過氧化氫驅動的氧化反應的制作方法
                            
            
                            
            文檔序號：555036閱讀：970來源：國知局
            
                            
                        
            
                        
            
                        
            
                            專利名稱：過氧化氫驅動的氧化反應的制作方法
            
技術領域：
            本發明涉及一種進行氧化反應的方法。
            
背景技術：
             單加氧酶催化廣泛底物的氧化反應。為催化該反應，單加氧酶通常需要輔助因子及至少一種電子傳遞配偶蛋白(electron-transfer partnerprotein)(還原酶)。然而，單加氧酶能夠使用過氧化氫(H2O2)作為氧化劑，這是因為H2O2可作為分子氧和兩個電子的來源。使用H2O2驅動氧化反應被稱為“過氧化物旁路(peroxide shunt)”。
            

            發明內容
            
與過氧化物酶相比，單加氧酶對H2O2通常具有高Km值(例如大約20mM)。因此，當使用過氧化物旁路進行氧化反應時，需要高濃度H2O2以取得單加氧酶活性的可觀水平。例如，使用50mM H2O2的單加氧酶活性的起始速率大大低于使用天然輔助因子NAD(P)H及生理電子傳遞配偶體時單加氧酶活性的起始速率。
            
本發明提供一種進行氧化反應的更有效的方法，該氧化反應使用過氧化物旁路，通過反應過程中不允許過高水平H2O2存在減少單加氧酶的氧化性損傷而進行。
            
H2O2以低于或等于其在單加氧酶催化的氧化反應中使用的速率同時產生，結果導致氧化反應效率提高及產物量增加。可使用多種方法以所需速率產生H2O2，例如電化學反應、酶或前體的使用。
            
因此，本發明提供一種進行氧化反應的方法，該氧化反應由單加氧酶催化并使用過氧化氫作為氧化劑，所述反應中單加氧酶的氧化損傷反應水平低，所述方法包括在氧化反應的同時產生過氧化氫，其中過氧化氫以低于或等于其在該反應中使用的速率產生。
            
本發明還提供一種單加氧酶催化及使用過氧化氫作為氧化劑進行氧化反應的方法，所述反應中單加氧酶的氧化損傷反應水平低，所述方法包括在H2O2或羥基螯合劑的存在下進行反應，所述螯合劑可控制H2O2或羥基濃度。
            
序列描述SEQ ID NO1顯示了惡臭假單胞菌(Pseudomonas putida)的細胞色素P450Cam的核苷酸序列。
            
SEQ ID NO2顯示了惡臭假單胞菌的細胞色素P450Cam的氨基酸序列。
            
SEQ ID NO3顯示了巨大芽孢桿菌(Bacillus megaterium)的細胞色素P450BM-3的核苷酸序列。
            
SEQ ID NO4顯示了巨大芽孢桿菌的細胞色素P450BM-3的氨基酸序列。開始的472個氨基酸殘基形成血紅素結構域。最后585個氨基酸殘基形成還原酶結構域。所有1048個氨基酸殘基形成全酶。
            
在此使用的本領域中指明一個突變體的常規是參照序列中某一位置的天然氨基酸殘基，在其后為突變體中該位置的氨基酸，例如F87是指野生型序列的87位置上的苯丙氨酸，F87A是指在變異體中已改變為丙氨酸的野生型序列87位置上的苯丙氨酸。氨基酸殘基編號從起始甲硫氨酸殘基之后的氨基酸殘基開始。
            
實例中使用的突變體為F87A(單突變；SEQ ID NO5和SEQ IDNO6)和F87V L188Q A74G(三重突變；SEQ ID NO7和SEQ IDNO8)。
            
SEQ ID NO5顯示了巨大芽孢桿菌的細胞色素P450BM-3的F87A突變體的氨基酸序列。
            
SEQ ID NO6顯示了巨大芽孢桿菌的細胞色素P450BM-3的F87A突變體的核苷酸序列。
            
SEQ ID NO7顯示了巨大芽孢桿菌的細胞色素P450BM-3的F87V L188Q A74G突變體的氨基酸序列。
            
SEQ ID NO8顯示了巨大芽孢桿菌的細胞色素P450BM-3的F87V L188Q A74G突變體的核苷酸序列。
            
SEQ ID NO9顯示了珊瑚諾卡氏菌(Nocardia coralline)的B-276烯烴環氧化酶1亞基的核苷酸序列。
            
SEQ ID NO10顯示了珊瑚諾卡氏菌的B-276烯烴環氧化酶1亞基的氨基酸序列。
            
SEQ ID NO11顯示了珊瑚諾卡氏菌的B-276烯烴環氧化酶2亞基的核苷酸序列。
            
SEQ ID NO12顯示了珊瑚諾卡氏菌的B-276烯烴環氧化酶2亞基的氨基酸序列。
            
SEQ ID NO13顯示了黃色桿菌屬種(Xanthobacta sp.)的Py2烯烴單加氧酶α亞基的核苷酸序列。
            
SEQ ID NO14顯示了黃色桿菌屬種的Py2烯烴單加氧酶α亞基的氨基酸序列。
            
SEQ ID NO15顯示了黃色桿菌屬種的Py2烯烴單加氧酶β亞基的核苷酸序列。
            
SEQ ID NO16顯示了黃色桿菌屬種的Py2烯烴單加氧酶β亞基的氨基酸序列。
            
SEQ ID NO17顯示了黃色桿菌屬種的Py2烯烴單加氧酶γ亞基的核苷酸序列。
            
SEQ ID NO18顯示了黃色桿菌屬種的Py2烯烴單加氧酶γ亞基的氨基酸序列。
            
SEQ ID NO19顯示了莢膜甲基球菌(Methylococcus capsulatas)可溶性甲烷單加氧酶的α亞基的核苷酸序列。
            
SEQ ID NO20顯示了莢膜甲基球菌可溶性甲烷單加氧酶α亞基的氨基酸序列。
            
SEQ ID NO21顯示了莢膜甲基球菌可溶性甲烷單加氧酶β亞基的核苷酸序列。
            
SEQ ID NO22顯示了莢膜甲基球菌可溶性甲烷單加氧酶β亞基的氨基酸序列。
            
SEQ ID NO23顯示了莢膜甲基球菌可溶性甲烷單加氧酶γ亞基的核苷酸序列。
            
SEQ ID NO24顯示了莢膜甲基球菌可溶性甲烷單加氧酶γ亞基的氨基酸序列。
            
SEQ ID NO25顯示了食油假單胞菌(Pseudomonas oleovorans)Gpo1烷羥化酶(AlkB基因)的核苷酸序列。
            
SEQ ID NO26顯示了食油假單胞菌Gpo1烷羥化酶的氨基酸序列。
            
SEQ ID NO27顯示了洋蔥伯克霍爾德氏菌(Burkholderiacepacia)甲苯2-單加氧酶α亞基的核苷酸序列。
            
SEQ ID NO28顯示了洋蔥伯克霍爾德氏菌甲苯2-單加氧酶α亞基的氨基酸序列。
            
SEQ ID NO29顯示了洋蔥伯克霍爾德氏菌甲苯2-單加氧酶β亞基的核苷酸序列。
            
SEQ ID NO30顯示了洋蔥伯克霍爾德氏菌甲苯2-單加氧酶β亞基的氨基酸序列。
            
SEQ ID NO31顯示了洋蔥伯克霍爾德氏菌甲苯2-單加氧酶γ亞基的核苷酸序列。
            
SEQ ID NO32顯示了洋蔥伯克霍爾德氏菌甲苯2-單加氧酶γ亞基的氨基酸序列。
            
SEQ ID NO33顯示了嗜熱脂肪芽孢桿菌(Bacillusstearothermophilus)苯酚羥化酶(pheA)基因的核苷酸序列。
            
SEQ ID NO34顯示了嗜熱脂肪芽孢桿菌苯酚羥化酶基因的氨基酸序列。
            
SEQ ID NO35顯示了向日葵(Helianthus annuus)硬脂酰-ACP去飽和酶的核苷酸序列。
            
SEQ ID NO36顯示了向日葵硬脂酰-ACP去飽和酶的氨基酸序列。
            
具體實施例方式
            
應該理解本發明不限于具體實施方案。還應該理解本發明所公開的方法的不同應用可以適合本領域的特定需要。還應該理解在此使用的術語僅為描述本發明具體實施方案使用，而不旨在限制。
            
另外，除非內容中清楚指出，否則本說明書及附加權利要求書中所使用的單數形式“一”、“一種”、“該”包括復數指示物。因此，例如“一種底物”包括兩種或更多種底物，“一種酶”包括引用兩種或更多種酶，等等。
            
本發明引用的所有出版物、專利及專利出版物，無論在上文還是下文，均通過援引的方式完整地納入本申請。
            
本發明方法能夠使多種底物被氧化。所述底物包括但不限于烷類、芳香族化合物、萜類化合物、烯烴及脂肪酸。
            
合適的烷類包括但不限于甲烷、乙烷、丙烷、丁烷、戊烷、己烷、庚烷、正辛烷、正壬烷、正癸烷、正十二烷及正十六烷。烷類氧化產生醇。甲烷氧化成甲醇在技術上及經濟上非常重要。中鏈醇(如正辛醇)為合成的中間產物，而較長鏈的醇(如正十二醇)用于脂肪酸衍生物的合成。
            
合適的芳香族化合物包括但不限于苯、甲苯、二甲苯、氯苯、苯酚及其取代物。酚及兒茶酚產物用于香味和香料化合物的合成。
            
合適的萜類化合物包括但不限于一萜類，如檸檬烯、蒎烯、松油烯及羅勒烯，例如朱欒倍半萜和香橙烯的倍半萜，以及包括甾族化合物類的三萜類。這些產物為用于合成的中間產物、優良香味和香料化學品及藥品。
            
合適的烯烴包括但不限于簡單分子如丙烯、己-1-烯、己-2-烯及苯乙烯，以及復雜分子中的碳-碳雙鍵。烯烴對于單對映異構物的選擇性環氧化作用在合成中非常重要。光學純的氧化丙烯及氧化苯乙烯在合成中是非常有用的中間產物。
            
羥化脂肪酸為聚合體的前體。
            
單加氧酶根據本發明用于進行氧化反應的酶為單加氧酶。本領域普通技術人員使用本領域標準技術能夠確定酶是否為單加氧酶。通常情況下，可使用蛋白質晶體學對輔基進行表征，尤其是非血紅素鐵酶的輔基，這是因為輔基通常不具有發色團。另外，本領域普通技術人員通常會使用序列對比尋找保守基序如活性部位、和鐵含量以及亞基組分。
            
單加氧酶對H2O2的Km值優選為至少15nM、至少20nM、至少25nM、至少30nM、至少35nM、至少40nM、至少45nM或至少50nM。
            
單加氧酶的實例包括但不限于細胞色素P450單加氧酶及非血紅素二鐵單加氧酶。合適的非血紅素二鐵單加氧酶包括但不限于甲烷單加氧酶(Colby et al.，Biochem.J.，1977；165395-402；Dalton，Adv.Appl.Microbiol.，1980；2671-87；Fox et al.，J.Biol.Chem.，1989；26410023-10033；Fox et al.，Methods Enzymol.，1990；188191-202；McDonald et al.，Appl.Environ.Microbiol.，1997；631898-1904)，烷羥化酶(van Beilen et al.，Enzyme Microb.Technol.，1994；16904-911)，甲苯單加氧酶(Luykx et al.，Biochem.Biophys.Res.Commun.，2003；312373-379；Pikus et al.，Biochemistry，1996；359106-9119；Newman & Wackett，Biochemistry，1995；3414066-14076)，烯烴單加氧酶(Gallagher et al.，Eur.J.Biochem.，1997；247635-641；Lange & Que，Curr.Opin.Chem.Biol.，1998；2159-172；Zhou et al.，FEBS Lett.，1998；430181-185)，酚單加氧酶(Divari et al.，Eur.J.Biochem.，2003；2702244-2253)以及甾族去飽和酶(Shanklin et al.，Biochemistry，1994；3312787-12794)。非血紅素二鐵單加氧酶通常為真核或原核來源，優選地為細菌、真菌、酵母、植物或動物來源。優選的序列示于SEQ ID NO1至SEQ ID NO36。
            
本發明方法使用的酶優選地為細胞色素P450酶，通常為真核或原核來源。細胞色素P450單加氧酶通常以亞鐵-一氧化碳絡合物的446-450nm亞鐵血紅素索雷譜帶為表征。酶通常為細菌、真菌、酵母、植物或動物來源，因此可來自假單胞菌屬(Pseudomonas)細菌。酶可為P450的天然存在形式，例如P450cam，巨大芽孢桿菌的P450BM-3，假單胞菌屬種(Pseudomonas sp.)的P450terp，紅色糖多孢菌(Saccharopollyspora erythraea)的P450eryF以及灰色鏈霉菌(Streptomyces griseus)菌株的P450 105 D1(CYP105)。
            
或者，酶可為P450天然存在形式的突變體。突變體保留天然存在的酶的必需生物活性，即催化使用H2O2的氧化反應的能力。突變體可在酶的活性部位具有一個或多個突變。
            
“活性部位”的氨基酸是在催化過程中界定或定義結合底物的部位的氨基酸或界定或定義底物在到達催化部位之前所必須經過的部位的氨基酸。因此，這類氨基酸在進入催化部位過程中或催化過程中通常與底物相互作用。所述相互作用通常通過靜電相互作用(在帶電荷基團或極性基團之間)、疏水性相互作用、氫鍵或范德華力產生。
            
可通過本領域普通技術人員使用的常規方法鑒定活性部位的氨基酸。這些方法包括標記研究，其中使酶結合修飾(“標記”)接觸底物的氨基酸的底物。另外，可獲得酶與結合的底物的晶體結構以推導出活性部位的氨基酸。
            
單加氧酶可具有1個、2個、3個、4個、5至10個、10至20個或更多其它突變，例如置換、插入或缺失。可在天然存在的酶的氨基酸序列進行氨基酸置換，例如從1、2、3、4或5至10、20或30個置換。例如可根據表1進行保守性置換。第二列中同一格的氨基酸及優選在第三列中同一行的氨基酸可互相置換。
            
表1保守性氨基酸置換  
            
     
            
突變可位于活性部位內或活性部位外。通常突變在影響或接觸活性部位一個或多個氨基酸位置或定位的“第二個域”殘基中。插入通常在N端和/或C端，因此酶可為嵌合蛋白的一部分。缺失通常包括與催化無關的的氨基酸的缺失，例如那些位于活性部位之外的氨基酸(由此而來的酶為天然產生酶的突變片段)。因此，單加氧酶可僅包括氧化活性所需的那些氨基酸。
            
活性部位的突變在底物結合于活性部位時通常改變底物的位置和/或構象。突變可使血紅素基團更易接近待氧化的底物上的位點。因此，突變可以為對具有較小或較大、或較多或較少極性的側鏈的氨基酸進行的置換。
            
突變通常增加蛋白質的的穩定性，或使蛋白質更易于被純化。它們通常通過去除蛋白質的半胱氨酸殘基(如通過在P450cam的334位置，或在同系物的相同位置半胱氨酸的置換，優選地置換為丙氨酸)以防止蛋白質的二聚體形成。它們通常使得蛋白質可以可溶形式制備，例如通過引入缺失或多組氨酸標簽，或通過N端膜錨定序列的突變進行。突變通常抑制蛋白質寡聚化，例如蛋白質表面疏水部位之間接觸產生的寡聚化。
            
突變可影響酶使用H2O2的方式，從而可改進反應的效率。例如，使用正亮氨酸置換P450BM-3的血紅素結構域的所有甲硫氨酸殘基可使該酶的過氧化物酶活性提高兩倍(Cirino et al.，Biotechnol.Bioeng.，2003；83(6)729-734)。另外，已報道直接進化研究發現酶突變體對過氧化物具有更大抵抗力(Cirino & Arnold，Angew.Chem.Int.Ed.，2003；423299-3301)。
            
因此，在氨基酸同一性基礎上突變酶通常與天然存在的酶具有至少70％同源性。
            
本發明所述的突變蛋白質(即描述為另一蛋白質的突變體)通常在至少20個、優選地至少30個、例如至少40個、60個、或100個或更多連續氨基酸中與相關蛋白質具有至少70％的同源性，或至少80％或90％及更優選地至少95％、97％或99％的同源性。連續氨基酸可包括活性部位。或者可以不通過連續氨基酸測量此同源性，而僅通過活性部位的氨基酸測量。
            
可使用已知方法測量同源性。例如，UWGCG程序包提供了BESTFIT程序，可用于計算同源性(例如在其缺省設置使用)(Devereux et al.(1984)Nucleic Acids Research 12，p387-395)。PILEUP和BLAST算法可用于計算同源性或排列序列(通常在其缺省設置)，例如Altschul S.F.(1993)J Mol Evol 36290-300；Altschul，S，F et al.(1990)J Mol Biol 215403-10中所述。
            
進行BLAST分析的軟件可通過國家生物技術信息中心(http://www.ncbi.nlm.nih.gov/)公開獲得。該算法涉及首先通過鑒定查詢序列中長度W的短字碼鑒定高評分序列對(HSPs)，此短字碼當與數據庫序列中相同長度字碼進行對比時可匹配或滿足一些陽性值域值評分T。T被稱為鄰近字碼評分域值(neighbourhood word scorethreshold)(Altschul et al.，supra)。這些起始鄰近字碼采樣數作為起動搜索的種子以發現含有它們的HSPs。字碼采樣數沿每一序列向兩個方向延伸直至累積對比得分可被增加。每一方向字碼采樣數的延伸在以下情況下被停止累積對比得分從其最大獲得值下降數量X；由于一個或多個負評分殘基對比的累積，累積得分到達零或更低；或到達任一序列的末端。BLAST算法參數W、T及X決定對比的靈敏度和速度。BLAST程序使用以下項目作為缺省值字碼長度(W)11、BLOSUM62評分矩陣(見Henikoff and Henikoff(1992)Proc.Natl.Acad.Sci.USA 8910915-10919)對比(B)50、期望值(E)10、M＝5、N＝4，以及對比兩條鏈。
            
BLLAST算法實行了兩條序列之間相似性的統計學分析，見于例如Karlin and Altschul(1993)Proc.Natl.Acad.Sci.USA 905873-5787。BLAST算法提供的一個相似性的量度是最小總概率(P(N))，它表示兩條核苷酸或氨基酸序列之間的隨機發生的匹配的概率。例如，在第一條序列與第二條序列對比中如果最小總概率小于約1，優選地小于約0.1，更優選地小于約0.01，最優選地小于約0.001，則此序列被認為與另一條序列相似。
            
突變體包括上述序列的片段。所述片段保留有單加氧酶活性。片段長度可為至少300、至少400或至少450個氨基酸。按照下列更詳細描述，所述片段可用于產生嵌合酶。
            
突變體還包括含有天然存在的酶的片段或部分的嵌合蛋白。一個或多個氨基酸可備選地或額外地添加至上述多肽。可于天然存在的酶或其變異體或片段的N端或C端提供延伸。延伸可以很短，例如1至10氨基酸長度。或者，延伸可較長。可將載體蛋白融合至上述氨基酸序列。因此，合并有上述一種酶的融合蛋白可用于本發明。
            
天然存在的酶或其變異體還可被化學修飾。許多側鏈修飾為本領域已知，并可用于上述酶的側鏈。所述修飾包括例如糖基化、磷酸化、通過與乙醛反應然后使用NaBH4還原的還原烴基化進行的氨基酸修飾、甲基乙酰亞胺酯脒基化(amidination)或使用乙酸酐的酰化作用。所述修飾優選為糖基化。
            
通常使用本領域已知的方法將本發明討論的突變引入酶中，例如酶的位點定向誘變，PCR和基因改組方法或使用位點定向誘變周期中多重誘變寡核苷酸。因此，可使用定向或隨機方法引入突變。因此，誘變方法可產生編碼一個或多個不同突變的一個或多個多核苷酸。通常產生可用于產生突變酶文庫的突變寡核苷酸文庫。
            
可使用本領域已知方法通過合成或重組的手段制備酶。單加氧酶的氨基酸序列可被修飾成包括非天然存在的氨基酸或增加酶的穩定性。當通過合成方法產生酶時，可于產生過程中引入所述氨基酸。蛋白質或肽還可以在合成或重組產生之后被修飾。
            
還可以使用D-氨基酸產生酶。在這種情況下，以由C至N的相反方向序列連接氨基酸。這是本領域產生所述蛋白質或肽的常規方法。
            
可以通過重組表達載體的多肽的原位表達在細胞中產生酶。表達載體任選地攜帶誘導型啟動子以控制多肽的表達。可以在重組表達之后通過任何蛋白質液相層析系統純化大規模產生酶。優選的蛋白質液相層析系統包括快速蛋白液相色譜(FPLC)、AKTA系統、Bio-Cad系統，Bio-Cad BioLogic系統及Gilsion高效液相色譜(HPLC)系統。
            
氧化反應本發明方法涉及由單加氧酶催化的高效率氧化反應。高效率氧化反應為一種在其發生時沒有可觀察到的酶轉化減少或產物產量減少或單加氧酶失活的反應。優選地，在反應1小時、2小時、6小時、12小時、1天或5天之后單加氧酶顯示有反應開始所表現活性的至少70％、至少80％、至少90％、至少95％、或100％。
            
通常情況下，本發明方法在體外進行，例如在無細胞系統中進行。
            
通過“過氧化物旁路”驅動反應。在單加氧酶(a)、底物(b)和H2O2(c)存在下進行本發明反應。通常在需氧條件下進行反應，并不需要任何輔助因子。(c)的產生在下文更為詳細描述。在此系統中，電子的流動通常為(c)→(a)→(b)。
            
在此方法中，(a)和(b)的濃度通常為10-8至10-2M，優選地為10-6至10-4M。(a)∶(b)的濃度比例通常為0.1∶10至1∶10，優選地為1∶0.5至1∶2，或1∶0.8至1∶1.2。優選地，(b)的濃度較(a)的濃度大。(a)的優選濃度為當其與底物反應時產生可通過可用的分析方法例如GC、HPLC檢測到足夠產物。這通常為uM數量等級。
            
通常在單加氧酶具有功能的溫度和/或pH情況下實施該方法，例如當酶具有峰值活性的至少20％、50％、80％或更多時。通常pH為2-10，例如5-9或6-8，優選地為7至7.8或7.4。可使用合適的緩沖劑如基于磷酸或醋酸的系統維持pH。通常溫度為0-80℃，例如25-75℃，30-60℃，或50-80℃。優選地，溫度為20-40℃。
            
通常在所述方法中發生至少20個轉化數/分鐘，例如至少50、100、200、300、500或更多轉化數(按照每納摩爾酶形成的產物納摩爾數測量轉化數)。
            
通常，H2O2產生速率為少于或等于每分鐘每mg單加氧酶1ug、2ug或3ug。通常，整個反應中H2O2的濃度低于或等于0.1、0.5或1mM。通常，反應持續至少60分鐘，至少240分鐘，至少6小時或至少10小時。
            
如果單加氧酶底物在反應條件下為液體，可在單加氧酶底物中實施本發明方法。還可以在溶劑中實施本發明方法。合適的溶劑包括但不限于水、水緩沖溶液混合水/有機和水緩沖液/有機溶劑系統。優選地，有機溶劑為碳氫化合物例如己烷、苯、乙腈、低脂肪族醇、酮及二噁烷、二甲基酰胺及二甲基亞砜及其混合物。溶劑通常為在其中反應物和產物為高溶解性和維持單加氧酶穩定性和活性的溶劑。
            
可以在溶液中具有所有組分的均質系統中進行反應。通常，在攪拌釜反應器中將單加氧酶和底物混合在一起，以分批、半分批或連續模式進行反應。
            
或者，在實施本發明方法之前可于合適的固體支持物例如二氧化硅上固定化單加氧酶。可將固定化單加氧酶充填入固定床反應器并使底物經過酶。在一個實施方案中，可于與單加氧酶相同或不同的材料上固定化產生H2O2的酶(下文較詳細討論)。固定化酶的方法為本領域已知。所述方法實例包括但不限于共價結合至不溶的有機或無機支持物，于凝膠中捕獲及吸附至離子交換樹脂或其它吸附性材料。(G.F.Bickerstaff ed.，″Immobilization of Enzymes and Cells，″HumanaPress，Totowa.New Jersey，1997)。
            
在另一個實施方案中，在“入口”側的膜允許底物緩慢從“反應物”側進入，然后在“出口”側的親水膜允許親水化合物流出至流動反應池的“產物”側。在此情況下，可于膜外產生H2O2，并允許其通過膜流至流動或不流動的酶。
            
在一個實施方案中，優選地通過下面較詳細描述的方法中的一種產生H2O2。在另一個實施方案中，H2O2或羥基螯合劑在氧化反應過程中用于螯合過量的H2O2或羥基。該螯合劑可為一種螯合試劑。在一個實施方案中，螯合劑為EDTA。EDTA抑制羥基的產生，例如痕量鐵(或銅)與H2O2反應產生的羥基。
            
電化學反應產生H2O2可以使用本發明方法通過一種電化學反應產生H2O2。電化學反應通常為一種將電流引入液體，優選地為溶劑的方法。電化學反應通常為在電流通過的電極處發生的氧化反應或還原反應。電極為能夠導電的固體，通常為基于碳的或金屬，其使得外部電源或接收器與優選為溶液的液體接觸。電極可為帶正電荷(負極)或帶負電荷(正極)。兩個或更多電極可形成電化學池，從電化學池外部電線可從每一電極連接至外部電裝置。氧化反應或還原反應可在一個電極發生，而氧化還原反應可于電化學池中發生或直接在液體中發生。
            
使用電化學反應產生H2O2為具有能效的。通常通過由分子氧至過氧化氫的受控電化學還原產生H2O2。負極的表面積及超電勢是由分子氧至過氧化氫的兩個電子還原的關鍵考慮因素。通常使用基于碳的負極，并可使用已知降低該反應超電勢的化合物修飾該負極。有效及有效率地實施該方法的電極材料及改性劑為本領域已知。通常可以通過施加于負極的電勢控制O2的還原并由此控制過氧化氫的產生。施加于負極的電勢根據負極以及對負極進行的任何修飾而改變。
            
本發明方法中使用的電化學反應可為分子氧的超聲電化學還原。該方法為本領域已知(Compton et al.，Electroanalysis，1997；9(7)509-522)。
            
酶催化產生H2O2在本發明方法中可由酶產生H2O2。酶優選地為氧化酶。合適氧化酶的實例包括但不限于葡萄糖氧化酶(E.C.1.1.3.4)、仲醇氧化酶(E.C.1.1.3.18)、甲醇氧化酶(E.C.1.1.3.31)、草酸氧化酶(E.C.1.2.3.4)、芳香基-乙醛氧化酶(E.C.1.2.3.9)、一氧化碳氧化酶(E.C.1.2.3.10)、胺氧化酶(E.C.1.4.3.4)、乙醇胺氧化酶(E.C.1.4.3.8)、硝基乙烷氧化酶(E.C.1.7.3.1)及亞硫酸鹽氧化酶。葡萄糖氧化酶(E.C.1.1.3.4)催化D-葡萄糖轉化為D-葡萄糖-1，5-內酯和H2O2。仲醇氧化酶(E.C.1.1.3.18)催化仲醇轉化為酮和H2O2。甲醇氧化酶(E.C.1.1.3.31)催化甲醇轉化為甲醛和H2O2。草酸氧化酶(E.C.1.2.3.4)催化草酸轉化為二氧化碳和H2O2。芳香基-乙醛氧化酶(E.C.1.2.3.9)催化芳族醛轉化為芳香酸和H2O2。一氧化碳氧化酶(E.C.1.2.3.10)催化CO和H2O轉化為二氧化碳和H2O2。胺氧化酶(E.C.1.4.3.4)催化RCH2NH2和H2O轉化為RCHO和NH3和H2O2。乙醇胺氧化酶(E.C.1.4.3.8)催化乙醇胺和H2O轉化為羥乙醛和H2O2。硝基乙烷氧化酶(E.C.1.7.3.1)催化硝基乙烷和H2O轉化為乙醛和H2O2。亞硫酸鹽氧化酶(E.C.1.8.3.1)催化亞硫酸鹽和H2O2轉化為硫酸鹽和H2O2。氧化酶(如葡萄糖氧化酶)可商購。另外，可使用本領域已知方法從已知微生物中提取氧化酶。
            
氧化酶的底物在本領域內為人熟知。除底物外，產生H2O2的反應還涉及水。通常在反應中還包括H2O2活化金屬。合適的金屬包括但不限于鈰、鉻、鈷、銅、鐵、錳、鉬、銀、鈦、鎢、釩及其混合物。含有上述金屬的金屬硅酸鹽可被制備并用于本發明方法。產生所述金屬硅酸鹽的方法為本領域已知(Neumann et al.，Journal of Catalysis，1997；166206-127)。金屬硅酸鹽優選地為四面體配位鈦如硅酸鹽-1(TS-1)、硅酸鹽-2(TS-2)、沸石-β、ZSM-48的硅類似物和MCM-41(Murugaveland Roesky，Angew.Chem.Int.Ed.Engl.，1997；36(5)477-479)。
            
在本發明的一個優選的實施方案中，含金屬固體或金屬硅酸鹽用作固定化產生H2O2的酶的支持物。在另一個優選的實施方案中，還于相同或不同的金屬硅酸鹽支持物上固定化單加氧酶。
            
優選地，首先將氧化酶與其它反應成分混合，然后通過添加氧化酶底物起動反應。例如，將單加氧酶、單加氧酶底物和氧化酶全部混合，然后添加氧化酶。在優選的實施方案中，將P450BM3、辛烷和葡萄糖氧化酶混合一起，然后添加葡萄糖。通常可通過控制添加氧化酶底物的速率來實現H2O2產生的控制。
            
由前體產生H2O2在本發明的方法中可由前體產生H2O2。通過添加前體至水中產生H2O2在本領域內為人熟知。前體包括但不限于過硼酸鹽、過碳酸鹽、過磷酸鹽及過亞硝酸鹽。優選的前體為鈉鹽。可通過使用化合物如四乙酰基乙二胺增強前體產生H2O2的性質。添加至含單加氧酶和底物的溶液中的前體量可使與底物的酶促反應最大化，并使H2O2引起的酶失活最小化。優選地，所產生H2O2的濃度不超過酶的Km值，但足夠產生酶的反應性種類。
            
實施例實施例1在此實驗中，在P450BM3血紅素結構域存在下使辛烷與電化學產生的H2O2反應。于100mL玻璃燒杯中室溫使用三電極結構進行該實驗。網狀玻璃碳(RVC)負極、鉑網正極及銀/氯化銀(Ag/AgCl)參照電極裝在同一容器中。將RVC負極短暫浸漬于1mM 2-氨基蒽醌乙醇溶液，然后取出并使其在空氣中干燥。使用含氧飽和的水Tris緩沖液、0.2MKCl、0.5mM辛烷、及3u M P450BM3F87V L188Q A74G血紅素結構域的反應溶液(50mM，pH 7.4)。將反應溶液攪拌至平衡(5-10分鐘)，然后施加相對于Ag/AgCl的-0.55V的電勢2小時，整個過程中持續攪拌溶液。氣相色譜(GC)分析顯示溶劑氯仿、辛烷、2-辛醇、3-辛醇及4-辛醇和內標準1-壬醇的存在。2-辛醇、3-辛醇及4-辛醇的相對比例為1∶1.1∶0.7。所形成的辛醇總濃度為141uM，表示每個酶轉化數為47。
            
使用1.43uM野生型P450BM3血紅素結構域進行了相似的實驗。所形成的辛醇的總濃度為8.4uM，表示每個酶轉化數為6。2-辛醇、3-辛醇及4-辛醇的相對比例為1∶1.7∶2.0。
            
實施例2在此實驗中，在P450BM3全酶存在下使辛烷與酶作用產生的H2O2發生反應。向玻璃瓶中添加由水Tris緩沖液(50mM，pH 7.4)、0.5mM辛烷、1.6μM P450BM3F87V L188Q A74G全酶及葡萄糖氧化酶(1.5U)組成的溶液(總體積為5mL)。平衡(5分鐘)之后，通過添加葡萄糖(1×10-6摩爾)起動反應。連續添加葡萄糖(1×10-6摩爾)，每5分鐘一次，直至1小時(總共12次添加的當量為1.2×10-5摩爾)。在此時間內持續攪拌反應物，1.5小時之后停止。GC分析顯示溶劑氯仿、辛烷、2-辛醇、3-辛醇和4-辛醇及內標準1-壬醇的存在。2-辛醇、3-辛醇和4-辛醇的相對比例為1∶1.1∶0.8。所形成的辛醇的總濃度為17uM，表示每個酶轉化數為10。
            
實施例3  
            
在此實驗中，在P450BM3全酶的存在下，使辛烷與過硼酸鈉產生的H2O2反應。向玻璃瓶中添加由水Tris緩沖液(40mM，pH 7.4)、0.5mM辛烷、1.3μM P450BM3F87V L188Q A74G全酶組成的溶液(總體積為5mL)。平衡(5分鐘)之后，通過添加NaBO3.4H2O(1×10-4摩爾)起動反應，連續攪拌1小時。GC分析顯示溶劑氯仿、辛烷、2-辛醇、3-辛醇和4-辛醇及內標準1-壬醇的存在。2-辛醇、3-辛醇和4-辛醇的相對比例為1∶1.8∶1.1。所形成的辛醇的總濃度為77uM，表示每個酶轉化數為59。
            
對于實施例1至3，當溶液中無P450酶時觀察不到壬醇產物。
            
實施例4在此實驗中，在P450BM3血紅素結構域的存在下，使蒎烯與過硼酸鈉產生的H2O2反應。向玻璃瓶中添加由水Tris緩沖液(40mM，pH7.4)、0.63mM蒎烯及3.7uM野生型P450BM3血紅素結構域組成的溶液(總體積為5mL)。平衡(5分鐘)之后，通過添加7.8×10-6摩爾NaBO3.4H2O起動反應，持續攪拌1小時。GC分析顯示順式/反式2，3-環氧化物(32％)、(+)-反式馬鞭草烯醇(16％)、(+)-順式馬鞭草烯醇(6％)、(+)-馬鞭草烯酮/(+)-桃金娘烯醇(13％)、桃金娘烯醛(4％)以及未鑒定的其它氧化產物(29％)的存在。所形成的產物的總濃度為80uM，表示每個酶轉化數為22。
            
實施例5在此實驗中，在過硼酸鈉產生的H2O2存在的條件下，使酚單加氧酶與苯酚反應。向玻璃瓶中添加由水Tris緩沖液(40mM，pH 7.4)、0.63mM苯酚及3.7uM野生型酚單加氧酶組成的溶液(總體積為5mL)。平衡(5分鐘)之后，通過添加7.8×10-6摩爾NaBO3.4H2O起動反應，持續攪拌1小時。GC分析顯示了氧化產物的存在。
            
實施例6在此實驗中，在葡萄糖氧化酶產生的H2O2的存在下，使P450BM3與棕櫚酸反應。向玻璃瓶中添加由水Tris緩沖液(50mM，pH 7.4)、0.5mM棕櫚酸、1.6μM P450BM3全酶及葡萄糖氧化酶(1.5U)組成的溶液(總體積為5mL)。平衡(5分鐘)之后，通過添加葡萄糖(1×10-6摩爾)起動反應。連續添加葡萄糖(1×10-6摩爾)，每5分鐘一次，直至1小時(總共12次添加的當量為1.2×10-5摩爾)。在此時間內持續攪拌反應物，1.5小時之后停止。GC分析顯示了氧化產物的存在。
            
實施例7使植物CYP74C與13S-氫過氧化亞麻酸反應形成化合物3Z-己烯醛(芳香劑)。H2O2由過硼酸鈉產生。向玻璃瓶中添加由水Tris緩沖液(40mM，pH 7.4)、0.63mM 13S-氫過氧化亞麻酸及3.7μM野生植物CYP74C組成的溶液(總體積為5mL)。平衡(5分鐘)之后，通過添加7.8×10-6摩爾NaBO3.4H2O起動反應，持續攪拌1小時。GC分析顯示了氧化產物的存在。
            
序列表<110>ISIS創新有限公司<120>過氧化氫氧化反應<130>N.91079A SA<160>36<170>PatentIn version 3.2<210>1<211>1248<212>DNA<213>惡臭假單胞菌<220>  
            
<221>CDS<222>(1)..(1248)<400>1atg acg act gaa acc ata caa agc aac gcc aat ctt gcc cct ctg cca48Met Thr Thr Glu Thr Ile Gln Ser Asn Ala Asn Leu Ala Pro Leu Pro1   5   10  15ccc cat gtg cca gag cac ctg gta ttc gac ttc gac atg tac aat ccg96Pro His Val Pro Glu His Leu Val Phe Asp Phe Asp Met Tyr Asn Pro20  25  30tcg aat ctg tct gcc ggc gtg cag gag gcc tgg gca gtt ctg caa gaa144Ser Asn Leu Ser Ala Gly Val Gln Glu Ala Trp Ala Val Leu Gln Glu35  40  45tca aac gta ccg gat ctg gtg tgg act cgc tgc aac ggc gga cac tgg192Ser Asn Val Pro Asp Leu Val Trp Thr Arg Cys Asn Gly Gly His Trp50  55  60atc gcc act cgc ggc caa ctg atc cgt gag gcc tat gaa gat tac cgc240Ile Ala Thr Arg Gly Gln Leu Ile Arg Glu Ala Tyr Glu Asp Tyr Arg65  70  75  80cac ttt tcc agc gag tgc ccg ttc atc cct cgt gaa gcc ggc gaa gcc288  
            
His Phe Ser Ser Glu Cys Pro Phe Ile Pro Arg Glu Ala Gly Glu Ala85  90  95tac gac ttc att ccc acc tcg atg gat ccg ccc gag cag cgc cag ttt336Tyr Asp Phe Ile Pro Thr Ser Met Asp Pro Pro Glu Gln Arg Gln Phe100 105 110cgt gcg ctg gcc aac caa gtg gtt ggc atg ccg gtg gtg gat aag ctg384Arg Ala Leu Ala Asn Gln Val Val Gly Met Pro Val Val Asp Lys Leu115 120 125gag aac cgg atc cag gag ctg gcc tgc tcg ctg atc gag agc ctg cgc432Glu Asn Arg Ile Gln Glu Leu Ala Cys Ser Leu Ile Glu Ser Leu Arg130 135 140ccg caa gga cag tgc aac ttc acc gag gac tac gcc gaa ccc ttc ccg480Pro Gln Gly Gln Cys Asn Phe Thr Glu Asp Tyr Ala Glu Pro Phe Pro145 150 155 160ata cgc atc ttc atg ctg ctc gca ggt cta ccg gaa gaa gat atc ccg528Ile Arg Ile Phe Met Leu Leu Ala Gly Leu Pro Glu Glu Asp Ile Pro165 170 175cac ttg aaa tac cta acg gat cag atg acc cgt ccg gat ggc agc atg576His Leu Lys Tyr Leu Thr Asp Gln Met Thr Arg Pro Asp Gly Ser Met180 185 190acc ttc gca gag gcc aag gag gcg ctc tac gac tat ctg ata ccg atc624Thr Phe Ala Glu Ala Lys Glu Ala Leu Tyr Asp Tyr Leu Ile Pro Ile195 200 205atc gag caa cgc agg cag aag ccg gga acc gac gct atc agc atc gtt672Ile Glu Gln Arg Arg Gln Lys Pro Gly Thr Asp Ala Ile Ser Ile Val210 215 220gcc aac ggc cag gtc aat ggg cga ccg atc acc agt gac gaa gcc aag720Ala Asn Gly Gln Val Asn Gly Arg Pro Ile Thr Ser Asp Glu Ala Lys225 230 235 240agg atg tgt ggc ctg tta ctg gtc ggc ggc ctg gat acg gtg gtc aat768Arg Met Cys Gly Leu Leu Leu Val Gly Gly Leu Asp Thr Val Val Asn245 250 255ttc ctc agc ttc agc atg gag ttc ctg gcc aaa agc ccg gag cat cgc816  
            
Phe Leu Ser Phe Ser Met Glu Phe Leu Ala Lys Ser Pro Glu His Arg260 265 270cag gag ctg atc gag cgt ccc gag cgt att cca gcc gct tgc gag gaa864Gln Glu Leu Ile Glu Arg Pro Glu Arg Ile Pro Ala Ala Cys Glu Glu275 280 285cta ctc cgg cgc ttc tcg ctg gtt gcc gat ggc cgc atc ctc acc tcc912Leu Leu Arg Arg Phe Ser Leu Val Ala Asp Gly Arg Ile Leu Thr Ser290 295 300gat tac gag ttt cat ggc gtg caa ctg aag aaa ggt gac cag atc ctg960Asp Tyr Glu Phe His Gly Val Gln Leu Lys Lys Gly Asp Gln Ile Leu305 310 315 320cta ccg cag atg ctg tct ggc ctg gat gag cgc gaa aac gcc tgc ccg1008Leu Pro Gln Met Leu Ser Gly Leu Asp Glu Arg Glu Asn Ala Cys Pro325 330 335atg cac gtc gac ttc agt cgc caa aag gtt tca cac acc acc ttt ggc1056Met His Val Asp Phe Ser Arg Gln Lys Val Ser His Thr Thr Phe Gly340 345 350cac ggc agc cat ctg tgc ctt ggc cag cac ctg gcc cgc cgg gaa atc1104His Gly Ser His Leu Cys Leu Gly Gln His Leu Ala Arg Arg Glu Ile355 360 365atc gtc acc ctc aag gaa tgg ctg acc agg att cct gac ttc tcc att1152Ile Val Thr Leu Lys Glu Trp Leu Thr Arg Ile Pro Asp Phe Ser Ile370 375 380gcc ccg ggt gcc cag att cag cac aag agc ggc atc gtc agc ggc gtg1200Ala Pro Gly Ala Gln Ile Gln His Lys Ser Gly Ile Val Ser Gly Val385 390 395 400cag gca ctc cct ctg gtc tgg gat ccg gcg act acc aaa gcg gta taa1248Gln Ala Leu Pro Leu Val Trp Asp Pro Ala Thr Thr Lys Ala Val405 410 415<210>2<211>415<212>PRT<213>惡臭假單胞菌  
            
<400>2Met Thr Thr Glu Thr Ile Gln Ser Asn Ala Asn Leu Ala Pro Leu Pro1   5   10  15Pro His Val Pro Glu His Leu Val Phe Asp Phe Asp Met Tyr Asn Pro20  25  30Ser Asn Leu Ser Ala Gly Val Gln Glu Ala Trp Ala Val Leu Gln Glu35  40  45Ser Asn Val Pro Asp Leu Val Trp Thr Arg Cys Asn Gly Gly His Trp50  55  60Ile Ala Thr Arg Gly Gln Leu Ile Arg Glu Ala Tyr Glu Asp Tyr Arg65  70  75  80His Phe Ser Ser Glu Cys Pro Phe Ile Pro Arg Glu Ala Gly Glu Ala85  90  95Tyr Asp Phe Ile Pro Thr Ser Met Asp Pro Pro Glu Gln Arg Gln Phe100 105 110Arg Ala Leu Ala Asn Gln Val Val Gly Met Pro Val Val Asp Lys Leu115 120 125Glu Asn Arg Ile Gln Glu Leu Ala Cys Ser Leu Ile Glu Ser Leu Arg130 135 140Pro Gln Gly Gln Cys Asn Phe Thr Glu Asp Tyr Ala Glu Pro Phe Pro145 150 155 160Ile Arg Ile Phe Met Leu Leu Ala Gly Leu Pro Glu Glu Asp Ile Pro165 170 175His Leu Lys Tyr Leu Thr Asp Gln Met Thr Arg Pro Asp Gly Ser Met180 185 190Thr Phe Ala Glu Ala Lys Glu Ala Leu Tyr Asp Tyr Leu Ile Pro Ile195 200 205Ile Glu Gln Arg Arg Gln Lys Pro Gly Thr Asp Ala Ile Ser Ile Val210 215 220  
            
Ala Asn Gly Gln Val Asn Gly Arg Pro Ile Thr Ser Asp Glu Ala Lys225 230 235 240Arg Met Cys Gly Leu Leu Leu Val Gly Gly Leu Asp Thr Val Val Asn245 250 255Phe Leu Ser Phe Ser Met Glu Phe Leu Ala Lys Ser Pro Glu His Arg260 265 270Gln Glu Leu Ile Glu Arg Pro Glu Arg Ile Pro Ala Ala Cys Glu Glu275 280 285Leu Leu Arg Arg Phe Ser Leu Val Ala Asp Gly Arg Ile Leu Thr Ser290 295 300Asp Tyr Glu Phe His Gly Val Gln Leu Lys Lys Gly Asp Gln Ile Leu305 310 315 320Leu Pro Gln Met Leu Ser Gly Leu Asp Glu Arg Glu Asn Ala Cys Pro325 330 335Met His Val Asp Phe Ser Arg Gln Lys Val Ser His Thr Thr Phe Gly340 345 350His Gly Ser His Leu Cys Leu Gly Gln His Leu Ala Arg Arg Glu Ile355 360 365Ile Val Thr Leu Lys Glu Trp Leu Thr Arg Ile Pro Asp Phe Ser Ile370 375 380Ala Pro Gly Ala Gln Ile Gln His Lys Ser Gly Ile Val Ser Gly Val385 390 395 400Gln Ala Leu Pro Leu Val Trp Asp Pro Ala Thr Thr Lys Ala Val405 410 415<210>3<211>4957<212>DNA<213>巨大芽孢桿菌<220>  
            
<221>CDS<222>(1541)..(4690)<400>3agatctttat gaagacatag ctgcagaaga aaaagcaaga gctacatatc aatggttaat60tgatatatca gatgatcccg atttaaacga cagcttacga tttttacgag aaagagagat120tgttcactca cagcggttcc gcgaggccgt ggagatttta aaagatgaca gagacaggaa180gaaaatcttt taactagtaa aaaaacatcc cccttggcga atgcaaacga aaggagggat240gttttttgtt gtgactgcgt tgattatgcg ctagaactgc agtgacaaga aacaaccttt300aatttccctt caacatcttt ccaaactcgc gtataactgt attcacctcc aatagattca360ccggttgcca gtgccccatt taacgctact tttgtaacgg taacggcaag ttcttgaaac420agtttaactt cttgttccaa cacttccatg cccgctatat caagactttt tgaacgatga480acatttatat cttcttcttt tgacaaccat tgcccaaggt gattcacaaa aataagctca540tctgaaagta attcttctaa tagctctatg ttattagaaa gcatggctga gcgaagcatt600tcttcgtatt ctataactct tgcttgattc atttttaatc ctcctttacg ccttgtgtaa660ctcttttcta tttccacgtt gcttttcctt taaacttctt tcattaataa ttcgtgctaa720attatgttaa tagaggggat aagtggacta attttctgta agcactaaat attctgaaat780actctgttaa ttacctttaa atggtataaa attagaatga aagaaccttt tctttccact840tttctagtta tctttttact attaagatgc agttttttat acttgtaatt gtagcggaat900gaacgttcat tccgtttttg aaaagaggtg ataaagtgga atctactcca acaaaacaaa960aagcgatttt ttctgcttcg cttctgctgt ttgcagaaag agggtttgat gcaaccacga1020tgccaatgat tgcagagaat gccaaagtag gagcaggaac aatttatcgc tactttaaaa1080ataaagaaag ccttgtaaat gaattattcc aacagcacgt aaacgagttt ttacagtgca1140ttgaaagcgg tctggcaaac gagagagatg gataccgaga tgggtttcat catatctttg1200  
            
aaggtatggt gacatttact aaaaaccatc ctcgtgctct tggatttatt aaaactcata1260gccaaggaac ttttttaaca gaagagagcc gcttagcata tcaaaagctg gtggaatttg1320tttgtacgtt cttcagagaa ggacaaaagc aaggtgtgat tagaaatctt cctgaaaatg1380cgctaattgc tattttattt ggaagtttca tggaagtata tgaaatgatt gaaaatgact1440acttatcttt aactgatgaa cttcttaccg gtgtagaaga gagtctgtgg gcagcactta1500gcagacaatc atgaaactta acaagtgaaa gagggataac atg aca att aaa gaa  1555Met Thr Ile Lys Glu1   5atg cct cag cca aaa acg ttt gga gag ctt aaa aat tta ccg tta tta  1603Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn Leu Pro Leu Leu10  15  20aac aca gat aaa ccg gtt caa gct ttg atg aaa att gcg gat gaa tta  1651Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile Ala Asp Glu Leu25  30  35gga gaa atc ttt aaa ttc gag gcg cct ggt cgt gta acg cgc tac tta   1699Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val Thr Arg Tyr Leu40  45  50tca agt cag cgt cta att aaa gaa gca tgc gat gaa tca cgc ttt gat  1747Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu Ser Arg Phe Asp55  60  65aaa aac tta agt caa gcg ctt aaa ttt gta cgt gat ttt gca gga gac  1795Lys Asn Leu Ser Gln Ala Leu Lys Phe Val Arg Asp Phe Ala Gly Asp70  75  80  85ggg tta ttt aca agc tgg acg cat gaa aaa aat tgg aaa aaa gcg cat  1843Gly Leu Phe Thr Ser Trp Thr His Glu Lys Asn Trp Lys Lys Ala His90  95  100aat atc tta ctt cca agc ttc agt cag cag gca atg aaa ggc tat cat  189lAsn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met Lys Gly Tyr His105 110 115gcg atg atg gtc gat atc gcc gtg cag ctt gtt caa aag tgg gag cgt  1939Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln Lys Trp Glu Arg  
            
120 125 110cta aat gca gat gag cat att gaa gta ccg gaa gac atg aca cgt tta1987Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp Met Thr Arg Leu135 140 145acg ctt gat aca att ggt ctt tgc ggc ttt aac tat cgc ttt aac agc2035Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr Arg Phe Asn Ser150 155 160 165ttt tac cga gat cag cct cat cca ttt att aca agt atg gtc cgt gca2083Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser Met Val Arg Ala170 175 180ctg gat gaa gca atg aac aag ctg cag cga gca aat cca gac gac cca2131Leu Asp Glu Ala Met Asn Lys Leu Gln Arg Ala Asn Pro Asp Asp Pro185 190 195gct tat gat gaa aac aag cgc cag ttt caa gaa gat atc aag gtg atg2179Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp Ile Lys Val Met200 205 210aac gac cta gta gat aaa att att gca gat cgc aaa gca agc ggt gaa2227Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys Ala Ser Gly Glu215 220 225caa agc gat gat tta tta acg cat atg cta aac gga aaa gat cca gaa2275Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly Lys Asp Pro Glu230 235 240 245acg ggt gag ccg ctt gat gac gag aac att cgc tat caa att att aca2323Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr Gln Ile Ile Thr250 255 260ttc tta att gcg gga cac gaa aca aca agt ggt ctt tta tca ttt gcg2371Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu Leu Ser Phe Ala265 270 275ctg tat ttc tta gtg aaa aat cca cat gta tta caa aaa gca gca gaa2419Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln Lys Ala Ala Glu280 285 290gaa gca gca cga gtt cta gta gat cct gtt cca agc tac aaa caa gtc2467Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser Tyr Lys Gln Val  
            
295 300 305aaa cag ctt aaa tat gtc ggc atg gtc tta aac gaa gcg ctg cgc tta2515Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu Ala Leu Arg Leu310 315 320 325tgg cca act gct cct gcg ttt tcc cta tat gca aaa gaa gat acg gtg2563Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys Glu Asp Thr Val330 335 340ctt gga gga gaa tat cct tta gaa aaa ggc gac gaa cta atg gtt ctg2611Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu Leu Met Val Leu345 350 355att cct cag ctt cac cgt gat aaa aca att tgg gga gac gat gtg gaa2659Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly Asp Asp Val Glu360 365 370gag ttc cgt cca gag cgt ttt gaa aat cca agt gcg att ccg cag cat2707Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala Ile Pro Gln His375 380 385gcg ttt aaa ccg ttt gga aac ggt cag cgt gcg tgt atc ggt cag cag2755Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys Ile Gly Gln Gln390 395 400 405ttc gct ctt cat gaa gca acg ctg gta ctt ggt atg atg cta aaa cac2803Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met Met Leu Lys His410 415 420ttt gac ttt gaa gat cat aca aac tac gag ctg gat att aaa gaa act2851Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp Ile Lys Glu Thr425 430 435tta acg tta aaa cct gaa ggc ttt gtg gta aaa gca aaa tcg aaa aaa2899Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala Lys Ser Lys Lys440 445 450att ccg ctt ggc ggt att cct tca cct agc act gaa cag tct gct aaa2947Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu Gln Ser Ala Lys455 460 465aaa gta cgc aaa aag gca gaa aac gct cat aat acg ccg ctg ctt gtg2995Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr Pro Leu Leu Val  
            
470 475 480 485cta tac ggt tca aat atg gga aca gct gaa gga acg gcg cgt gat tta3043Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr Ala Arg Asp Leu490 495 500gca gat att gca atg agc aaa gga ttt gca ccg cag gtc gca acg ctt3091Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln Val Ala Thr Leu505 510 515gat tca cac gcc gga aat ctt ccg cgc gaa gga gct gta tta att gta3139Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala Val Leu Ile Val520 525 530acg gcg tct tat aac ggt cat ccg cct gat aac gca aag caa ttt gtc3187Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala Lys Gln Phe Val535 540 545gac tgg tta gac caa gcg tct gct gat gaa gta aaa ggc gtt cgc tac3235Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys Gly Val Arg Tyr550 555 560 565tcc gta ttt gga tgc ggc gat aaa aac tgg gct act acg tat caa aaa3283Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr Thr Tyr Gln Lys570 575 580gtg cct gct ttt atc gat gaa acg ctt gcc gct aaa ggg gca gaa aac3331Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys Gly Ala Glu Asn585 590 595atc gct gac cgc ggt gaa gca gat gca agc gac gac ttt gaa ggc aca3379Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp Phe Glu Gly Thr600 605 610tat gaa gaa tgg cgt gaa cat atg tgg agt gac gta gca gcc tac ttt3427Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val Ala Ala Tyr Phe615 620 625aac ctc gac att gaa aac agt gaa gat aat aaa tct act ctt tca ctt3475Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser Thr Leu Ser Leu630 635 640 645caa ttt gtc gac agc gcc gcg gat atg ccg ctt gcg aaa atg cac ggt3523Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala Lys Met His Gly  
            
650 655 660gcg ttt tca acg aac gtc gta gca agc aaa gaa ctt caa cag cca ggc3571Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu Gln Gln Pro Gly665 670 675agt gca cga agc acg cga cat ctt gaa att gaa ctt cca aaa gaa gct3619Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu Pro Lys Glu Ala680 685 690tct tat caa gaa gga gat cat tta ggt gtt att cct cgc aac tat gaa3667Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro Arg Asn Tyr Glu695 700 705gga ata gta aac cgt gta aca gca agg ttc ggc cta gat gca tca cag3715Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu Asp Ala Ser Gln710 715 720 725caa atc cgt ctg gaa gca gaa gaa gaa aaa tta gct cat ttg cca ctc3763Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala His Leu Pro Leu730 735 740gct aaa aca gta tcc gta gaa gag ctt ctg caa tac gtg gag ctt caa3811Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr Val Glu Leu Gln745 750 755gat cct gtt acg cgc acg cag ctt cgc gca atg gct gct aaa acg gtc3859Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala Ala Lys Thr Val760 765 770tgc ccg ccg cat aaa gta gag ctt gaa gcc ttg ctt gaa aag caa gcc3907Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu Glu Lys Gln Ala775 780 785tac aaa gaa caa gtg ctg gca aaa cgt tta aca atg ctt gaa ctg ctt3955Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met Leu Glu Leu Leu790 795 800 805gaa aaa tac ccg gcg tgt gaa atg aaa ttc agc gaa ttt atc gcc ctt4003Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu Phe Ile Ala Leu810 815 820ctg cca agc ata cgc ccg cgc tat tac tcg att tct tca tca cct cgt4051Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser Ser Ser Pro Arg  
            
825 830 835gtc gat gaa aaa caa gca agc atc acg gtc agc gtt gtc tca gga gaa4099Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val Val Ser Gly Glu840 845 850gcg tgg agc gga tat gga gaa tat aaa gga att gcg tcg aac tat ctt4147Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala Ser Asn Tyr Leu855 860 865gcc gag ctg caa gaa gga gat acg att acg tgc ttt att tcc aca ccg4195Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe Ile Ser Thr Pro870 875 880 885cag tca gaa ttt acg ctg cca aaa gac cct gaa acg ccg ctt atc atg4243Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr Pro Leu Ile Met890 895 900gtc gga ccg gga aca ggc gtc gcg ccg ttt aga ggc ttt gtg cag gcg4291Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly Phe Val Gln Ala905 910 915cgc aaa cag cta aaa gaa caa gga cag tca ctt gga gaa gca cat tta4339Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly Glu Ala His Leu920 925 930tac ttc ggc tgc cgt tca cct cat gaa gac tat ctg tat caa gaa gag4387Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu Tyr Gln Glu Glu935 940 945ctt gaa aac gcc caa agc gaa ggc atc att acg ctt cat acc gct ttt4435Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu His Thr Ala Phe950 955 960 965tct cgc atg cca aat cag ccg aaa aca tac gtt cag cac gta atg gaa4483Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln His Val Met Glu970 975 980caa gac ggc aag aaa ttg att gaa ctt ctt gat caa gga gcg cac ttc4531Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln Gly Ala His Phe985 990 995tat att tgc gga gac gga agc caa atg gca cct gcc gtt gaa gca4576Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala Val Glu Ala  
            
100010051010acg ctt atg aaa agc tat gct gac gtt cac caa gtg agt gaa gca   4621Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val Ser Glu Ala101510201025gac gct cgc tta tgg ctg cag cag cta gaa gaa aaa ggc cga tac   4666Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys Gly Arg Tyr103010351040gca aaa gac gtg tgg gct ggg taa attaaaaaga ggctaggata aaagtagttt  4720Ala Lys Asp Val Trp Ala Gly1045agttggttga aggaagatcc gaacgatgaa tcgttcggat ctttttattg gtagagtaaa 4780cgtagatttc atctatttag tgacttgtag cggttgattg gagggcaagg tgaagactcc 4840aatcaaccgc ggtgtcacat gcaagccata cgaaattcat ttctcccatt tattcgtctt 4900ttgtccccac ttaattttta tagcgcctta acgtttcttc tgcgtgacag cagatct4957<210>4<211>1049<212>PRT<213>巨大芽孢桿菌<400>4Met Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys1   5   10  15Asn Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys20  25  30Ile Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg35  40  45Val Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp50  55  60Glu Ser Arg Phe Asp Lys Asn Leu Ser Gln Ala Leu Lys Phe Val Arg65  70  75  80  
            
Asp Phe Ala Gly Asp Gly Leu Phe Thr Ser Trp Thr His Glu Lys Asn85  90  95Trp Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala100 105 110Met Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val115 120 125Gln Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu130 135 140Asp Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn145 150 155 160Tyr Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr165 170 175Ser Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Leu Gln Arg Ala180 185 190Asn Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu195 200 205Asp Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg210 215 220Lys Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn225 230 235 240Gly Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg245 250 255Tyr Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly260 265 270Leu Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu275 280 285Gln Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro290 295 300Ser Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn305 310 315 320  
            
Glu Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala325 330 335Lys Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp340 345 350Glu Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp355 360 365Gly Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser370 375 380Ala Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala385 390 395 400Cys Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly405 410 415Met Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu420 425 430Asp Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys435 440 445Ala Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr450 455 460Glu Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn465 470 475 480Thr Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly485 490 495Thr Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro500 505 510Gln Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly515 520 525Ala Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn530 535 540Ala Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val  
            
545 550 555 560Lys Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala565 570 575Thr Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala580 585 590Lys Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp595 600 605Asp Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp610 615 620Val Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys625 630 635 640Ser Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu645 650 655Ala Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu660 665 670Leu Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu675 680 685Leu Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile690 695 700Pro Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly705 710 715 720Leu Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu725 730 735Ala His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln740 745 750Tyr Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met755 760 765Ala Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu770 775 780  
            
Leu Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr785 790 795 800Met Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser805 810 815Glu Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile820 825 830Ser Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser835 840 845Val Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile850 855 860Ala Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys865 870 875 880Phe Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu885 890 895Thr Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg900 905 910Gly Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu915 920 925Gly Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr930 935 940Leu Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr945 950 955 960Leu His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val965 970 975Gln His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp980 985 990Gln Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro995 10001005Ala Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln101010151020  
            
Val Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu102510301035Lys Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly10401045<210>5<211>3147<212>DNA<213>人工序列<220>  
            
<223>細胞色素P450BM-3突變體<220>  
            
<221>CDS<222>(1)..(3147)<400>5aca att aaa gaa atg cct cag cca aaa acg ttt gga gag ctt aaa aat48Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn1   5   10  15tta ccg tta tta aac aca gat aaa ccg gtt caa gct ttg atg aaa att96Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile20  25  30gcg gat gaa tta gga gaa atc ttt aaa ttc gag gcg cct ggt cgt gta144Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val35  40  45acg cgc tac tta tca agt cag cgt cta att aaa gaa gca tgc gat gaa192Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu50  55  60tca cgc ttt gat aaa aac tta agt caa gcg ctt aaa ttt gta cgt gat240Ser Arg Phe Asp Lys Asn Leu Ser Gln Ala Leu Lys Phe Val Arg Asp65  70  75  80ttt gca gga gac ggg tta gct aca agc tgg acg cat gaa aaa aat tgg288Phe Ala Gly Asp Gly Leu Ala Thr Ser Trp Thr His Glu Lys Asn Trp  
            
85  90  95aaa aaa gcg cat aat atc tta ctt cca agc ttc agt cag cag gca atg336Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met100 105 110aaa ggc tat cat gcg atg atg gtc gat atc gcc gtg cag ctt gtt caa384Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln115 120 125aag tgg gag cgt cta aat gca gat gag cat att gaa gta ccg gaa gac432Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp130 135 140atg aca cgt tta acg ctt gat aca att ggt ctt tgc ggc ttt aac tat480Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr145 150 155 160cgc ttt aac agc ttt tac cga gat cag cct cat cca ttt att aca agt528Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser165 170 175atg gtc cgt gca ctg gat gaa gca atg aac aag ctg cag cga gca aat576Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Leu Gln Arg Ala Asn180 185 190cca gac gac cca gct tat gat gaa aac aag cgc cag ttt caa gaa gat624Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp195 200 205atc aag gtg atg aac gac cta gta gat aaa att att gca gat cgc aaa672Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys210 215 220gca agc ggt gaa caa agc gat gat tta tta acg cat atg cta aac gga720Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly225 230 235 240aaa gat cca gaa acg ggt gag ccg ctt gat gac gag aac att cgc tat768Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr245 250 255caa att att aca ttc tta att gcg gga cac gaa aca aca agt ggt ctt816Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu  
            
260 265 270tta tca ttt gcg ctg tat ttc tta gtg aaa aat cca cat gta tta caa864Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln275 280 285aaa gca gca gaa gaa gca gca cga gtt cta gta gat cct gtt cca agc912Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser290 295 300tac aaa caa gtc aaa cag ctt aaa tat gtc ggc atg gtc tta aac gaa960Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu305 310 315 320gcg ctg cgc tta tgg cca act gct cct gcg ttt tcc cta tat gca aaa1008Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys325 330 335gaa gat acg gtg ctt gga gga gaa tat cct tta gaa aaa ggc gac gaa1056Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu340 345 350cta atg gtt ctg att cct cag ctt cac cgt gat aaa aca att tgg gga1104Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly355 360 365gac gat gtg gaa gag ttc cgt cca gag cgt ttt gaa aat cca agt gcg1152Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala370 375 380att ccg cag cat gcg ttt aaa ccg ttt gga aac ggt cag cgt gcg tgt1200Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys385 390 395 400atc ggt cag cag ttc gct ctt cat gaa gca acg ctg gta ctt ggt atg1248Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met405 410 415atg cta aaa cac ttt gac ttt gaa gat cat aca aac tac gag ctg gat1296Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp420 425 430att aaa gaa act tta acg tta aaa cct gaa ggc ttt gtg gta aaa gca1344Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala  
            
435 440 445aaa tcg aaa aaa att ccg ctt ggc ggt att cct tca cct agc act gaa1392Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu450 455 460cag tct gct aaa aaa gta cgc aaa aag gca gaa aac gct cat aat acg1440Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr465 470 475 480ccg ctg ctt gtg cta tac ggt tca aat atg gga aca gct gaa gga acg1488Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr485 490 495gcg cgt gat tta gca gat att gca atg agc aaa gga ttt gca ccg cag1536Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln500 505 510gtc gca acg ctt gat tca cac gcc gga aat ctt ccg cgc gaa gga gct1584Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala515 520 525gta tta att gta acg gcg tct tat aac ggt cat ccg cct gat aac gca1632Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala530 535 540aag caa ttt gtc gac tgg tta gac caa gcg tct gct gat gaa gta aaa1680Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys545 550 555 560ggc gtt cgc tac tcc gta ttt gga tgc ggc gat aaa aac tgg gct act1728Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr565 570 575acg tat caa aaa gtg cct gct ttt atc gat gaa acg ctt gcc gct aaa1776Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys580 585 590ggg gca gaa aac atc gct gac cgc ggt gaa gca gat gca agc gac gac1824Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp595 600 605ttt gaa ggc aca tat gaa gaa tgg cgt gaa cat atg tgg agt gac gta1872Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val  
            
610 615 620gca gcc tac ttt aac ctc gac att gaa aac agt gaa gat aat aaa tct1920Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser625 630 635 640act ctt tca ctt caa ttt gtc gac agc gcc gcg gat atg ccg ctt gcg1968Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala645 650 655aaa atg cac ggt gcg ttt tca acg aac gtc gta gca agc aaa gaa ctt2016Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu660 665 670caa cag cca ggc agt gca cga agc acg cga cat ctt gaa att gaa ctt2064Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu675 680 685cca aaa gaa gct tct tat caa gaa gga gat cat tta ggt gtt att cct2112Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro690 695 700cgc aac tat gaa gga ata gta aac cgt gta aca gca agg ttc ggc cta2160Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu705 710 715 720gat gca tca cag caa atc cgt ctg gaa gca gaa gaa gaa aaa tta gct2208Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala725 730 735cat ttg cca ctc gct aaa aca gta tcc gta gaa gag ctt ctg caa tac2256His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr740 745 750gtg gag ctt caa gat cct gtt acg cgc acg cag ctt cgc gca atg gct2304Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala755 760 765gct aaa acg gtc tgc ccg ccg cat aaa gta gag ctt gaa gcc ttg ctt2352Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu770 775 780gaa aag caa gcc tac aaa gaa caa gtg ctg gca aaa cgt tta aca atg2400Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met  
            
785 790 795 800ctt gaa ctg ctt gaa aaa tac ccg gcg tgt gaa atg aaa ttc agc gaa2448Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu805 810 815ttt atc gcc ctt ctg cca agc ata cgc ccg cgc tat tac tcg att tct2496Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser820 825 830tca tca cct cgt gtc gat gaa aaa caa gca agc atc acg gtc agc gtt2544Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val835 840 845gtc tca gga gaa gcg tgg agc gga tat gga gaa tat aaa gga att gcg2592Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala850 855 860tcg aac tat ctt gcc gag ctg caa gaa gga gat acg att acg tgc ttt2640Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe865 870 875 880att tcc aca ccg cag tca gaa ttt acg ctg cca aaa gac cct gaa acg2688Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr885 890 895ccg ctt atc atg gtc gga ccg gga aca ggc gtc gcg ccg ttt aga ggc2736Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly900 905 910ttt gtg cag gcg cgc aaa cag cta aaa gaa caa gga cag tca ctt gga2784Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly915 920 925gaa gca cat tta tac ttc ggc tgc cgt tca cct cat gaa gac tat ctg2832Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu930 935 940tat caa gaa gag ctt gaa aac gcc caa agc gaa ggc atc att acg ctt2880Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu945 950 955 960cat acc gct ttt tct cgc atg cca aat cag ccg aaa aca tac gtt cag2928His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln  
            
965 970 975cac gta atg gaa caa gac ggc aag aaa ttg att gaa ctt ctt gat caa2976His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln980 985 990gga gcg cac ttc tat att tgc gga gac gga agc caa atg gca cct gcc3024Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala995 10001005gtt gaa gca acg ctt atg aaa agc tat gct gac gtt cac caa gtg3069Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val101010151020agt gaa gca gac gct cgc tta tgg ctg cag cag cta gaa gaa aaa3114Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys102510301035ggc cga tac gca aaa gac gtg tgg gct ggg taa3147Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly10401045<210>6<211>1048<212>PRT<213>人工序列<220>  
            
<223>細胞色素P450BM-3突變體<400>6Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn1   5   10  15Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile20  25  30Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val35  40  45Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu50  55  60  
            
Ser Arg Phe Asp Lys Asn Leu Ser Gln Ala Leu Lys Phe Val Arg Asp65  70  75  80Phe Ala Gly Asp Gly Leu Ala Thr Ser Trp Thr His Glu Lys Asn Trp85  90  95Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met100 105 110Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln115 120 125Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp130 135 140Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr145 150 155 160Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser165 170 175Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Leu Gln Arg Ala Asn180 185 190Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp195 200 205Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys2l0 215 220Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly225 230 235 240Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr245 250 255Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu260 265 270Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln275 280 285Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser  
            
290 295 300Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu305 310 315 320Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys325 330 335Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu340 345 350Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly355 360 365Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala370 375 380Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys385 390 395 400Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met405 410 415Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp420 425 430Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala435 440 445Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu450 455 460Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr465 470 475 480Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr485 490 495Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln500 505 510Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala515 520 525  
            
Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala530 535 540Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys545 550 555 560Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr565 570 575Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys580 585 590Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp595 600 605Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val610 615 620Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser625 630 635 640Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala645 650 655Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu660 665 670Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu675 680 685Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro690 695 700Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu705 710 715 720Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala725 730 735His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr740 745 750Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala755 760 765  
            
Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu770 775 780Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met785 790 795 800Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu805 810 815Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser820 825 830Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val835 840 845Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala850 855 860Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe865 870 875 880Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr885 890 895Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly900 905 910Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly915 920 925Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu930 935 940Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu945 950 955 960His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln965 970 975His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln980 985 990  
            
Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala995 10001005Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val101010151020Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys102510301035Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly10401045<210>7<211>3147<212>DNA<213>人工序列<220>  
            
<223>細胞色素P450BM-3突變體<220>  
            
<221>CDS<222>(1)..(3147)<400>7aca att aaa gaa atg cct cag cca aaa acg ttt gga gag ctt aaa aat48Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn1   5   10  15tta ccg tta tta aac aca gat aaa ccg gtt caa gct ttg atg aaa att96Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile20  25  30gcg gat gaa tta gga gaa atc ttt aaa ttc gag gcg cct ggt cgt gta144Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val35 40 45acg cgc tac tta tca agt cag cgt cta att aaa gaa gca tgc gat gaa192Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu50 55 60tca cgc ttt gat aaa aac tta agt caa ggg ctt aaa ttt gta cgt gat240Ser Arg Phe Asp Lys Asn Leu Ser Gln Gly Leu Lys Phe Val Arg Asp  
            
65  70  75  80ttt gca gga gac ggg tta gtt aca agc tgg acg cat gaa aaa aat tgg288Phe Ala Gly Asp Gly Leu Val Thr Ser Trp Thr His Glu Lys Asn Trp85 90 95aaa aaa gcg cat aat atc tta ctt cca agc ttc agt cag cag gca atg336Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met100 105 110aaa ggc tat cat gcg atg atg gtc gat atc gcc gtg cag ctt gtt caa384Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln115 120 125aag tgg gag cgt cta aat gca gat gag cat att gaa gta ccg gaa gac432Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp130 135 140atg aca cgt tta acg ctt gat aca att ggt ctt tgc ggc ttt aac tat480Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr145 150 155 160cgc ttt aac agc ttt tac cga gat cag cct cat cca ttt att aca agt528Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser165 170 175atg gtc cgt gca ctg gat gaa gca atg aac aag cag cag cga gca aat576Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Gln Gln Arg Ala Asn180 185 190cca gac gac cca gct tat gat gaa aac aag cgc cag ttt caa gaa gat624Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp195 200 205atc aag gtg atg aac gac cta gta gat aaa att att gca gat cgc aaa672Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys210 215 220gca agc ggt gaa caa agc gat gat tta tta acg cat atg cta aac gga720Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly225 230 235 240aaa gat cca gaa acg ggt gag ccg ctt gat gac gag aac att cgc tat768Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr  
            
245 250 255caa att att aca ttc tta att gcg gga cac gaa aca aca agt ggt ctt816Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu260 265 270tta tca ttt gcg ctg tat ttc tta gtg aaa aat cca cat gta tta caa864Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln275 280 285aaa gca gca gaa gaa gca gca cga gtt cta gta gat cct gtt cca agc912Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser290 295 300tac aaa caa gtc aaa cag ctt aaa tat gtc ggc atg gtc tta aac gaa960Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu305 310 315 320gcg ctg cgc tta tgg cca act gct cct gcg ttt tcc cta tat gca aaa1008Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys325 330 335gaa gat acg gtg ctt gga gga gaa tat cct tta gaa aaa ggc gac gaa1056Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu340 345 350cta atg gtt ctg att cct cag ctt cac cgt gat aaa aca att tgg gga1104Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly355 360 365gac gat gtg gaa gag ttc cgt cca gag cgt ttt gaa aat cca agt gcg1152Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala370 375 380att ccg cag cat gcg ttt aaa ccg ttt gga aac ggt cag cgt gcg tgt1200Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys385 390 395 400atc ggt cag cag ttc gct ctt cat gaa gca acg ctg gta ctt ggt atg1248Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met405 410 415atg cta aaa cac ttt gac ttt gaa gat cat aca aac tac gag ctg gat1296Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp  
            
420 425 430att aaa gaa act tta acg tta aaa cct gaa ggc ttt gtg gta aaa gca1344Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala435 440 445aaa tcg aaa aaa att ccg ctt ggc ggt att cct tca cct agc act gaa1392Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu450 455 460cag tct gct aaa aaa gta cgc aaa aag gca gaa aac gct cat aat acg1440Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr465 470 475 480ccg ctg ctt gtg cta tac ggt tca aat atg gga aca gct gaa gga acg1488Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr485 490 495gcg cgt gat tta gca gat att gca atg agc aaa gga ttt gca ccg cag1536Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln500 505 510gtc gca acg ctt gat tca cac gcc gga aat ctt ccg cgc gaa gga gct1584Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala515 520 525gta tta att gta acg gcg tct tat aac ggt cat ccg cct gat aac gca1632Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala530 535 540aag caa ttt gtc gac tgg tta gac caa gcg tct gct gat gaa gta aaa1680Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys545 550 555 560ggc gtt cgc tac tcc gta ttt gga tgc ggc gat aaa aac tgg gct act1728Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr565 570 575acg tat caa aaa gtg cct gct ttt atc gat gaa acg ctt gcc gct aaa1776Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys580 585 590ggg gca gaa aac atc gct gac cgc ggt gaa gca gat gca agc gac gac1824Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp  
            
595 600 605ttt gaa ggc aca tat gaa gaa tgg cgt gaa cat atg tgg agt gac gta1872Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val610 615 620gca gcc tac ttt aac ctc gac att gaa aac agt gaa gat aat aaa tct1920Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser625 630 635 640act ctt tca ctt caa ttt gtc gac agc gcc gcg gat atg ccg ctt gcg1968Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala645 650 655aaa atg cac ggt gcg ttt tca acg aac gtc gta gca agc aaa gaa ctt2016Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu660 665 670caa cag cca ggc agt gca cga agc acg cga cat ctt gaa att gaa ctt2064Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu675 680 685cca aaa gaa gct tct tat caa gaa gga gat cat tta ggt gtt att cct2112Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro690 695 700cgc aac tat gaa gga ata gta aac cgt gta aca gca agg ttc ggc cta2160Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu705 710 715 720gat gca tca cag caa atc cgt ctg gaa gca gaa gaa gaa aaa tta gct2208Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala725 730 735cat ttg cca ctc gct aaa aca gta tcc gta gaa gag ctt ctg caa tac2256His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr740 745 750gtg gag ctt caa gat cct gtt acg cgc acg cag ctt cgc gca atg gct2304Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala755 760 765gct aaa acg gtc tgc ccg ccg cat aaa gta gag ctt gaa gcc ttg ctt2352Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu  
            
770 775 780gaa aag caa gcc tac aaa gaa caa gtg ctg gca aaa cgt tta aca atg2400Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met785 790 795 800ctt gaa ctg ctt gaa aaa tac ccg gcg tgt gaa atg aaa ttc agc gaa2448Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu805 810 815ttt atc gcc ctt ctg cca agc ata cgc ccg cgc tat tac tcg att tct2496Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser820 825 830tca tca cct cgt gtc gat gaa aaa caa gca agc atc acg gtc agc gtt2544Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val835 840 845gtc tca gga gaa gcg tgg agc gga tat gga gaa tat aaa gga att gcg2592Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala850 855 860tcg aac tat ctt gcc gag ctg caa gaa gga gat acg att acg tgc ttt2640Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe865 870 875 880att tcc aca ccg cag tca gaa ttt acg ctg cca aaa gac cct gaa acg2688Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr885 890 895ccg ctt atc atg gtc gga ccg gga aca ggc gtc gcg ccg ttt aga ggc2736Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly900 905 910ttt gtg cag gcg cgc aaa cag cta aaa gaa caa gga cag tca ctt gga2784Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly915 920 925gaa gca cat tta tac ttc ggc tgc cgt tca cct cat gaa gac tat ctg2832Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu930 935 940tat caa gaa gag ctt gaa aac gcc caa agc gaa ggc atc att acg ctt2880Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu  
            
945 950 955 960cat acc gct ttt tct cgc atg cca aat cag ccg aaa aca tac gtt cag2928His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln965 970 975cac gta atg gaa caa gac ggc aag aaa ttg att gaa ctt ctt gat caa2976His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln980 985 990gga gcg cac ttc tat att tgc gga gac gga agc caa atg gca cct gcc3024Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala99510001005gtt gaa gca acg ctt atg aaa agc tat gct gac gtt cac caa gtg3069Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val101010151020agt gaa gca gac gct cgc tta tgg ctg cag cag cta gaa gaa aaa3114Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys102510301035ggc cga tac gca aaa gac gtg tgg gct ggg taa3147Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly10401045<210>8<211>1048<212>PRT<213>人工序列<220>  
            
<223>細胞色素P450BM-3突變體<400>8Thr Ile Lys Glu Met Pro Gln Pro Lys Thr Phe Gly Glu Leu Lys Asn1   5   10  15Leu Pro Leu Leu Asn Thr Asp Lys Pro Val Gln Ala Leu Met Lys Ile20  25  30  
            
Ala Asp Glu Leu Gly Glu Ile Phe Lys Phe Glu Ala Pro Gly Arg Val35  40  45Thr Arg Tyr Leu Ser Ser Gln Arg Leu Ile Lys Glu Ala Cys Asp Glu50  55  60Ser Arg Phe Asp Lys Asn Leu Ser Gln Gly Leu Lys Phe Val Arg Asp65  70  75  80Phe Ala Gly Asp Gly Leu Val Thr Ser Trp Thr His Glu Lys Asn Trp85  90  95Lys Lys Ala His Asn Ile Leu Leu Pro Ser Phe Ser Gln Gln Ala Met100 105 110Lys Gly Tyr His Ala Met Met Val Asp Ile Ala Val Gln Leu Val Gln115 120 125Lys Trp Glu Arg Leu Asn Ala Asp Glu His Ile Glu Val Pro Glu Asp130 135 140Met Thr Arg Leu Thr Leu Asp Thr Ile Gly Leu Cys Gly Phe Asn Tyr145 150 155 160Arg Phe Asn Ser Phe Tyr Arg Asp Gln Pro His Pro Phe Ile Thr Ser165 170 175Met Val Arg Ala Leu Asp Glu Ala Met Asn Lys Gln Gln Arg Ala Asn180 185 190Pro Asp Asp Pro Ala Tyr Asp Glu Asn Lys Arg Gln Phe Gln Glu Asp195 200 205Ile Lys Val Met Asn Asp Leu Val Asp Lys Ile Ile Ala Asp Arg Lys210 215 220Ala Ser Gly Glu Gln Ser Asp Asp Leu Leu Thr His Met Leu Asn Gly225 230 235 240Lys Asp Pro Glu Thr Gly Glu Pro Leu Asp Asp Glu Asn Ile Arg Tyr245 250 255Gln Ile Ile Thr Phe Leu Ile Ala Gly His Glu Thr Thr Ser Gly Leu260 265 270  
            
Leu Ser Phe Ala Leu Tyr Phe Leu Val Lys Asn Pro His Val Leu Gln275 280 285Lys Ala Ala Glu Glu Ala Ala Arg Val Leu Val Asp Pro Val Pro Ser290 295 300Tyr Lys Gln Val Lys Gln Leu Lys Tyr Val Gly Met Val Leu Asn Glu305 310 315 320Ala Leu Arg Leu Trp Pro Thr Ala Pro Ala Phe Ser Leu Tyr Ala Lys325 330 335Glu Asp Thr Val Leu Gly Gly Glu Tyr Pro Leu Glu Lys Gly Asp Glu340 345 350Leu Met Val Leu Ile Pro Gln Leu His Arg Asp Lys Thr Ile Trp Gly355 360 365Asp Asp Val Glu Glu Phe Arg Pro Glu Arg Phe Glu Asn Pro Ser Ala370 375 380Ile Pro Gln His Ala Phe Lys Pro Phe Gly Asn Gly Gln Arg Ala Cys385 390 395 400Ile Gly Gln Gln Phe Ala Leu His Glu Ala Thr Leu Val Leu Gly Met405 410 415Met Leu Lys His Phe Asp Phe Glu Asp His Thr Asn Tyr Glu Leu Asp420 425 430Ile Lys Glu Thr Leu Thr Leu Lys Pro Glu Gly Phe Val Val Lys Ala435 440 445Lys Ser Lys Lys Ile Pro Leu Gly Gly Ile Pro Ser Pro Ser Thr Glu450 455 460Gln Ser Ala Lys Lys Val Arg Lys Lys Ala Glu Asn Ala His Asn Thr465 470 475 480Pro Leu Leu Val Leu Tyr Gly Ser Asn Met Gly Thr Ala Glu Gly Thr485 490 495Ala Arg Asp Leu Ala Asp Ile Ala Met Ser Lys Gly Phe Ala Pro Gln  
            
500 505 510Val Ala Thr Leu Asp Ser His Ala Gly Asn Leu Pro Arg Glu Gly Ala515 520 525Val Leu Ile Val Thr Ala Ser Tyr Asn Gly His Pro Pro Asp Asn Ala530 535 540Lys Gln Phe Val Asp Trp Leu Asp Gln Ala Ser Ala Asp Glu Val Lys545 550 555 560Gly Val Arg Tyr Ser Val Phe Gly Cys Gly Asp Lys Asn Trp Ala Thr565 570 575Thr Tyr Gln Lys Val Pro Ala Phe Ile Asp Glu Thr Leu Ala Ala Lys580 585 590Gly Ala Glu Asn Ile Ala Asp Arg Gly Glu Ala Asp Ala Ser Asp Asp595 600 605Phe Glu Gly Thr Tyr Glu Glu Trp Arg Glu His Met Trp Ser Asp Val610 615 620Ala Ala Tyr Phe Asn Leu Asp Ile Glu Asn Ser Glu Asp Asn Lys Ser625 630 635 640Thr Leu Ser Leu Gln Phe Val Asp Ser Ala Ala Asp Met Pro Leu Ala645 650 655Lys Met His Gly Ala Phe Ser Thr Asn Val Val Ala Ser Lys Glu Leu660 665 670Gln Gln Pro Gly Ser Ala Arg Ser Thr Arg His Leu Glu Ile Glu Leu675 680 685Pro Lys Glu Ala Ser Tyr Gln Glu Gly Asp His Leu Gly Val Ile Pro690 695 700Arg Asn Tyr Glu Gly Ile Val Asn Arg Val Thr Ala Arg Phe Gly Leu705 710 715 720Asp Ala Ser Gln Gln Ile Arg Leu Glu Ala Glu Glu Glu Lys Leu Ala725 730 735  
            
His Leu Pro Leu Ala Lys Thr Val Ser Val Glu Glu Leu Leu Gln Tyr740 745 750Val Glu Leu Gln Asp Pro Val Thr Arg Thr Gln Leu Arg Ala Met Ala755 760 765Ala Lys Thr Val Cys Pro Pro His Lys Val Glu Leu Glu Ala Leu Leu770 775 780Glu Lys Gln Ala Tyr Lys Glu Gln Val Leu Ala Lys Arg Leu Thr Met785 790 795 800Leu Glu Leu Leu Glu Lys Tyr Pro Ala Cys Glu Met Lys Phe Ser Glu805 810 815Phe Ile Ala Leu Leu Pro Ser Ile Arg Pro Arg Tyr Tyr Ser Ile Ser820 825 830Ser Ser Pro Arg Val Asp Glu Lys Gln Ala Ser Ile Thr Val Ser Val835 840 845Val Ser Gly Glu Ala Trp Ser Gly Tyr Gly Glu Tyr Lys Gly Ile Ala850 855 860Ser Asn Tyr Leu Ala Glu Leu Gln Glu Gly Asp Thr Ile Thr Cys Phe865 870 875 880Ile Ser Thr Pro Gln Ser Glu Phe Thr Leu Pro Lys Asp Pro Glu Thr885 890 895Pro Leu Ile Met Val Gly Pro Gly Thr Gly Val Ala Pro Phe Arg Gly900 905 910Phe Val Gln Ala Arg Lys Gln Leu Lys Glu Gln Gly Gln Ser Leu Gly915 920 925Glu Ala His Leu Tyr Phe Gly Cys Arg Ser Pro His Glu Asp Tyr Leu930 935 940Tyr Gln Glu Glu Leu Glu Asn Ala Gln Ser Glu Gly Ile Ile Thr Leu945 950 955 960His Thr Ala Phe Ser Arg Met Pro Asn Gln Pro Lys Thr Tyr Val Gln965 970 975  
            
His Val Met Glu Gln Asp Gly Lys Lys Leu Ile Glu Leu Leu Asp Gln980 985 990Gly Ala His Phe Tyr Ile Cys Gly Asp Gly Ser Gln Met Ala Pro Ala99510001005Val Glu Ala Thr Leu Met Lys Ser Tyr Ala Asp Val His Gln Val101010151020Ser Glu Ala Asp Ala Arg Leu Trp Leu Gln Gln Leu Glu Glu Lys102510301035Gly Arg Tyr Ala Lys Asp Val Trp Ala Gly10401045<210>9<211>1032<212>DNA<213>珊瑚諾卡氏菌<220>  
            
<221>CDS<222>(1)..(1032)<400>9atg acg aca gag gcg acg gtg gcc cga ccg gtg gag ctc gaa ggt cac48Met Thr Thr Glu Ala Thr Val Ala Arg Pro Val Glu Leu Glu Gly His1   5  10  15cgg aca ttc acc tgg ttc acg ccc gcc agg cga aag ccg acg gag tac96Arg Thr Phe Thr Trp Phe Thr Pro Ala Arg Arg Lys Pro Thr Glu Tyr20 25  30gag ctc tac acc gtg ggt caa cag tcc act ccg gac gag tgg ctg cat144Glu Leu Tyr Thr Val Gly Gln Gln Ser Thr Pro Asp Glu Trp Leu His35  40  45gtg gac tgg ccg ctg cgc ttc gac gac ggc cgc gcc ccg tgg gag gag192Val Asp Trp Pro Leu Arg Phe Asp Asp Gly Arg Ala Pro Trp Glu Glu50  55  60  
            
gag tcg agt gcg gta cgg acc tcg gag tgg tcg gct tac cgc gac cca240Glu Ser Ser Ala Val Arg Thr Ser Glu Trp Ser Ala Tyr Arg Asp Pro65  70  75  80cac caa ctg tgg cag cgt ccc tac gtc agc acg tgc aac cag gac cag288His Gln Leu Trp Gln Arg Pro Tyr Val Ser Thr Cys Asn Gln Asp Gln85  90  95cag gcc ctc gcg cgg ctg gtc ccc gtc ctg acc atg ggg tcg gcg gcg336Gln Ala Leu Ala Arg Leu Val Pro Val Leu Thr Met Gly Ser Ala Ala100 105 110atc acg ccc atc tgg tcg cag aag atc ctc gcc agg tcc tac gcc gcc384Ile Thr Pro Ile Trp Ser Gln Lys Ile Leu Ala Arg Ser Tyr Ala Ala115 120 125tgg cca ttc gtc gag tac ggg ctc ttc ctg agc ctg gcc tac gcc gtg432Trp Pro Phe Val Glu Tyr Gly Leu Phe Leu Ser Leu Ala Tyr Ala Val130 135 140cgc cag gcc atg tcc gac acg gtc cag ttc agc gtg gtg ttc cag gcc480Arg Gln Ala Met Ser Asp Thr Val Gln Phe Ser Val Val Phe Gln Ala145 150 155 160gtg gac cgc atg cgg ctg ctc cag gac atc gtc cac cac ctg gac cac528Val Asp Arg Met Arg Leu Leu Gln Asp Ile Val His His Leu Asp His165 170 175ctg cag gag tcg ccg gaa ttc agc gac gcc ggg gcc cgc gag gcc tgg576Leu Gln Glu Ser Pro Glu Phe Ser Asp Ala Gly Ala Arg Glu Ala Trp180 185 190atg tcc gac tcc acc ctg gtc ccg atc cgg gaa gtg atc gag cgc atc624Met Ser Asp Ser Thr Leu Val Pro Ile Arg Glu Val Ile Glu Arg Ile195 200 205gcc gcc agc cag gac tgg gtg gag atc ctg gtc gcc ggc acg ctc gtc672Ala Ala Ser Gln Asp Trp Val Glu Ile Leu Val Ala Gly Thr Leu Val210 215 220ttc gag cct ctg gtc ggc cac ctg gcg aag gcc gag ttg ttc agc cgc720Phe Glu Pro Leu Val Gly His Leu Ala Lys Ala Glu Leu Phe Ser Arg225 230 235 240  
            
cgt gcg cca atg ttc ggg gac ggg acc acg ccg gcg gtg ctg gcg tcg768Arg Ala Pro Met Phe Gly Asp Gly Thr Thr Pro Ala Val Leu Ala Ser245 250 255gcc ctg ctg gac agc ggc agg cac ctc gaa tcg gtc cag gcg ctc gtc816Ala Leu Leu Asp Ser Gly Arg His Leu Glu Ser Val Gln Ala Leu Val260 265 270cgc ctc gtc tgc caa gac ccc gtc cat ggc gac cag aac cag gcg act864Arg Leu Val Cys Gln Asp Pro Val His Gly Asp Gln Asn Gln Ala Thr275 280 285gtg cgg cgg tgg atc gag gaa tgg cag ccg cgg tgc aag gcg gcg gcc912Val Arg Arg Trp Ile Glu Glu Trp Gln Pro Arg Cys Lys Ala Ala Ala290 295 300cag tcc ttc ctg ccg acg ttc tcc gac tgc ggc atc gac gcc aag gaa960Gln Ser Phe Leu Pro Thr Phe Ser Asp Cys Gly Ile Asp Ala Lys Glu305 310 315 320agc gcc aac gcg ctg tcc cgg gcg ctg gcg aac cag cgg gcc gcc gtc1008Ser Ala Asn Ala Leu Ser Arg Ala Leu Ala Asn Gln Arg Ala Ala Val325 330 335gag ggc gcc ggc atc acg gca tga1032Glu Gly Ala Gly Ile Thr Ala340<210>10<211>343<212>PRT<213>珊瑚諾卡氏菌<400>10Met Thr Thr Glu Ala Thr Val Ala Arg Pro Val Glu Leu Glu Gly His1   5   10  15Arg Thr Phe Thr Trp Phe Thr Pro Ala Arg Arg Lys Pro Thr Glu Tyr20 25 30Glu Leu Tyr Thr Val Gly Gln Gln Ser Thr Pro Asp Glu Trp Leu His35  40  45  
            
Val Asp Trp Pro Leu Arg Phe Asp Asp Gly Arg Ala Pro Trp Glu Glu50  55  60Glu Ser Ser Ala Val Arg Thr Ser Glu Trp Ser Ala Tyr Arg Asp Pro65  70  75  80His Gln Leu Trp Gln Arg Pro Tyr Val Ser Thr Cys Asn Gln Asp Gln85 90 95Gln Ala Leu Ala Arg Leu Val Pro Val Leu Thr Met Gly Ser Ala Ala100 105 110Ile Thr Pro Ile Trp Ser Gln Lys Ile Leu Ala Arg Ser Tyr Ala Ala115 120 125Trp Pro Phe Val Glu Tyr Gly Leu Phe Leu Ser Leu Ala Tyr Ala Val130 135 140Arg Gln Ala Met Ser Asp Thr Val Gln Phe Ser Val Val Phe Gln Ala145 150 155 160Val Asp Arg Met Arg Leu Leu Gln Asp Ile Val His His Leu Asp His165 170 175Leu Gln Glu Ser Pro Glu Phe Ser Asp Ala Gly Ala Arg Glu Ala Trp180 185 190Met Ser Asp Ser Thr Leu Val Pro Ile Arg Glu Val Ile Glu Arg Ile195 200 205Ala Ala Ser Gln Asp Trp Val Glu Ile Leu Val Ala Gly Thr Leu Val210 215 220Phe Glu Pro Leu Val Gly His Leu Ala Lys Ala Glu Leu Phe Ser Arg225 230 235 240Arg Ala Pro Met Phe Gly Asp Gly Thr Thr Pro Ala Val Leu Ala Ser245 250 255Ala Leu Leu Asp Ser Gly Arg His Leu Glu Ser Val Gln Ala Leu Val260 265 270Arg Leu Val Cys Gln Asp Pro Val His Gly Asp Gln Asn Gln Ala Thr  
            
275 280 285Val Arg Arg Trp Ile Glu Glu Trp Gln Pro Arg Cys Lys Ala Ala Ala290 295 300Gln Ser Phe Leu Pro Thr Phe Ser Asp Cys Gly Ile Asp Ala Lys Glu305 310 315 320Ser Ala Asn Ala Leu Ser Arg Ala Leu Ala Asn Gln Arg Ala Ala Val325 330 335Glu Gly Ala Gly Ile Thr Ala340<210>11<211>1506<212>DNA<213>珊瑚諾卡氏菌<220>  
            
<221>CDS<222>(1)..(1506)<400>11atg gca tcg aac ccc acc cag ctc cac gag aag tcg aag tcc tac gac48Met Ala Ser Asn Pro Thr Gln Leu His Glu Lys Ser Lys Ser Tyr Asp15   10  15tgg gac ttc acc tcc gtc gag cgg cgc ccc aag ttc gag acg aag tac96Trp Asp Phe Thr Ser Val Glu Arg Arg Pro Lys Phe Glu Thr Lys Tyr20  25  30aag atg ccc aag aag ggc aag gac ccg ttc cgc gtc ctg atc cgt gac144Lys Met Pro Lys Lys Gly Lys Asp Pro Phe Arg Val Leu Ile Arg Asp35  40  45tac atg aag atg gaa gcg gag aag gac gac cgg acc cat ggc ttc ctc192Tyr Met Lys Met Glu Ala Glu Lys Asp Asp Arg Thr His Gly Phe Leu50  55  60gac ggc gcc gtg cgg acg cgt gag gcc acc agg att gag ccg cgg ttc240Asp Gly Ala Val Arg Thr Arg Glu Ala Thr Arg Ile Glu Pro Arg Phe65  70  75  80  
            
gct gag gcc atg aag atc atg gtg ccg cag ctg acc aac gcc gag tac288Ala Glu Ala Met Lys Ile Met Val Pro Gln Leu Thr Asn Ala Glu Tyr85  90  95cag gcg gtg gcg ggc tgc gga atg atc atc tcg gcc gtc gag aac cag336Gln Ala Val Ala Gly Cys Gly Met Ile Ile Ser Ala Val Glu Asn Gln100 105 110gag ctc cgt cag ggc tac gcc gct cag atg ctc gat gag gtg cgg cac384Glu Leu Arg Gln Gly Tyr Ala Ala Gln Met Leu Asp Glu Val Arg His115 120 125gcg cag ctc gag atg acg cta cgc aac tac tac gcg aag cac tgg tgc432Ala Gln Leu Glu Met Thr Leu Arg Asn Tyr Tyr Ala Lys His Trp Cys130 135 140gat ccc tcc ggc ttc gac atc ggt cag cgc ggc ctg tac cag cac ccc480Asp Pro Ser Gly Phe Asp Ile Gly Gln Arg Gly Leu Tyr Gln His Pro145 150 155 160gcg ggg ctg gtg tcc atc ggc gag ttc cag cac ttc aat act ggt gac528Ala Gly Leu Val Ser Ile Gly Glu Phe Gln His Phe Asn Thr Gly Asp165 170 175ccg ctt gac gtc atc atc gat ctc aac atc gtg gcc gag acg gcg ttc576Pro Leu Asp Val Ile Ile Asp Leu Asn Ile Val Ala Glu Thr Ala Phe180 185 190acg aac atc ctg ctg gtg gcc act cca cag gtc gcc gtg gcc aac ggg624Thr Asn Ile Leu Leu Val Ala Thr Pro Gln Val Ala Val Ala Asn Gly195 200 205gac aac gcg atg gcc agc gtg ttc ctc tcg atc cag tcg gac gag gcc672Asp Asn Ala Met Ala Ser Val Phe Leu Ser Ile Gln Ser Asp Glu Ala210 215 220agg cac atg gcc aac ggg tac ggc tcg gtc atg gcg ctg ctg gag aac720Arg His Met Ala Asn Gly Tyr Gly Ser Val Met Ala Leu Leu Glu Asn225 230 235 240gag gac aac ctc ccg ctg ctc aac cag tct ctc gat cgg cac ttc tgg768Glu Asp Asn Leu Pro Leu Leu Asn Gln Ser Leu Asp Arg His Phe Trp245 250 255  
            
cgt gcc cac aag gcc ttg gac aac gcg gtc gga tgg tgt tcg gag tat816Arg Ala His Lys Ala Leu Asp Asn Ala Val Gly Trp Cys Ser Glu Tyr260 265 270ggc gcc cgc aag cgg cca tgg agc tac aag gcc cag tgg gag gaa tgg864Gly Ala Arg Lys Arg Pro Trp Ser Tyr Lys Ala Gln Trp Glu Glu Trp275 280 285gtc gtc gac gac ttc gtg ggc ggc tac atc gac cga ctc agc gag ttc912Val Val Asp Asp Phe Val Gly Gly Tyr Ile Asp Arg Leu Ser Glu Phe290 295 300ggc gtt cag gct ccg gcc tgc ctt ggc gcg gcc gcc gac gag gtc aag960Gly Val Gln Ala Pro Ala Cys Leu Gly Ala Ala Ala Asp Glu Val Lys305 310 315 320tgg tcg cac cac acg ctc ggt cag gtg ctg tcg gcg gtg tgg ccg ctg1008Trp Ser His His Thr Leu Gly Gln Val Leu Ser Ala Val Trp Pro Leu325 330 335aac ttc tgg cgc tcg gac gcc atg gga ccg gcg gac ttc gag tgg ttc1056Asn Phe Trp Arg Ser Asp Ala Met Gly Pro Ala Asp Phe Glu Trp Phe340 345 350gag aac cac tac ccg ggc tgg agc gcg gcc tac cag ggt tac tgg gag1104Glu Asn His Tyr Pro Gly Trp Ser Ala Ala Tyr Gln Gly Tyr Trp Glu355 360 365ggc tac aag gcg ctc gcc gac cca gca ggc gga cgc atc atg ctc cag1152Gly Tyr Lys Ala Leu Ala Asp Pro Ala Gly Gly Arg Ile Met Leu Gln370 375 380gag ctg ccg ggt ctg ccg ccg atg tgt cag gtg tgc cag gtg ccg tgc1200Glu Leu Pro Gly Leu Pro Pro Met Cys Gln Val Cys Gln Val Pro Cys385 390 395 400gtg atg ccg cgg ctg gat atg aac gcc gcg cgg atc atc gag ttc gag1248Val Met Pro Arg Leu Asp Met Asn Ala Ala Arg Ile Ile Glu Phe Glu405 410 415ggg cag aaa atc gcg ctg tgc agc gaa ccc tgc cag cgg atc ttc acc1296Gly Gln Lys Ile Ala Leu Cys Ser Glu Pro Cys Gln Arg Ile Phe Thr420 425 430  
            
aac tgg ccg gag gcg tac cgc cac cgc aag caa tac tgg gcc cgc tac1344Asn Trp Pro Glu Ala Tyr Arg His Arg Lys Gln Tyr Trp Ala Arg Tyr435 440 445cac gga tgg gac ctg gcg gac gtc atc gtt gat ctc ggc tac atc cgc1392His Gly Trp Asp Leu Ala Asp Val Ile Val Asp Leu Gly Tyr Ile Arg450 455 460ccg gac ggc aag acc ctc atc ggc cag ccg ctg ctc gag atg gag cgg1440Pro Asp Gly Lys Thr Leu Ile Gly Gln Pro Leu Leu Glu Met Glu Arg465 470 475 480ctg tgg acc atc gac gac atc cgg gcc ctt cag tac gaa gtc aag gac1488Leu Trp Thr Ile Asp Asp Ile Arg Ala Leu Gln Tyr Glu Val Lys Asp485 490 495ccg ttg cag gag gcg tga1506Pro Leu Gln Glu Ala500<210>12<211>501<212>PRT<213>珊瑚諾卡氏菌<400>12Met Ala Ser Asn Pro Thr Gln Leu His Glu Lys Ser Lys Ser Tyr Asp1   5   10  15Trp Asp Phe Thr Ser Val Glu Arg Arg Pro Lys Phe Glu Thr Lys Tyr20  25  30Lys Met Pro Lys Lys Gly Lys Asp Pro Phe Arg Val Leu Ile Arg Asp35 40 45Tyr Met Lys Met Glu Ala Glu Lys Asp Asp Arg Thr His Gly Phe Leu50 55 60Asp Gly Ala Val Arg Thr Arg Glu Ala Thr Arg Ile Glu Pro Arg Phe65  70  75  80Ala Glu Ala Met Lys Ile Met Val Pro Gln Leu Thr Asn Ala Glu Tyr  
            
85 90 95Gln Ala Val Ala Gly Cys Gly Met Ile Ile Ser Ala Val Glu Asn Gln100 105 110Glu Leu Arg Gln Gly Tyr Ala Ala Gln Met Leu Asp Glu Val Arg His115 120 125Ala Gln Leu Glu Met Thr Leu Arg Asn Tyr Tyr Ala Lys His Trp Cys130 135 140Asp Pro Ser Gly Phe Asp Ile Gly Gln Arg Gly Leu Tyr Gln His Pro145 150 155 160Ala Gly Leu Val Ser Ile Gly Glu Phe Gln His Phe Asn Thr Gly Asp165 170 175Pro Leu Asp Val Ile Ile Asp Leu Asn Ile Val Ala Glu Thr Ala Phe180 185 190Thr Asn Ile Leu Leu Val Ala Thr Pro Gln Val Ala Val Ala Asn Gly195 200 205Asp Asn Ala Met Ala Ser Val Phe Leu Ser Ile Gln Ser Asp Glu Ala210 215 220Arg His Met Ala Asn Gly Tyr Gly Ser Val Met Ala Leu Leu Glu Asn225 230 235 240Glu Asp Asn Leu Pro Leu Leu Asn Gln Ser Leu Asp Arg His Phe Trp245 250 255Arg Ala His Lys Ala Leu Asp Asn Ala Val Gly Trp Cys Ser Glu Tyr260 265 270Gly Ala Arg Lys Arg Pro Trp Ser Tyr Lys Ala Gln Trp Glu Glu Trp275 280 285Val Val Asp Asp Phe Val Gly Gly Tyr Ile Asp Arg Leu Ser Glu Phe290 295 300Gly Val Gln Ala Pro Ala Cys Leu Gly Ala Ala Ala Asp Glu Val Lys305 310 315 320  
            
Trp Ser His His Thr Leu Gly Gln Val Leu Ser Ala Val Trp Pro Leu325 330 335Asn Phe Trp Arg Ser Asp Ala Met Gly Pro Ala Asp Phe Glu Trp Phe340 345 350Glu Asn His Tyr Pro Gly Trp Ser Ala Ala Tyr Gln Gly Tyr Trp Glu355 360 365Gly Tyr Lys Ala Leu Ala Asp Pro Ala Gly Gly Arg Ile Met Leu Gln370 375 380Glu Leu Pro Gly Leu Pro Pro Met Cys Gln Val Cys Gln Val Pro Cys385 390 395 400Val Met Pro Arg Leu Asp Met Asn Ala Ala Arg Ile Ile Glu Phe Glu405 410 415Gly Gln Lys Ile Ala Leu Cys Ser Glu Pro Cys Gln Arg Ile Phe Thr420 425 430Asn Trp Pro Glu Ala Tyr Arg His Arg Lys Gln Tyr Trp Ala Arg Tyr435 440 445His Gly Trp Asp Leu Ala Asp Val Ile Val Asp Leu Gly Tyr Ile Arg450 455 460Pro Asp Gly Lys Thr Leu Ile Gly Gln Pro Leu Leu Glu Met Glu Arg465 470 475 480Leu Trp Thr Ile Asp Asp Ile Arg Ala Leu Gln Tyr Glu Val Lys Asp485 490 495Pro Leu Gln Glu Ala500<210>13<211>1494<212>DNA<213>黃色桿菌屬種<220>  
            
<221>CDS<222>(1)..(1494)<400>13atg gcg ctc ttg aat cgg gac gat tgg tac gac atc gcg cgc gat gtc48Met Ala Leu Leu Asn Arg Asp Asp Trp Tyr Asp Ile Ala Arg Asp Val1   5   10  15gac tgg acg ctc agc tat gtc gac cgc gcg gtc gcc ttt ccc gag gag96Asp Trp Thr Leu Ser Tyr Val Asp Arg Ala Val Ala Phe Pro Glu Glu20  25  30tgg aaa ggc gaa aag gac att tgc ggc acg gcc tgg gac gat tgg gac144Trp Lys Gly Glu Lys Asp Ile Cys Gly Thr Ala Trp Asp Asp Trp Asp35  40  45gag ccc ttc cgg gtc tcc ttc cgc gaa tat gtg atg gtc cag cgc gac192Glu Pro Phe Arg Val Ser Phe Arg Glu Tyr Val Met Val Gln Arg Asp50  55  60aag gaa gcg agc gtc ggc gcc atc cgc gag gcc atg gtc cgc gcc aag240Lys Glu Ala Ser Val Gly Ala Ile Arg Glu Ala Met Val Arg Ala Lys65  70  75  80gcc tat gag aag ctc gac gac ggc cac aag gcc acc tcg cac ctg cac288Ala Tyr Glu Lys Leu Asp Asp Gly His Lys Ala Thr Ser His Leu His85  90  95atg ggc acc atc acc atg gtg gag cac atg gcg gtc acc atg cag agc336Met Gly Thr Ile Thr Met Val Glu His Met Ala Val Thr Met Gln Ser100 105 110cgg ttc gtg cgc ttc gcg ccg tcc gcc cgc tgg cgc agc ctc ggg gcg384Arg Phe Val Arg Phe Ala Pro Ser Ala Arg Trp Arg Ser Leu Gly Ala115 120 125ttc ggc atg ctg gac gag acc cgc cac acc cag ctc gac ctg cgc ttc432Phe Gly Met Leu Asp Glu Thr Arg His Thr Gln Leu Asp Leu Arg Phe130 135 140agc cac gat ctg ctc aac gat tcc ccg agc ttc gac tgg agc cag cgg480Ser His Asp Leu Leu Asn Asp Ser Pro Ser Phe Asp Trp Ser Gln Arg145 150 155 160  
            
gcg ttc cac acc gac gaa tgg gcg gtt ctc gcc acc cgc aac ctg ttc528Ala Phe His Thr Asp Glu Trp Ala Val Leu Ala Thr Arg Asn Leu Phe165 170 175gac gac atc atg ctc aac gcc gac tgc gtg gag gcg gcg ctc gcc acc576Asp Asp Ile Met Leu Asn Ala Asp Cys Val Glu Ala Ala Leu Ala Thr180 185 190agc ctg acg ctg gag cac ggc ttc acc aac atc cag ttc gtg gcg ctc624Ser Leu Thr Leu Glu His Gly Phe Thr Asn Ile Gln Phe Val Ala Leu195 200 205gcc tcc gac gcc atg gaa gcc ggc gac gtg aac ttc tcc aac ctc ttg672Ala Ser Asp Ala Met Glu Ala Gly Asp Val Asn Phe Ser Asn Leu Leu210 215 220tcg agc atc cag acc gac gag gcg cgg cac gcc cag ttg ggc ttt ccc720Ser Ser Ile Gln Thr Asp Glu Ala Arg His Ala Gln Leu Gly Phe Pro225 230 235 240acc ctc gac gtg atg atg aag cac gac ccc aag cgc gcc cag cag atc768Thr Leu Asp Val Met Met Lys His Asp Pro Lys Arg Ala Gln Gln Ile245 250 255ctg gac gtc gcc ttc tgg cgc tcc tac cgc atc ttc cag gcg gtg acc816Leu Asp Val Ala Phe Trp Arg Ser Tyr Arg Ile Phe Gln Ala Val Thr260 265 270ggc gtc tcc atg gac tac tac acg ccg gtc gcc aag cgg cag atg tcg864Gly Val Ser Met Asp Tyr Tyr Thr Pro Val Ala Lys Arg Gln Met Ser275 280 285ttc aag gag ttc atg ctg gag tgg atc gtc aag cat cat gag cgc atc912Phe Lys Glu Phe Met Leu Glu Trp Ile Val Lys His His Glu Arg Ile290 295 300ctg cgc gac tac ggc ctc cag aag ccc tgg tac tgg gac acg ttc gag960Leu Arg Asp Tyr Gly Leu Gln Lys Pro Trp Tyr Trp Asp Thr Phe Glu305 310 315 320aag acc ctc gat cac ggc cac cac gcg ctg cac atc ggc acc tgg ttc1008Lys Thr Leu Asp His Gly His His Ala Leu His Ile Gly Thr Trp Phe325 330 335  
            
tgg cgc ccg acc ctg ttc tgg gat ccc aat ggc ggc gtc tcg cgc gag1056Trp Arg Pro Thr Leu Phe Trp Asp Pro Asn Gly Gly Val Ser Arg Glu340 345 350gag cgg cgc tgg ctg aac cag aag tat ccg aac tgg gaa gag agc tgg1104Glu Arg Arg Trp Leu Asn Gln Lys Tyr Pro Asn Trp Glu Glu Ser Trp355 360 365ggc gtc ctg tgg gac gag atc atc tcc aac atc aat gcg ggc aac att1152Gly Val Leu Trp Asp Glu Ile Ile Ser Asn Ile Asn Ala Gly Asn Ile370 375 380gaa aag acc ttg ccc gag acg ctg ccg atg ctg tgc aac gtc acc aac1200Glu Lys Thr Leu Pro Glu Thr Leu Pro Met Leu Cys Asn Val Thr Asn385 390 395 400ctg ccc atc ggc tcg cac tgg gac cgc ttc cac ctg aag ccc gag cag1248Leu Pro Ile Gly Ser His Trp Asp Arg Phe His Leu Lys Pro Glu Gln405 410 415ctc gtc tac aag ggg cgg ctc tac acc ttc gac agc gac gtc tcc aag1296Leu Val Tyr Lys Gly Arg Leu Tyr Thr Phe Asp Ser Asp Val Ser Lys420 425 430tgg atc ttc gag ctc gat ccg gag cgc tat gcc ggc cac acc aac gtg1344Trp Ile Phe Glu Leu Asp Pro Glu Arg Tyr Ala Gly His Thr Asn Val435 440 445gtc gac cgc ttc atc ggc ggg cag atc cag ccc atg acc atc gag ggc1392Val Asp Arg Phe Ile Gly Gly Gln Ile Gln Pro Met Thr Ile Glu Gly450 455 460gtg ctc aac tgg atg ggc ctg acg ccc gaa gtc atg ggc aag gac gtg1440Val Leu Asn Trp Met Gly Leu Thr Pro Glu Val Met Gly Lys Asp Val465 470 475 480ttc aac tac cgt tgg gcc ggc gat tac gcc gag aac cgg atc gcc gcc1488Phe Asn Tyr Arg Trp Ala Gly Asp Tyr Ala Glu Asn Arg Ile Ala Ala485 490 495gag taa1494Glu  
            
<210>14<211>497<212>PRT<213>黃色桿菌屬種<400>14Met Ala Leu Leu Asn Arg Asp Asp Trp Tyr Asp Ile Ala Arg Asp Val1   5   10  15Asp Trp Thr Leu Ser Tyr Val Asp Arg Ala Val Ala Phe Pro Glu Glu20  25  30Trp Lys Gly Glu Lys Asp Ile Cys Gly Thr Ala Trp Asp Asp Trp Asp35  40  45Glu Pro Phe Arg Val Ser Phe Arg Glu Tyr Val Met Val Gln Arg Asp50  55  60Lya Glu Ala Ser Val Gly Ala Ile Arg Glu Ala Met Val Arg Ala Lys65  70  75  80Ala Tyr Glu Lys Leu Asp Asp Gly His Lys Ala Thr Ser His Leu His85  90  95Met Gly Thr Ile Thr Met Val Glu His Met Ala Val Thr Met Gln Ser100 105 110Arg Phe Val Arg Phe Ala Pro Ser Ala Arg Trp Arg Ser Leu Gly Ala115 120 125Phe Gly Met Leu Asp Glu Thr Arg His Thr Gln Leu Asp Leu Arg Phe130 135 140Ser His Asp Leu Leu Asn Asp Ser Pro Ser Phe Asp Trp Ser Gln Arg145 150 155 160Ala Phe His Thr Asp Glu Trp Ala Val Leu Ala Thr Arg Asn Leu Phe165 170 175Asp Asp Ile Met Leu Asn Ala Asp Cys Val Glu Ala Ala Leu Ala Thr180 185 190  
            
Ser Leu Thr Leu Glu His Gly Phe Thr Asn Ile Gln Phe Val Ala Leu195 200 205Ala Ser Asp Ala Met Glu Ala Gly Asp Val Asn Phe Ser Asn Leu Leu210215 220Ser Ser Ile Gln Thr Asp Glu Ala Arg His Ala Gln Leu Gly Phe Pro225 230 235 240Thr Leu Asp Val Met Met Lys His Asp Pro Lys Arg Ala Gln Gln Ile245 250 255Leu Asp Val Ala Phe Trp Arg Ser Tyr Arg Ile Phe Gln Ala Val Thr260 265 270Gly Val Ser Met Asp Tyr Tyr Thr Pro Val Ala Lys Arg Gln Met Ser275 280 285Phe Lys Glu Phe Met Leu Glu Trp Ile Val Lys His His Glu Arg Ile290 295 300Leu Arg Asp Tyr Gly Leu Gln Lys Pro Trp Tyr Trp Asp Thr Phe Glu305 310 315 320Lys Thr Leu Asp His Gly His His Ala Leu His Ile Gly Thr Trp Phe325 330 335Trp Arg Pro Thr Leu Phe Trp Asp Pro Asn Gly Gly Val Ser Arg Glu340 345 350Glu Arg Arg Trp Leu Asn Gln Lys Tyr Pro Asn Trp Glu Glu Ser Trp355 360 365Gly Val Leu Trp Asp Glu Ile Ile Ser Asn Ile Asn Ala Gly Asn Ile370 375 380Glu Lys Thr Leu Pro Glu Thr Leu Pro Met Leu Cys Asn Val Thr Asn385 390 395 400Leu Pro Ile Gly Ser His Trp Asp Arg Phe His Leu Lys Pro Glu Gln405 410 415Leu Val Tyr Lys Gly Arg Leu Tyr Thr Phe Asp Ser Asp Val Ser Lys420 425 430  
            
Trp Ile Phe Glu Leu Asp Pro Glu Arg Tyr Ala Gly His Thr Asn Val435 440 445Val Asp Arg Phe Ile Gly Gly Gln Ile Gln Pro Met Thr Ile Glu Gly450 455 460Val Leu Asn Trp Met Gly Leu Thr Pro Glu Val Met Gly Lys Asp Val465 470 475 480Phe Asn Tyr Arg Trp Ala Gly Asp Tyr Ala Glu Asn Arg Ile Ala Ala485 490 495Glu<210>15<211>1026<212>DNA<213>黃色桿菌屬種<220>  
            
<221>CDS<222>(1)..(1026)<400>15atg aca cag cag cgc ccc acc cgc acg cgc gag cgc aag aag acc tgg48Met Thr Gln Gln Arg Pro Thr Arg Thr Arg Glu Arg Lys Lys Thr Trp1   5   10  15acg gct ttc ggc aat ctc gga cgc aag ccg acc gac tac gag gtc gtc96Thr Ala Phe Gly Asn Leu Gly Arg Lys Pro Thr Asp Tyr Glu Val Val20  25  30acc cac aac atg aac cac acc atg cgc ggc acg ccc ctg gag ctg tcg144Thr His Asn Met Asn His Thr Met Arg Gly Thr Pro Leu Glu Leu Ser35  40  45ccg acg gtg cac gcc aat gtg tgg ctc aag aag aac cgc gac gag atc192Pro Thr Val His Ala Asn Val Trp Leu Lys Lys Asn Arg Asp Glu Ile50  55  60  
            
gcg ctc aag gtc gac agc tgg gat ctg ttc cgc gat ccc gac cgc acc240Ala Leu Lys Val Asp Ser Trp Asp Leu Phe Arg Asp Pro Asp Arg Thr65  70  75  80acc tac gac acc tac gtc aag atg cag gac gac cag gag acc tat gtc288Thr Tyr Asp Thr Tyr Val Lys Met Gln Asp Asp Gln Glu Thr Tyr Val85  90  95gac aac ctg ctc ctg tcc tac acc ggc gag ggc cgc tac gac gag gag336Asp Asn Leu Leu Leu Ser Tyr Thr Gly Glu Gly Arg Tyr Asp Glu Glu100 105 110ctt tcc tcg cgc agc ctc gac ctc ctg tcc gcg ggg ctg acg ccg acc384Leu Ser Ser Arg Ser Leu Asp Leu Leu Ser Ala Gly Leu Thr Pro Thr115 120 125cgc tat ctg ggc cat ggg ctg cag atg ctc gcg gcc tat atc cag cag432Arg Tyr Leu Gly His Gly Leu Gln Met Leu Ala Ala Tyr Ile Gln Gln130 135 140ctc gcc ccg tcg gcc tat gtg ggc aat tgc gcg gtg ttc cag acc tcc480Leu Ala Pro Ser Ala Tyr Val Gly Asn Cys Ala Val Phe Gln Thr Ser145 150 155 160gac gcg ctg cgc cgc gtg cag cgc gtc gcc tac cgc acc cgc cag ctc528Asp Ala Leu Arg Arg Val Gln Arg Val Ala Tyr Arg Thr Arg Gln Leu165 170 175gcc gac gcc cat ccg gcc cgc ggc ttc ggc tcc ggc gac cgg gcg gtg576Ala Asp Ala His Pro Ala Arg Gly Phe Gly Ser Gly Asp Arg Ala Val180 185 190tgg gag aag tcc ccg gac tgg cag ccc atc cgc aag gcc atc gag gag624Trp Glu Lys Ser Pro Asp Trp Gln Pro Ile Arg Lys Ala Ile Glu Glu195 200 205ctg ctc gtc acc ttc gaa tgg gac aag gcg ctc gcc ggc acc aat ttc672Leu Leu Val Thr Phe Glu Trp Asp Lys Ala Leu Ala Gly Thr Asn Phe210 215 220gtg gtg aag ccg atc ctc gac gag ctg ttc ctc aac cac ctg gcg cgc720Val Val Lys Pro Ile Leu Asp Glu Leu Phe Leu Asn His Leu Ala Arg225 230 235 240  
            
ctg ctc cac gtg gag ggc gac gag ctc gac agc ctc gtg ctg cgg aac768Leu Leu His Val Glu Gly Asp Glu Leu Asp Ser Leu Val Leu Arg Asn245 250 255ctt cac ggc gac gcc cag cgc cac gcc cgc tgg acg gcc gcg ctc ggc816Leu His Gly Asp Ala Gln Arg His Ala Arg Trp Thr Ala Ala Leu Gly260 265 270cgc ttc gcc gtc gag cag aac gtg aac aac cgc acg gtc ctg cgc gac864Arg Phe Ala Val Glu Gln Asn Val Asn Asn Arg Thr Val Leu Arg Asp275 280 285gcc atc gcc ggc tgg cac gag acc ggc gag gcg gtc ctc gcc gcg ggc912Ala Ile Ala Gly Trp His Glu Thr Gly Glu Ala Val Leu Ala Ala Gly290 295 300gcc ggg atg ctt gcg agc cgc gcc ccc agc gcg gat gcg gcc aag atc960Ala Gly Met Leu Ala Ser Arg Ala Pro Ser Ala Asp Ala Ala Lys Ile305 310 315 320gcc gac gag gtc cgc gcc acg ctc gcg cag ctg cac gcc aat gcg ggc1008Ala Asp Glu Val Arg Ala Thr Leu Ala Gln Leu His Ala Asn Ala Gly325 330 335ctc ggg cac gat gcc tga1026Leu Gly His Asp Ala340<210>16<211>341<212>PRT<213>黃色桿菌屬種<400>16Met Thr Gln Gln Arg Pro Thr Arg Thr Arg Glu Arg Lys Lys Thr Trp1   5   10  15Thr Ala Phe Gly Asn Leu Gly Arg Lys Pro Thr Asp Tyr Glu Val Val20  25  30Thr His Asn Met Asn His Thr Met Arg Gly Thr Pro Leu Glu Leu Ser35  40  45  
            
Pro Thr Val His Ala Asn Val Trp Leu Lys Lys Asn Arg Asp Glu Ile50  55  60Ala Leu Lys Val Asp Ser Trp Asp Leu Phe Arg Asp Pro Asp Arg Thr65  70  75  80Thr Tyr Asp Thr Tyr Val Lys Met Gln Asp Asp Gln Glu Thr Tyr Val85  90  95Asp Asn Leu Leu Leu Ser Tyr Thr Gly Glu Gly Arg Tyr Asp Glu Glu100 105 110Leu Ser Ser Arg Ser Leu Asp Leu Leu Ser Ala Gly Leu Thr Pro Thr115 120 125Arg Tyr Leu Gly His Gly Leu Gln Met Leu Ala Ala Tyr Ile Gln Gln130 135 140Leu Ala Pro Ser Ala Tyr Val Gly Asn Cys Ala Val Phe Gln Thr Ser145 150 155 160Asp Ala Leu Arg Arg Val Gln Arg Val Ala Tyr Arg Thr Arg Gln Leu165 170 175Ala Asp Ala His Pro Ala Arg Gly Phe Gly Ser Gly Asp Arg Ala Val180 185 190Trp Glu Lys Ser Pro Asp Trp Gln Pro Ile Arg Lys Ala Ile Glu Glu195 200 205Leu Leu Val Thr Phe Glu Trp Asp Lys Ala Leu Ala Gly Thr Asn Phe210 215 220Val Val Lys Pro Ile Leu Asp Glu Leu Phe Leu Asn His Leu Ala Arg225 230 235 240Leu Leu His Val Glu Gly Asp Glu Leu Asp Ser Leu Val Leu Arg Asn245 250 255Leu His Gly Asp Ala Gln Arg His Ala Arg Trp Thr Ala Ala Leu Gly260 265 270Arg Phe Ala Val Glu Gln Asn Val Asn Asn Arg Thr Val Leu Arg Asp  
            
275 280 285Ala Ile Ala Gly Trp His Glu Thr Gly Glu Ala Val Leu Ala Ala Gly290 295 300Ala Gly Met Leu Ala Ser Arg Ala Pro Ser Ala Asp Ala Ala Lys Ile305 310 315 320Ala Asp Glu Val Arg Ala Thr Leu Ala Gln Leu His Ala Asn Ala Gly325 330 335Leu Gly His Asp Ala340<210>17<211>267<212>DNA<213>黃色桿菌屬種<220>  
            
<221>CDS<222>(1)..(267)<400>17atg tct ttg ttc ccc atc gtg ggc cgc ttc gtg ggg gat ttc gtc ccc48Met Ser Leu Phe Pro Ile Val Gly Arg Phe Val Gly Asp Phe Val Pro1   5   10  15cac ctg gtg gcg gtg gac acc tct gac acc atc gat cag atc gcc gag96His Leu Val Ala Val Asp Thr Ser Asp Thr Ile Asp Gln Ile Ala Glu20  25  30aag gtg gcg gtc cac acg gtc ggg cgg cgc ttg ccg ccc gat ccc acc144Lys Val Ala Val His Thr Val Gly Arg Arg Leu Pro Pro Asp Pro Thr35  40  45gcc acc ggc tat gag gtg ctc ctc gac ggc gag acc ctg gac ggg ggc192Ala Thr Gly Tyr Glu Val Leu Leu Asp Gly Glu Thr Leu Asp Gly Gly50  55  60gcc acc ctg gag gcc atc atg acc aag cgc gag atg ctg ccc ctg cag240Ala Thr Leu Glu Ala Ile Met Thr Lys Arg Glu Met Leu Pro Leu Gln  
            
65  70  75  80tgg ttc gac gtg agg ttc aag aag tga  267Trp Phe Asp Val Arg Phe Lys Lys85<210>18<211>88<212>PRT<213>黃色桿菌屬種<400>18Met Ser Leu Phe Pro Ile Val Gly Arg Phe Val Gly Asp Phe Val Pro1   5   10  15His Leu Val Ala Val Asp Thr Ser Asp Thr Ile Asp Gln Ile Ala Glu20  25  30Lys Val Ala Val His Thr Val Gly Arg Arg Leu Pro Pro Asp Pro Thr35  40  45Ala Thr Gly Tyr Glu Val Leu Leu Asp Gly Glu Thr Leu Asp Gly Gly50  55  60Ala Thr Leu Glu Ala Ile Met Thr Lys Arg Glu Met Leu Pro Leu Gln65  70  75  80Trp Phe Asp Val Arg Phe Lys Lys85<210>19<211>1584<212>DNA<213>莢膜甲基球菌<220>  
            
<221>CDS<222>(1)..(1584)<400>19  
            
atg gca ctt agc acc gca acc aag gcc gcg acg gac gcg ctg gct gcc48Met Ala Leu Ser Thr Ala Thr Lys Ala Ala Thr Asp Ala Leu Ala Ala1   5   10  15aat cgg gca ccc acc agc gtg aat gca cag gaa gtg cac cgt tgg ctc96Asn Arg Ala Pro Thr Ser Val Asn Ala Gln Glu Val His Arg Trp Leu20  25  30cag agc ttc aac tgg gat ttc aag aac aac cgg acc aag tac gcc acc144Gln Ser Phe Asn Trp Asp Phe Lys Asn Asn Arg Thr Lys Tyr Ala Thr35  40  45aag tac aag atg gcg aac gag acc aag gaa cag ttc aag ctg atc gcc192Lys Tyr Lys Met Ala Asn Glu Thr Lys Glu Gln Phe Lys Leu Ile Ala50  55  60aag gaa tat gcg cgc atg gag gca gtc aag gac gaa agg cag ttc ggt240Lys Glu Tyr Ala Arg Met Glu Ala Val Lys Asp Glu Arg Gln Phe Gly65  70  75  80agc ctg cag gat gcg ctg acc cgc ctc aac gcc ggt gtt cgc gtt cat288Ser Leu Gln Asp Ala Leu Thr Arg Leu Asn Ala Gly Val Arg Val His85  90  95ccg aag tgg aac gag acc atg aaa gtg gtt tcg aac ttc ctg gaa gtg336Pro Lys Trp Asn Glu Thr Met Lys Val Val Ser Asn Phe Leu Glu Val100 105 110ggc gaa tac aac gcc atc gcc gct acc ggg atg ctg tgg gat tcc gcc384Gly Glu Tyr Asn Ala Ile Ala Ala Thr Gly Met Leu Trp Asp Ser Ala115 120 125cag gcg gcg gaa cag aag aac ggc tat ctg gcc cag gtg ttg gat gaa432Gln Ala Ala Glu Gln Lys Asn Gly Tyr Leu Ala Gln Val Leu Asp Glu130 135 140atc cgc cac acc cac cag tgt gcc tac gtc aac tac tac ttc gcg aag480Ile Arg His Thr His Gln Cys Ala Tyr Val Asn Tyr Tyr Phe Ala Lys145 150 155 160aac ggc cag gac ccg gcc ggt cac aac gat gct cgc cgc acc cgt acc528Asn Gly Gln Asp Pro Ala Gly His Asn Asp Ala Arg Arg Thr Arg Thr165 170 175  
            
atc ggt ccg ctg tgg aag ggc atg aag cgc gtg ttt tcc gac ggc ttc576Ile Gly Pro Leu Trp Lys Gly Met Lys Arg Val Phe Ser Asp Gly Phe180 185 190att tcc ggc gac gcc gtg gaa tgc tcc ctc aac ctg cag ctg gtg ggt624Ile Ser Gly Asp Ala Val Glu Cys Ser Leu Asn Leu Gln Leu Val Gly195 200 205gag gcc tgc ttc acc aat ccg ctg atc gtc gca gtg acc gaa tgg gct672Glu Ala Cys Phe Thr Asn Pro Leu Ile Val Ala Val Thr Glu Trp Ala210 215 220gcc gcc aac ggc gat gaa atc acc ccg acg gtg ttc ctg tcg atc gag720Ala Ala Asn Gly Asp Glu Ile Thr Pro Thr Val Phe Leu Ser Ile Glu225 230 235 240acc gac gaa ctg cgc cac atg gcc aac ggt tac cag acc gtc gtt tcc768Thr Asp Glu Leu Arg His Met Ala Asn Gly Tyr Gln Thr Val Val Ser245 250 255atc gcc aac gat ccg gct tcc gcc aag tat ctc aac acg gac ctg aac816Ile Ala Asn Asp Pro Ala Ser Ala Lys Tyr Leu Asn Thr Asp Leu Asn260 265 270aac gcc ttc tgg acc cag cag aag tac ttc acg ccg gtg ttg ggc atg864Asn Ala Phe Trp Thr Gln Gln Lys Tyr Phe Thr Pro Val Leu Gly Met275 280 285ctg ttc gag tat ggc tcc aag ttc aag gtc gag ccg tgg gtc aag acg912Leu Phe Glu Tyr Gly Ser Lys Phe Lys Val Glu Pro Trp Val Lys Thr290 295 300tgg gac cgc tgg gtg tac gag gac tgg ggc ggc atc tgg atc ggc cgt960Trp Asp Arg Trp Val Tyr Glu Asp Trp Gly Gly Ile Trp Ile Gly Arg305 310 315 320ctg ggc aag tac ggg gtg gag tcg ccg cgc agc ctc aag gac gcc aag1008Leu Gly Lys Tyr Gly Val Glu Ser Pro Arg Ser Leu Lys Asp Ala Lys325 330 335cag gac gct tac tgg gct cac cac gac ctg tat ctg ctg gct tat gcg1056Gln Asp Ala Tyr Trp Ala His His Asp Leu Tyr Leu Leu Ala Tyr Ala340 345 350  
            
ctg tgg ccg acc ggc ttc ttc cgt ctg gcg ctg ccg gat cag gaa gaa1104Leu Trp Pro Thr Gly Phe Phe Arg Leu Ala Leu Pro Asp Gln Glu Glu355 360 365atg gag tgg ttc gag gcc aac tac ccc ggc tgg tac gac cac tac ggc1152Met Glu Trp Phe Glu Ala Asn Tyr Pro Gly Trp Tyr Asp His Tyr Gly370 375 380aag atc tac gag gaa tgg cgc gcc cgc ggt tgc gag gat ccg tcc tcg1200Lys Ile Tyr Glu Glu Trp Arg Ala Arg Gly Cys Glu Asp Pro Ser Ser385 390 395 400ggc ttc atc ccg ctg atg tgg ttc atc gaa aac aac cat ccc atc tac1248Gly Phe Ile Pro Leu Met Trp Phe Ile Glu Asn Asn His Pro Ile Tyr405 410 415atc gat cgc gtg tcg caa gtg ccg ttc tgc ccg agc ttg gcc aag ggc1296Ile Asp Arg Val Ser Gln Val Pro Phe Cys Pro Ser Leu Ala Lys Gly420 425 430gcc agc acc ctg cgc gtg cac gag tac aac ggc gag atg cac acc ttc1344Ala Ser Thr Leu Arg Val His Glu Tyr Asn Gly Glu Met His Thr Phe435 440 445agc gac cag tgg ggc gag cgc atg tgg ctg gcc gag ccg gag cgc tac1392Ser Asp Gln Trp Gly Glu Arg Met Trp Leu Ala Glu Pro Glu Arg Tyr450 455 460gag tgc cag aac atc ttc gaa cag tac gaa gga cgc gaa ctg tcg gaa1440Glu Cys Gln Asn Ile Phe Glu Gln Tyr Glu Gly Arg Glu Leu Ser Glu465 470 475 480gtg atc gcc gaa ctg cac ggg ctg cgc agt gat ggc aag acc ctg atc1488Val Ile Ala Glu Leu His Gly Leu Arg Ser Asp Gly Lys Thr Leu Ile485 490 495gcc cag ccg cat gtc cgt ggc gac aag ctg tgg acg ttg gac gat atc1536Ala Gln Pro His Val Arg Gly Asp Lys Leu Trp Thr Leu Asp Asp Ile500 505 510aaa cgc ctg aac tgc gtc ttc aag aac ccg gtg aag gca ttc aat tga1584Lys Arg Leu Asn Cys Val Phe Lys Asn Pro Val Lys Ala Phe Asn515 520 525  
            
<210>20<21l>527<212>PRT<213>莢膜甲基球菌<400>20Met Ala Leu Ser Thr Ala Thr Lys Ala Ala Thr Asp Ala Leu Ala Ala1   5   10  15Asn Arg Ala Pro Thr Ser Val Asn Ala Gln Glu Val His Arg Trp Leu20  25  30Gln Ser Phe Asn Trp Asp Phe Lys Asn Asn Arg Thr Lys Tyr Ala Thr35  40  45Lys Tyr Lys Met Ala Asn Glu Thr Lys Glu Gln Phe Lys Leu Ile Ala50  55  60Lys Glu Tyr Ala Arg Met Glu Ala Val Lys Asp Glu Arg Gln Phe Gly65  70  75  80Ser Leu Gln Asp Ala Leu Thr Arg Leu Asn Ala Gly Val Arg Val His85  90  95Pro Lys Trp Asn Glu Thr Met Lys Val Val Ser Asn Phe Leu Glu Val100 105 110Gly Glu Tyr Asn Ala Ile Ala Ala Thr Gly Met Leu Trp Asp Ser Ala115 120 125Gln Ala Ala Glu Gln Lys Asn Gly Tyr Leu Ala Gln Val Leu Asp Glu130 135 140Ile Arg His Thr His Gln Cys Ala Tyr Val Asn Tyr Tyr Phe Ala Lys145 150 155 160Asn Gly Gln Asp Pro Ala Gly His Asn Asp Ala Arg Arg Thr Arg Thr165 170 175Ile Gly Pro Leu Trp Lys Gly Met Lys Arg Val Phe Ser Asp Gly Phe180 185 190Ile Ser Gly Asp Ala Val Glu Cys Ser Leu Asn Leu Gln Leu Val Gly  
            
195 200 205Glu Ala Cys Phe Thr Asn Pro Leu Ile Val Ala Val Thr Glu Trp Ala210 215 220Ala Ala Asn Gly Asp Glu Ile Thr Pro Thr Val Phe Leu Ser Ile Glu225 230 235 240Thr Asp Glu Leu Arg His Met Ala Asn Gly Tyr Gln Thr Val Val Ser245 250 255Ile Ala Asn Asp Pro Ala Ser Ala Lys Tyr Leu Asn Thr Asp Leu Asn260 265 270Asn Ala Phe Trp Thr Gln Gln Lys Tyr Phe Thr Pro Val Leu Gly Met275 280 285Leu Phe Glu Tyr Gly Ser Lys Phe Lys Val Glu Pro Trp Val Lys Thr290 295 300Trp Asp Arg Trp Val Tyr Glu Asp Trp Gly Gly Ile Trp Ile Gly Arg305 310 315 320Leu Gly Lys Tyr Gly Val Glu Ser Pro Arg Ser Leu Lys Asp Ala Lys325 330 335Gln Asp Ala Tyr Trp Ala His His Asp Leu Tyr Leu Leu Ala Tyr Ala340 345 350Leu Trp Pro Thr Gly Phe Phe Arg Leu Ala Leu Pro Asp Gln Glu Glu355 360 365Met Glu Trp Phe Glu Ala Asn Tyr Pro Gly Trp Tyr Asp His Tyr Gly370 375 380Lys Ile Tyr Glu Glu Trp Arg Ala Arg Gly Cys Glu Asp Pro Ser Ser385 390 395 400Gly Phe Ile Pro Leu Met Trp Phe Ile Glu Asn Asn His Pro Ile Tyr405 410 415Ile Asp Arg Val Ser Gln Val Pro Phe Cys Pro Ser Leu Ala Lys Gly420 425 430  
            
Ala Ser Thr Leu Arg Val His Glu Tyr Asn Gly Glu Met His Thr Phe435 440 445Ser Asp Gln Trp Gly Glu Arg Met Trp Leu Ala Glu Pro Glu Arg Tyr450 455 460Glu Cys Gln Asn Ile Phe Glu Gln Tyr Glu Gly Arg Glu Leu Ser Glu465 470 475 480Val Ile Ala Glu Leu His Gly Leu Arg Ser Asp Gly Lys Thr Leu Ile485 490 495Ala Gln Pro His Val Arg Gly Asp Lys Leu Trp Thr Leu Asp Asp Ile500 505 510Lys Arg Leu Asn Cys Val Phe Lys Asn Pro Val Lys Ala Phe Asn515 520 525<210>21<211>1170<212>DNA<213>莢膜甲基球菌<220>  
            
<221>CDS<222>(1)..(1170)<400>21atg agc atg tta gga gaa aga cgc cgc ggt ctg acc gat ccg gaa atg48Met Ser Met Leu Gly Glu Arg Arg Arg Gly Leu Thr Asp Pro Glu Met1   5   10  15gcg gcc gtc att ttg aag gcg ctt cct gaa gct ccg ctg gac ggc aac96Ala Ala Val Ile Leu Lys Ala Leu Pro Glu Ala Pro Leu Asp Gly Asn20  25  30aac aag atg ggt tat ttc gtc acc ccc cgc tgg aaa cgc ttg acg gaa144Asn Lys Met Gly Tyr Phe Val Thr Pro Arg Trp Lys Arg Leu Thr Glu35  40  45tat gaa gcc ctg acc gtt tat gcg cag ccc aac gcc gac tgg atc gcc192Tyr Glu Ala Leu Thr Val Tyr Ala Gln Pro Asn Ala Asp Trp Ile Ala  
            
50  55  60ggc ggc ctg gac tgg ggc gac tgg acc cag aaa ttc cac ggc ggc cgc240Gly Gly Leu Asp Trp Gly Asp Trp Thr Gln Lys Phe His Gly Gly Arg65  70  75  80cct tcc tgg ggc aac gag acc acg gag ctg cgc acc gtc gac tgg ttc288Pro Ser Trp Gly Asn Glu Thr Thr Glu Leu Arg Thr Val Asp Trp Phe85  90  95aag cac cgt gac ccg ctc cgc cgt tgg cat gcg ccg tac gtc aag gac336Lys His Arg Asp Pro Leu Arg Arg Trp His Ala Pro Tyr Val Lys Asp100 105 110aag gcc gag gaa tgg cgc tac acc gac cgc ttc ctg cag ggt tac tcc384Lys Ala Glu Glu Trp Arg Tyr Thr Asp Arg Phe Leu Gln Gly Tyr Ser115 120 125gcc gac ggt cag atc cgg gcg atg aac ccg acc tgg cgg gac gag ttc432Ala Asp Gly Gln Ile Arg Ala Met Asn Pro Thr Trp Arg Asp Glu Phe130 135 140atc aac cgg tat tgg ggc gcc ttc ctg ttc aac gaa tac gga ttg ttc480Ile Asn Arg Tyr Trp Gly Ala Phe Leu Phe Asn Glu Tyr Gly Leu Phe145 150 155 160aac gct cat tcg cag ggc gcc cgg gag gcg ctg tcg gac gta acc cgc528Asn Ala His Ser Gln Gly Ala Arg Glu Ala Leu Ser Asp Val Thr Arg165 170 175gtc agc ctg gct ttc tgg ggc ttc gac aag atc gac atc gcc cag atg576Val Ser Leu Ala Phe Trp Gly Phe Asp Lys Ile Asp Ile Ala Gln Met180 185 190atc caa ctc gaa cgg ggt ttc ctc gcc aag atc gta ccc ggt ttc gac624Ile Gln Leu Glu Arg Gly Phe Leu Ala Lys Ile Val Pro Gly Phe Asp195 200 205gag tcc aca gcg gtg ccg aag gcc gaa tgg acg aac ggg gag gtc tac672Glu Ser Thr Ala Val Pro Lys Ala Glu Trp Thr Asn Gly Glu Val Tyr210 215 220aag agc gcc cgt ctg gcc gtg gaa ggg ctg tgg cag gag gtg ttc gac720Lys Ser Ala Arg Leu Ala Val Glu Gly Leu Trp Gln Glu Val Phe Asp  
            
225 230 235 240tgg aac gag agc gct ttc tcg gtg cac gcc gtc tat gac gcg ctg ttc768Trp Asn Glu Ser Ala Phe Ser Val His Ala Val Tyr Asp Ala Leu Phe245 250 255ggt cag ttc gtc cgc cgc gag ttc ttt cag cgg ctg gct ccc cgc ttc816Gly Gln Phe Val Arg Arg Glu Phe Phe Gln Arg Leu Ala Pro Arg Phe260 265 270ggc gac aat ctg acg cca ttc ttc atc aac cag gcc cag aca tac ttc864Gly Asp Asn Leu Thr Pro Phe Phe Ile Asn Gln Ala Gln Thr Tyr Phe275 280 285cag atc gcc aag cag ggc gta cag gat ctg tat tac aac tgt ctg ggt912Gln Ile Ala Lys Gln Gly Val Gln Asp Leu Tyr Tyr Asn Cys Leu Gly290 295 300gac gat ccg gag ttc agc gat tac aac cgt acc gtg atg cgc aac tgg960Asp Asp Pro Glu Phe Ser Asp Tyr Asn Arg Thr Val Met Arg Asn Trp305 310 315 320acc ggc aag tgg ctg gag ccc acg atc gcc gct ctg cgc gac ttc atg1008Thr Gly Lys Trp Leu Glu Pro Thr Ile Ala Ala Leu Arg Asp Phe Met325 330 335ggg ctg ttt gcg aag ctg ccg gcg ggc acc act gac aag gaa gaa atc1056Gly Leu Phe Ala Lys Leu Pro Ala Gly Thr Thr Asp Lys Glu Glu Ile340 345 350acc gcg tcc ctg tac cgg gtg gtc gac gac tgg atc gag gac tac gcc1104Thr Ala Ser Leu Tyr Arg Val Val Asp Asp Trp Ile Glu Asp Tyr Ala355 360 365agc gcg atc gac ttc aag gcg gac cgc gat cag atc gtt aaa gcg gtt1152Ser Ala Ile Asp Phe Lys Ala Asp Arg Asp Gln Ile Val Lys Ala Val370 375 380ctg gca gga ttg aaa taa1170Leu Ala Gly Leu Lys385<210>22  
            
<211>389<212>PRT<213>莢膜甲基球菌<400>22Met Ser Met Leu Gly Glu Arg Arg Arg Gly Leu Thr Asp Pro Glu Met1   5   10  15Ala Ala Val Ile Leu Lys Ala Leu Pro Glu Ala Pro Leu Asp Gly Asn20  25  30Asn Lys Met Gly Tyr Phe Val Thr Pro Arg Trp Lys Arg Leu Thr Glu35  40  45Tyr Glu Ala Leu Thr Val Tyr Ala Gln Pro Asn Ala Asp Trp Ile Ala50  55  60Gly Gly Leu Asp Trp Gly Asp Trp Thr Gln Lys Phe His Gly Gly Arg65  70  75  80Pro Ser Trp Gly Asn Glu Thr Thr Glu Leu Arg Thr Val Asp Trp Phe85  90  95Lys His Arg Asp Pro Leu Arg Arg Trp His Ala Pro Tyr Val Lys Asp100 105 110Lys Ala Glu Glu Trp Arg Tyr Thr Asp Arg Phe Leu Gln Gly Tyr Ser115 120 125Ala Asp Gly Gln Ile Arg Ala Met Asn Pro Thr Trp Arg Asp Glu Phe130 135 140Ile Asn Arg Tyr Trp Gly Ala Phe Leu Phe Asn Glu Tyr Gly Leu Phe145 150 155 160Asn Ala His Ser Gln Gly Ala Arg Glu Ala Leu Ser Asp Val Thr Arg165 170 175Val Ser Leu Ala Phe Trp Gly Phe Asp Lys Ile Asp Ile Ala Gln Met180 185 190Ile Gln Leu Glu Arg Gly Phe Leu Ala Lys Ile Val Pro Gly Phe Asp195 200 205  
            
Glu Ser Thr Ala Val Pro Lys Ala Glu Trp Thr Asn Gly Glu Val Tyr210 215 220Lys Ser Ala Arg Leu Ala Val Glu Gly Leu Trp Gln Glu Val Phe Asp225 230 235 240Trp Asn Glu Ser Ala Phe Ser Val His Ala Val Tyr Asp Ala Leu Phe245 250 255Gly Gln Phe Val Arg Arg Glu Phe Phe Gln Arg Leu Ala Pro Arg Phe260 265 270Gly Asp Asn Leu Thr Pro Phe Phe Ile Asn Gln Ala Gln Thr Tyr Phe275 280 285Gln Ile Ala Lys Gln Gly Val Gln Asp Leu Tyr Tyr Asn Cys Leu Gly290 295 300Asp Asp Pro Glu Phe Ser Asp Tyr Asn Arg Thr Val Met Arg Asn Trp305 310 315 320Thr Gly Lys Trp Leu Glu Pro Thr Ile Ala Ala Leu Arg Asp Phe Met325 330 335Gly Leu Phe Ala Lys Leu Pro Ala Gly Thr Thr Asp Lys Glu Glu Ile340 345 350Thr Ala Ser Leu Tyr Arg Val Val Asp Asp Trp Ile Glu Asp Tyr Ala355 360 365Ser Ala Ile Asp Phe Lys Ala Asp Arg Asp Gln Ile Val Lys Ala Val370 375 380Leu Ala Gly Leu Lys385<210>23<211>513<212>DNA<213>莢膜甲基球菌  
            
<220>  
            
<221>CDS<222>(1)..(513)<400>23atg gcg aaa ctg ggt ata cac agc aac gac acc cgc gac gcc tgg gtg48Met Ala Lys Leu Gly Ile His Ser Asn Asp Thr Arg Asp Ala Trp Val1   5   10  15aac aag atc gcg cag ctc aac acc ctg gaa aaa gcg gcc gag atg ctg96Asn Lys Ile Ala Gln Leu Asn Thr Leu Glu Lys Ala Ala Glu Met Leu20  25  30aag cag ttc cgg atg gac cac acc acg ccg ttc cgc aac agc tac gaa144Lys Gln Phe Arg Met Asp His Thr Thr Pro Phe Arg Asn Ser Tyr Glu35  40  45ctg gac aac gac tac ctc tgg atc gag gcc aag ctc gaa gag aag gtc192Leu Asp Asn Asp Tyr Leu Trp Ile Glu Ala Lys Leu Glu Glu Lys Val50  55  60gcc gtc ctc aag gca cgc gcc ttc aac gag gtg gac ttc cgt cat aag240Ala Val Leu Lys Ala Arg Ala Phe Asn Glu Val Asp Phe Arg His Lys65  70  75  80acc gct ttc ggc gag gat gcc aag tcc gtt ctg gac ggc acc gtc gcg288Thr Ala Phe Gly Glu Asp Ala Lys Ser Val Leu Asp Gly Thr Val Ala85  90  95aag atg aac gcg gcc aag gac aag tgg gag gcg gag aag atc cat atc336Lys Met Asn Ala Ala Lys Asp Lys Trp Glu Ala Glu Lys Ile His Ile100 105 110ggt ttc cgc cag gcc tac aag ccg ccg atc atg ccg gtg aac tat ttc384Gly Phe Arg Gln Ala Tyr Lys Pro Pro Ile Met Pro Val Asn Tyr Phe115 120 125ctg gac ggc gag cgt cag ttg ggg acc cgg ctg atg gaa ctg cgc aac432Leu Asp Gly Glu Arg Gln Leu Gly Thr Arg Leu Met Glu Leu Arg Asn130 135 140ctc aac tac tac gac acg ccg ctg gaa gaa ctg cgc aaa cag cgc ggt480Leu Asn Tyr Tyr Asp Thr Pro Leu Glu Glu Leu Arg Lys Gln Arg Gly145 150 155 160  
            
gtg cgg gtg gtg cat ctg cag tcg ccg cac tga 513Val Arg Val Val His Leu Gln Ser Pro His165 170<210>24<211>170<212>PRT<213>莢膜甲基球菌<400>24Met Ala Lys Leu Gly Ile His Ser Asn Asp Thr Arg Asp Ala Trp Val1   5   10  15Asn Lys Ile Ala Gln Leu Asn Thr Leu Glu Lys Ala Ala Glu Met Leu20  25  30Lys Gln Phe Arg Met Asp His Thr Thr Pro Phe Arg Asn Ser Tyr Glu35  40  45Leu Asp Asn Asp Tyr Leu Trp Ile Glu Ala Lys Leu Glu Glu Lys Val50  55  60Ala Val Leu Lys Ala Arg Ala Phe Asn Glu Val Asp Phe Arg His Lys65  70  75  80Thr Ala Phe Gly Glu Asp Ala Lys Ser Val Leu Asp Gly Thr Val Ala85  90  95Lys Met Asn Ala Ala Lys Asp Lys Trp Glu Ala Glu Lys Ile His Ile100 105 110Gly Phe Arg Gln Ala Tyr Lys Pro Pro Ile Met Pro Val Asn Tyr Phe115 120 125Leu Asp Gly Glu Arg Gln Leu Gly Thr Arg Leu Met Glu Leu Arg Asn130 135 140Leu Asn Tyr Tyr Asp Thr Pro Leu Glu Glu Leu Arg Lys Gln Arg Gly145 150 155 160Val Arg Val Val His Leu Gln Ser Pro His165 170  
            
<210>25<211>1206<212>DNA<213>食油假單胞菌<220>  
            
<221>CDS<222>(1)..(1206)<400>25atg ctt gag aaa cac aga gtt ctg gat tcc gct cca gag tac gta gat48Met Leu Glu Lys His Arg Val Leu Asp Ser Ala Pro Glu Tyr Val Asp1   5   10  15aaa aag aaa tat ctc tgg ata cta tca act ttg tgg ccg gct act ccg96Lys Lys Lys Tyr Leu Trp Ile Leu Ser Thr Leu Trp Pro Ala Thr Pro20  25  30atg atc gga atc tgg ctt gca aat gaa act ggt tgg ggg att ttt tat144Met Ile Gly Ile Trp Leu Ala Asn Glu Thr Gly Trp Gly Ile Phe Tyr35  40  45ggg ctg gta ttg ctc gta tgg tac ggc gca ctt cca ttg ctt gat gcg192Gly Leu Val Leu Leu Val Trp Tyr Gly Ala Leu Pro Leu Leu Asp Ala50  55  60atg ttt ggt gag gac ttt aat aat ccg cct gaa gaa gtg gtg ccg aaa240Met Phe Gly Glu Asp Phe Asn Asn Pro Pro Glu Glu Val Val Pro Lys65  70  75  80cta gag aag gag cgg tac tat cga gtt ttg aca tat cta aca gtt cct288Leu Glu Lys Glu Arg Tyr Tyr Arg Val Leu Thr Tyr Leu Thr Val Pro85  90  95atg cat tac gct gca tta att gtg tca gca tgg tgg gtc gga act cag336Met His Tyr Ala Ala Leu Ile Val Ser Ala Trp Trp Val Gly Thr Gln100 105 110cca atg tct tgg ctt gaa att ggt gcg ctt gcc ttg tca ctg ggt atc384Pro Met Ser Trp Leu Glu Ile Gly Ala Leu Ala Leu Ser Leu Gly Ile115 120 125  
            
gtg aac gga cta gcg ctc aat aca gga cac gaa ctc ggt cac aag aag432Val Asn Gly Leu Ala Leu Asn Thr Gly His Glu Leu Gly His Lys Lys130 135 140gag act ttt gat cgt tgg atg gcc aaa att gtg ttg gct gtc gta ggg480Glu Thr Phe Asp Arg Trp Met Ala Lys Ile Val Leu Ala Val Val Gly145 150 155 160tac ggt cac ttc ttt att gag cat aat aag ggt cat cac cgt gat gtc528Tyr Gly His Phe Phe Ile Glu His Asn Lys Gly His His Arg Asp Val165 170 175gct aca ccg atg gat cct gca aca tcc cgg atg gga gaa agc att tat576Ala Thr Pro Met Asp Pro Ala Thr Ser Arg Met Gly Glu Ser Ile Tyr180 185 190aag ttt tca atc cgt gag atc cca gga gca ttt att cgt gct tgg ggg624Lys Phe Ser Ile Arg Glu Ile Pro Gly Ala Phe Ile Arg Ala Trp Gly195 200 205ctt gag gaa caa cgc ctt tcg cgc cgt ggc caa agc gtt tgg agt ttc672Leu Glu Glu Gln Arg Leu Ser Arg Arg Gly Gln Ser Val Trp Ser Phe210 215 220gat aat gaa atc ctc caa cca atg atc atc aca gtt att ctt tac gcc720Asp Asn Glu Ile Leu Gln Pro Met Ile Ile Thr Val Ile Leu Tyr Ala225 230 235 240gtt ctc ctt gcc ttg ttt gga cct aag atg ctg gtg ttc ctg ccg att768Val Leu Leu Ala Leu Phe Gly Pro Lys Met Leu Val Phe Leu Pro Ile245 250 255caa atg gct ttc ggt tgg tgg cag ctg acc agt gcg aac tat att gaa816Gln Met Ala Phe Gly Trp Trp Gln Leu Thr Ser Ala Asn Tyr Ile Glu260 265 270cat tac ggc ttg ctc cgt caa aaa atg gag gac ggt cga tat gag cat864His Tyr Gly Leu Leu Arg Gln Lys Met Glu Asp Gly Arg Tyr Glu His275 280 285caa aag ccg cac cat tct tgg aat agt aat cac atc gtc tct aat cta912Gln Lys Pro His His Ser Trp Asn Ser Asn His Ile Val Ser Asn Leu290 295 300  
            
gtg ctg ttc cac ctt cag cgg cac tcg gat cac cac gcg cat cca aca960Val Leu Phe His Leu Gln Arg His Ser Asp His His Ala His Pro Thr305 310 315 320cgt tct tat cag rca crt cgg gat ttt ccc ggc ctg ccg gct ctt ccg1008Arg Ser Tyr Gln Ser Leu Arg Asp Phe Pro Gly Leu Pro Ala Leu Pro325 330 335acg ggt tac cct ggt gca ttt ttg atg gcg atg att cct cag tgg ttt1056Thr Gly Tyr Pro Gly Ala Phe Leu Met Ala Met Ile Pro Gln Trp Phe340 345 350aga tca gtt atg gat ccc aag gta gta gat tgg gct ggt ggt gac ctt1104Arg Ser Val Met Asp Pro Lys Val Val Asp Trp Ala Gly Gly Asp Leu355 360 365aat aag atc caa att gat gat tcg atg cga gaa acc tat ttg aaa aaa1152Asn Lys Ile Gln Ile Asp Asp Ser Met Arg Glu Thr Tyr Leu Lys Lys370 375 380ttt ggc act agt agt gct ggt cat agt tcg agt acc tct gcg gta gca1200Phe Gly Thr Ser Ser Ala Gly His Ser Ser Ser Thr Ser Ala Val Ala385 390 395 400tcg tag1206Ser<210>26<211>401<212>PRT<213>食油假單胞菌<400>26Met Leu Glu Lys His Arg Val Leu Asp Ser Ala Pro Glu Tyr Val Asp1   5   10  15Lys Lys Lys Tyr Leu Trp Ile Leu Ser Thr Leu Trp Pro Ala Thr Pro20  25  30Met Ile Gly Ile Trp Leu Ala Asn Glu Thr Gly Trp Gly Ile Phe Tyr35  40  45  
            
Gly Leu Val Leu Leu Val Trp Tyr Gly Ala Leu Pro Leu Leu Asp Ala50  55  60Met Phe Gly Glu Asp Phe Asn Asn Pro Pro Glu Glu Val Val Pro Lys65  70  75  80Leu Glu Lys Glu Arg Tyr Tyr Arg Val Leu Thr Tyr Leu Thr Val Pro85  90  95Met His Tyr Ala Ala Leu Ile Val Ser Ala Trp Trp Val Gly Thr Gln100 105 110Pro Met Ser Trp Leu Glu Ile Gly Ala Leu Ala Leu Ser Leu Gly Ile115 120 125Val Asn Gly Leu Ala Leu Asn Thr Gly His Glu Leu Gly His Lys Lys130 135 140Glu Thr Phe Asp Arg Trp Met Ala Lys Ile Val Leu Ala Val Val Gly145 150 155 160Tyr Gly His Phe Phe Ile Glu His Asn Lys Gly His His Arg Asp Val165 170 175Ala Thr Pro Met Asp Pro Ala Thr Ser Arg Met Gly Glu Ser Ile Tyr180 185 190Lys Phe Ser Ile Arg Glu Ile Pro Gly Ala Phe Ile Arg Ala Trp Gly195 200 205Leu Glu Glu Gln Arg Leu Ser Arg Arg Gly Gln Ser Val Trp Ser Phe210 215 220Asp Asn Glu Ile Leu Gln Pro Met Ile Ile Thr Val Ile Leu Tyr Ala225 230 235 240Val Leu Leu Ala Leu Phe Gly Pro Lys Met Leu Val Phe Leu Pro Ile245 250 255Gln Met Ala Phe Gly Trp Trp Gln Leu Thr Ser Ala Asn Tyr Ile Glu260 265 270His Tyr Gly Leu Leu Arg Gln Lys Met Glu Asp Gly Arg Tyr Glu His  
            
275 280 285Gln Lys Pro His His Ser Trp Asn Ser Asn His Ile Val Ser Asn Leu290 295 300Val Leu Phe His Leu Gln Arg His Ser Asp His His Ala His Pro Thr305 310 315 320Arg Ser Tyr Gln Ser Leu Arg Asp Phe Pro Gly Leu Pro Ala Leu Pro325 330 335Thr Gly Tyr Pro Gly Ala Phe Leu Met Ala Met Ile Pro Gln Trp Phe340 345 350Arg Ser Val Met Asp Pro Lys Val Val Asp Trp Ala Gly Gly Asp Leu355 360 365Asn Lys Ile Gln Ile Asp Asp Ser Met Arg Glu Thr Tyr Leu Lys Lys370 375 380Phe Gly Thr Ser Ser Ala Gly His Ser Ser Ser Thr Ser Ala Val Ala385 390 395 400Ser<210>27<211>1560<212>DNA<213>洋蔥伯克霍爾德氏菌<220>  
            
<221>CDS<222>(1)..(1560)<400>27atg gac act tct gtg cag aag aag aaa ctc ggt tta aag aat cgc tac48Met Asp Thr Ser Val Gln Lys Lys Lys Leu Gly Leu Lys Asn Arg Tyr1   5   10  15gca gcg atg acc cgc ggt ctt ggc tgg cag acc agc tac cag ccg atg96Ala Ala Met Thr Arg Gly Leu Gly Trp Gln Thr Ser Tyr Gln Pro Met  
            
20  25  30gag aaa gtg ttt ccg tac gac aag tac gaa ggc atc aag atc cac gat144Glu Lys Val Phe Pro Tyr Asp Lys Tyr Glu Gly Ile Lys Ile His Asp35  40  45tgg gat aaa tgg gaa gac ccc ttc cgc ctg acc atg gac gcc tac tgg192Trp Asp Lys Trp Glu Asp Pro Phe Arg Leu Thr Met Asp Ala Tyr Trp50  55  60aaa tat cag ggc gag aag gaa aaa aag ctt tac gcc gtc atc gac gct240Lys Tyr Gln Gly Glu Lys Glu Lys Lys Leu Tyr Ala Val Ile Asp Ala65  70  75  80ttc gcg cag aac aac ggg cag ttg agc att tcc gac gcg cga tat gtc288Phe Ala Gln Asn Asn Gly Gln Leu Ser Ile Ser Asp Ala Arg Tyr Val85  90  95aac gca ctc aag gtg ttt atc cag ggt gtg aca ccg ttg gag tat atg336Asn Ala Leu Lys Val Phe Ile Gln Gly Val Thr Pro Leu Glu Tyr Met100 105 110gca cac cga ggt ttt gcc cac att ggt cgg cat ttt acg ggt gaa ggg384Ala His Arg Gly Phe Ala His Ile Gly Arg His Phe Thr Gly Glu Gly115 120 125gca cgt gtt gct tgc cag atg cag tcc atc gac gag ctg cgt cac ttc432Ala Arg Val Ala Cys Gln Met Gln Ser Ile Asp Glu Leu Arg His Phe130 135 140cag acc gaa atg cat gct ctc tcg cac tac aac aag tat ttt aac ggt480Gln Thr Glu Met His Ala Leu Ser His Tyr Asn Lys Tyr Phe Asn Gly145 150 155 160ctg cac aac tcc atc cat tgg tac gac cgg gtt tgg tat ttg tcg gtg528Leu His Asn Ser Ile His Trp Tyr Asp Arg Val Trp Tyr Leu Ser Val165 170 175ccc aag tca ttt ttt gaa gac gcg gcc acc ggt gga ccg ttc gag ttt576Pro Lys Ser Phe Phe Glu Asp Ala Ala Thr Gly Gly Pro Phe Glu Phe180 185 190ctt acc gcg gtg agc ttt tcg ttc gaa tat gtg ttg acc aac ctg ctg624Leu Thr Ala Val Ser Phe Ser Phe Glu Tyr Val Leu Thr Asn Leu Leu  
            
195 200 205ttt gtc ccc ttc atg tcg ggt gct gct tac aac ggg gac atg tct acg672Phe Val Pro Phe Met Ser Gly Ala Ala Tyr Asn Gly Asp Met Ser Thr210 215 220gtc act ttc ggt ttt tcg gcg caa agt gac gaa tcg cgc cac atg aca720Val Thr Phe Gly Phe Ser Ala Gln Ser Asp Glu Ser Arg His Met Thr225 230 235 240ctc ggc atc gaa tgc atc aag ttc atg cta gaa cag gat ccg gac aac768Leu Gly Ile Glu Cys Ile Lys Phe Met Leu Glu Gln Asp Pro Asp Asn245 250 255gtg ccc atc gtg cag cgc tgg atc gac aag tgg ttc tgg cgc ggc tat816Val Pro Ile Val Gln Arg Trp Ile Asp Lys Trp Phe Trp Arg Gly Tyr260 265 270cgg ctg ttg agc atc gtg gcc atg atg cag gac tac atg ctg ccc aac864Arg Leu Leu Ser Ile Val Ala Met Met Gln Asp Tyr Met Leu Pro Asn275 280 285cgg gtg atg agc tgg cgc gag agc tgg gag atg tac gtc gag cag aac912Arg Val Met Ser Trp Arg Glu Ser Trp Glu Met Tyr Val Glu Gln Asn290 295 300ggc ggc gcg ctg ttc aag gat ctt gcg cgt tat ggc atc cgc aag ccc960Gly Gly Ala Leu Phe Lys Asp Leu Ala Arg Tyr Gly Ile Arg Lys Pro305 310 315 320aag ggc tgg gac cag gct tgc gaa ggc aag gac cac atc agc cat cag1008Lys Gly Trp Asp Gln Ala Cys Glu Gly Lys Asp His Ile Ser His Gln325 330 335acc ttc gcc gta ttc tat aac tat aac gcc gcg gcc ccc atc cac acc1056Thr Phe Ala Val Phe Tyr Asn Tyr Asn Ala Ala Ala Pro Ile His Thr340 345 350tgg gtt ccc aca aaa gaa gaa atg gga tgg ctg tcg gag aag tac ccc1104Trp Val Pro Thr Lys Glu Glu Met Gly Trp Leu Ser Glu Lys Tyr Pro355 360 365gag acg ttc gac aag tat tac cgt ccg cgt tgg gac tac tgg cgc gag1152Glu Thr Phe Asp Lys Tyr Tyr Arg Pro Arg Trp Asp Tyr Trp Arg Glu  
            
370 375 380cag gcc gcc aag ggc aac cgt ttc tac aac aag acg ctg ccg atg ctc1200Gln Ala Ala Lys Gly Asn Arg Phe Tyr Asn Lys Thr Leu Pro Met Leu385 390 395 400tgc act acc tgc cag att ccg atg ata ttc acc gag cct ggc gac gca1248Cys Thr Thr Cys Gln Ile Pro Met Ile Phe Thr Glu Pro Gly Asp Ala405 410 415acc aag atc tgc tat cgc gag tcg gcc tac ctc ggc gac aag tat cac1296Thr Lys Ile Cys Tyr Arg Glu Ser Ala Tyr Leu Gly Asp Lys Tyr His420 425 430ttc tgc agc gac cac tgc aag gag att ttt gac aac gaa ccc gaa aag1344Phe Cys Ser Asp His Cys Lys Glu Ile Phe Asp Asn Glu Pro Glu Lys435 440 445ttc gtg cag tca tgg ctt ccg ccg cag caa gtg tat caa gga aac tgt1392Phe Val Gln Ser Trp Leu Pro Pro Gln Gln Val Tyr Gln Gly Asn Cys450 455 460ttc aag ccg gat gcc gat ccg acc aag gag ggt ttt gat ccc ttg atg1440Phe Lys Pro Asp Ala Asp Pro Thr Lys Glu Gly Phe Asp Pro Leu Met465 470 475 480gcc ttg ctc gac tac tac aac ctg aat gta ggc cgg gac aac ttc gat1488Ala Leu Leu Asp Tyr Tyr Asn Leu Asn Val Gly Arg Asp Asn Phe Asp485 490 495ttc gag gga tcg gaa gac caa aag aac ttt gct gcc tgg cgt gga gag1536Phe Glu Gly Ser Glu Asp Gln Lys Asn Phe Ala Ala Trp Arg Gly Glu500 505 510gtc ttg caa gga gaa gcc aaa tga1560Val Leu Gln Gly Glu Ala Lys515<210>28<211>519<212>PRT<213>洋蔥伯克霍爾德氏菌  
            
<400>28Met Asp Thr Ser Val Gln Lys Lys Lys Leu Gly Leu Lys Asn Arg Tyr1   5   10  15Ala Ala Met Thr Arg Gly Leu Gly Trp Gln Thr Ser Tyr Gln Pro Met20  25  30Glu Lys Val Phe Pro Tyr Asp Lys Tyr Glu Gly Ile Lys Ile His Asp35  40  45Trp Asp Lys Trp Glu Asp Pro Phe Arg Leu Thr Met Asp Ala Tyr Trp50  55  60Lys Tyr Gln Gly Glu Lys Glu Lys Lys Leu Tyr Ala Val Ile Asp Ala65  70  75  80Phe Ala Gln Asn Asn Gly Gln Leu Ser Ile Ser Asp Ala Arg Tyr Val85  90  95Asn Ala Leu Lys Val Phe Ile Gln Gly Val Thr Pro Leu Glu Tyr Met100 105 110Ala His Arg Gly Phe Ala His Ile Gly Arg His Phe Thr Gly Glu Gly115 120 125Ala Arg Val Ala Cys Gln Met Gln Ser Ile Asp Glu Leu Arg His Phe130 135 140Gln Thr Glu Met His Ala Leu Ser His Tyr Asn Lys Tyr Phe Asn Gly145 150 155 160Leu His Asn Ser Ile His Trp Tyr Asp Arg Val Trp Tyr Leu Ser Val165 170 175Pro Lys Ser Phe Phe Glu Asp Ala Ala Thr Gly Gly Pro Phe Glu Phe180 185 190Leu Thr Ala Val Ser Phe Ser Phe Glu Tyr Val Leu Thr Asn Leu Leu195 200 205Phe Val Pro Phe Met Ser Gly Ala Ala Tyr Asn Gly Asp Met Ser Thr210 215 220  
            
Val Thr Phe Gly Phe Ser Ala Gln Ser Asp Glu Ser Arg His Met Thr225 230 235 240Leu Gly Ile Glu Cys Ile Lys Phe Met Leu Glu Gln Asp Pro Asp Asn245 250 255Val Pro Ile Val Gln Arg Trp Ile Asp Lys Trp Phe Trp Arg Gly Tyr260 265 270Arg Leu Leu Ser Ile Val Ala Met Met Gln Asp Tyr Met Leu Pro Asn275 280 285Arg Val Met Ser Trp Arg Glu Ser Trp Glu Met Tyr Val Glu Gln Asn290 295 300Gly Gly Ala Leu Phe Lys Asp Leu Ala Arg Tyr Gly Ile Arg Lys Pro305 310 315 320Lys Gly Trp Asp Gln Ala Cys Glu Gly Lys Asp His Ile Ser His Gln325 330 335Thr Phe Ala Val Phe Tyr Asn Tyr Asn Ala Ala Ala Pro Ile His Thr340 345 350Trp Val Pro Thr Lys Glu Glu Met Gly Trp Leu Ser Glu Lys Tyr Pro355 360 365Glu Thr Phe Asp Lys Tyr Tyr Arg Pro Arg Trp Asp Tyr Trp Arg Glu370 375 380Gln Ala Ala Lys Gly Asn Arg Phe Tyr Asn Lys Thr Leu Pro Met Leu385 390 395 400Cys Thr Thr Cys Gln Ile Pro Met Ile Phe Thr Glu Pro Gly Asp Ala405 410 415Thr Lys Ile Cys Tyr Arg Glu Ser Ala Tyr Leu Gly Asp Lys Tyr His420 425 430Phe Cys Ser Asp His Cys Lys Glu Ile Phe Asp Asn Glu Pro Glu Lys435 440 445Phe Val Gln Ser Trp Leu Pro Pro Gln Gln Val Tyr Gln Gly Asn Cys450 455 460  
            
Phe Lys Pro Asp Ala Asp Pro Thr Lys Glu Gly Phe Asp Pro Leu Met465 470 475 480Ala Leu Leu Asp Tyr Tyr Asn Leu Asn Val Gly Arg Asp Asn Phe Asp485 490 495Phe Glu Gly Ser Glu Asp Gln Lys Asn Phe Ala Ala Trp Arg Gly Glu500 505 510Val Leu Gln Gly Glu Ala Lys515<210>29<211>996<212>DNA<213>洋蔥伯克霍爾德氏菌<220>  
            
<221>CDS<222>(1)..(996)<400>29atg acc atc gat ttg aag acg cgg gaa atc aaa cca ctg cgt cac acc48Met Thr Ile Asp Leu Lys Thr Arg Glu Ile Lys Pro Leu Arg His Thr1   5   10  15tac acg cac gtg gct caa tac atc ggg gcc gat aaa gcc gct tcg cgc96Tyr Thr His Val Ala Gln Tyr Ile Gly Ala Asp Lys Ala Ala Ser Arg20  25  30tat cag gaa ggc act gta ggt gct caa ccc gca gcg aat ttt cat tac144Tyr Gln Glu Gly Thr Val Gly Ala Gln Pro Ala Ala Asn Phe His Tyr35  40  45cgg ccc acg tgg gat ccc gag cat gaa ctg ttc gac acg tcg cgt acc192Arg Pro Thr Trp Asp Pro Glu His Glu Leu Phe Asp Thr Ser Arg Thr50  55  60gcg att caa atg aag gac tgg tat gcg ctg aaa gac ccg cgt cag ttc240Ala Ile Gln Met Lys Asp Trp Tyr Ala Leu Lys Asp Pro Arg Gln Phe65  70  75  80  
            
tac tac gcg tcg tgg acg atg acc cga gcg cgg cag caa gac gcg atg288Tyr Tyr Ala Ser Trp Thr Met Thr Arg Ala Arg Gln Gln Asp Ala Met85  90  95gaa tcc aac ttc gag ttt gtc gag tcg cgc ggc atg atc gat ctc gtt336Glu Ser Asn Phe Glu Phe Val Glu Ser Arg Gly Met Ile Asp Leu Val100 105 110tcc gat gag gtt cga caa cgg gcg ctt tcc gtt ctc gtg cct ttg cgt384Ser Asp Glu Val Arg Gln Arg Ala Leu Ser Val Leu Val Pro Leu Arg115 120 125cac gcg gcc tgg ggc gcg aac atg aac aac tcc cag atc tgt gcc cta432His Ala Ala Trp Gly Ala Asn Met Asn Asn Ser Gln Ile Cys Ala Leu130 135 140ggt tat ggc acg acc ttc act gcg ccg gct atg ttc cac gca atg gac480Gly Tyr Gly Thr Thr Phe Thr Ala Pro Ala Met Phe His Ala Met Asp145 150 155 160aat ctg ggt gta gcg cag tat ctc aca cga ctg gcg ctg gta atg tct528Asn Leu Gly Val Ala Gln Tyr Leu Thr Arg Leu Ala Leu Val Met Ser165 170 175gga ccc gat ctt ctt gac gaa gcc aag caa gcc tgg atg acg agt ccc576Gly Pro Asp Leu Leu Asp Glu Ala Lys Gln Ala Trp Met Thr Ser Pro180 185 190gat tgg caa ccg ttg cgt cgt tat gtg gaa aac act ctg gtg ctg caa624Asp Trp Gln Pro Leu Arg Arg Tyr Val Glu Asn Thr Leu Val Leu Gln195 200 205gat ccg gtg gaa ctg ttc atc gcc caa aat ctg gcg ctc gac ggt ctt672Asp Pro Val Glu Leu Phe Ile Ala Gln Asn Leu Ala Leu Asp Gly Leu210 215 220ctt tat ccc atg atc tac ggc gct ttc gtc gac gat tac atc gca ctc720Leu Tyr Pro Met Ile Tyr Gly Ala Phe Val Asp Asp Tyr Ile Ala Leu225 230 235 240aac ggt ggt agc gca gtg gca atg cta acc act ttc atg ccc gag tgg768Asn Gly Gly Ser Ala Val Ala Met Leu Thr Thr Phe Met Pro Glu Trp245 250 255  
            
cat gac gaa tcc agt cgc tgg gtc gat gcg gta gta aag acc atg gcg816His Asp Glu Ser Ser Arg Trp Val Asp Ala Val Val Lys Thr Met Ala260 265 270acg gaa tcg gag gat aac aaa gcg ctg ctc att cac tgg ttg cgt acc864Thr Glu Ser Glu Asp Asn Lys Ala Leu Leu Ile His Trp Leu Arg Thr275 280 285tgg gaa gat cag gcg gcg tca gcg ttg ttg cct gtc gct gaa atg gct912Trp Glu Asp Gln Ala Ala Ser Ala Leu Leu Pro Val Ala Glu Met Ala290 295 300ttg gcg gaa aac ggc cac gac gcc ttg gaa gaa gta agg cag caa ctt960Leu Ala Glu Asn Gly His Asp Ala Leu Glu Glu Val Arg Gln Gln Leu305 310 315 320cgt gcc cgc gtt gcg aag gcc ggg att gtt ctg taa 996Arg Ala Arg Val Ala Lys Ala Gly Ile Val Leu325 330<210>30<211>331<212>PRT<213>洋蔥伯克霍爾德氏菌<400>30Met Thr Ile Asp Leu Lys Thr Arg Glu Ile Lys Pro Leu Arg His Thr1   5   10  15Tyr Thr His Val Ala Gln Tyr Ile Gly Ala Asp Lys Ala Ala Ser Arg20  25  30Tyr Gln Glu Gly Thr Val Gly Ala Gln Pro Ala Ala Asn Phe His Tyr35  40  45Arg Pro Thr Trp Asp Pro Glu His Glu Leu Phe Asp Thr Ser Arg Thr50  55  60Ala Ile Gln Met Lys Asp Trp Tyr Ala Leu Lys Asp Pro Arg Gln Phe65  70  75  80Tyr Tyr Ala Ser Trp Thr Met Thr Arg Ala Arg Gln Gln Asp Ala Met  
            
85  90  95Glu Ser Asn Phe Glu Phe Val Glu Ser Arg Gly Met Ile Asp Leu Val100 105 110Ser Asp Glu Val Arg Gln Arg Ala Leu Ser Val Leu Val Pro Leu Arg115 120 125His Ala Ala Trp Gly Ala Asn Met Asn Asn Ser Gln Ile Cys Ala Leu130 135 140Gly Tyr Gly Thr Thr Phe Thr Ala Pro Ala Met Phe His Ala Met Asp145 150 155 160Asn Leu Gly Val Ala Gln Tyr Leu Thr Arg Leu Ala Leu Val Met Ser165 170 175Gly Pro Asp Leu Leu Asp Glu Ala Lys Gln Ala Trp Met Thr Ser Pro180 185 190Asp Trp Gln Pro Leu Arg Arg Tyr Val Glu Asn Thr Leu Val Leu Gln195 200 205Asp Pro Val Glu Leu Phe Ile Ala Gln Asn Leu Ala Leu Asp Gly Leu210 215 220Leu Tyr Pro Met Ile Tyr Gly Ala Phe Val Asp Asp Tyr Ile Ala Leu225 230 235 240Asn Gly Gly Ser Ala Val Ala Met Leu Thr Thr Phe Met Pro Glu Trp245 250 255His Asp Glu Ser Ser Arg Trp Val Asp Ala Val Val Lys Thr Met Ala260 265 270Thr Glu Ser Glu Asp Asn Lys Ala Leu Leu Ile His Trp Leu Arg Thr275 280 285Trp Glu Asp Gln Ala Ala Ser Ala Leu Leu Pro Val Ala Glu Met Ala290 295 300Leu Ala Glu Asn Gly His Asp Ala Leu Glu Glu Val Arg Gln Gln Leu305 310 315 320  
            
Arg Ala Arg Val Ala Lys Ala Gly Ile Val Leu325 330<210>31<211>357<212>DNA<213>洋蔥伯克霍爾德氏菌<220>  
            
<221>CDS<222>(1)..(357)<400>31atg agc gtt gtt gcc ctc aaa ccc tac aag ttc ccg gca cga gac gcg48Met Ser Val Val Ala Leu Lys Pro Tyr Lys Phe Pro Ala Arg Asp Ala1   5   10  15cgc gaa aac ttt ccg gcg ccg ttg ctg ttt atc ggc tgg gaa gac cat96Arg Glu Asn Phe Pro Ala Pro Leu Leu Phe Ile Gly Trp Glu Asp His20  25  30ctg ttg ttt gcg gca cct gtt gcc ttg ccc ctg ccg tcg gac acg ttg144Leu Leu Phe Ala Ala Pro Val Ala Leu Pro Leu Pro Ser Asp Thr Leu35  40  45ttc ggt gcg ctg tgc acc cag gtg ttg ccc ggc act tat ggc tat cac192Phe Gly Ala Leu Cys Thr Gln Val Leu Pro Gly Thr Tyr Gly Tyr His50  55  60ccc gat ttc tca aag atc gac tgg agc cag gtg cag tgg ttt aag tcc240Pro Asp Phe Ser Lys Ile Asp Trp Ser Gln Val Gln Trp Phe Lys Ser65  70  75  80ggc cag ccg tgg cat ccc gac ccg gcg aag tcg ctg gct gaa aac ggt288Gly Gln Pro Trp His Pro Asp Pro Ala Lys Ser Leu Ala Glu Asn Gly85  90  95ctg acg cac aaa gac gtg atc cgc ttt cgc acg cct ggc ttg aac ggt336Leu Thr His Lys Asp Val Ile Arg Phe Arg Thr Pro Gly Leu Asn Gly100 105 110ctg agc ggt tcc tgc aat tga357  
            
Leu Ser Gly Ser Cys Asn115<210>32<211>118<212>PRT<213>洋蔥伯克霍爾德氏菌<400>32Met Ser Val Val Ala Leu Lys Pro Tyr Lys Phe Pro Ala Arg Asp Ala1   5   10  15Arg Glu Asn Phe Pro Ala Pro Leu Leu Phe Ile Gly Trp Glu Asp His20  25  30Leu Leu Phe Ala Ala Pro Val Ala Leu Pro Leu Pro Ser Asp Thr Leu35  40  45Phe Gly Ala Leu Cys Thr Gln Val Leu Pro Gly Thr Tyr Gly Tyr His50  55  60Pro Asp Phe Ser Lys Ile Asp Trp Ser Gln Val Gln Trp Phe Lys Ser65  70  75  80Gly Gln Pro Trp His Pro Asp Pro Ala Lys Ser Leu Ala Glu Asn Gly85  90  95Leu Thr His Lys Asp Val Ile Arg Phe Arg Thr Pro Gly Leu Asn Gly100 105 110Leu Ser Gly Ser Cys Asn115<210>33<211>1143<212>DNA<213>嗜熱脂肪芽孢桿菌<220>  
            
<221>CDS  
            
<222>(1)..(1143)<400>33atg gaa aaa aat aaa atg tta ata gaa gaa aag ttg gac act gct gct48Met Glu Lys Asn Lys Met Leu Ile Glu Glu Lys Leu Asp Thr Ala Ala1   5   10  15ctt ctt gct aag gcg gag gaa ata ggc cgg att gct gag gaa gag gcg96Leu Leu Ala Lys Ala Glu Glu Ile Gly Arg Ile Ala Glu Glu Glu Ala20  25  30ggt gaa gcg gac cgc aat gcc tgt ttc tcc gac cgg gtg gct agg gcc144Gly Glu Ala Asp Arg Asn Ala Cys Phe Ser Asp Arg Val Ala Arg Ala35  40  45att aaa gaa gct gga ttc cac aag ctc atg cgt ccc aag cag tac gga192Ile Lys Glu Ala Gly Phe His Lys Leu Met Arg Pro Lys Gln Tyr Gly50  55  60gga ctg caa gta gac ttg cga act tac ggg gag att gtc cgc aca gtg240Gly Leu Gln Val Asp Leu Arg Thr Tyr Gly Glu Ile Val Arg Thr Val65  70  75  80gcc cgg tac agt gtt gcc gca gga tgg ctg acc tat ttt tat tcc atg288Ala Arg Tyr Ser Val Ala Ala Gly Trp Leu Thr Tyr Phe Tyr Ser Met85  90  95cat gag gtt tgg gct gca tat ctg cct cca aaa ggc aga gaa gaa att336His Glu Val Trp Ala Ala Tyr Leu Pro Pro Lys Gly Arg Glu Glu Ile100 105 110ttt gga caa gga ggg ctg ttg gca gac gtc gtt gcc cct gtt ggc cgg384Phe Gly Gln Gly Gly Leu Leu Ala Asp Val Val Ala Pro Val Gly Arg115 120 125gtg gag aag gac ggg gac ggc tac cgt ctc tat ggg cag tgg aac ttc432Val Glu Lys Asp Gly Asp Gly Tyr Arg Leu Tyr Gly Gln Trp Asn Phe130 135 140tgt agc ggt gtc ctc cat agt gac tgg atc gga ctt ggc gcc atg atg480Cys Ser Gly Val Leu His Ser Asp Trp Ile Gly Leu Gly Ala Met Met145 150 155 160gag ctg cct gac ggc aat agt cct gag tac tgt ttg tta gtg ctg cct528  
            
Glu Leu Pro Asp Gly Asn Ser Pro Glu Tyr Cys Leu Leu Val Leu Pro165 170 175aag tcg gat gtc cag atc gta gaa aat tgg gat acc atg ggc ctc cgc576Lys Ser Asp Val Gln Ile Val Glu Asn Trp Asp Thr Met Gly Leu Arg180 185 190gct tcg gga agc aac ggg gta tta gtt gaa ggt gct tat gtt cca tta624Ala Ser Gly Ser Asn Gly Val Leu Val Glu Gly Ala Tyr Val Pro Leu195 200 205cac cgg atc ttt ccg gct ggc cgg gtg atg gct cat ccg ctt ttc ttg672His Arg Ile Phe Pro Ala Gly Arg Val Met Ala His Pro Leu Phe Leu210 215 220ctt ggg ttc cct tta gta tct tta ggc ggc gac gaa cga ttg gtg tca720Leu Gly Phe Pro Leu Val Ser Leu Gly Gly Asp Glu Arg Leu Val Ser225 230 235 240ctt ttc caa gaa cgc act gag aag cgc att cgt gtc ttc aaa ggc ggc768Leu Phe Gln Glu Arg Thr Glu Lys Arg Ile Arg Val Phe Lys Gly Gly245 250 255gcg aaa gaa aag gat tct gcc gct agc cag cgg ctg tta gcc gag atg816Ala Lys Glu Lys Asp Ser Ala Ala Ser Gln Arg Leu Leu Ala Glu Met260 265 270aaa aca gaa tta aat gca atg gaa ggc att gtg gaa caa tat atc cgc864Lys Thr Glu Leu Asn Ala Met Glu Gly Ile Val Glu Gln Tyr Ile Arg275 280 285cag ctt gag gct tgc caa aaa gaa gga aag acg gtg atg aac gat atg912Gln Leu Glu Ala Cys Gln Lys Glu Gly Lys Thr Val Met Asn Asp Met290 295 300gag cga gag cag cta ttc gca tgg cgt gga tat gtg gca aaa gcg tcc960Glu Arg Glu Gln Leu Phe Ala Trp Arg Gly Tyr Val Ala Lys Ala Ser305 310 315 320gcc aat att gcc gtc aga aca ctg tta act ctt gga ggc aat tcg atc1008Ala Asn Ile Ala Val Arg Thr Leu Leu Thr Leu Gly Gly Asn Ser Ile325 330 335ttt aaa ggc gat ccg gta gaa ctg ttc aca aga gat ttg cta gcg gtg1056  
            
Phe Lys Gly Asp Pro Val Glu Leu Phe Thr Arg Asp Leu Leu Ala Val340 345 350gcc gca cat cct aac tcc ctg tgg gag gat gcg atg gct gca tat gga1104Ala Ala His Pro Asn Ser Leu Trp Glu Asp Ala Met Ala Ala Tyr Gly355 360 365aga acg ata ttc ggg ctg cca ggg gac cca gtc tgg taa1143Arg Thr Ile Phe Gly Leu Pro Gly Asp Pro Val Trp370 375 380<210>34<211>380<212>PRT<213>嗜熱脂肪芽孢桿菌<400>34Met Glu Lys Asn Lys Met Leu Ile Glu Glu Lys Leu Asp Thr Ala Ala1   5   10  15Leu Leu Ala Lys Ala Glu Glu Ile Gly Arg Ile Ala Glu Glu Glu Ala20 25 30Gly Glu Ala Asp Arg Asn Ala Cys Phe Ser Asp Arg Val Ala Arg Ala35  40  45Ile Lys Glu Ala Gly Phe His Lys Leu Met Arg Pro Lys Gln Tyr Gly50  55  60Gly Leu Gln Val Asp Leu Arg Thr Tyr Gly Glu Ile Val Arg Thr Val65  70  75  80Ala Arg Tyr Ser Val Ala Ala Gly Trp Leu Thr Tyr Phe Tyr Ser Met85  90  95His Glu Val Trp Ala Ala Tyr Leu Pro Pro Lys Gly Arg Glu Glu Ile100 105 110Phe Gly Gln Gly Gly Leu Leu Ala Asp Val Val Ala Pro Val Gly Arg115 120 125Val Glu Lys Asp Gly Asp Gly Tyr Arg Leu Tyr Gly Gln Trp Asn Phe  
            
130 135 140Cys Ser Gly Val Leu His Ser Asp Trp Ile Gly Leu Gly Ala Met Met145 150 155 160Glu Leu Pro Asp Gly Asn Ser Pro Glu Tyr Cys Leu Leu Val Leu Pro165 170 175Lys Ser Asp Val Gln Ile Val Glu Asn Trp Asp Thr Met Gly Leu Arg180 185 190Ala Ser Gly Ser Asn Gly Val Leu Val Glu Gly Ala Tyr Val Pro Leu195 200 205His Arg Ile Phe Pro Ala Gly Arg Val Met Ala His Pro Leu Phe Leu210 215 220Leu Gly Phe Pro Leu Val Ser Leu Gly Gly Asp Glu Arg Leu Val Ser225 230 235 240Leu Phe Gln Glu Arg Thr Glu Lys Arg Ile Arg Val Phe Lys Gly Gly245 250 255Ala Lys Glu Lys Asp Ser Ala Ala Ser Gln Arg Leu Leu Ala Glu Met260 265 270Lys Thr Glu Leu Asn Ala Met Glu Gly Ile Val Glu Gln Tyr Ile Arg275 280 285Gln Leu Glu Ala Cys Gln Lys Glu Gly Lys Thr Val Met Asn Asp Met290 295 300Glu Arg Glu Gln Leu Phe Ala Trp Arg Gly Tyr Val Ala Lys Ala Ser305 310 315 320Ala Asn Ile Ala Val Arg Thr Leu Leu Thr Leu Gly Gly Asn Ser Ile325 330 335Phe Lys Gly Asp Pro Val Glu Leu Phe Thr Arg Asp Leu Leu Ala Val340 345 350Ala Ala His Pro Asn Ser Leu Trp Glu Asp Ala Met Ala Ala Tyr Gly355 360 365  
            
Arg Thr Ile Phe Gly Leu Pro Gly Asp Pro Val Trp370 375 380<210>35<211>1191<212>DNA<213>向日葵<220>  
            
<221>CDS<222>(1)..(1191)<400>35atg gcg att cgc atc aat acg gcg acg ttt caa tca gac ctg tac cgt48Met Ala Ile Arg Ile Asn Thr Ala Thr Phe Gln Ser Asp Leu Tyr Arg1   5   10  15tca ttc gcg ttt cct caa ccg aaa cct ctc aga tct ccc aaa ttc gcc96Ser Phe Ala Phe Pro Gln Pro Lys Pro Leu Arg Ser Pro Lys Phe Ala20  25  30atg gct tcc acc att gga tcc gct aca acg aag gtt gaa agc acc aaa144Met Ala Ser Thr Ile Gly Ser Ala Thr Thr Lys Val Glu Ser Thr Lys35  40  45aag ccc ttt acc cct cca agg gag gtt cac caa cag gtg cta cac tca192Lys Pro Phe Thr Pro Pro Arg Glu Val His Gln Gln Val Leu His Ser50  55  60atg ccg cca caa aag atc gaa atc ttc aaa tcc atg gag ggt tgg gcc240Met Pro Pro Gln Lys Ile Glu Ile Phe Lys Ser Met Glu Gly Trp Ala65  70  75  80gaa aat aac ata ttg gtt cac cta aag cct gtc gaa aaa tgc tgg caa288Glu Asn Asn Ile Leu Val His Leu Lys Pro Val Glu Lys Cys Trp Gln85  90  95gca cag gat ttc cta cca gat ccc gca tct gac gga ttt atg gaa caa336Ala Gln Asp Phe Leu Pro Asp Pro Ala Ser Asp Gly Phe Met Glu Gln100 105 110gtg gag gaa tta cgg gct cgg gct aag gag att ccg gat gat tac ttt384  
            
Val Glu Glu Leu Arg Ala Arg Ala Lys Glu Ile Pro Asp Asp Tyr Phe115 120 125gtt gtt ttg gtt gga gat atg att act gaa gaa gca ctg cct act tac432Val Val Leu Val Gly Asp Met Ile Thr Glu Glu Ala Leu Pro Thr Tyr130 135 140caa aca atg ctt aat act ctt gat ggt gtg cgt gat gag acc ggg gct480Gln Thr Met Leu Asn Thr Leu Asp Gly Val Arg Asp Glu Thr Gly Ala145 150 155 160acc cta ctt ctt ggg cta gtc tgg act cgg gct tgg acc gct gaa gaa528Thr Leu Leu Leu Gly Leu Val Trp Thr Arg Ala Trp Thr Ala Glu Glu165 170 175aac agg cac ggt gat ctt cta cat cag tat ctg tat ctt agt ggg cgg576Asn Arg His Gly Asp Leu Leu His Gln Tyr Leu Tyr Leu Ser Gly Arg180 185 190gtc gac atg agg cag att cag aag aca att cag tac ctc att ggg tct624Val Asp Met Arg Gln Ile Gln Lys Thr Ile Gln Tyr Leu Ile Gly Ser195 200 205gga atg gac ccc cgg acc gaa aac agt cct tac ctt ggg ttc atc tac672Gly Met Asp Pro Arg Thr Glu Asn Ser Pro Tyr Leu Gly Phe Ile Tyr210 215 220act tca ttt caa gag cgt gcc acc ttc atc tct cac gga aac aca gcc720Thr Ser Phe Gln Glu Arg Ala Thr Phe Ile Ser His Gly Asn Thr Ala225 230 235 240cgg cac gca aag gag cat ggt gac gtg aag ctg gct caa atg tgc ggt768Arg His Ala Lys Glu His Gly Asp Val Lys Leu Ala Gln Met Cys Gly245 250 255ata att gca gct gat gaa aaa agg cac gaa acc gcc tac aca aaa ata816Ile Ile Ala Ala Asp Glu Lys Arg His Glu Thr Ala Tyr Thr Lys Ile260 265 270gta gaa aaa ctc ttc gaa att gac ccg gac ggc act gtt ctc gct ttt864Val Glu Lys Leu Phe Glu Ile Asp Pro Asp Gly Thr Val Leu Ala Phe275 280 285gcc gac atg atg agg aaa aag atc tcc atg cct gca cac ttg atg tac912  
            
Ala Asp Met Met Arg Lys Lys Ile Ser Met Pro Ala His Leu Met Tyr290 295 300gat ggg cgt gat gat aac ctc ttc gaa aat ttc tca gct gtt gcc caa960Asp Gly Arg Asp Asp Asn Leu Phe Glu Asn Phe Ser Ala Val Ala Gln305 310 315 320agg ctc ggt gtg tac act gcg aag gac tat gca gac att ctg gag ttt1008Arg Leu Gly Val Tyr Thr Ala Lys Asp Tyr Ala Asp Ile Leu Glu Phe325 330 335ctg gtg ggc cgg tgg aag gtg gcg gat tta acc ggg ctt tct ggt gaa1056Leu Val Gly Arg Trp Lys Val Ala Asp Leu Thr Gly Leu Ser Gly Glu340 345 350ggg cgt aaa gcc caa gac tat gtg tgc ggg ctg gcc cca aga atc aga1104Gly Arg Lys Ala Gln Asp Tyr Val Cys Gly Leu Ala Pro Arg Ile Arg355 360 365agg ctt gag gag agg aac tcg gca agg gcg aag gaa agt gtg aac gtt1152Arg Leu Glu Glu Arg Asn Ser Ala Arg Ala Lys Glu Ser Val Asn Val370 375 380ccg ttc agc tgg atc ttt gat aga gaa gtg aag ctc tga1191Pro Phe Ser Trp Ile Phe Asp Arg Glu Val Lys Leu385 390 395<210>36<211>396<212>PRT<213>向日葵<400>36Met Ala Ile Arg Ile Asn Thr Ala Thr Phe Gln Ser Asp Leu Tyr Arg1   5   10  15Ser Phe Ala Phe Pro Gln Pro Lys Pro Leu Arg Ser Pro Lys Phe Ala20  25  30Met Ala Ser Thr Ile Gly Ser Ala Thr Thr Lys Val Glu Ser Thr Lys35  40  45Lys Pro Phe Thr Pro Pro Arg Glu Val His Gln Gln Val Leu His Ser  
            
50  55  60Met Pro Pro Gln Lys Ile Glu Ile Phe Lys Ser Met Glu Gly Trp Ala65  70  75  80Glu Asn Asn Ile Leu Val His Leu Lys Pro Val Glu Lys Cys Trp Gln85  90  95Ala Gln Asp Phe Leu Pro Asp Pro Ala Ser Asp Gly Phe Met Glu Gln100 105 110Val Glu Glu Leu Arg Ala Arg Ala Lys Glu Ile Pro Asp Asp Tyr Phe115 120 125Val Val Leu Val Gly Asp Met Ile Thr Glu Glu Ala Leu Pro Thr Tyr130 135 140Gln Thr Met Leu Asn Thr Leu Asp Gly Val Arg Asp Glu Thr Gly Ala145 150 155 160Thr Leu Leu Leu Gly Leu Val Trp Thr Arg Ala Trp Thr Ala Glu Glu165 170 175Asn Arg His Gly Asp Leu Leu His Gln Tyr Leu Tyr Leu Ser Gly Arg180 185 190Val Asp Met Arg Gln Ile Gln Lys Thr Ile Gln Tyr Leu Ile Gly Ser195 200 205Gly Met Asp Pro Arg Thr Glu Asn Ser Pro Tyr Leu Gly Phe Ile Tyr210 215 220Thr Ser Phe Gln Glu Arg Ala Thr Phe Ile Ser His Gly Asn Thr Ala225 230 235 240Arg His Ala Lys Glu His Gly Asp Val Lys Leu Ala Gln Met Cys Gly245 250 255Ile Ile Ala Ala Asp Glu Lys Arg His Glu Thr Ala Tyr Thr Lys Ile260 265 270Val Glu Lys Leu Phe Glu Ile Asp Pro Asp Gly Thr Val Leu Ala Phe275 280 285  
            
Ala Asp Met Met Arg Lys Lys Ile Ser Met Pro Ala His Leu Met Tyr290 295 300Asp Gly Arg Asp Asp Asn Leu Phe Glu Asn Phe Ser Ala Val Ala Gln305 310 315 320Arg Leu Gly Val Tyr Thr Ala Lys Asp Tyr Ala Asp Ile Leu Glu Phe325 330 335Leu Val Gly Arg Trp Lys Val Ala Asp Leu Thr Gly Leu Ser Gly Glu340 345 350Gly Arg Lys Ala Gln Asp Tyr Val Cys Gly Leu Ala Pro Arg Ile Arg355 360 365Arg Leu Glu Glu Arg Asn Ser Ala Arg Ala Lys Glu Ser Val Asn Val370 375 380Pro Phe Ser Trp Ile Phe Asp Arg Glu Val Lys Leu385 390 39權利要求
            
1.一種進行氧化反應的方法，該氧化反應由單加氧酶催化并使用過氧化氫作為氧化劑，其中單加氧酶的氧化損傷反應水平低，所述方法包括在氧化反應同時產生過氧化氫，其中所述過氧化氫以低于或等于其在該反應中使用的速率產生。
            
2.根據權利要求1的方法，其中所述的單加氧酶對于H2O2具有至少15nM的Km值。
            
3.根據權利要求1或2的方法，其中所述的單加氧酶為P450酶。
            
4.根據前述權利要求任一項的方法，其中所述的H2O2產生速率為小于或等于每mg酶3ug。
            
5.根據前述權利要求任一項的方法，其中所述整個反應過程中H2O2的濃度小于或等于1mM。
            
6.根據前述權利要求任一項的方法，其中所述反應持續至少240分鐘。
            
7.根據前述權利要求任一項的方法，其中所述的H2O2由電化學反應產生。
            
8.根據權利要求1至6任一項的方法，其中所述的H2O2由酶反應產生。
            
9.根據權利要求8的方法，其中所述的酶為葡萄糖氧化酶。
            
10.根據權利要求1至6任一項的方法，其中所述的H2O2由H2O2前體產生。
            
11.根據權利要求10的方法，其中所述的H2O2前體為過硼酸鹽、過碳酸鹽或過磷酸鹽。
            
12.根據前述權利要求任一項的方法，其中所述的由單加氧酶氧化的底物為烷、芳香族化合物、萜類化合物、烯烴或脂肪酸。
            
13.電極用于產生H2O2以驅動權利要求7所限定的氧化反應的用途。
            
14.酶用于產生H2O2以驅動權利要求8或9所限定的氧化反應的用途。
            
15.過硼酸鹽、過碳酸鹽或過磷酸鹽用于產生H2O2以驅動權利要求10所限定的氧化反應的用途。
            
16.一種進行氧化反應的方法，該氧化反應由單加氧酶催化并使用過氧化氫作為氧化劑，其中單加氧酶氧化損傷反應水平低，所述方法包括在H2O2或羥基螯合劑的存在下進行反應，所述螯合劑可控制H2O2或羥基濃度。
            
17.根據權利要求16的方法，其中所述螯合劑為EDTA。
            
全文摘要
            
本發明提供一種進行由單加氧酶催化及使用過氧化氫作為氧化劑的氧化反應的方法，其中發生單加氧酶的低水平氧化損傷反應，所述方法包括在氧化反應同時產生過氧化氫，其中過氧化氫以低于或等于其在該反應中使用的速率產生，或所述方法包括在H
            
文檔編號C12P7/00GK1934261SQ200580005724 
            
公開日2007年3月21日 申請日期2005年2月23日 優先權日2004年2月23日
            
發明者H·A·O·希爾, L·L·旺格, B·D·弗萊明 申請人:Isis創新有限公司
            


                        
            
                        
            
                        
            
                            
                        
            

                        
                            
                    
            
                    
                    


                     
                        
                        
                    

                
            

                
                
            
                    
            
                        網友詢問留言
                        已有0條留言
                    
            
                    
            
                        
            
                            
                            
              

                                								

						

            • 還沒有人留言評論。精彩留言會獲得點贊！
              • 


                            
            
                        
            
                        
            
                         1 
                    
            
                        
                        
            
                            
            
                              
                            
            
                            
                        
            
                    
            
                
            
            
            
            
            
                
                     
            
            
          
            
      
            
    
            
      

        

  
婷婷六月激情在线综合激情,亚洲国产大片,久久中文字幕综合婷婷,精品久久久久久中文字幕,亚洲一区二区三区高清不卡,99国产精品热久久久久久夜夜嗨
,欧美日韩亚洲综合在线一区二区,99国产精品电影,伊人精品线视天天综合,精品伊人久久久大香线蕉欧美

            

亚洲精品1区
国产成人一级
91精品国产欧美一区二区
亚洲精品乱码久久久久久下载
国产精品久久久久久久伊一
九色国产
国产精品九九视频
伊人久久成人爱综合网
欧美日韩亚洲区久久综合
欧美日本一道免费一区三区
夜夜爽一区二区三区精品
欧美日韩高清一区二区三区
国产成人av在线
国产精品对白交换绿帽视频
国产视频亚洲
国产在线欧美精品
国产精品综合网
国产日韩精品欧美一区色
国产日韩精品欧美一区喷
欧美日韩在线观看区一二
国产区精品
欧美视频日韩视频
中文字幕天天躁日日躁狠狠躁97
视频一二三区
欧美高清在线精品一区二区不卡
国产精品揄拍一区二区久久
99久久综合狠狠综合久久aⅴ
亚洲乱码视频在线观看
日韩在线第二页
亚洲精品无码专区在线播放
成人亚洲网站www在线观看
欧美三级一区二区
99久久精品免费看国产高清
91麻豆国产在线观看
最新日韩欧美不卡一二三区
成人在线观看不卡
日韩国产在线
在线亚洲精品
亚洲午夜久久久久中文字幕
国产精品成人久久久久久久
精品国产一区二区在线观看
欧美精品国产一区二区三区
中文在线播放
亚洲第一页在线视频
国产午夜精品福利久久
九色国产
精品国产九九
国产永久视频
久久精品人人做人人综合试看
国产一区二区三区免费观看
亚洲精品国产电影
9999热视频
国产精品资源在线
麻豆久久婷婷国产综合五月
国产精品免费一级在线观看
亚洲国产一区二区三区青草影视
中文在线播放
国产成人综合在线
国产在线观看色
国产亚洲三级
国产片一区二区三区
久久99精品久久久久久牛牛影视
亚洲欧美日韩国产
四虎永久免费网站
国产一毛片
国产精品视频在
九九热在线精品
99精品福利视频
色婷婷色99国产综合精品
97成人精品视频在线播放
精品久久久久久中文字幕
亚洲欧美一区二区三区孕妇
亚洲欧美成人网
日韩高清在线二区
国产尤物在线观看
在线不卡一区二区
91网站在线看
韩国精品福利一区二区
欧美日韩国产成人精品
99热精品久久
国产精品免费视频一区
高清视频一区
精品九九久久
欧美日韩在线观看免费
91欧美激情一区二区三区成人
99福利视频
亚洲国产精品91
久热国产在线
精品久久久久久中文字幕女
国产精品久久久久久久久99热
成人自拍视频网
国产精品视频久久久久久
久久影院国产
国产玖玖在线观看
99精品在线免费
亚洲欧美一区二区三区导航
久久久久久久综合
国产欧美日韩精品高清二区综合区
国产精品视频自拍
亚洲一级片免费
久久久久久九九
国产欧美自拍视频
视频一区二区在线观看
欧美日韩一区二区三区久久
中文在线亚洲
伊人热人久久中文字幕
日韩欧美亚洲国产一区二区三区
欧美亚洲国产成人高清在线
欧美日韩国产码高清综合人成
国产性大片免费播放网站
亚洲午夜综合网
91精品久久一区二区三区
国产无套在线播放
国产精品视频网站
国产成人亚洲精品老王
91在线网站
国产视频97
欧美黑人欧美精品刺激
国产一区二区三区免费在线视频
久久久国产精品免费看
99re6久精品国产首页
久久精品91
国产成人一级
国产成人精品曰本亚洲
日本福利在线观看
伊人成综合网
久久综合一本
国产综合久久久久久
久久精品成人免费看
久久福利
91精品国产91久久久久久麻豆
亚洲精品成人在线
亚洲伊人久久精品
欧美日本二区
国产永久视频
国产一区二
一区二区福利
国产一毛片
亚洲精品1区
毛片一区二区三区
伊人久久大香线蕉综合影
国产欧美在线观看一区
亚洲国产欧洲综合997久久
国产一区二区免费视频
国产91精品对白露脸全集观看
久久亚洲国产伦理
欧美成人伊人久久综合网
亚洲性久久久影院
久久99国产精一区二区三区!
91精品国产欧美一区二区
欧美日韩亚洲区久久综合
日韩精品一二三区
久久久夜色精品国产噜噜
国产在线精品福利91香蕉
久久久久久久亚洲精品
97se色综合一区二区二区
91国语精品自产拍在线观看性色
91久久国产综合精品女同我
日韩中文字幕a
国产成人亚洲日本精品
久久国产精品-国产精品
久久国产经典视频
久久国产精品伦理
亚洲第一页在线视频
国产精品久久久久三级
日韩毛片网
久久免费高清视频
麻豆国产在线观看一区二区
91麻豆国产福利在线观看
国产成人精品男人的天堂538
一区二区三区中文字幕
免费在线视频一区
欧美日韩国产成人精品
国产综合网站
国产资源免费观看
亚洲精品亚洲人成在线播放
精品久久久久久中文字幕专区
亚洲人成人毛片无遮挡
国产一起色一起爱
国产香蕉精品视频在
九九热免费观看
日韩亚洲欧美一区
九九热精品在线观看
精品久久久久久中文字幕专区
亚洲欧美自拍偷拍
国产精品每日更新
久久久久国产一级毛片高清板
久久天天躁狠狠躁夜夜中文字幕
久久精品片
日韩在线毛片
国产成人精品本亚洲
国产成人精品一区二区三区
九九热在线观看
国产r级在线观看
国产欧美日韩精品高清二区综合区
韩国电影一区二区
国产精品毛片va一区二区三区
五月婷婷伊人网
久久一区二区三区免费
一本色道久久综合狠狠躁篇
亚洲综合色站
国产尤物在线观看
亚洲一区亚洲二区
免费在线视频一区
欧洲精品视频在线观看
日韩中文字幕a
中文字幕日本在线mv视频精品
91精品在线免费视频
精品国产免费人成在线观看
精品a级片
中文字幕日本在线mv视频精品
日韩在线精品视频
婷婷丁香色
91精品国产高清久久久久
国产成人精品日本亚洲直接
五月综合视频
欧美日韩在线亚洲国产人
精液呈暗黄色
亚洲乱码一区
久久精品中文字幕不卡一二区
亚洲天堂精品在线
激情婷婷综合
国产免费久久精品久久久
国产精品亚洲二区在线
久久免费播放视频
五月婷婷丁香综合
在线亚洲欧美日韩
久久免费精品高清麻豆
精品久久久久久中文字幕
亚洲一区网站
国产精品福利社
日韩中文字幕免费
亚洲综合丝袜
91精品在线播放
国产精品18
亚洲日日夜夜
伊人久久大香线蕉综合影
亚洲精品中文字幕乱码影院
亚洲一区二区黄色
亚洲第一页在线视频
一区二区在线观看视频
国产成人福利精品视频
亚洲高清二区
国内成人免费视频
精品亚洲性xxx久久久
国产精品合集一区二区三区
97av免费视频
国产一起色一起爱
国产区久久
国产资源免费观看
99精品视频免费
国产成人一级
国产精品九九免费视频
欧美91精品久久久久网免费
99热国产免费
久久精品色
98精品国产综合久久
久久精品播放
中文字幕视频免费
国产欧美日韩一区二区三区在线
精品久久蜜桃
国产小视频精品
一本色道久久综合狠狠躁篇
91在线免费观看
亚洲精品区
伊人成综合网
伊人热人久久中文字幕
伊人黄色片
99国产精品热久久久久久夜夜嗨
久久免费精品视频
亚洲一区二区三区高清不卡
久久久久国产一级毛片高清板
国产片一区二区三区
久久狠狠干
99久久婷婷国产综合精品电影
国产99区
国产精品成人久久久久
久久狠狠干
青青国产在线观看
亚洲高清国产拍精品影院
国产精品一区二区av
九九热在线免费视频
伊人久久国产
国产精品久久久久久久久久一区
在线观看免费视频一区
国产精品自在在线午夜区app
国产精品综合色区在线观看
国产毛片久久久久久国产毛片
97国产免费全部免费观看
国产精品每日更新
国产尤物视频在线
九九视频这里只有精品99
一本一道久久a久久精品综合
久久综合给会久久狠狠狠
国产成人精品男人的天堂538
欧美一区二区高清
毛片一区二区三区
国产欧美日韩在线观看一区二区三区
在线国产二区
欧美不卡网
91在线精品中文字幕
在线国产福利
国内精品91久久久久
91亚洲福利
日韩欧美国产中文字幕
91久久精品国产性色也91久久
亚洲性久久久影院
欧美精品1区
国产热re99久久6国产精品
九九热免费观看
国产精品欧美日韩
久久久久国产一级毛片高清板
久久国产经典视频
日韩欧美亚洲国产一区二区三区
欧美亚洲综合另类在线观看
国产精品自在在线午夜区app
97中文字幕在线观看
视频一二三区
精品国产一区在线观看
国产欧美日韩在线一区二区不卡
欧美一区二三区
伊人成人在线观看
国内精品91久久久久
97在线亚洲
国产在线不卡一区
久久久全免费全集一级全黄片
国产精品v欧美精品∨日韩
亚洲毛片网站
在线不卡一区二区
99re热在线视频
久久激情网
国产毛片一区二区三区精品
久久亚洲综合色
中文字幕视频免费
国产视频亚洲
婷婷伊人久久
国产一区二区免费播放
久久99国产精品成人欧美
99国产在线视频
国产成人免费视频精品一区二区
国产不卡一区二区三区免费视
国产码欧美日韩高清综合一区
久久精品国产主播一区二区
国产一区电影
久久精品国产夜色
国产精品国产三级国产
日韩一区二区三区在线
久久97久久97精品免视看
久久国产免费一区二区三区
伊人久久大香线蕉综合电影网
99re6久精品国产首页
久久激情网
亚洲成人高清在线
国产精品网址
国产成人精品男人的天堂538
香蕉国产综合久久猫咪
国产专区中文字幕
91麻豆精品国产高清在线
久久国产经典视频
国产精品成人va在线观看
国产精品爱啪在线线免费观看
日本精品久久久久久久久免费
亚洲综合一区二区三区
久久五月网
精品国产网红福利在线观看
久久综合亚洲伊人色
亚洲国产精品久久久久久网站
在线日韩国产
99国产精品热久久久久久夜夜嗨
国产综合精品在线
国产区福利
精品亚洲综合久久中文字幕
国产制服丝袜在线
毛片在线播放网站
在线观看免费视频一区
国产精品久久久精品三级
亚洲国产电影在线观看
最新日韩欧美不卡一二三区
狠狠综合久久综合鬼色
日本精品1在线区
国产日韩一区二区三区在线播放
欧美日韩精品在线播放
亚洲欧美日韩国产一区二区三区精品
久久综合久久网
婷婷六月激情在线综合激情
亚洲乱码一区
国产专区91
97av视频在线观看
精品久久久久久中文字幕
久久五月视频
国产成人福利精品视频
国产精品网址
中文字幕视频在线
精品一区二区三区免费视频
伊人手机在线视频
亚洲精品中文字幕乱码
国产在线视频www色
色噜噜国产精品视频一区二区
精品亚洲成a人在线观看
国产香蕉尹人综合在线
成人免费一区二区三区在线观看
国产不卡一区二区三区免费视
欧美精品久久天天躁
国产专区中文字幕
久久精品国产免费中文
久久精品国产免费一区
久久无码精品一区二区三区
国产欧美另类久久久精品免费
欧美精品久久天天躁
亚洲精品在线视频
国产视频91在线
91精品福利一区二区三区野战
日韩中文字幕免费
国产精品99一区二区三区
欧美成人高清性色生活
国产精品系列在线观看
亚洲国产福利精品一区二区
国产成人在线小视频
国产精品久久久久免费
99re热在线视频
久久久久久久综合
一区二区国产在线播放
成人国产在线视频
亚洲精品乱码久久久久
欧美日韩一区二区综合
精品久久久久免费极品大片
中文字幕视频二区
激情粉嫩精品国产尤物
国产成人精品一区二区视频
久久精品中文字幕首页
亚洲高清在线
国产精品亚洲一区二区三区
伊人久久艹
中文在线亚洲
国产精品一区二区在线播放
国产精品九九免费视频
亚洲二区在线播放
亚洲狠狠婷婷综合久久久久网站
亚洲欧美日韩网站
日韩成人精品
亚洲国产一区二区三区青草影视
91精品国产福利在线观看
国产精品久久久久久久久99热
国产一区二区精品尤物
久碰香蕉精品视频在线观看
亚洲日日夜夜
在线不卡一区二区
国产午夜亚洲精品
九九热在线视频观看这里只有精品
伊人手机在线视频
91免费国产精品
日韩欧美中字
91精品国产91久久久久
国产全黄三级播放
视频一区二区三区免费观看
国产开裆丝袜高跟在线观看
国产成人欧美
激情综合丝袜美女一区二区
国产成人亚洲综合无
欧美精品一区二区三区免费观看
欧美亚洲国产日韩
日韩亚州
国产欧美日韩精品高清二区综合区
亚洲午夜国产片在线观看
精品久久久久久中文字幕
欧美精品1区
久久伊人久久亚洲综合
亚洲欧美日韩精品
国产成人精品久久亚洲高清不卡
久久福利影视
国产精品99精品久久免费
久久久久免费精品视频
国产日产亚洲精品
亚洲国产午夜电影在线入口
精品无码一区在线观看
午夜国产精品视频
亚洲一级片免费
伊人久久大香线蕉综合影
国产精品久久影院
久碰香蕉精品视频在线观看
www.欧美精品
在线小视频国产
亚洲国产天堂久久综合图区
欧美一区二区三区不卡
日韩美女福利视频
九九精品免视频国产成人
不卡国产00高中生在线视频
亚洲第一页在线视频
欧美日韩在线播放成人
99re视频这里只有精品
国产精品91在线
精品乱码一区二区三区在线
国产区久久
91麻豆精品国产自产在线观看一区
日韩精品成人在线
九九热在线观看
国产精品久久不卡日韩美女
欧美一区二区三区综合色视频
欧美精品免费一区欧美久久优播
国产精品网址
国产专区中文字幕
国产精品欧美亚洲韩国日本久久
日韩美香港a一级毛片
久久精品123
欧美一区二区三区免费看
99r在线视频
亚洲精品国产字幕久久vr
国产综合激情在线亚洲第一页
91免费国产精品
日韩免费小视频
亚洲国产精品综合一区在线
国产亚洲第一伦理第一区
在线亚洲精品
国产精品一区二区制服丝袜
国产在线成人精品
九九精品免视频国产成人
亚洲国产网
欧美日韩亚洲一区二区三区在线观看
在线亚洲精品
欧美一区二区三区高清视频
国产成人精品男人的天堂538
欧美日韩在线观看区一二
亚洲欧美一区二区久久
久久精品中文字幕首页
日本高清www午夜视频
久久精品国产免费
久久999精品
亚洲国产精品欧美综合
88国产精品视频一区二区三区
91久久偷偷做嫩草影院免费看
国产精品夜色视频一区二区
欧美日韩导航
国产成人啪精品午夜在线播放
一区二区视频在线免费观看
99久久精品国产自免费
精液呈暗黄色
久久99国产精品
日本精品久久久久久久久免费
精品国产97在线观看
99re视频这里只有精品
国产视频91在线
999av视频
亚洲美女视频一区二区三区
久久97久久97精品免视看
亚洲国产成人久久三区
99久久亚洲国产高清观看
日韩毛片在线视频
综合激情在线
91福利一区二区在线观看
一区二区视频在线免费观看
激情粉嫩精品国产尤物
国产成人精品曰本亚洲78
国产成人精品本亚洲
国产精品成人免费视频
国产成人啪精品视频免费软件
久久精品国产亚洲妲己影院
国产精品成人久久久久久久
久久大香线蕉综合爱
欧美一区二区三区高清视频
99热国产免费
在线观看欧美国产
91精品视频在线播放
国产精品福利社
欧美精品一区二区三区免费观看
国产一区二区免费视频
国产午夜精品一区二区
精品视频在线观看97
91精品福利久久久
国产一区福利
国产综合激情在线亚洲第一页
国产精品久久久久久久久久久不卡
九色国产
在线日韩国产
黄网在线观看
亚洲一区小说区中文字幕
中文字幕丝袜
日本二区在线观看
日本国产一区在线观看
欧美日韩一区二区三区久久
欧美精品亚洲精品日韩专
国产日产亚洲精品
久久综合九色综合欧美播
亚洲国产欧美无圣光一区
欧美视频区
亚洲乱码视频在线观看
久久无码精品一区二区三区
九九热精品免费视频
久久99精品久久久久久牛牛影视
国产精品成久久久久三级
国产一区福利
午夜国产精品视频
日本二区在线观看
99久久网站
国产亚洲天堂
精品国产一区二区三区不卡
亚洲国产日韩在线一区
国产成人综合在线观看网站
久久免费高清视频
欧美在线导航
午夜精品久久久久久99热7777
欧美久久综合网
国产小视频精品
国产尤物在线观看
亚洲国产精品综合一区在线
欧美一区二区三区不卡视频
欧美黑人欧美精品刺激
日本福利在线观看
久久国产偷
国产手机精品一区二区
国产热re99久久6国产精品
国产高清啪啪
欧美亚洲国产成人高清在线
国产在线第三页
亚洲综合一区二区三区
99r在线视频
99精品久久久久久久婷婷
国产精品乱码免费一区二区
国产在线精品福利91香蕉
国产尤物视频在线
五月婷婷亚洲
中文字幕久久综合伊人
亚洲精品一级毛片
99国产精品电影
在线视频第一页
久久99国产精品成人欧美
国产白白视频在线观看2
成人精品一区二区www
亚洲成人网在线观看
麻豆91在线视频
色综合合久久天天综合绕视看
久久精品国产免费高清
国产不卡一区二区三区免费视
欧美国产中文
99精品欧美
九九在线精品
国产中文字幕在线免费观看
国产一区中文字幕在线观看
国产成人一级
国产精品一区二区制服丝袜
国产一起色一起爱
亚洲精品成人在线
亚洲欧美精品在线
国产欧美自拍视频
99精品久久久久久久婷婷
久99视频
国产热re99久久6国产精品
视频一区亚洲
国产精品视频分类
国产精品成在线观看
99re6久精品国产首页
亚洲在成人网在线看
亚洲国产日韩在线一区
久久国产三级
日韩国产欧美
欧美在线一区二区三区
国产精品美女一级在线观看
成人午夜免费福利视频
亚洲天堂精品在线
91精品国产手机
欧美日韩视频在线播放
狠狠综合久久综合鬼色
九一色视频
青青视频国产
亚洲欧美自拍一区
中文字幕天天躁日日躁狠狠躁97
日韩免费大片
996热视频
伊人成综合网
亚洲天堂欧美
日韩精品亚洲人成在线观看
久久综合给会久久狠狠狠
日韩精品亚洲人成在线观看
日韩国产欧美
亚洲成aⅴ人片在线影院八
亚洲精品1区
99久久精品免费
国产精品高清在线观看
国产精品久久久免费视频
在线亚洲欧美日韩
91在线看视频
国产精品96久久久久久久
欧美日韩国产成人精品
91在线亚洲
热久久亚洲
国产精品美女免费视频观看
日韩在线毛片
亚洲永久免费视频
九九免费在线视频
亚洲一区网站
日本高清二区视频久二区
精品国产美女福利在线
伊人久久艹
国产精品久久久久三级
欧美成人精品第一区二区三区
99久久精品国产自免费
在线观看日韩一区
国产中文字幕一区
成人免费午夜视频
欧美日韩另类在线
久久99国产精品成人欧美
色婷婷中文网
久久天天躁夜夜躁狠狠躁2020
欧美成人伊人久久综合网
国产精品福利资源在线
国产伦精品一区二区三区高清
国产精品亚洲综合色区韩国
亚洲一区欧美日韩
色综合视频
国语自产精品视频在线区
国产高清a
成人国内精品久久久久影
国产在线精品香蕉综合网一区
国产不卡在线看
国产成人精品精品欧美
国产欧美日韩综合精品一区二区三区
韩国电影一区二区
国产在线视频www色
91中文字幕在线一区
国产人成午夜免视频网站
亚洲综合一区二区三区
色综合视频一区二区观看
久久五月网
九九热精品在线观看
国产一区二区三区国产精品
99久热re在线精品996热视频
亚洲国产网
在线视频亚洲一区
日韩字幕一中文在线综合
国产高清一级毛片在线不卡
精品国产色在线
国产高清视频一区二区
精品日本久久久久久久久久
亚洲国产午夜精品乱码
成人免费国产gav视频在线
日韩欧美一区二区在线观看
欧美曰批人成在线观看
韩国电影一区二区
99re这里只有精品6
日韩精品一区二区三区视频
99re6久精品国产首页
亚洲欧美一区二区三区导航
欧美色图一区二区三区
午夜精品视频在线观看
欧美激情在线观看一区二区三区
亚洲热在线
成人国产精品一区二区网站
亚洲一级毛片在线播放
亚洲一区小说区中文字幕
亚洲午夜久久久久影院
国产自产v一区二区三区c
国产精品视频免费
久久调教视频
国产成人91激情在线播放
国产精品欧美亚洲韩国日本久久
久久亚洲日本不卡一区二区
91中文字幕网
成人国产在线视频
国产视频91在线
欧美成人精品第一区二区三区
国产精品福利在线
久久综合九色综合精品
欧美一区二区三区精品
久久国产综合尤物免费观看
久久99青青久久99久久
日韩精品免费
久久国产精品999
91亚洲视频在线观看
国产精品igao视频
色综合区
在线亚洲欧国产精品专区
国产一区二区三区在线观看视频
亚洲精品成人在线
一区二区国产在线播放
中文在线亚洲
亚洲精品第一国产综合野
国产一区二区精品久久
一区二区三区四区精品视频
99热精品久久
中文字幕视频二区
国产成人精品男人的天堂538
99精品影视
美女福利视频一区二区
久久午夜夜伦伦鲁鲁片
综合久久久久久久综合网
国产精品国产欧美综合一区
国产99视频在线观看
国产亚洲女在线精品
婷婷影院在线综合免费视频
国产亚洲3p一区二区三区
91成人爽a毛片一区二区
亚洲一区二区高清
国产欧美亚洲精品第二区首页
欧美日韩导航
亚洲高清二区
欧美激情观看一区二区久久
日韩毛片在线播放
亚洲欧美日韩高清中文在线
亚洲日本在线播放
国产精品一区二区制服丝袜
精品国产一区二区三区不卡
国产不卡在线看
国产欧美网站
四虎永久在线观看视频精品
国产黄色片在线观看
夜夜综合
一本色道久久综合狠狠躁篇
欧美亚洲综合另类在线观看
国产91在线看
伊人久久国产
欧美一区二区在线观看免费网站
国产精品久久久久三级
久久福利
日韩中文字幕a
亚洲午夜久久久久影院
91在线高清视频
国产亚洲一区二区三区啪
久久人精品
国产精品亚洲午夜一区二区三区
综合久久久久久
久久伊人一区二区三区四区
国产综合久久久久久
日韩一区精品视频在线看
国产精品日韩欧美制服
日本精品1在线区
99re视频
无码av免费一区二区三区试看
国产视频1区
日韩欧美中文字幕一区
日本高清中文字幕一区二区三区a
亚洲国产欧美无圣光一区
国产在线视频一区二区三区
欧美国产第一页
在线亚洲欧美日韩
日韩中文字幕第一页
在线不卡一区二区
伊人久久青青
国产精品一区二区在线播放
www.五月婷婷
麻豆久久婷婷国产综合五月
亚洲精品区
久久国产欧美另类久久久
99在线视频免费
伊人久久中文字幕久久cm
久久精品成人免费看
久久这里只有精品首页
88国产精品视频一区二区三区
中文字幕日本在线mv视频精品
国产在线精品成人一区二区三区
伊人精品线视天天综合
亚洲一区二区黄色
国产尤物视频在线
亚洲精品99久久久久中文字幕
国产一区二区三区免费观看
伊人久久大香线蕉综合电影网
国产成人精品区在线观看
日本精品一区二区三区视频
日韩高清在线二区
久久免费播放视频
一区二区成人国产精品
国产精品免费精品自在线观看
亚洲精品视频二区
麻豆国产精品有码在线观看
精品日本一区二区
亚洲欧洲久久
久久中文字幕综合婷婷
中文字幕视频在线
国产成人精品综合在线观看
91精品国产91久久久久福利
精液呈暗黄色
香蕉国产综合久久猫咪
国产专区精品
亚洲精品无码不卡
国产永久视频
亚洲成a人片在线播放观看国产
一区二区国产在线播放
亚洲一区二区黄色
欧美日韩在线观看视频
亚洲精品另类
久久国产综合尤物免费观看
国产一区二区三区国产精品
高清视频一区
国产精品igao视频
国产精品资源在线
久久综合精品国产一区二区三区
www.五月婷婷
精品色综合
99热国产免费
麻豆福利影院
亚洲伊人久久大香线蕉苏妲己
久久电影院久久国产
久久精品伊人
在线日韩理论午夜中文电影
亚洲国产欧洲综合997久久
伊人国产精品
久草国产精品
欧美一区精品二区三区
亚洲成人高清在线
91免费国产精品
日韩精品福利在线
国产一线在线观看
国产不卡在线看
久久99青青久久99久久
亚洲精品亚洲人成在线播放
99久久免费看国产精品
国产日本在线观看
青草国产在线视频
麻豆久久婷婷国产综合五月
国产中文字幕一区
91久久精品国产性色也91久久
国产一区a
国产欧美日韩成人
国产亚洲女在线精品
一区二区美女
中文字幕在线2021一区
在线小视频国产
久久这里只有精品首页
国产在线第三页
欧美日韩中文字幕
在线亚洲+欧美+日本专区
精品国产一区二区三区不卡
久久这里精品
欧美在线va在线播放
精液呈暗黄色
91精品国产手机
91在线免费播放
欧美视频亚洲色图
欧美国产日韩精品
日韩高清不卡在线
精品视频免费观看
欧美日韩一区二区三区四区
国产欧美亚洲精品第二区首页
亚洲韩精品欧美一区二区三区
国产精品视频免费
在线精品小视频
久久午夜夜伦伦鲁鲁片
国产无套在线播放
久热这里只精品99re8久
欧美久久久久
久久香蕉国产线看观看精品蕉
国产成人精品男人的天堂538
亚洲人成网站色7799在线观看
日韩在线第二页
一本色道久久综合狠狠躁篇
国产一区二区三区不卡在线观看
亚洲乱码在线
在线观看欧美国产
久久福利青草精品资源站免费
国产玖玖在线观看
在线亚洲精品
亚洲成aⅴ人在线观看
精品91在线
欧美一区二三区
日韩中文字幕视频在线
日本成人一区二区
日韩免费专区
国内精品在线观看视频
久久国产综合尤物免费观看
国产精品系列在线观看
一本一道久久a久久精品综合
亚洲免费播放
久久精品国产免费
久久人精品
亚洲毛片网站
亚洲成a人一区二区三区
韩国福利一区二区三区高清视频
亚洲精品天堂在线
一区二区三区中文字幕
亚洲国产色婷婷精品综合在线观看
亚洲国产成人久久笫一页
999国产视频
国产精品香港三级在线电影
欧美日韩一区二区三区四区
日韩国产欧美
国产精品99一区二区三区
午夜国产精品理论片久久影院
亚洲精品中文字幕麻豆
亚洲国产高清视频
久久免费手机视频
日韩a在线观看
五月婷婷亚洲
亚洲精品中文字幕麻豆
中文字幕丝袜
www国产精品
亚洲天堂精品在线
亚洲乱码一区
国产日韩欧美三级
久久999精品
伊人热人久久中文字幕
久热国产在线视频
国产欧美日韩在线观看一区二区三区
国产一二三区在线
日韩国产欧美
91精品国产91久久久久
亚洲一区小说区中文字幕
精品一区二区免费视频
国产精品视频免费
国产精品亚洲综合色区韩国
亚洲国产精品成人午夜在线观看
欧美国产日韩精品
中文字幕精品一区二区精品