鹽(鈉) 0.04 0.04 0.04 0.04 0.04 0.04香料 0.35 0.35 0.35 0.35 0.35 0.35水平衡至100％產物pH為約7。
在實施例13-16中，用表2-36中所列的熱酶變異體代替Thr111Asn，得到基本上類似的結果。
在實施例17-18中，用表2-36中所列的熱酶變異體的任何組合代替Thr111Asn和Thr217Gly+Ser222Glu，得到基本上類似的結果。
2.餐具洗滌組合物在本發明的另一實施方案中，餐具洗滌組合物含有一種或多種本發明的酶變異體。本文使用的“餐具洗滌組合物”指的是用于洗滌餐具的組合物的所有形式，其包括，但不限制于，顆粒和液體形式。本發明的餐具洗滌組合物的實施方案由如下實施例說明。
實施例19-24餐具洗滌組合物實施例組分 192021222324Gly139Asn 0.05 0.50 0.02 0.40 0.10 0.03Ser207Glu+Tyr210Ser- - - - 0.40 0.02+Gly223AsnC12-C14N-甲基葡糖酰 0.90 0.90 0.90 0.90 0.90 0.90胺C12乙氧基(1)硫酸鹽12.00 12.00 12.00 12.00 12.00 12.002-甲基十一烷酸 4.50 4.50 4.50 4.50 4.50 4.50C12乙氧基(2)羧酸鹽4.50 4.50 4.50 4.50 4.50 4.50C12醇乙氧基化物(4)3.00 3.00 3.00 3.00 3.00 3.00C12胺氧化物 3.00 3.00 3.00 3.00 3.00 3.00枯烯磺酸鈉 2.00 2.00 2.00 2.00 2.00 2.00乙醇 4.00 4.00 4.00 4.00 4.00 4.00Mg++(作為氯化鎂) 0.20 0.20 0.20 0.20 0.20 0.20Ca++(作為氯化鈣) 0.40 0.40 0.40 0.40 0.40 0.40水 平衡至100％產物pH調節至7。
在實施例19-22中，用表2-36中所列的熱酶變異體代替Gly139Asn，得到基本上類似的結果。
在實施例23-24中，用表2-36中所列的熱酶變異體的任何組合代替Gly139Asn和Ser207Glu+Tyr210Ser+Gly223Asn，得到基本上類似的結果。
3.織物洗滌組合物在本發明的另一實施方案中，織物洗滌組合物含有一種或多種本發明的酶變異體。本文使用的“織物洗滌組合物”指的是用于洗滌織物的組合物的所有形式，其包括，但不限制于，顆粒、液體和條狀形式。優選的織物洗滌組合物是液體形式。a.顆粒織物洗滌組合物本發明的顆粒織物洗滌組合物含有有效量的一種或多種本發明的酶變異體，按組合物活性酶重量計優選為約0.001％-約10％，更優選約0.005％-約5％，還優選約0.01％-約1％。除了含有一種或多種酶變異體之外，顆粒織物洗滌組合物通常含有至少一種表面活性劑、一種或多種助劑和，在某些情況下，漂白劑。
本發明的顆粒織物洗滌組合物的實施方案由如下實施例說明。
實施例25-28顆粒織物洗滌組合物實施例組分 25 26 27 28Ala168Asn 0.10 0.20 0.03 0.05Tyr210Gly+Thr212Pro+Pro- - 0.02 0.05214Gly+Ser222AspC13直鏈烷基苯磺酸鹽 22.00 22.00 22.00 22.00磷酸鹽(作為三聚磷酸鈉) 23.00 23.00 23.00 23.00碳酸鈉 23.00 23.00 23.00 23.00硅酸鈉 14.00 14.00 14.00 14.00沸石 8.20 8.20 8.20 8.20螯合劑(二亞乙三胺五乙酸) 0.40 0.40 0.40 0.40硫酸鈉 5.50 5.50 5.50 5.50水 平衡至100％在實施例25-26中，用表2-36中所列的熱酶變異體代替Ala168Asn，得到基本上類似的結果。
在實施例27-28中，用表2-36中所列的熱酶變異體的任何組合代替Ala168Asn和Tyr210Gly+Thr212Pro+Pro214Gly+Ser222Asp，得到基本上類似的結果。
實施例29-32顆粒織物洗滌組合物實施例組分 29 30 31 32Thr195Pro0.10 0.20 0.03 0.05Trp208Phe+Tyr213Met+Thr215 - - 0.02 0.05Pro+Ser220Glu+Thr224GlyC12烷基苯磺酸鹽 12.00 12.00 12.00 12.00沸石A(1-10微米) 26.00 26.00 26.00 26.002-丁基辛酸 4.00 4.00 4.00 4.00C12-C14仲(2，3)烷基硫酸鈉 5.00 5.00 5.00 5.00檸檬酸鈉 5.00 5.00 5.00 5.00熒光增白劑 0.10 0.10 0.10 0.10硫酸鈉 17.00 17.00 17.00 17.00水和少量組分 平衡至100％在實施例29-30中，用表2-36中所列的熱酶變異體代替Thr195Pro，得到基本上類似的結果。
在實施例31-32中，用表2-36中所列的熱酶變異體的任何組合代替Thr195Pro和Trp208Phe+Tyr213Met+Thr215Pro+Ser220Glu+Thr224Gly，得到基本上類似的結果。
實施例33-36顆粒織物洗滌組合物實施例組分 33 34 35 36Gly67Ser+Gly72Ser 0.100.20 0.030.05Tyr171Thr - - 0.020.05C13直鏈烷基苯磺酸鹽 22.00 22.00 22.00 22.00磷酸鹽(作為三聚磷酸鈉) 23.00 23.00 23.00 23.00碳酸鈉 23.00 23.00 23.00 23.00硅酸鈉 14.00 14.00 14.00 14.00沸石 8.208 20 8.208.20螯合劑(二亞乙三胺五乙酸) 0.400.40 0.400.40硫酸鈉 5.505.50 5.505.50水 平衡至100％在實施例33-34中，用表2-36中所列的熱酶變異體代替Gly67Ser+Gly72Ser，得到基本上類似的結果。
在實施例35-36中，用表2-36中所列的熱酶變異體的任何組合代替Gly67Ser+Gly72Ser和Tyr171Thr，得到基本上類似的結果。
實施例37-40顆粒織物洗滌組合物實施例組分 37 38 39 40Ser192Asp+Phe193Leu+Thr195As 0.100.200.030.05nTyr213Ser+Thr217Gly+Gly223Glu - - 0.020.05C12烷基苯磺酸鹽 12.0012.00 12.00 12.00沸石A(1-10微米) 26.0026.00 26.00 26002-丁基辛酸4.00 4.004.004.00C12-C14仲(2，3)烷基硫酸鈉 5.00 5.005.005.00檸檬酸鈉 5.00 5.005.005.00熒光增白劑0.10 0.100.100.10硫酸鈉17.0017.00 17.00 17.00水和少量組分平衡至100％在實施例37-38中，用表2-36中所列的熱酶變異體代替Ser192Asp+Phe193Leu+Thr195Asn，得到基本上類似的結果。
在實施例39-40中，用表2-36中所列的熱酶變異體的任何組合代替Ser192Asp+Phe193Leu+Thr195Asn和Tyr213Ser+Thr217Gly+Gly223Glu，得到基本上類似的結果。
實施例41-42顆粒織物洗滌組合物實施例組分41 42直鏈烷基苯磺酸鹽11.410.70動物脂烷基硫酸鹽1.802.40C14-15烷基硫酸鹽3.003.10C14-15醇7倍乙氧基化 4.004.00動物脂醇11倍乙氧基化1.801.80分散劑 0.070.1硅氧烷液體 0.800.80檸檬酸三鈉 14.00 15.00檸檬酸 3.002.50沸 32.50 32.10馬來酸丙烯酸共聚物 5.005.00二亞乙三胺五亞甲基膦酸 1.000.20Leu104Asp+Gly139Pro+Tyr171Asp 0.300.30+Ser191Asp+Ser222Glu+Thr224Ser脂酶0.360.40淀粉酶 0.300.30硅酸鈉 2.002.50硫酸鈉 3.505.20聚乙烯吡咯烷酮 0.300.50過硼酸鹽0.5 1苯酚磺酸鹽 0.1 0.2過氧化酶0.1 0.1少量組分至100 至100實施例43-44顆粒織物洗滌組合物實施例組分 43 44直鏈C12烷基苯磺酸鈉 6.58.0硫酸鈉 15.0 18.0沸石A26.0 22.0次氮基三乙酸鈉 5.05.0聚乙烯吡咯烷酮 0.50.7四乙酰基乙二胺 3.03.0硼酸 4.0-過硼酸鹽 0.51苯酚磺酸鹽 0.10.2Pro205Gly+Trp208Leu+Thr217Gln+Leu0.40.4221Gln+Gly223Gln+Thr224Glu填料(例如，硅酸鹽；碳酸鹽；香料；水) 至100 至100實施例45壓型顆粒織物洗滌組合物組分 重量％烷基硫酸鹽 8.0烷基乙氧基硫酸鹽 2.0C25和C45醇3和7倍乙氧基化的混合物 6.0多羥基脂肪酸酰胺 2.5沸石 17.0層狀硅酸鹽/檸檬酸鹽16.0碳酸鹽 7.0馬來酸丙烯酸共聚物 5.0污垢解脫聚合物 0.4羧甲基纖維素 0.4聚(4-乙烯基吡啶)-N-氧化物 0.1乙烯基咪唑和乙烯基吡咯烷酮共聚物 0.1PEG20000.2Asn66Ser+Ser70Glu+Gly72Asn 0.5脂酶 0.2纖維素酶 0.2四乙酰基乙二胺 6.0過碳酸鹽 22.0乙二胺二琥珀酸 0.3泡沫抑制劑 3.54，4’-雙(2-嗎啉基-4-苯胺基-s-三嗪-6-基氨基) 0.25芪-2，2’-二磺酸二鈉4，4’-雙(2-磺基苯乙烯基)聯苯二鈉 0.05水、香料和少量組分 至100實施例46顆粒織物洗滌組合物組分 重量％直鏈烷基苯磺酸鹽 7.6C16-C18烷基硫酸鹽1.3C14-15醇7倍乙氧基化物 4.0椰子烷基二甲基羥基乙基銨氯化物 1.4分散劑 0.07硅氧烷液體 0.8檸檬酸三鈉 5.0沸石4A 15.0馬來酸丙烯酸共聚物 4.0二亞乙三胺五亞甲基膦酸 0.4過硼酸鹽 15.0四乙酰基乙二胺 5.0綠土 10.0聚(環氧乙烷)(MW300000) 0.3Ser109Glu+Thr113Gly0.4脂酶 0.2淀粉酶 0.3纖維素酶 0.2硅酸鈉 3.0碳酸鈉 10.0羧甲基纖維素 0.2增白劑 0.2水、香料和少量組分 至100實施例47顆粒織物洗滌組合物組分 重量％直鏈烷基苯磺酸鹽 6.92動物脂烷基硫酸鹽 2.05C14-15醇7倍乙氧基化物 4.4C12-15烷基乙氧基硫酸鹽-3倍乙氧基化 0.16沸石 20.2檸檬酸鹽 5.5碳酸鹽 15.4硅酸鹽 3.4馬來酸丙烯酸共聚物 4.0羧甲基纖維素酶 0.31污垢解脫聚合物 0.30Gly135Gln+Val138Asp+Gly139Ser 0.20脂酶 0.36纖維素酶 0.13過硼酸鹽四水合物 11.64過硼酸鹽單水合物 8.7四乙酰基乙二胺 5.0二亞乙三胺五甲基膦酸 0.38硫酸鎂 0.40增白劑 0.19香料、硅氧烷、泡沫抑制劑 0.85少量組分 至100b.液體織物洗滌組合物本發明的液體織物洗滌組合物含有有效量的一種或多種本發明的酶變異體，按組合物活性酶重量計優選為約0.005％-約5％，更優選為約0.01％-約1％。該液體織物洗滌組合物通常還含有一種陰離子表面活性劑、脂肪酸、水溶性洗滌劑助劑和水。
本發明的液體織物洗滌組合物的實施方案由如下實施例說明。
實施例48-52液體織物洗滌組合物實施例組分48 49 50 51 52Asn166Asp+Thr167Asn+0.05 0.03 0.30 0.03 0.10Asn170Ser+Tyr171ThrSer191Glu+Ser192Asp+- - - 0.01 0.20Phe193Asp+Thr195ProC12-C14烷基硫酸鈉 20.00 20.00 20.00 20.00 20.002-丁基辛酸 5.00 5.00 5.00 5.00 5.00檸檬酸鈉1.00 1.00 1.00 1.00 1.00C10醇乙氧基化物(3) 13.00 13.00 13.00 13.00 13.00單乙醇胺2.50 2.50 2.50 2.50 2.50水/丙二醇/乙醇(100∶1∶1) 平衡至100％在實施例48-50中，用表2-36所列的熱酶變異體代替Asn166Asp+Thr167Asn+Asn170Ser+Tyr171Thr，得到基本上類似的結果。
在實施例51-52中，用表2-36中所列的熱酶變異體的任何組合代替Asn166Asp+Thr167Asn+Asn170Ser+Tyr171Thr和Ser191Glu+Ser192Asp+Phe193Asp+Thr195Pro，得到基本上類似的結果。
實施例53-57液體織物洗滌組合物實施例組分 53 54 55 56 57Pro205Gln+Gly206Ser+Trp208V0.05 0.03 0.30 0.03 0.10al+Tyr210Leu+Thr212Asn+Tyr218Met+Ser220GluAsn68Asp+Val138Asp+Ala164 - - - 0.01 0.20Gly+Ala168Glu+Thr217GlyC12-C14烷基硫酸鈉20.00 20.00 20.00 20.00 20.002-丁基辛酸 5.00 5.00 5.00 5.00 5.00檸檬酸鈉 1.00 1.00 1.00 1.00 1.00C10醇乙氧基化物(3)13.00 13.00 13.00 13.00 13.00單乙醇胺 2.50 2.50 2.50 2.50 2.50水/丙二醇/乙醇(100∶1∶1)平衡至100％在實施例53-55中，用表2-36中所列的熱酶變異體代替Pro205Gln+Gly206Ser+Trp208Val+Tyr210Leu+Thr212Asn+Tyr218Met+Ser220Glu，得到基本上類似的結果。
在實施例56-57中，用表2-36中所列的熱酶變異體的任何組合代替Pro205Gln+Gly206Ser+Trp208Val+Tyr210Leu+Thr212Asn+Tyr218Met+Ser220Glu和Asn68Asp+Val138Asp+Ala164Gly+Ala168Glu+Thr217Gly，得到基本上類似的結果。
實施例58-59液體織物洗滌組合物實施例組分 58 59C12-14烯基琥珀酸 3.08.0檸檬酸單水合物 10.0 15.0C12-15烷基硫酸鈉 8.08.0C12-15醇2倍乙氧基化硫酸鈉- 3.0C12-15醇7倍乙氧基化物- 8.0C12-15醇5倍乙氧基化物8.0-二亞乙三胺五(亞甲基膦酸) 0.2-油酸 1.8-乙醇 4.04.0丙二醇 2.02.0Leu134Cys+Thr137Glu+Val138Gly0.20 2聚乙烯吡咯烷酮 1.02.0泡沫抑制劑 0.15 0.15氫氧化鈉 至pH7.5過硼酸鹽 0.51苯酚磺酸鹽 0.10.2過氧化酶 0.40.1水和少量組分 至100份在實施例58和59中，用表2-36中所列的熱酶變異體代替Leu134Cys+Thr137Glu+Val138Gly，得到基本上類似的結果。
實施例60-62液體織物洗滌組合物實施例組分 6061 62檸檬酸 7.10 3.003.00脂肪酸 2.00 - 2.00乙醇 1.93 3.203.20硼酸 2.22 3.503.50單乙醇胺 0.71 1.091.091，2-丙二醇7.89 8.008.00枯烯磺酸鈉 1.80 3.003.00甲酸鈉 0.08 0.080.08氫氧化鈉 6.70 3.803.80含硅消泡劑 1.16 1.181.18Gly67Glu+Ser70Asp+Gly72Ser+Thr73Ser0.0145- -Ile209Ala+Ala219Pro- 0.0145 -Leu216Asn - - 0.0145脂酶 0.200 0.200 0.200纖維素酶 - 7.507.50污垢解脫聚合物 0.29 0.150.15消泡劑 0.06 0.085 0.085增白劑36 0.095 - -增白劑3- 0.050.05C12烷基苯磺酸 9.86 - -C12-15烷基多乙氧基化(2.5)硫酸鹽13.80 18.00 18.00C12葡糖酰胺 - 5.005.00C12-13烷基聚乙氧基化物(9) 2.00 2.002.00水、香料和少量組分 平衡至100％c.條狀織物洗滌組合物本發明的適合于手洗臟織物的條狀織物洗滌組合物含有有效量的一種或多種本發明的酶變異體，按組合物活性酶重量計優選為約0.001％-約10％，更優選為約0.01％-約1％。
本發明的條狀織物洗滌組合物的實施方案由如下實施例說明。
實施例63-66條狀織物洗滌組合物實施例組分 63 64 65 66Trp112Cys+Thr217Gly 0.3- 0.10.02Val103Ala+Thr212Ser - - 0.40.03C12-C16烷基硫酸鈉 20.0 20.0 20.0 20.00C12-C14N-甲基葡糖酰胺 5.05.05.05.00C11-C13烷基苯磺酸鈉10.0 10.0 10.0 10.00碳酸鈉 25.0 25.0 25.0 25.00焦磷酸鈉 7.07.07.07.00三聚磷酸鈉 7.07.07.07.00沸石A(0.1-10μ) 5.05.05.05.00羧甲基纖維素 0.20.20.20.20聚丙烯酸酯(MW1400) 0.20.20.20.20椰子烷基單乙醇胺 5.05.05.05.00增白劑、香料 0.20.20.20.20硫酸鈣 1.01.01.01.00硫酸鎂 1.01.01.01.00水 4.04.04.04.00填料*平衡至100％*可以選自常規材料，例如碳酸鈣、滑石、粘土、硅酸鹽等等。
在實施例63-64中，用表2-36中所列的熱酶變異體代替Trp112Cys+Thr217Gly，得到基本上類似的結果。
在實施例65-66中，用表2-36中所列的熱酶變異體的任何組合代替Trp112Cys+Thr217Gly和Val103Ala+Thr212Ser，得到基本上類似的結果。
實施例67-70條狀織物洗滌組合物實施例組分67 6869 70Ser109Glu 0.3- 0.1 0.02Pro169Glu - 0.3 0.4 0.03C12-C16烷基硫酸鈉 20.0 20.0 20.020.0C12-C14N-甲基葡糖酰胺 5.05.0 5.0 5.00C11-C13烷基苯磺酸鈉 10.0 10.0 10.010.00碳酸鈉 25.0 25.0 25.025.00焦磷酸鈉7.07.0 7.0 7.00三聚磷酸鈉 7.07.0 7.0 7.00沸石A(0.1-10μ) 5.05.0 5.0 5.00羧甲基纖維素0.20.2 0.2 0.20聚丙烯酸酯(MW1400) 0.20.2 0.2 0.20椰子烷基單乙醇胺5.05.0 5.0 5.00增白劑、香料0.20.2 0.2 0.20硫酸鈣 1.01.0 1.0 1.00硫酸鎂 1.01.0 1.0 1.00水 4.04.0 4.0 4.00填料*平衡至100％*可以選自常規材料，例如碳酸鈣、滑石、粘土、硅酸鹽等等。
在實施例67中，用表2-36中所列的熱酶變異體代替Ser109Glu，得到基本上類似的結果。
在實施例68中，用表2-36中所列的熱酶變異體代替Pro169Glu，得到基本上類似的結果。
在實施例69-70中，用表2-36中所列的熱酶變異體的任何組合代替Ser109Glu和Pro169Glu，得到基本上類似的結果。B.其它洗滌組合物除了上述硬表面洗滌、餐具洗滌和織物洗滌組合物之外，本發明的一種或多種酶變異體可加入各種需要水解不溶解底物的其它洗滌組合物中。其它洗滌組合物包括，但不限制于，口腔洗滌組合物、假牙洗滌組合物和接觸透鏡洗滌組合物。
1.口腔洗滌組合物在本發明的另一實施方案中，在組合物中包含一種或多種可藥用量的本發明的酶變異體以從牙齒或假牙中除去蛋白質污垢。本文使用的“口腔洗滌組合物”指的是潔齒粉、牙膏、牙凝膠、牙粉、漱口劑、口腔噴霧劑、口腔凝膠、口香糖、錠劑、香囊、片劑、生物凝膠、預防膏、假牙處理溶液等等。按組合物的重量計，洗滌組合物優選含有約0.0001-約20％，更優選約0.001％-約10％，還優選約0.01％-約5％的一種或多種本發明的酶變異體和可藥用的載體。本文使用的“可藥用的”指的是該術語描述的藥物、藥劑或惰性成分適合于與人體和低級動物的組織接觸，而沒有不適宜的毒性、不適性、不穩定性、刺激性、過敏反應等等，與有益效果/風險比率相當。
口腔洗滌組合物的可藥用的口腔洗滌載體組分按組合物重量計通常占約50％-約99.99％，優選約65％-約99.99％，更優選約65％-約99％。
可包括在本發明的口腔洗滌組合物中的可藥用的載體組分和選擇性的組分是本領域技術人員已知的。用于口腔洗滌組合物的各種組合物類型、載體組分和選擇性組分在1992年3月17日授權的Seibel的US5096700；1991年7月2日授權的Sampathkumar的US5028414和1991年7月2日授權的Benedict、Bush和Sunberg的US5028415中，列為本文參考文獻。
本發明的口腔洗滌組合物的實施方案在如下實施例中說明。
實施例71-74潔牙粉組合物實施例組分 71 72 73 74Tyr218Ala 2.000 3.500 1.500 2.000山梨糖醇(70％水溶液) 35.000 35.000 35.000 35.000PEG-6*1.000 1.000 1.000 1.000氧化硅牙用磨料**20.000 20.000 20.000 20.000氟化鈉 0.243 0.243 0.243 0.243二氧化鈦 0.500 0.500 0.500 0.500糖精鈉 0.286 0.286 0.286 0.286烷基硫酸鈉(27.9％水溶液) 4.000 4.000 4.000 4.000香料 1.040 1.040 1.040 1.040羧乙烯基聚合物***0.300 0.300 0.300 0.300Carrageenan****0.800 0.800 0.800 0.800水 平衡至100％*PEG-6＝分子量600的聚乙二醇。**由J.M.Huber提供的沉淀的氧化硅Zeodent119***由B.F.Goodrich Chemical Company提供的Carbopol****由Hercules Chemical Company提供的lota Carrageenan。
在實施例71-74中，用表2-36中所列的熱酶變異體代替Tyr218Ala，得到基本上類似的結果。
實施例75-78漱口劑組合物實施例組分 75 76 77 78Ala164His+Ser220Glu3.007.501.005.00SDA40醇8.008.008.008.00香料 0.080.080.080.08乳化利 0.080.080.080.08氟化鈉 0.050.050.050.05甘油 10.00 10.00 10.00 10.00增甜劑 0.020.020.020.02苯甲酸 0.050.050.050.05氫氧化鈉 0.200.200.200.20染料 0.040.040.040.04水 平衡至100％在實施例75-78中，用表2-36中所列的熱酶變異體代替Ala164His+Ser220Glu，得到基本上類似的結果。
實施例79-82錠劑組合物實施例組分 79 80 81 82Leu221Val+Thr224Gln0.010.03 0.10 0.02山梨糖醇 17.50 17.50 17.50 17.50甘露糖醇 17.50 17.50 17.50 17.50淀粉 13.60 13.60 13.60 13.60增甜劑 1.201.20 1.20 1.20香料 11.70 11.70 11.70 11.70色素 0.100.10 0.10 0.10玉米糖漿 平衡至100％在實施例79-82中，用表2-36中所列的熱酶變異體代替Leu221Val+Thr224Gln，得到基本上類似的結果。
實施例83-86咀嚼膠組合物實施例組分83 84 85 86Ser191Asp+Phe193Ile+Thr195Gly 0.03 0.02 0.10 0.05山梨糖醇結晶38.44 38.40 38.40 38.40Palija-T膠基料*20.00 20.00 20.00 20.00山梨糖醇(70％水溶液)22.00 22.00 22.00 22.00甘露糖醇10.00 10.00 10.00 10.00甘油7.56 7.56 7.56 7.56香料1.00 1.00 1.00 1.00*L.A.Dreyfus Company提供。
在實施例83-86中，用表2-36中所列的熱酶變異體代替Ser191Asp+Phe193Ile+Thr195Gly，得到基本上類似的結果。
2.假牙洗滌組合物在本發明的另一實施方案中，用于在口腔外洗滌假牙的假牙洗滌組合物含有一種或多種本發明的酶變異體。該假牙洗滌組合物含有有效量的一種或多種酶變異體，優選按組合物重量計，約0.0001-約50％，更優選約0.001％-約35％，還優選約0.01％-約20％的一種或多種本發明的酶變異體，和假牙洗滌載體。各種假牙洗滌組合物配方，如泡騰片等等是現有技術中已知的(參見例如Young的US5055305，列為本文參考文獻)，通常適合于加入一種或多種酶變異體以便從假牙除去蛋白質污垢。
本發明的假牙洗滌組合物的實施方案由如下實施例說明。
實施例87-90雙層泡騰假牙洗滌片實施例組分878889 90酸性層Gly135Gln+Gly139Asn+1.0 1.5 0.01 0.05Asn140Gln+Ser141Asp酒石酸 24.0 24.0 24.00 24.00碳酸鈉 4.0 4.0 4.00 4.00氨基磺酸10.0 10.0 10.00 10.00PEG200004.0 4.0 4.00 4.00碳酸氫鈉24.5 24.5 24.50 24.50過硫酸鉀15.0 15.0 15.00 15.00酸性焦磷酸鈉7.0 7.0 7.00 7.00熱解氧化硅 2.0 2.0 2.00 2.00TAED*7.0 7.0 7.00 7.00蓖麻酰基磺基琥珀酸鹽0.5 0.5 0.50 0.50香料1.0 1.0 1.00 1.00堿性層過硼酸鈉單水合物32.0 32.0 32.00 32.00碳酸氫鈉19.0 19.0 19.00 19.00EDTA3.0 3.0 3.00 3.00三聚磷酸鈉 12.0 12.0 12.00 12.00PEG200002.0 2.0 2.00 2.0過硫酸鉀26.0 26.0 26.00 26.00碳酸鈉 2.0 2.0 2.00 2.00熱解氧化硅 2.0 2.0 2.00 2.00染料/香料 2.0 2.0 2.00 2.00*四乙酰基乙二胺在實施例87-90中，用表2-36中所列的熱酶變異體代替Gly135Gln+Gly139Asn+Asn140Gln+Ser141Asp，得到基本上類似的結果。
2.接觸透鏡洗滌組合物在本發明的另一實施方案中，接觸透鏡洗滌組合物含有一種或多種本發明的酶變異體。該接觸透鏡洗滌組合物含有有效量的一種或多種酶變異體，優選按組合物重量計，約0.01-約50％，更優選約0.01％-約20％，還優選約1％-約5％的一種或多種本發明的酶變異體，和接觸透鏡洗滌載體。各種接觸透鏡洗滌組合物配方，例如片劑、液體等等在現有技術中是已知的(參見例如1989年9月5日授權的Davies，Meaken和Rees，US4863627；1988年5月24日再次授權的Huth，Lam和Kirai，USRe32672；有1986年9月2日授權的Schafer，US4609493；1987年9月1日授權的Ogunbiyi和Smith，US4690793；1986年9月30日授權的Ogunbiyi，Riedhammer和Smiht，US4614549；和1981年8月25日授權的Ogata，US4285738；均列為本文參考文獻)，通常適合于加入本發明的一種或多種酶變異體以便從接觸透鏡除去蛋白質污垢。
本發明的接觸透鏡洗滌組合物的實施方案由如下實施例說明。
實施例91-94酶解接觸透鏡洗滌溶液實施例組分91 92 93 94Leu221Gln 0.010.50.12.0葡萄糖 50.00 50.0 50.0 50.0非離子表面活性劑(聚氧乙烯- 2.002.02.02.0聚氧丙烯共聚物)陰離子表面活性劑(聚氧乙烯- 1.001.01.01.0烷基苯基醚硫酸酯鈉)氯化鈉 1.001.01.01.0硼砂0.300.30.30.3水 平衡至100％在實施例91-94中，用表2-36中所列的熱酶變異體代替Leu221Gln，得到基本上類似的結果。
盡管描述了本發明的具體實施方案，但顯然本領域的技術人員可作出本發明的各種改變和改進而不會違背本發明的精神和范圍。在所附的權利要求中，本發明的范圍覆蓋了所有這些改進。
權利要求
1.具有熱酶(thermitase)野生型氨基酸序列的修飾氨基酸序列的熱酶變異體，野生型氨基酸序列包括第一環區域、第二環區域、第三環區域、第四環區域、第五環區域和第六環區域；其中修飾的氨基酸序列在一個或多個環區域內的一個或多個位點上含有取代基；其中A.當取代發生在第一環區域中時，取代發生在位點66、69、70、72或73的一個或多個上；其中a.當取代發生在位點66時，取代氨基酸是Asn；b.當取代發生在位點69時，取代氨基酸是Gln；c.當取代發生在位點70時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser；d.當取代發生在位點72時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser；e.當取代發生在位點73時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser；B.當取代發生在第二環區域中時，取代發生在位點103、104、105、106、108、110、112、113或114的一個或多個上；其中a.當取代發生在位點103時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr；b.當取代發生在位點104時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Gly、His、Ile、Met、Pro或Ser；c.當取代發生在位點105時，取代氨基酸是Glu；d.當取代發生在位點106時，取代氨基酸是Gln；e.當取代發生在位點108時，取代氨基酸是Asn、Gln、Pro或Ser；f.當取代發生在位點110時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser；g.當取代發生在位點112時，取代氨基酸是Asn、Asp、Cys、Gln、Glu、His、Ile、Met、Phe、Pro、Thr或Tyr；h.當取代發生在位點113時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser；和i.當取代發生在位點114時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr；C.當取代發生在第三環區域中時，取代發生位點134、135、136、137、138、139或141的一個或多個上；其中a.當取代發生在位點134時，取代氨基酸是Ala、Asn、Cys、Gln、Gly、His、Met、Pro、Ser、Thr或Val；b.當取代發生在位點135時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser；c.當取代發生在位點136時，取代氨基酸是Pro；d.當取代發生在位點137時，取代氨基酸是Asn、Asp、Gln、Glu或Ser；e.當取代發生在位點138時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Met、Pro、Ser或Thr；f.當取代發生在位點139時，取代氨基酸是Asn、Gln、Pro或Ser；和g.當取代發生在位點141時，取代氨基酸是Asp或Glu；D.當取代發生在第四環區域中時，取代發生在位點167、169或171一個或多個上；其中a.當取代發生在位點167時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser；b當取代發生在位點169時，取代氨基酸是Asn、Asp、Gln、Glu、Gly或Ser；和c.當取代發生在位點171時，取代氨基酸是His、Ile、Leu、Met或Pro；E.當取代發生在第五環區域中時，取代發生在位點193一個或多個上；其中a.當取代發生在位點193時，取代氨基酸是Asn、Cys、Gln、His、Ile、Met、Thr或Tyr；F.當取代發生在第六環區域中時，取代發生在位點204、205、206、207、208、209、210、211、212、213、214、215、216、217、220、221、223或224的一個或多個上，其中a.當取代發生在位點204時，取代氨基酸是Asn、Asp、Gln、Glu、Gly、His、Pro、Ser或Thr；b.當取代發生在位點205時，取代氨基酸是Asn、Asp、Gln、Glu、Gly或Ser；c.當取代發生在位點206時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser；d.當取代發生在位點207時，取代氨基酸是Asp或Glu；e.當取代發生在位點208時，取代氨基酸是Asp或Glu；f.當取代發生在位點209時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Leu、Met、Pro、Ser、Thr或Val；g.當取代發生在位點210時，取代氨基酸是Asp、His、Ile、Leu、Met或Pro；h.當取代發生在位點211時，取代氨基酸是Asp或Glu；i.當取代發生在位點212時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser；j.當取代發生在位點213時，取代氨基酸是Asp、Glu、His、Ile、Leu、Met、Phe、Pro或Val；i.當取代發生在位點214時，取代氨基酸是Asn、Gln、Gly或Ser；j.當取代發生在位點215時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser；m.當取代發生在位點216時，取代氨基酸是Asp或Glu；n.當取代發生在位點217時，取代氨基酸是Asp、Glu或Pro；o.當取代發生在位點220時，取代氨基酸是Asp或Glu；p.當取代發生在位點221時，取代氨基酸是Ala、Asn、Asp、Cys、Gln、Glu、Gly、His、Ile、Met、Pro、Ser、Thr或Val；q.當取代發生在位點222時，取代氨基酸是Asp或Glu；r.當取代發生在位點223時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser；s.當取代發生在位點224時，取代氨基酸是Asn、Asp、Gln、Glu、Pro或Ser；從而，與野生型熱酶相比較，熱酶變異體對不溶解底物具有降低的吸附作用和增加的水解作用。
2.權利要求1的熱酶變異體，其中一個或多個取代基發生在第一環區域。
3.權利要求1的熱酶變異體，其中一個或多個取代基發生在第二環區域。
4.權利要求1的熱酶變異體，其中一個或多個取代基發生在第三環區域。
5.權利要求1的熱酶變異體，其中一個或多個取代基發生在第四環區域。
6.權利要求1的熱酶變異體，其中一個或多個取代基發生在第五環區域。
7.權利要求1的熱酶變異體，其中一個或多個取代基發生在第六環區域。
8.一種選自硬表面洗滌組合物、餐具洗滌組合物、口腔洗滌組合物、假牙洗滌組合物、接觸透鏡洗滌組合物和織物洗滌組合物的洗滌組合物，其特征在于其中洗滌組合物含有權利要求1-7的任何一項的熱酶變異體和洗滌組合物載體；洗滌組合物優選是硬表面洗滌組合物或洗滌組合物是織物洗滌組合物；組合物優選含有按組合物重量計至少約5％的表面活性劑和至少約5％助劑；組合物還優選含有選自溶劑、緩沖劑、酶、污垢解脫劑、粘土去污劑、分散劑、增白劑、泡沫抑制劑、織物柔軟劑、增泡劑、酶穩定劑、漂白劑、染料、香料和它們的混合物的洗滌組合物材料。
9.編碼權利要求1-7的任何一項的熱酶變異體的DNA序列。
全文摘要
本發明涉及具有野生型熱酶氨基酸序列的修飾氨基酸序列的熱酶(thermitase)變異體,野生型氨基酸序列包括第一環區域、第二環區域、第三環區域、第四環區域、第五環區域和第六環區域;其中,修飾的氨基酸序列在一個或多個環區域中的特定標記的位點上含有與野生型熱酶不同的氨基酸(即取代),從而與野生型熱酶相比,熱酶變異體具有對不溶解底物降低的吸附作用和增加的水解作用。本發明還涉及編碼這種熱酶變異體的DNA序列。本發明還涉及含有這種熱酶變異體的用于洗滌各種表面的組合物。
文檔編號C11D3/386GK1183118SQ96193608
公開日1998年5月27日 申請日期1996年3月6日 優先權日1995年3月9日
發明者P·F·布羅德三世, B·L·巴爾內特, D·N·魯賓 申請人:普羅格特-甘布爾公司