包含枯草杆菌蛋白酶变体的组合物和方法技术

本发明专利技术提供由丝氨酸蛋白酶制备的丝氨酸蛋白酶变体。具体地讲，本发明专利技术提供与参照丝氨酸蛋白酶相比具有一个或多个置换的丝氨酸蛋白酶变体。此外，本发明专利技术提供包含这些丝氨酸蛋白酶变体的组合物。在一些实施例中，本发明专利技术提供包含这些丝氨酸蛋白酶变体中的至少一种的清洁组合物。

【技术实现步骤摘要】
本申请是申请日为2011年5月5日、专利技术名称为“包含枯草杆菌蛋白酶变体的组合物和方法”的中国专利申请201180022692.4(国际申请号PCT/US2011/035389)的分案申请。相关专利申请的交叉引用本专利申请要求以下美国临时申请的权利：于2010年5月6日提交的No.61/332,151；于2010年5月6日提交的No.61/332,006；于2010年10月12日提交的No.61/392,364以及于2010年10月12日提交的No.61/392,188，所有这些临时申请全文均以引用方式并入本文。
本专利技术提供丝氨酸蛋白酶变体。具体地讲，本专利技术提供与参照丝氨酸蛋白酶相比具有一个或多个置换的丝氨酸蛋白酶变体。此外，本专利技术提供包含这些丝氨酸蛋白酶变体的组合物。在一些实施例中，本专利技术提供清洁组合物，其包含这些丝氨酸蛋白酶变体中的至少一种。专利技术背景尽管丝氨酸蛋白酶在工业酶领域中已久为人所知，但对于适于特定条件和用途的工程化蛋白酶仍存在需要。
技术实现思路
本专利技术提供丝氨酸蛋白酶变体。具体地讲，本专利技术提供与参照丝氨酸蛋白酶相比具有一个或多个置换的丝氨酸蛋白酶变体。此外，本专利技术提供包含这些丝氨酸蛋白酶变体的组合物。在一些实施例中，本专利技术提供清洁组合物，其包含这些丝氨酸蛋白酶变体中的至少一种。本专利技术提供分离的枯草杆菌蛋白酶变体，其中该枯草杆菌蛋白酶变体为具有蛋白水解活性的成熟形式并包含具有选自以下的氨基酸置换的组合的氨基酸序列：X1R,X2W,X2M,X2R,X2A,X2S,X3R,X4R,X4C,X4S,X8A,X9F,X9W,X9A,X10S,X10M,X10H,X10A,X12R,X12F,X14K,X14F,X14Q,X15R,X15F,X16S,X17R,X17M,X17F,X18R,X18K,X20F,X20R,X20K,X22Y,X22A,X22R,X22V,X22Q,X22W,X22L,X23F,X23S,X23A,X24W,X24R,X24H,X24F,X24Q,X24L,X25V,X25F,X25R,X26F,X27V,X27F,X27L,X27R,X28N,X28E,X28A,X29T,X30E,X31F,X33D,X33G,X33S,X34P,X35M,X36F,X36R,X36T,X38L,X38F,X38R,X40L,X40W,X40N,X40R,X40T,X40H,X42I,X43D,X43I,X43R,X43M,X43F,X43W,X43S,X43A,X45T,X46R,X48R,X50C,X51H,X51W,X51F,X52F,X52E,X52N,X55Y,X57R,X59R,X59A,X59F,X60A,X60Q,X60P,X62E,X62Q,X63I,X63V,X63T,X63P,X63D,X63M,X63H,X63Q,X63E,X63A,X63S,X64F,X64T,X68C,X68A,X69N,X69P,X69W,X69T,X71G,X72C,X74C,X75F,X75A,X75R,X75E,X76D,X78I,X78R,X78N,X79W,X79Q,X81R,X82V,X82T,X82F,X82M,X82R,X85M,X86L,X86I,X86W,X89P,X89T,X89V,X89G,X89W,X89H,X89F,X89L,X89I,X91N,X91F,X92F,X94N,X99G,X99F,X99M,X99T,X99P,X100I,X100S,X100N,X100Q,X101N,X101A,X101G,X101P,X101F,X101E,X101T,X101D,X102H,X102N,X102E,X102T,X102A,X103G,X103D,X103N,X104D,X104E,X104I,X104L,X105Q,X105E,X105T,X106F,X106V,X106G,X106E,X106T,X106D,X106A,X107F,X107M,X108G,X108I,X109M,X111V,X111I,X112V,X112L,X112Q,X114G,X115K,X115R,X116A,X116K,X116L,X117F,X118I,X118R,X119C,X120F,X120A,X120R,X121E,X121F,X123G,X123E,X124S,X128F,X128H,X128I,X128L,X128Q,X128N,X128M,X128D,X129E,X132E,X132A,X138G,X144R,X147L,X148I,X158E,X158E,X159E,X159C,X160D,X166D,X166E,X167W,X175V,X177C,X181A,X182R,X183F,X183I,X183D,X183R,X183M,X185E,X185I,X185V,X186H,X186K,X188R,X188E,X188D,X192H,X192W,X194V,X194F,X194E,X197F,X198L,X198F,X203E,X203C,X208S,X209N,X209F,X209E,X209S,X209H,X209G,X209T,X209L,X210R,X210V,X210L,X211R,X211Q,X212I,X212M,X212F,X213A,X214F,X215F,X215N,X215D,X215H,X215E,X216F,X216A,X217N,X217E,X217D,X218P,X218D,X218E,X224A,X224G,X227I,X230E,X231I,X231C,X233C,X234F,X235F,X236F,X236N,X238L,X238K,X238R,X239K,X239S,X239T,X239G,X239H,X239R,X239N,X239F,X240R,X241R,X242L,X242R,X243R,X243F,X244R,X246S,X248I,X248V,X248R,X249R,X249T,X250I,X251S,X251R,X252I,X252F,X252H,X252R,X253F,X253I,X253R,X254C,X256N,X258R,X260V,X260I,X262H,X262D,X263F,X265F,X267N,X267V,X267M,X269I,X269R,X270C,X271F,X271V,X271I,X271P,X271H,X271M,X271T,X271L,X271A,X272F,X272R,X272F,X273I,X273F，和X274G，其中该枯草杆菌蛋白酶变体的氨基酸位置通过与如SEQ ID NO:1所示解淀粉芽孢杆菌(B.amyloliquefaciens)枯草杆菌蛋白酶BPN’的氨基酸序列的对应关系进行编号。本专利技术还提供分离的枯草杆菌蛋白酶变体，其中该枯草杆菌蛋白酶变体为具有蛋白水解活性的成熟形式并包含具有选自以下的氨基酸置换的组合的氨基酸序列：X16S,X18R,X20R,X22A,X24R,X43R/D,X45T,X76D,X101A,X103G,X104L本文档来自技高网...

【技术保护点】
一种分离的枯草杆菌蛋白酶变体，其中所述枯草杆菌蛋白酶变体为具有蛋白水解活性的成熟形式并包含具有选自以下的氨基酸置换的组合的氨基酸序列：G020R‑N043R,N062E‑A158E,S103G‑A158E,S128N‑A158E,A016S‑A158E,V104L‑A158E,E089P‑A158E,L111V‑A158E,T022A‑A158E,S101A‑A158E,L148I‑A158E,P129E‑A158E,T022A‑E089P,A016S‑E089P,N062E‑E089P,N062E‑E271F,A158E‑E271F,R186H‑E271F,P129E‑E271F,L111V‑E271F,Y209E‑E271F,A016S‑E271F,S188D‑E271F,T022A‑E271F,G159E‑E271F,V104L‑E271F,S101A‑E271F,E089P‑E271F,S128N‑E271F,S103G‑E271F,L148I‑E271F,H249R‑E271F,N062E‑G159E,A016S‑G159E,S128N‑G159E,L148I‑G159E,L111V‑G159E,E089P‑G159E,T022A‑G159E,P129E‑G159E,S103G‑G159E,V104L‑G159E,A158E‑G159E,S101A‑G159E,A158E‑H249R,L111V‑H249R,P129E‑H249R,N062E‑H249R,A016S‑H249R,R186H‑H249R,L148I‑H249R,G159E‑H249R,S101A‑H249R,S188D‑H249R,V104L‑H249R,Y209E‑H249R,T022A‑H249R,S128N‑H249R,S103G‑H249R,E089P‑H249R,T022A‑L111V,S101A‑L111V,A016S‑L111V,V104L‑L111V,N062E‑L111V,S103G‑L111V,E089P‑L111V,A016S‑L148I,N062E‑L148I,T022A‑L148I,P129E‑L148I,V104L‑L148I,S103G‑L148I,S128N‑L148I,S101A‑L148I,E089P‑L148I,L111V‑L148I,A016S‑N062E,T022A‑N062E,N062E‑P129E,T022A‑P129E,S128N‑P129E,A016S‑P129E,S101A‑P129E,V104L‑P129E,E089P‑P129E,S103G‑P129E,L111V‑P129E,N062E‑R186H,S128N‑R186H,S101A‑R186H,T022A‑R186H,A016S‑R186H,A158E‑R186H,E089P‑R186H,P129E‑R186H,G159E‑R186H,S103G‑R186H,V104L‑R186H,L111V‑R186H,L148I‑R186H,N062E‑S101A,T022A‑S101A,A016S‑S101A,E089P‑S101A,N062E‑S103G,T022A‑S103G,A016S‑S103G,S101A‑S103G,E089P‑S103G,N062E‑S128N,A016S‑S128N,T022A‑S128N,S101A‑S128N,V104L‑S128N,E089P‑S128N,S103G‑S128N,L111V‑S128N,L111V‑S188D,N062E‑S188D,A016S‑S188D,L148I‑S188D,T022A‑S188D,S128N‑S188D,S101A‑S188D,V104L‑S188D,E089P‑S188D,P129E‑S188D,G159E‑S188D,R186H‑S188D,S103G‑S188D,A158E‑S188D,A016S‑T022A,A016S‑V104L,T022A‑V104L,S101A‑V104L,N062E‑V104L,S103G‑V104L,E089P‑V104L,G159E‑Y209E,L111V‑Y209E,S101A‑Y209E,A016S‑Y209E,S128N‑Y209E,L148I‑Y209E,P129E‑Y209E,N062E‑Y209E,T022A‑Y209E,S103G‑Y209E,A158E‑Y209E,S188D‑Y209E,V104L‑Y209E,E089P‑Y209E,R186H‑Y209E,N018R‑W241R,G020R‑W241R,S024R‑W241R,S009A‑W241R,G020R‑W241R,V004R‑W241R,N043R‑W241R,S...

【技术特征摘要】
2010.05.06 US 61/332,006;2010.05.06 US 61/332,151;1.一种分离的枯草杆菌蛋白酶变体，其中所述枯草杆菌蛋白酶变体为具有蛋白水解活性的成熟形式并包含具有选自以下的氨基酸置换的组合的氨基酸序列：G020R-N043R,N062E-A158E,S103G-A158E,S128N-A158E,A016S-A158E,V104L-A158E,E089P-A158E,L111V-A158E,T022A-A158E,S101A-A158E,L148I-A158E,P129E-A158E,T022A-E089P,A016S-E089P,N062E-E089P,N062E-E271F,A158E-E271F,R186H-E271F,P129E-E271F,L111V-E271F,Y209E-E271F,A016S-E271F,S188D-E271F,T022A-E271F,G159E-E271F,V104L-E271F,S101A-E271F,E089P-E271F,S128N-E271F,S103G-E271F,L148I-E271F,H249R-E271F,N062E-G159E,A016S-G159E,S128N-G159E,L148I-G159E,L111V-G159E,E089P-G159E,T022A-G159E,P129E-G159E,S103G-G159E,V104L-G159E,A158E-G159E,S101A-G159E,A158E-H249R,L111V-H249R,P129E-H249R,N062E-H249R,A016S-H249R,R186H-H249R,L148I-H249R,G159E-H249R,S101A-H249R,S188D-H249R,V104L-H249R,Y209E-H249R,T022A-H249R,S128N-H249R,S103G-H249R,E089P-H249R,T022A-L111V,S101A-L111V,A016S-L111V,V104L-L111V,N062E-L111V,S103G-L111V,E089P-L111V,A016S-L148I,N062E-L148I,T022A-L148I,P129E-L148I,V104L-L148I,S103G-L148I,S128N-L148I,S101A-L148I,E089P-L148I,L111V-L148I,A016S-N062E,T022A-N062E,N062E-P129E,T022A-P129E,S128N-P129E,A016S-P129E,S101A-P129E,V104L-P129E,E089P-P129E,S103G-P129E,L111V-P129E,N062E-R186H,S128N-R186H,S101A-R186H,T022A-R186H,A016S-R186H,A158E-R186H,E089P-R186H,P129E-R186H,G159E-R186H,S103G-R186H,V104L-R186H,L111V-R186H,L148I-R186H,N062E-S101A,T022A-S101A,A016S-S101A,E089P-S101A,N062E-S103G,T022A-S103G,A016S-S103G,S101A-S103G,E089P-S103G,N062E-S128N,A016S-S128N,T022A-S128N,S101A-S128N,V104L-S128N,E089P-S128N,S103G-S128N,L111V-S128N,L111V-S188D,N062E-S188D,A016S-S188D,L148I-S188D,T022A-S188D,S128N-S188D,S101A-S188D,V104L-S188D,E089P-S188D,P129E-S188D,G159E-S188D,R186H-S188D,S103G-S188D,A158E-S188D,A016S-T022A,A016S-V104L,T022A-V104L,S101A-V104L,N062E-V104L,S103G-V104L,E089P-V104L,G159E-Y209E,L111V-Y209E,S101A-Y209E,A016S-Y209E,S128N-Y209E,L148I-Y209E,P129E-Y209E,N062E-Y209E,T022A-Y209E,S103G-Y209E,A158E-Y209E,S188D-Y209E,V104L-Y209E,E089P-Y209E,R186H-Y209E,N018R-W241R,G020R-W241R,S024R-W241R,S009A-W241R,G020R-W241R,V004R-W241R,N043R-W241R,S078R-W241R,T022R-W241R,G115R-W241R,A001R-W241R,S212F-W241R,L082R-W241R,N018R-V244R,S024R-V244R,S078R-V244R,G020R-V244R,S212F-V244R,S009A-V244R,L082R-V244R,A001R-V244R,N043R-V244R,T022R-V244R,V004R-V244R,G115R-V244R,W241R-V244R,S242R-V244R,A001R-V004R,S009A-T022R,N018R-T022R,G020R-T022R,V004R-T022R,A001R-T022R,S024R-S242R,N018R-S242R,V004R-S242R,G020R-S242R,S212F-S242R,L082R-S242R,S078R-S242R,A001R-S242R,S009A-S242R,T022R-S242R,G115R-S242R,N043R-S242R,W241R-S242R,N018R-S212F,T022R-S212F,V004R-S212F,S024R-S212F,A001R-S212F,G115R-S212F,G020R-S212F,S009A-S212F,N043R-S212F,S078R-S212F,L082R-S212F,S009A-S078R,G020R-S078R,S024R-S078R,T022R-S078R,N018R-S078R,V004R-S078R,A001R-S078R,N043R-S078R,T022R-S024R,G020R-S024R,N018R-S024R,A001R-S024R,V004R-S024R,S009A-S024R,V004R-S009A,A001R-S009A,S242R-N269R,S024R-N269R,G020R-N269R,T022R-N269R,H249R-N269R,S212F-N269R,N043R-N269R,V244R-N269R,A001R-N269R,N018R-N269R,S078R-N269R,S009A-N269R,G115R-N269R,W241R-N269R,V004R-N269R,L082R-N269R,N018R-N043R,G020R-N043R,V004R-N043R,T022R-N043R,S009A-N043R,A001R-N043R,S024R-N043R,S009A-N018R,V004R-N018R,A001R-N018R,S024R-L082R,S009A-L082R,N018R-L082R,A001R-L082R,S078R-L082R,G020R-L082R,T022R-L082R,V004R-L082R,N043R-L082R,N043R-H249R,G020R-H249R,V004R-H249R,N018R-H249R,S009A-H249R,S212F-H249R,T022R-H249R,S024R-H249R,G115R-H249R,A001R-H249R,L082R-H249R,S242R-H249R,W241R-H249R,V244R-H249R,S078R-H249R,N018R-G115R,G020R-G115R,T022R-G115R,S078R-G115R,S009A-G115R,V004R-G115R,A001R-G115R,L082R-G115R,N043R-G115R,S024R-G115R,S009A-G020R,N018R-G020R,V004R-G020R,A001R-G020R,S009A-E271L,G020R-E271L,S024R-E271L,V244R-E271L,W241R-E271L,N043R-E271L,T022R-E271L,H249R-E271L,S212F-E271L,G115R-E271L,S242R-E271L,S078R-E271L,V004R-E271L,N269R-E271L,A001R-E271L,N018R-E271L，和L082R-E271L，并且其中所述变体的总净电荷相对于迟缓芽孢杆菌(Bacillus lentus)枯草杆菌蛋白酶GG36蛋白酶的总净电荷为0、+1、+2、+3、+4、+5、-1、-2、-3、-4或-5，并且其中所述总净电荷是通过一种或多种选自以下的置换来获得的：N43D,R45T,N62E,N76D,S101D,P129E,A158E,G159D,G159E,S166D,S188D,A230E,N18R,G20K,G20R,T22R,S24R,N43R,G118R,Q245R,H249R,N269R,E271F，和E271L，并且其中所述蛋白酶变体的氨基酸位置根据SEQ ID NO:1所示解淀粉芽孢杆菌(Bacillus amyloliquefaciens)枯草杆菌蛋白酶BPN’的氨基酸序列中对应的氨基酸位置的编号方式来进行编号。2.一种分离的枯草杆菌蛋白酶变体，其中所述枯草杆菌蛋白酶变体为具有蛋白水解活性的成熟形式并包含具有选自以下的氨基酸置换的组合的氨基酸序列：G020R-N043R,G020K-N062E,S024F-N116L,G020K-S024F,S024R-A174T,S024R-G118R,S024R-K235F,S024R-P086R,S024R-P086W,S078R-G118R,T033S-G118R,T033S-K235F,Y209A-W241R,G020R-N076D,N018R-Q245R,S024R-R045T,A232V-Q245R,G118R-A172V,G118R-A194T,I008T-S024R,K235F-N243F,N018R-S103A,N018R-V104I,P086W-G118R,P086W-N243F,P086W-Y209A,S024C-T033S,S024R-A232V,S024R-N243F,S024R-P239Q,S024R-S101G,S024R-S141G,S024R-T033S,S024R-T274I,S024R-Y209A,S078R-P086W,S101G-A232V,T033S-L148F,T033S-P086W,T033S-P201S,T033S-S078R,T033S-W241R,T033S-Y209A,A230E-H249R,A232V-H249R,G118R-K235F,N076D-Q245R,P086W-K235F,S024R-R247H,S024R-V104A,S078R-K235F,S101G-H249R,S103A-A232V,T033S-A048T,T033S-P239T,T033S-T253A,T143A-Y209A,Y209A-K235F,N018R-R045T,Y209A-N243F,S024R-A272P,S024R-R269C,S101G-V104I,V104I-A232V,N076D-H249R，和S024R-N076D，并且其中所述变体的总净电荷相对于迟缓芽孢杆菌(Bacillus lentus)枯草杆菌蛋白酶GG36蛋白酶的总净电荷为0、+1、+2、+3、+4、+5、-1、-2、-3、-4或-5，并且其中所述总净电荷是通过一种或多种选自以下的置换来获得的：N43D,R45T,N62E,N76D,S101D,P129E,A158E,G159D,G159E,S166D,S188D,A230E,N18R,G20K,G20R,T22R,S24R,N43R,G118R,Q245R,H249R,N269R,E271F，和E271L，并且其中所述蛋白酶变体的氨基酸位置根据SEQ ID NO:1所示解淀粉芽孢杆菌(Bacillus amyloliquefaciens)枯草杆菌蛋白酶BPN’的氨基酸序列中对应的氨基酸位置的编号方式来进行编号。3.一种分离的枯草杆菌蛋白酶变体，其中所述枯草杆菌蛋白酶变体为具有蛋白水解活性的成熟形式并包含具有选自以下的氨基酸置换的组合的氨基酸序列：G020R-N076D,S024R-R045T,A230E-H249R,N018R-R045T,N018R-Q245R,S101G-A232V,S024R-A232V,A232V-Q245R,S024R-S101G,N018R-V104I,N018R-S103A,S101G-H249R,A232V-H249R,S103A-A232V,N076D-Q245R,S101G-V104I,V104I-A232V,N076D-H249R,S024R-N076D,S024F-N116L,G020K-S024F,G020K-N062E,T033S-G118R,S024R-P086W,S024R-G118R,S024R-P086R,Y209A-W241R,S024R-W241R,S024R-K235F,G118R-Y209A,S078R-G118R,T033S-K235F,S024R-A174T,P086W-Y209A,I008T-S024R,P086W-G118R,T033S-W241R,S024R-N243F,S024R-Y209A,T033S-P086W,S024R-T033S,P086W-N243F,T033S-P201S,S024R-P239Q,S078R-P086W,K235F-N243F,G118R-A172V,T033S-L148F,T033S-S078R,T033S-N243F,S024C-T033S,G118R-A194T,T033S-Y209A,S024R-S141G,S024R-T274I,P086W-K235F,A015T-T033S,Y209A-K235F,S024R-R247H,S078R-K235F,S024R-V104A,T033S-A048T,G118R-K235F,T033S-T253A,T143A-Y209A,T033S-P239T,Y209A-N243F,S024R-A272P，和S024R-R269C，并且其中所述变体的总净电荷相对于迟缓芽孢杆菌(Bacillus lentus)枯草杆菌蛋白酶GG36蛋白酶的总净电荷为0、+1、+2、+3、+4、+5、-1、-2、-3、-4或-5，并且其中所述总净电荷是通过一种或多种选自以下的置换来获得的：N43D,R45T,N62E,N76D,S101D,P129E,A158E,G159D,G159E,S166D,S188D,A230E,N18R,G20K,G20R,T22R,S24R,N43R,G118R,Q245R,H249R,N269R,E271F，和E271L，并且其中所述蛋白酶变体的氨基酸位置根据SEQ ID NO:1所示解淀粉芽孢杆菌(Bacillus amyloliquefaciens)枯草杆菌蛋白酶BPN’的氨基酸序列中对应的氨基酸位置的编号方式来进行编号。4.一种分离的枯草杆菌蛋白酶变体，其中所述枯草杆菌蛋白酶变体为具有蛋白水解活性的成熟形式并包含具有选自以下的氨基酸置换的组合的氨基酸序列：G020K-G023A-N043W-G118R-S128I-P129E-G159D-S188D,G020K-G023A-N062E-N116L,G020K-G023A-N062E-N116L-S188D,G020K-G023A-N062E-N116L-S188D-T213A,G020K-G100S-N116L-A158E-S166D-N243F,G020K-N043W-N062E-N116L-S188D,G020K-N062E,G020K-N062E-N116L,G020K-N062E-N116L-S188D,G020K-N062E-N116L-T213A,G020K-N062E-S188D-T213A,G020K-S024F-N062E-N116L-G118R-S188D,G020K-S024F-N062E-N116L-S188D,G020K-S024F-N062E-S188D-T213A,G023A-N062E-N116L-S188D,G023A-S024F-N062E-N116L-T213A,N043R-N076D-S101A-S103A-V104I-A158E-S188D-L217E-A232V-Q245R-N248D-H249R-E271F,N043R-S101A-S103A-V104I-A158E-S188D-L217E-A232V-Q245R-N248D-H249R,N043W-N062E-N116L,N043W-S101D-S212M-N243F,N062E-N116L-G118R-S188D,N062E-N116L-G118R-T213A,N062E-N116L-S188D-T213A,S024F-G118R-S128I-P129E-G159D,S024F-N062E-N116L-S188D,S024R-S101G-S103A-V104I-A158E-N183D-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-A158E-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-A158E-S188D-L217E-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-P129Q-A158E-N183D-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-P129Q-A158E-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-P129Q-A158E-S188D-L217E-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-P129Q-S130A-A158E-N183D-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-S128L-P129Q-A158E-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-S130A-A158E-N183D-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-S130A-A158E-S188D-A232V-Q245R-N248D-H249R,S101D-S103N-N116L-S144R-A215D,S101G-S103A-V104I-A158E-S188D-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-S130A-A158E-S188D-A232V-Q245R-N248D-H249R,T022A-N043R-S101G-S103A-V104I-G159D-S188D-L217E-A232V-Q245R-N248D-E271F,T022A-N043R-S103A-V104I-G118R-S128I-P129E-G159D-S188D-A232V-N248D-E271F,T022A-N043R-S103A-V104I-S128I-P129E-G159D-S188D-A232V-Q245R-N248D,T022A-S024R-S103A-V104I-G118R-S128I-P129E-G159D-S188D-A232V-N248D,T022A-S101G-S103A-V104I-G159D-L217E-A232V-Q245R-N248D-E271F,T022A-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F,T022L-T038F-A048R-N062E-G100S-R186K,T033S-N043W-N218D-P239G-N243F,V026F-A048R-S105T-T213A-N218D-T224A,V026F-L031F-S078N-G102A-S160D,V026F-V051W-V104L-S106E,V104L-S105T-T213A-L217E-S256N,A016S-N062E-A158E-H249R,A016S-N062E-A158E-R186H-E271F,A016S-N062E-A158E-R186H-H249R,A016S-N062E-V104L-A158E-R186H-E271F,A016S-S024R-S101G-S103A-V104I-A158E-S188D-A232V-Q245R-N248D-H249R,G020K-G023A-N062E-S188D,G020K-G023A-S024F-N062E-G118R-S188D-T213A,G020K-N043W-N062E-N116L-S188D-T213A-P239G,G020K-S024F-N062E-S188D-P239G,G023A-N062E-N116L-G118R,G023A-N062E-N116L-G118R-S188D-P239G,G023A-S024F-N062E-N116L-G118R,H017R-T022A-N076D-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F,N043R-N076D-S101A-S103A-V104I-A158E-S188D-A232V-Q245R-N248D-H249R-E271F,N062E-A158E-G159E-H249R,N062E-A158E-H249R,N062E-A158E-R186H-E271F,N062E-A158E-R186H-H249R,N062E-A158E-R186H-H249R-E271F,N062E-A158E-S188D-H249R,N062E-A158E-S188D-H249R-E271F,N062E-G159E-H249R,N062E-G159E-H249R-E271F,N062E-G159E-R186H-H249R,N062E-G159E-S188D-H249R,N062E-R186H-S188D-H249R-E271F,N062E-S101A-A158E-H249R,N062E-S101A-A158E-R186H-E271F,N062E-S101A-A158E-R186H-H249R-E271F,N062E-S101A-G159E-H249R,N062E-S101A-R186H-H249R,N062E-S101A-S188D-H249R,N062E-S101A-S188D-H249R-E271F,N062E-S101A-V104L-A158E-R186H-E271F,N062E-S128N-A158E-G159E-E271F,N062E-V104L-A158E-S188D-H249R-E271F,N076D-S101G-S103A-V104I-A232V-M222Q-Q245R,S024F-N062E-N116L-P239G,S024F-N062E-S188D-T213A,S024F-N116L,S024R-N043R-N076D-S101A-S103A-V104I-A158E-S188D-L217E-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-A158E-G159E-A232V-N238R-Q245R-N248D,S024R-S101G-S103A-V104I-A158E-G159E-S188D-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-A158E-S188D-A232V-N238R-Q245R-N248D,S024R-S101G-S103A-V104I-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-G159E-S188D-A232V-N238R-Q245R-N248D,S024R-S101G-S103A-V104I-G159E-S188D-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-G159E-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-L148I-A158E-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-L148I-A158E-S188D-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-L148I-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-P129E-A158E-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-P129E-A158E-G159E-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-P129E-A158E-S188D-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-P129E-A158E-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-P129E-G159E-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-P129E-G159E-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-P129E-L148I-A158E-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-P129E-S188D-A232V-N238R-Q245R-N248D,S024R-S101G-S103A-V104I-P129E-S188D-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-P129E-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-S128N-A158E-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-S128N-P129E-A158E-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-S128N-P129E-A232V-Q245R-N248D,S101A-S103A-V104I-A158E-S188D-A232V-Q245R-N248D-H249R,S101A-S103A-V104I-A158E-S188D-A232V-Q245R-N248D-H249R-E271F-E271F,S101A-S103A-V104L-A158E-S188D-A232V-Q245R-N248D-H249R,S101A-S128N-A158E-R186H-E271F,S101A-S128N-A158E-Y209E-H249R,S101A-V104L-A158E-R186H-S188D-H249R,S101G-S103A-V104I-A158E-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-A158E-G159E-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-A158E-S188D-M222Q-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-A158E-S188D-M222S-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-A232V-M222Q-Q245R,S101G-S103A-V104I-P129E-A158E-S188D-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-P129E-G159E-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-P129E-S188D-A232V-N238R-Q245R-N248D,S101G-S103A-V104I-P129E-S188D-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-S188D-A232V-Q245R-N248D-H249R,S101G-S103G-V104I-A158E-S188D-A232V-Q245R-N248D-H249R,S101G-S103S-V104I-A158E-S188D-A232V-Q245R-N248D-H249R,S101S-S103A-V104I-A158E-S188D-A232V-Q245R-N248D-H249R,S101S-S103G-V104I-A158E-S188D-A232V-Q245R-N248D-H249R,S101 S-S103G-V104V-A158E-S188D-A232V-Q245R-N248D-H249R,S128N-A158E-G159E-E271F,S128N-A158E-H249R,S128N-A158E-R186H-E271F,S128N-A158E-R186H-H249R,S128N-A158E-R186H-S188D-E271F,S128N-A158E-S188D-E271F,S128N-A158E-S188D-H249R,S128N-A158E-S188D-Y209E-E271F,S128N-A158E-Y209E-,T022A-N043R-S103A-V104I-G118R-P129E-G159D-S188D-A232V-Q245R-N248D,T022A-N062E-A158E,T022A-S024R-S101D-S103A-V104I-G118R-G159D-S188D-A232V-N248D-E271F,T022A-S024R-S101D-S103A-V104I-G118R-P129E-G159D-S188D-A232V-Q245R-N248D,T022A-S024R-S101D-S103A-V104I-G159D-S188D-A232V-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-A158E-A232V-N238R-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-A158E-A232V-Q245R-N248D-H249R,T022A-S024R-S101G-S103A-V104I-A158E-G159E-S188D-A232V-N238R-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-A158E-G159E-S188D-A232V-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-A158E-G159E-S188D-A232V-Q245R-N248D-H249R,T022A-S024R-S101G-S103A-V104I-P129E-A158E-A232V-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-P129E-A158E-A232V-Q245R-N248D-H249R,T022A-S024R-S101G-S103A-V104I-P129E-A158E-G159E-A232V-Q245R-N248D-H249R,T022A-S024R-S101G-S103A-V104I-P129E-A158E-G159E-S188D-A232V-N238R-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-P129E-A158E-S188D-A232V-N238R-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-P129E-A158E-S188D-A232V-Q245R-N248D-H249R,T022A-S024R-S101G-S103A-V104I-P129E-A232V-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-P129E-A232V-Q245R-N248D-H249R,T022A-S024R-S101G-S103A-V104I-P129E-G159E-S188D-A232V-Q245R-N248D-H249R,T022A-S024R-S101G-S103A-V104I-P129E-S188D-A232V-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-S128N-A158E-S188D-A232V-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-S128N-P129E-A158E-A232V-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-S128N-P129E-S188D-A232V-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-S128N-S188D-A232V-Q245R-N248D,T022A-S024R-S103A-V104I-G118R-G159D-S188D-L217D-A232V-N248D,T022A-S024R-S103A-V104I-P129E-G159D-S188D-A232V-N248D-E271F,T022A-S101G-S103A-V104I-A158E-G159E-S188D-A232V-Q245R-N248D-H249R,T022A-S101G-S103A-V104I-A158E-S188D-A232V-Q245R-N248D,T022A-S101G-S103A-V104I-P129E-A158E-A232V-N238R-Q245R-N248D,T022A-S101G-S103A-V104I-P129E-A158E-G159E-A232V-N238R-Q245R-N248D,T022A-S101G-S103A-V104I-P129E-A158E-S188D-A232V-Q245R-N248D-H249R,T022A-S101G-S103A-V104I-P129E-G159E-A232V-N238R-Q245R-N248D,T022A-S101G-S103A-V104I-P129E-S188D-A232V-N238R-Q245R-N248D,T022A-S101G-S103A-V104I-S128N-P129E-A232V-N238R-Q245R-N248D,T022A-S101G-S103A-V104I-S128N-P129E-S188D-A232V-N238R-Q245R-N248D,T022A-S128N-A158E-H249R,V104L-S128N-A158E-H249R,V104L-S128N-A158E-R186H-E271F,V104L-S128N-A158E-R186H-H249R，和V104L-S128N-A158E-S188D-H249R，并且其中所述变体的总净电荷相对于迟缓芽孢杆菌(Bacillus lentus)枯草杆菌蛋白酶GG36蛋白酶的总净电荷为0、+1、+2、+3、+4、+5、-1、-2、-3、-4或-5，并且其中所述总净电荷是通过一种或多种选自以下的置换来获得的：N43D,R45T,N62E,N76D,S101D,P129E,A158E,G159D,G159E,S166D,S188D,A230E,N18R,G20K,G20R,T22R,S24R,N43R,G118R,Q245R,H249R,N269R,E271F，和E271L，并且其中所述蛋白酶变体的氨基酸位置根据SEQ ID NO:1所示解淀粉芽孢杆菌(Bacillus amyloliquefaciens)枯草杆菌蛋白酶BPN’的氨基酸序列中对应的氨基酸位置的编号方式来进行编号。5.一种分离的枯草杆菌蛋白酶变体，其中所述枯草杆菌蛋白酶变体为具有蛋白水解活性的成熟形式并包含具有选自以下的氨基酸置换的组合的氨基酸序列：A016S-N062E-A158E-H249R-E271F,A016S-N062E-S128N-R186H-E271F,A016S-N062E-V104L-R186H-S188D-E271F,A016S-S101A-S128N-R186H,A016S-S128N-A158E-R186H,A016S-V104L-A158E-R186H-E271F,A016S-V104L-S188D-H249R,A158E-R186H-H249R,A158E-R186H-S188D-H249R-E271F,G020K-G023A-N116L-S188D,G020K-G023A-S024F-N062E-N116L-S188D-T213A,G020K-N043W-N062E-N116L-P239G,G020K-S024F-N043W-N062E-N116L-T213A,G020K-S024F-N062E,G023A-N043W-N062E-N116L-G118R-T213A,G023A-T038F-S078N-G100S-S212M-A215D,G100S-N116L-A158E-T213A,G102A-S103N-S105T-A194E,H017R-T022A-N076D-S101G-S103A-V104I-G159D-A232V-Q245R-N248D-E271F,N043R-N076D-S101A-S103A-V104I-A158E-S166D-S188D-A232V-Q245R-N248D-H249R-E271F,N043R-S101A-S103A-V104I-A158E-S166D-S188D-A232V-Q245R-N248D-H249R,N043W-N062E-N116L-S188D,N062E-A158E-H249R-E271F,N062E-A158E-R186H-S188D-H249R,N062E-L148I-G159E,N062E-N116L-S188D,N062E-R186H-H249R,N062E-S078N-G102A-N116L-S144R-L250I,N062E-S101A-A158E-R186H-H249R,N062E-S101A-A158E-S188D-H249R,N062E-S101A-R186H,N062E-S101A-R186H-E271F,N062E-S101A-R186H-H249R-E271F,N062E-S101A-R186H-S188D-E271F,N062E-S101A-R186H-S188D-H249R,N062E-S101A-V104L-R186H-S188D-E271F,N062E-S128N-A158E,N062E-S128N-G159E-H249R,N062E-S188D-P239G,N062E-V104L-G159E-H249R,N076D-S101A-S103A-V104I-A158E-S166D-S188D-A232V-Q245R-N248D-H249R-E271F,N076D-S101A-S103A-V104I-A158E-S188D-A232V-Q245R-N248D-H249R,N076D-S101A-S103A-V104I-A158E-S188D-A232V-Q245R-N248D-H249R-E271F,N076D-S101G-S103A-V104I-A114V-A158E-S188D-A232V-Q245R-N248D-H249R-E271F,N076D-S101G-S103A-V104I-A232V-M222S-Q245R,S024F-A048R-G118R-S166D-L217E,S024F-N062E-N116L,S024F-N062E-N116L-T213A-P239G,S024F-N062E-S188D-P239G,S024F-S101D-G118R-A215D-L250I-A272F,S024R-K027R-S101G-S103A-V104I-S128L-P129Q-S130A-A158E-S188D-A232V-Q245R-N248D-H249R,S024R-N076D-S101A-S103A-V104I-A158E-S166D-S188D-A232V-Q245R-N248D-H249R-E271F,S024R-S101G-S103A-V104I-A158E-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-P129E-A158E-G159E-A232V-N238R-Q245R-N248D,S024R-S101G-S103A-V104I-P129E-A158E-G159E-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-P129E-G159E-A232V-N238R-Q245R-N248D,S024R-S101G-S103A-V104I-P129E-G159E-S188D-A232V-N238R-Q245R-N248D,S024R-S101G-S103A-V104I-P129E-L148I-A158E-S188D-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-P129E-L148I-S188D-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-P129Q-S130A-A158E-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-S128L-A158E-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-S128L-P129Q-S130A-A158E-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-S128N-G159E-S188D-A232V-Q245R-N248D,S078N-V104L-G118R-S128D,S101A-A158E-R186H-H249R,S101A-A158E-R186H-S188D-H249R,S101A-S103A-V104I-A158E-S166D-S188D-A232V-Q245R-N248D-H249R-E271F,S101A-S103A-V104I-A158E-S188D-A232V-Q245R-N248D-H249R-E271F,S101A-S103A-V104I-A158E-S188D-L217E-A232V-Q245R-N248D-H249R,S101A-S103A-V104I-A158E-S188D-L217E-A232V-Q245R-N248D-H249R-E271F,S101A-S103G-A158E-R186H-H249R,S101A-S103S-V104I-A158E-S188D-A232V-Q245R-N248D-H249R,S101A-S103S-V104I-G159E-A232V-Q245R-N248D-H249R,S101A-S128N-A158E-H249R,S101A-S128N-A158E-S188D-Y209E-E271F,S101A-S128N-A158E-Y209E,S101A-S128N-P129E-R186H-H249R,S101A-V104L-A158E-R186H-H249R,S101A-V104L-A158E-R186H-S188D-E271F,S101A-V104L-S128N-A158E-R186H-E271F,S101G-S103A-V104I-A158E-A232V-N238R-Q245R-N248D,S101G-S103A-V104I-A158E-G159E-S188D-A232V-N238R-Q245R-N248D,S101G-S103A-V104I-A158E-N183D-S188D-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-A158E-S166D-S188D-A232V-Q245R-N248D-H249R-E271F,S101G-S103A-V104I-A158E-S188D-A232V-Q245R-N248D,S101G-S103A-V104I-A232V-M222S-Q245R,S101G-S103A-V104I-P129E-A158E-G159E-A232V-N238R-Q245R-N248D,S101G-S103A-V104I-P129E-A158E-G159E-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-P129E-A158E-S188D-A232V-N238R-Q245R-N248D,S101G-S103A-V104I-P129E-G159E-A232V-N238R-Q245R-N248D,S101G-S103A-V104I-P129E-S188D-A232V-Q245R-N248D,S101G-S103A-V104I-P129Q-A158E-S188D-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-P129Q-S130A-A158E-S188D-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-S128L-P129Q-A158E-S188D-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-S128L-S130A-A158E-S188D-L217E-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-S128N-P129E-A232V-N238R-Q245R-N248D,S101G-S103A-V104I-S128N-P129E-A232V-Q245R-N248D,S101G-S103A-V104I-S130A-A158E-N183D-S188D-L217E-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-S188D-A232V-N238R-Q245R-N248D,S101G-S103A-V104L-A158E-S188D-A232V-Q245R-N248D-H249R,S101 S-S103A-V104I-G159E-A232V-Q245R-N248D-H249R,S101S-S103S-V104I-A158E-S188D-A232V-Q245R-N248D-H249R,S101 S-S103S-V104V-A158E-S188D-A232V-Q245R-N248D-H249R,T022A-N043R-N062E-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F,T022A-N043R-N076D-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F,T022A-N043R-S101D-S103A-V104I-G118R-P129E-G159D-S188D-A232V-N248D-E271F,T022A-N043R-S101G-S103A-V104I-G159D-A232V-Q245R-N248D-E271F,T022A-N043R-S103A-V104I-G118R-G159D-S188D-A232V-N248D-E271F,T022A-N043R-S103A-V104I-G118R-G159D-S188D-L217D-A232V-N248D-E271F,T022A-N043R-S103A-V104I-G118R-S128I-P129E-G159D-S188D-A232V-N248D,T022A-N043R-S103A-V104I-P129E-G159D-S188D-A232V-Q245R-N248D,T022A-N076D-S101G-S103A-V104I-G159D-A232V-Q245R-N248D-E271F,T022A-N076D-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F,T022A-S024R-N043R-S101D-S103A-V104I-G159D-S188D-A232V-Q245R-N248D,T022A-S024R-N043R-S103A-V104I-G159D-S188D-L217D-A232V-N248D-E271F,T022A-S024R-S101D-S103A-V104I-G118R-S128I-G159D-S188D-A232V-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-L148I-A158E-A232V-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-P129E-A158E-G159E-A232V-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-P129E-A158E-G159E-S188D-A232V-Q245R-N248D-H249R,T022A-S024R-S101G-S103A-V104I-P129E-L148I-A232V-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-S188D-A232V-Q245R-N248D-H249R,T022A-S024R-S103A-V104I-G118R-P129E-G159D-S188D-A232V-N248D-E271F,T022A-S024R-S103A-V104I-G159D-S188D-L217D-A232V-N248D-E271F,T022A-S024R-S103A-V104I-G159D-S188D-L217D-A232V-Q245R-N248D-E271F,T022A-S101A-A158E-R186H-H249R,T022A-S101G-S103A-V104I-A158E-G159E-A232V-N238R-Q245R-N248D,T022A-S101G-S103A-V104I-A158E-G159E-A232V-Q245R-N248D-H249R,T022A-S101G-S103A-V104I-G159D-A232V-Q245R-N248D-E271F,T022A-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-H249R-E271F,T022A-S101G-S103A-V104I-G159E-S188D-A232V-N238R-Q245R-N248D,T022A-S101G-S103A-V104I-G159E-S188D-A232V-Q245R-N248D-H249R,T022A-S101G-S103A-V104I-P129E-A158E-A232V-Q245R-N248D-H249R,T022A-S101G-S103A-V104I-P129E-A158E-G159E-A232V-Q245R-N248D-H249R,T022A-S101G-S103A-V104I-P129E-A232V-N238R-Q245R-N248D,T022A-S101G-S103A-V104I-P129E-G159E-A232V-Q245R-N248D-H249R,T022A-S101G-S103A-V104I-P129E-G159E-S188D-A232V-Q245R-N248D-H249R,T022A-S103A-V104I-G118R-G159D-S188D-L217D-A232V-Q245R-N248D,T022A-S103A-V104I-G159D-S188D-A232V-N248D,T022L-L031F-G102A-S128D-T224A-N243F,T022L-T038F-V121F-S160D-A272F,V026F-S078N-G159C-R186K-N243F,V104L-A158E-H249R,V104L-A158E-R186H-H249R,V104L-A158E-R186H-H249R-E271F,V104L-A158E-R186H-S188D-H249R,V104L-A158E-R186H-S188D-H249R-E271F,V104L-A158E-S188D-H249R-E271F,V104L-S128N-A158E-R186H-S188D-E271F,V104L-S128N-G159E-E271F,V104L-S128N-R186H-S188D-H249R，和V121F-N185E-T224A-P239G，并且其中所述变体的总净电荷相对于迟缓芽孢杆菌(Bacillus lentus)枯草杆菌蛋白酶GG36蛋白酶的总净电荷为0、+1、+2、+3、+4、+5、-1、-2、-3、-4或-5，并且其中所述总净电荷是通过一种或多种选自以下的置换来获得的：N43D,R45T,N62E,N76D,S101D,P129E,A158E,G159D,G159E,S166D,S188D,A230E,N18R,G20K,G20R,T22R,S24R,N43R,G118R,Q245R,H249R,N269R,E271F，和E271L，并且其中所述蛋白酶变体的氨基酸位置根据SEQ ID NO:1所示解淀粉芽孢杆菌(Bacillus amyloliquefaciens)枯草杆菌蛋白酶BPN’的氨基酸序列中对应的氨基酸位置的编号方式来进行编号。6.一种分离的枯草杆菌蛋白酶变体，其中所述枯草杆菌蛋白酶变体为具有蛋白水解活性的成熟形式并包含具有选自以下的氨基酸置换的组合的氨基酸序列：G118R-Y209A,G118R-Y209A-N243F,L021M-S024R-T033S,P005S-S078R-G118R-W241R,P086W-G118R-A133V,P086W-G118R-N243F,P086W-G118R-Y209A,P086W-Y209A-N243F,S024R-A174T,S024R-G118R,S024R-G118R-Y209A,S024R-G118R-Y209A-K235F,S024R-G118R-Y209A-N243F,S024R-K235F,S024R-P086R,S024R-P086S-S141G,S024R-P086W,S024R-P086W-G118R,S024R-P086W-G118R-V203I,S024R-P086W-N243F,S024R-P086W-Y209A,S024R-S078R-P086W,S024R-S078R-P086W-G118R-A270T,S024R-S078R-P086W-N243F,S024R-T033S-A133V,S024R-T033S-G118R,S024R-T033S-P086S-S085N-K235F,S024R-T033S-P086S-S087N-Y209A,S024R-T033S-P086W,S024R-T033S-P086W-G118R,S024R-T033S-S078R-P086W,S024R-T033S-S078R-P086W-G118R,S024R-T033S-S078R-Y209A,S024R-T033S-W241R,S024R-T033S-Y209A-K235F,S024R-Y209A-W241R,S078R-G118R,S078R-P086W-K235F,S078R-P086W-N243F,S078R-P086W-Y209A,S078R-Y209A-K235F,T033S-A172V-Y209A,T033S-G118R,T033S-G118R-G159D-Y209A,T033S-G118R-N243F,T033S-G118R-W241R,T033S-G118R-Y209A-N243F,T033S-K235F,T033S-P086W-G118R,T033S-P086W-G118R-Y209A-N243F,T033S-P086W-N243F,T033S-S078R-P086W,T033S-S078R-P086W-G118R-Y209A,T033S-S078R-Y209A,T033S-Y209A-N243F,Y209A-W241R,A001R-S009A-N043R,A001R-S101G-S103A-V104I-A232V-Q245R,A001T-N018R-S024R-N076D-N116A-T213A-H249R,A194F-G211Q-Q236N,E089I-N116A-N117F-T224A-H249R,G020R-G025R-N116A-Y167W,G020R-H249R-N269R,G020R-N043D-N076D-A230E-H249R,G020R-N043D-S078R-S101G-S103A-V104I-A232V-Q245R,G020R-N043D-S101G-S103A-V104I-A232V-Q245R,G020R-N043D-S101G-S103A-V104I-A232V-Q245R-N269R,G020R-N043R-A230E,G020R-N043R-A230E-S242R,G020R-N043R-E271L,G020R-N043R-H249R,G020R-N043R-H249R-E271L,G020R-N043R-N269R,G020R-N043R-R045T-A230E,G020R-N043R-R045T-S101A-N269R,G020R-N043R-R045T-S242R,G020R-N043R-S101A-N116A-T213A-A215F,G020R-N043R-S101A-N269R,G020R-N043R-S101G-S103A-V104I-A232V-Q245R,G020R-N043R-S212F,G020R-N043R-S212F-W241R,G020R-N043R-S242R,G020R-N043R-S242R-E271L,G020R-N043R-S242R-H249R,G020R-N043R-V244R,G020R-N043R-W241R,G020R-N062Q-E089I-R186K-S212M,G020R-N076D,G020R-N076D-S101G-S103A-V104I-A232V-Q245R,G020R-N076D-S101G-S103A-V104I-A232V-Q245R-N269R,G020R-N116A-N269R,G020R-P052N-N062Q-Y091F-Y192W,G020R-Q059A-S144R-Y192W-T224A,G020R-S024R-K27E-N043R-N076D-A230E,G020R-S024R-N043D-H249R,G020R-S024R-N043D-N076D-S078R-S101G-S103A-V104I-A232V-Q245R,G020R-S024R-N043D-S242R,G020R-S024R-N043R-R045T,G020R-S024R-N043R-R045T-H249R,G020R-S024R-N043R-R045T-N116A,G020R-S024R-N043R-R045T-N116A-T213A,G020R-S024R-N043R-R045T-S101A-N269R,G020R-S024R-N043R-R045T-S101A-T213A,G020R-S024R-N043R-S242R,G020R-S024R-N076D-H249R,G020R-S024R-N116A,G020R-S024R-N116A-A215F,G020R-S024R-N116A-T213A,G020R-S024R-N116A-T213A-A215F,G020R-S024R-P052N-Q059A-S216F,G020R-S024R-R045T-A230E-S242R,G020R-S024R-R045T-N116A-N269R,G020R-S024R-R045T-N269R,G020R-S024R-S101A-A215F,G020R-S024R-S101A-N116A,G020R-S024R-S101G-S103A-V104I-L217E-A232V-Q245R-H249R,G020R-S024R-S242R-H249R,G020R-S024R-T213A,G020R-S024R-T213A-A215F,G020R-S078R-S101A-S103A-V104I-N116A-N183D-T213A-A232V-Q245R,G020R-S078R-S101G-A232V-Q245R,G020R-S078R-S101G-S103A-V104I-A215F-A232V-Q245R,G020R-S078R-S101G-S103A-V104I-G211Q-T213A-A215F-A232V-Q245R,G020R-S078R-S101G-S103A-V104I-N116A-A232V-Q245R,G020R-S078R-S101G-S103A-V104I-N116A-G211Q-A232V-Q245R,G020R-S078R-S101G-S103A-V104I-N116A-N183D-A232V-Q245R,G020R-S087D-S101G-S103A-V104I-A232V-Q245R,G020R-S101A-N116A-N269R,G020R-S101A-S103A-V104I-A215F-A232V-Q245R,G020R-S101A-S103A-V104I-G118R-A232V-Q245R,G020R-S101A-S103A-V104L-A232V-Q245R,G020R-S101A-S103A-V104V-A232V-Q245R,G020R-S101A-S103S-V104I-A232V-Q245R,G020R-S101A-S103S-V104V-A232V-Q245R,G020R-S101G-I198L-A215F-A232V-Q245R,G020R-S101G-S103A-A232V-Q245R,G020R-S101G-S103A-V104I-A215F-A232V-Q245R,G020R-S101G-S103A-V104I-A232V-Q245R,G020R-S101G-S103A-V104I-A232V-Q245R-N269R,G020R-S101G-S103A-V104I-G211Q-T213A-A215F-A232V-Q245R,G020R-S101G-S103A-V104I-N116A-A215F-A232V-Q245R,G020R-S101G-S103A-V104I-T213A-A215F-A232V-Q245R,G020R-S101G-S103A-V104I-V150L-A232V-Q245R,G020R-S101G-S103A-V104L-A232V-Q245R,G020R-S101G-S103A-V104V-A232V-Q245R,G020R-S101G-S103G-V104I-A232V-Q245R,G020R-S101G-S103S-V104I-A232V-Q245R,G020R-S101G-S103S-V104L-A232V-Q245R,G020R-S101G-V104I-T213A-A215F-A232V-Q245R,G020R-S101S-S103A-V104I-A232V-Q245R,G020R-S101S-S103A-V104V-A232V-Q245R,G020R-S101S-S103G-V104I-A232V-Q245R,G020R-S101S-S103G-V104V-A232V-Q245R,G020R-S101S-S103S-V104I-A232V-Q245R,G020R-S101S-S103S-V104L-A232V-Q245R,G020R-S101S-S103S-V104V-A232V-Q245R,G020R-S103N-S216F-Q236N-N252R,G020R-S144R-N185I-L233C-Q236N,G020R-S212F-H249R,G020R-S242R-N269R,G020R-T022R-E271L,G020R-T022R-N043R,G020R-T022R-N043R-S212F,G020R-T022R-N043R-W241R,G020R-T022R-S024R-S242R,G020R-T022R-S078R-S242R,G020R-T022R-S078R-W241R,G020R-T022R-S212F-W241R,G020R-T022R-S242R,G020R-T022R-W241R,G020R-T022W-S078R-S101A-S103A-V104I-N116A-N183D-A215F-A232V-Q245R,G020R-T022W-S078R-S101A-S103A-V104I-N116A-N183D-A232V-Q245R,G020R-T022W-S078R-S101A-S103A-V104I-N116A-N183D-T213A-A232V-Q245R,G020R-T022W-S078R-S101A-S103A-V104I-N116S-T213A-A215F-A232V-Q245R,G020R-T022W-S078R-S101G-S103A-V104I-N116A-A232V-Q245R,G020R-T022W-S078R-S101G-S103A-V104I-N116A-N183D-A232V-Q245R,G020R-T022W-S078R-S101G-S103A-V104I-N116A-N183D-G211Q-A232V-Q245R,G020R-T022W-S078R-S101G-S103A-V104I-N116A-N183D-T213A-A232V-Q245R,G020R-T022W-S078R-S101G-S103A-V104I-N116A-T213A-A215F-A232V-Q245R,G020R-T022W-S101A-S103A-V104I-G211Q-A215F-A232V-Q245R,G020R-T022W-S101G-S103A-V104I-A232V-Q245R,G020R-T022W-S101G-S103A-V104I-G211Q-A232V-Q245R,G020R-T022W-S101G-S103A-V104I-I198L-G211Q-T213A-A232V-Q245R,G020R-T022W-S101G-S103A-V104I-N116A-N183D-T213A-A232V-Q245R,G020R-T038A-N043R-S101A,G020R-T22W-S078R-S101A-S103A-V104I-N116A-N183D-A232V,G020R-T22W-S078R-S101G-S103A-V104I-N116A-N183D-A232V-Q245R,G020R-T22W-S078R-S101G-S103A-V104I-N116A-T213A-A215F-A232V-Q245R,G020R-T22W-S101A-S103A-V104I-A215F-A232V-Q245R,G020R-T22W-S101A-S103A-V104I-A232V-Q245R,G020R-T22W-S101A-S103A-V104I-G211Q-T213A-A232V-Q245R,G020R-T22W-S101G-S103A-V104I-A232V-Q245R,G020R-T22W-S101G-S103A-V104I-G211Q-A232V-Q245R,G020R-T22W-S103A-V104I-A232V-Q245R,G023A-P052N-S144R-Y192W-S216F,G023A-P052N-Y192W-I198L-N252R,G023A-S024R-N117F-S212M-S216F,G023A-S078R-S216F-Q236N-H249R,G023A-Y091F-S101A-I198L-N252R,G023A-Y091F-V121F-Y192W-Q236N,G025R-E089I-N116A-P239S-A270C,G025R-G046R-V121F,G025R-N043A-E089I-N117F,G025R-S103N-R186K-A194F-T224A,G046R-A194F-S212M,G046R-E089I-Y091F-S101A-N116A,G046R-E089I-Y192W-L233C-A270C,G046R-N183D-N238L,G046R-Q059A-S103N-G211Q-S212M,G046R-Q059A-S106G-L217E-H249R,L148I-T213A-N252R,N018R-G020R-N043D,N018R-G020R-N043D-A230E-S242R,N018R-G020R-N043D-N076D-S101G-S103A-V104I-A232V-Q245R,N018R-G020R-N043D-R045T-A230E,N018R-G020R-N043R-N076D-H249R,N018R-G020R-N043R-N076D-S242R-H249R,N018R-G020R-N043R-R045T-S242R,N018R-G020R-N076D-S101G-S103A-V104I-A232V-Q245R,N018R-G020R-R045T,N018R-G020R-S024R-N043D-R045T-L233I-S242R,N018R-G020R-S024R-N043R-R045T-N076D-A230E,N018R-G020R-S024R-N076D-S087D-H249R,N018R-G020R-S024R-N076D-V150L-H249R,N018R-G020R-S024R-R045T-N076D-S101G-S103A-V104I-A232V-Q245R-N269R,N018R-N043D-N076D-S101G-S103A-V104I-A232V-Q245R-N269R,N018R-N043D-S078R-S101G-S103A-V104I-A232V-Q245R,N018R-N043D-S078R-S101G-S103A-V104I-L217E-A232V-Q245R,N018R-N043D-S101G-S103A-V104I-A232V-Q245R,N018R-N043D-S101G-S103A-V104I-A232V-Q245R-H249R,N018R-N043D-S101G-S103A-V104I-A232V-Q245R-N269R,N018R-N043R-N076D-A230E,N018R-N043R-R045T-H249R,N018R-N043R-R045T-S101G-S103A-V104I-A232V-Q245R,N018R-N043R-R045T-S242R-H249R,N018R-N043R-S101G-S103A-V104I-A232V-Q245R,N018R-N043R-S101G-S103A-V104I-A232V-Q245R-N269R,N018R-N043R-S242R-H249R,N018R-N076D-S078R-S101G-S103A-V104I-A232V-Q245R,N018R-N076D-S101G-V104I-A232V-Q245R,N018R-Q245R,N018R-R045T-N076D-S078R-S101G-S103A-V104I-A232V-Q245R,N018R-R045T-N076D-S101G-S103A-V104I-A232V-Q245R-N269R,N018R-R045T-S078R-S101G-S103A-V104I-A232V-Q245R,N018R-R045T-S101G-S103A-V104I-A232V-Q245R,N018R-S024R-N043D-A230E,N018R-S024R-N043D-N076D-H249R,N018R-S024R-N043D-N076D-H249R-N269R,N018R-S024R-N043D-N076D-S078R-H249R,N018R-S024R-N043D-R045T-H249R,N018R-S024R-N043D-S101G-S103A-V104I-A232V-Q245R,N018R-S024R-N043D-S101G-S103A-V104I-A232V-Q245R-N269R,N018R-S024R-N043R-N076D-H249R,N018R-S024R-N043R-N076D-H249R-N269R,N018R-S024R-N043R-N076D-S087D-H249R,N018R-S024R-N043R-N076D-V150L-H249R,N018R-S024R-N076D-G211Q-A215F-H249R,N018R-S024R-N076D-G211Q-T213A-A215F-H249R,N018R-S024R-N076D-G211Q-T213A-H249R,N018R-S024R-N076D-H249R,N018R-S024R-N076D-H249R-N269R,N018R-S024R-N076D-N116A-A215F-H249R,N018R-S024R-N076D-N116A-G211Q-A215F-H249R,N018R-S024R-N076D-N116A-G211Q-H249R,N018R-S024R-N076D-N116A-T213A-A215F-H249R,N018R-S024R-N076D-S078R-H249R,N018R-S024R-N076D-S078R-S087D-H249R,N018R-S024R-N076D-S078R-V150L-H249R,N018R-S024R-N076D-S087D-H249R-N269R,N018R-S024R-N076D-S087D-S242R-H249R,N018R-S024R-N076D-S087D-V150L-H249R,N018R-S024R-N076D-S101A-A215F-H249R,N018R-S024R-N076D-S101A-G211Q-T213A-A215F-H249R,N018R-S024R-N076D-S101A-I198L-G211Q-A215V-H249R,N018R-S024R-N076D-S101A-I198L-G211Q-T213A-H249R,N018R-S024R-N076D-S101A-N116A-G211Q-H249R,N018R-S024R-N076D-S101A-N116A-T213A-H249R,N018R-S024R-N076D-S101G-V104I-A232V-H249R,N018R-S024R-N076D-S242R-H249R,N018R-S024R-N076D-T213A-A215F-H249R,N018R-S024R-N076D-V104I-H249R,N018R-S024R-N076D-V150L-H249R,N018R-S024R-R045T-S242R,N018R-S024R-S101G-V104I,N018R-S024R-S101G-V104I-A232V,N018R-S024R-S242R,N018R-S024R-V104I-H249R,N018R-S024R-V244R,N018R-S078R-S101G-S103A-V104I-A232V-Q245R,N018R-S087D-S101G-S103A-V104I-A232V-Q245R,N018R-S101G-Q245R,N018R-S101G-S103A-H249R,N018R-S101G-S103A-Q245R,N018R-S101G-S103A-V104I-A232V-Q245R,N018R-S101G-S103A-V104I-V150L-A232V-Q245R,N018R-S101G-V104I-A232V-H249R,N018R-S101G-V104I-A232V-Q245R,N018R-S101G-V104I-H249R,N018R-S103A-A232V-H249R,N018R-S103A-V104I-H249R,N018R-T022R-S024R-N043D-N076D-H249R,N018R-T022R-S024R-N043R-N076D-H249R,N018R-T022R-S024R-N076D-H249R,N018R-T022R-S024R-N076D-S087D-H249R,N018R-T022R-S024R-N076D-V150L-H249R,N018R-T022W-S024R-N076D-N116A-G211Q-H249R,N018R-T022W-S024R-N076D-N116A-T213A-H249R,N018R-T022W-S024R-N076D-S101A-I198L-A215F-H249R,N018R-T022W-S024R-N076D-S101A-I198L-H249R,N018R-T022W-S024R-N076D-S101A-N116A-A232V-Q245R,N018R-T22K-N043D-S101G-S103A-V104I-A232V-Q245R,N018R-T22W-S024R-N076D-N116A-T213A-H249R,N018R-T22W-S024R-N076D-S101A-N116A-A232V-Q245R,N018R-V104I-A232V-H249R,N043A-N062Q-A194F-G211Q,N043A-T057R-N117F-S144R-N183D,N043D-N076D-S078R-S101G-S103A-V104I-A232V-Q245R-N269R,N043D-R045T-N076D-H249R,N043D-S078R-S101G-S103A-V104I-A232V-Q245R,N043D-S078R-S101G-S103A-V104I-A232V-Q245R-H249R,N043D-S101G-S103A-V104I-A232V-Q245R-N269R,N043R-N076D-S078R-S101G-S103A-V104I-A232V-Q245R,N043R-N076D-S101G-S103A-V104I-A232V-Q245R,N043R-N076D-S101G-S103A-V104I-A232V-Q245R-N269R,N043R-N076D-S242R-H249R,N043R-N116A-N269R,N043R-R045T-N076D-S078R-S101G-S103A-V104I-A232V-Q245R,N043R-R045T-S078R-S101G-S103A-V104I-A232V-Q245R,N043R-R045T-S101G-S103A-V104I-A232V-Q245R-N269R,N043R-S078R-S101G-S103A-V104I-A232V-Q245R,N043R-S087D-S101G-S103A-V104I-A232V-Q245R-N269R,N043R-S101A-N116A-A215F-N269R,N043R-S101A-N116A-N269R,N043R-S101A-N116A-T213A-A215F-N269R,N043R-S101A-N269R,N043R-S101A-T213A-N269R,N043R-S101G-S103A-V104I-A232V-Q245R,N043R-S101G-S103A-V104I-A232V-Q245R-E271L,N043R-S101G-S103A-V104I-A232V-Q245R-H249R,N043R-S101G-S103A-V104I-A232V-Q245R-N269R,N043R-S101G-S103A-V104I-A232V-S242R-Q245R,N043R-S101G-S103A-V104I-V150L-A232V-Q245R,N043R-S242R-H249R,N043R-T213A-A215F-N269R,N062Q-S103N-N117F-A194F,N062Q-S103N-V121F-S144R-H249R,N076D-S078R-S101G-S103A-V104I-A232V-Q245R,N076D-S078R-S101G-S103A-V104I-A232V-Q245R-N269R,N076D-S101G-S103A-V104I-A232V-Q245R,N076D-S101G-S103A-V104I-A232V-Q245R-N269R,N076D-S103A-V104I-Q109R,N076D-S103A-V104I-Q109R-Q245R,N076D-S103A-V104I-S212P-E271V,N076D-S103A-V104I-S212P-Q245L-E271V,N076D-S87R-S103A-V104I-S212P-E271V,N117F-A194F-T213A-A270C,N117F-T213A-A215F,P052N-S078R-S103N-L148I-T213A,P052N-S103N-N116A-L148I-Y192W,R019H-G020R-T022W-S078R-S101G-S103A-V104I-G211Q-A232V-Q245R,R045T-N076D-S078R-S101G-S103A-V104I-A232V-Q245R-N269R,R045T-S078R-S101G-S103A-V104I-A232V-Q245R,R045T-S078R-S101G-S103A-V104I-A232V-Q245R-N269R,S009A-G020R-N043R-S212F,S009A-G020R-N043R-W241R,S009A-G020R-S024R-N043R,S009A-N043R-S078R,S009A-N043R-S078R-S242R,S009A-N043R-S212F,S009A-N043R-W241R,S009A-T022R-N043R-S078R,S009A-T022R-S078R-S212F,S009A-T022R-S078R-S212F-W241R,S009A-T022R-S212F-W241R,S024R-A215F-N269R,S024R-A232V-Q245R,S024R-G025R-N183D-Y192W-P239S,S024R-N043A-N117F-A194F-G211Q,S024R-N043D-H249R,S024R-N043D-S101G-S103A-V104I-A232V-Q245R-H249R,S024R-N043D-S101G-S103A-V104I-A232V-Q245R-N269R,S024R-N043R-A215F,S024R-N043R-A230E,S024R-N043R-A230E-S242R,S024R-N043R-H249R,S024R-N043R-N076D-H249R,S024R-N043R-N076D-S078R-S101G-S103A-V104I-A232V-Q245R,S024R-N043R-N076D-S101G-S103A-V104I-A232V-Q245R,S024R-N043R-N116A-A215F,S024R-N043R-N116A-A215F-N269R,S024R-N043R-N116A-N269R,S024R-N043R-N116A-T213A-N269R,S024R-N043R-R045T-N076D-A230E-H249R,S024R-N043R-R045T-N269R,S024R-N043R-R045T-S101A-N116A-A215F-N269R,S024R-N043R-R045T-S101A-N116A-T213A-N269R,S024R-N043R-R045T-S242R,S024R-N043R-R045T-T213A-A215F-N269R,S024R-N043R-S101A-A215F,S024R-N043R-S101A-A215F-N269R,S024R-N043R-S101A-N116A,S024R-N043R-S101A-N116A-A215F-N269R,S024R-N043R-S101G-S103A-V104I-A232V-Q245R,S024R-N043R-S242R,S024R-N062Q-V104L-S106G-H249R,S024R-N076D-S078R-S101G-S103A-V104I-A232V-Q245R-N269R,S024R-N076D-V104I-A232V-Q245R,S024R-N116A-T213A-N269R,S024R-R045T,S024R-R045T-N076D-A230E-S242R-H249R,S024R-R045T-N076D-S078R-S101G-S103A-V104I-A232V-Q245R-N269R,S024R-R045T-S101G-S103A-V104I-A232V-Q245R-N269R,S024R-S078R-S212F,S024R-S078R-V104L-N116A-N183D,S024R-S087D-S101G-S103A-V104I-A232V-Q245R,S024R-S101A-H120F-A194F-H249R,S024R-S101A-N269R,S024R-S101G-S103A-V104I-A232V-Q245R,S024R-S101G-S103A-V104I-V150L-A232V-Q245R,S024R-S101G-V104I-Q245R,S024R-S103A-Q245R,S024R-S103A-V104I-A232V-H249R,S024R-S103A-V104I-H249R,S024R-S103A-V104I-Q245R,S024R-Y167W-T224A-H249R,S078R-S087D-S101G-S103A-V104I-A232V-Q245R,S078R-S101G-S103A-V104I-A232V-Q245R,S078R-S101G-S103A-V104I-A232V-Q245R-N269R,S078R-S101G-S103A-V104I-V150L-A232V-Q245R,S078R-S103N-S106G-Y167W-Q236N,S078R-V104L-T213A-A215F-T224A,S078R-Y091F-V121F-L233C-N252R,S087D-S101G-S103A-V104I-A232V-Q245R-N269R,S087R-S101G-S103A-V104I-Q109R-S212P-A232V-Q245R-E271V,S099F-S144R-Y167W-N252R,S101A-H120F-Y192W-A215F-T224A,S101A-S103A-V104I-T213A-A232V-Q245R-N269R,S101G-S103A-V104I-A232V-Q245R,S101G-S103A-V104I-A232V-Q245R-E271L,S101G-S103A-V104I-A232V-Q245R-H249R,S101G-S103A-V104I-A232V-Q245R-N269R,S101G-S103A-V104I-A232V-Q245R-N269R,S101G-S103A-V104I-A232V-S242R-Q245R,S101G-S103A-V104I-A232V-V244R-Q245R,S101G-S103A-V104I-G115R-A232V-Q245R,S101G-S103A-V104I-N116A-T213A-A232V-Q245R-N269R,S101G-S103A-V104I-Q109R-A232V-Q245R,S101G-S103A-V104I-Q109R-S212P-A232V-Q245R,S101G-S103A-V104I-Q109R-S212P-A232V-Q245R-E271V,S101G-S103A-V104I-S212F-A232V-Q245R,S101G-S103A-V104I-V150L-A232V-Q245R-H249R,S101G-S103A-V104I-V150L-A232V-Q245R-N269R,S105T-S128N-S144R-L148I-S212M,S106G-N117F-N238L,S144R-G211Q-N238L-P239S-H249R,T022R-N043R-S101G-S103A-V104I-A232V-Q245R,T022R-N076D-S101G-S103A-V104I-A232V-Q245R,T022R-S087D-S101G-S103A-V104I-A232V-Q245R,T022R-S101G-S103A-V104I-A232V-Q245R,T022W-S078R-Y167W-S212M-A270C,T057R-E089I-I198L,T057R-S099F-S105T-I198L-T213A,T057R-S099F-V121F-N185I-Y192W,T057R-Y167W-H249R,V004R-G020R-H249R,V004R-G020R-N043R,V004R-G020R-S024R,V004R-G020R-S024R-H249R,V004R-G020R-S024R-N043R,V004R-G020R-S024R-N043R-S242R,V004R-G020R-S024R-V244R,V004R-N043R-S101G-S103A-V104I-A232V-Q245R,V004R-S009A-G020R-N043R,V004R-S009A-G020R-N043R-S242R,V004R-S009A-G020R-S024R-S242R,V004R-S009A-G020R-S242R,V004R-S009A-N043R-W241R,V004R-S009A-S024R-N043R-W241R,V004R-S009A-S078R-W241R,V004R-S009A-W241R,V004R-S101G-S103A-V104I-A232V-Q245R,V104L-L217E-T224A-H249R-N252R,V104L-T213A-S216F,V121F-N252R-A270C,Y091F-S099F-S101A-S105T-Y167W,N043R-N076D-A230E-H249R,S024R-N043D-N076D-S242R-H249R,G020R-S024R-N043D-R045T,N043R-N076D-H249R,N043R-R045T-H249R,G020R-N043R-N076D,A230E-H249R,N043D-S242R-H249R,G020R-S024R-N043D-N076D-S242R-H249R,S024R-N043D-S242R-H249R,N018R-N043D-R045T-N076D-S242R-H249R,S024R-N043R-N076D,N018R-G020R-H249R,N018R-G020R-N043R,G020R-S024R-T38I-N043R-R045T-N076D-S242R-H249R,S024R-N043R-N076D-A230E-S242R,N018R-G020R-N043D-R045T-N076D-S242R,N018R-S024R-N043D-A230E-H249R,N018R-N043R-N076D-S242R-H249R,G020R-S024R-K27R-N043D-S242R-H249R,G020R-S242R-H249R,N018R-N043R-R045T-S242R,G020R-N043R-N076D-A230E-H249R,N018R-N043R-N076D-S242R,N018R-G020R-N043R-N076D,N018R-S024R-N043D-S242R,R045T-S242R-H249R,N018R-G020R-N043R-R045T-N076D-H249R，并且其中所述变体的总净电荷相对于迟缓芽孢杆菌(Bacillus lentus)枯草杆菌蛋白酶GG36蛋白酶的总净电荷为0、+1、+2、+3、+4、+5、-1、-2、-3、-4或-5，并且其中所述总净电荷是通过一种或多种选自以下的置换来获得的：N43D,R45T,N62E,N76D,S101D,P129E,A158E,G159D,G159E,S166D,S188D,A230E,N18R,G20K,G20R,T22R,S24R,N43R,G118R,Q245R,H249R,N269R,E271F，和E271L，并且其中所述蛋白酶变体的氨基酸位置根据SEQ ID NO:1所示解淀粉芽孢杆菌(Bacillus amyloliquefaciens)枯草杆菌蛋白酶BPN’的氨基酸序列中对应的氨基酸位置的编号方式来进行编号。7.一种分离的枯草杆菌蛋白酶变体，其中所述枯草杆菌蛋白酶变体为具有蛋白水解活性的成熟形式并包含具有选自以下的氨基酸置换的组合的氨基酸序列：A016S-S128N-R186H-E271F,N076D-S101G-S103A-V104I-A158E-S188D-M222S-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-S128L-P129Q-A158E-N183D-S188D-A232V-Q245R-N248D-H249R,S024R-S101G-S103A-V104I-S128L-P129Q-A158E-S188D-A232V-Q245R-N248D-H249R-E271G,S024R-S101G-S103A-V104I-S128L-P129Q-S130A-A158E-N183D-S188D-A232V-Q245R-N248D-H249R,S101G-S103A-V104I-P129Q-A158E-N183D-S188D-A232V-Q245R-N248D-H249R,T022A-N076D-S101G-S103A-V104I-G159D-A232V-Q245R-N248D-H249R-E271F,T022A-S101G-G102A-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F,A016S-A158E-H249R,A016S-A158E-H249R-E271F,A016S-A158E-R186H-H249R,A016S-L111V-S188D-,A016S-T022A-A158E-R186H-E271F,A048R-S078N-N116L-N185E-L217E-P239G,A048R-S128D-N185E-P239G,A158E-R186H-H249R-E271F,E089P-S101A-P129E-R186H,G020K-G023A-N043W-N116L-S188D-T213A-P239G,G020K-G023A-N062E-N116L-G118R-T213A,G020K-G023A-S024F-N062E-S188D-T213A-P239G,G020K-K027R-P129E-S166D-P239G,G020K-S024F-N062E-P239G,G020K-S024F-T033S-P129E-A194E,G023A-G100S-A194E-S212M,G023A-N043W-N116L-G118R-S188D,G023A-S024F-G118R-S188D-P239G,G023A-S024F-K027R-N062E,G023A-S024F-N043W-N062E-N116L-G118R,G023A-S024F-N116L-G118R-S188D-T213A,G023A-S024F-N116L-S188D-P239G,K027R-G100S-G118R-S160D-S188D-N243F,K027R-S101D-S103N-S105T-A272F,N043R-N076D-S101A-S103A-V104I-A158E-S166D-S188D-A232V-Q245R-N248D-H249R,N062E-S078N-N116L-T224A,N062E-V104L-A158E-R186H-S188D-H249R,N076D-S101A-S103A-V104I-A158E-S188D-L217E-A232V-Q245R-N248D-H249R-E271F,N076D-S101G-S103A-V104I-A158E-S166D-S188D-A232V-Q245R-N248D-H249R-E271F,S024F-G102A-R186K-T213A-L217E-N243F,S024F-N043W-G118R-S188D,S024F-N043W-V104L-V121F-P129E,S024F-N116L-G118R-S188D-P239G,S024F-S103N-V104L-G118R-S188D,S024R-N043R-S101A-S103A-V104I-A158E-S188D-A232V-Q245R-N248D-H249R,S024R-N043R-S101A-S103A-V104I-A158E-S188D-L217E-A232V-Q245R-N248D-H249R,S024R-N076D-S101G-S103A-V104I-A158E-S188D-A232V-Q245R-N248D-H249R-E271F,S024R-S101G-S103A-V104I-A158E-G159E-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-S128N-A158E-G159E-S188D-A232V-Q245R-N248D,S024R-S101G-S103A-V104I-S128N-P129E-S188D-A232V-Q245R-N248D,S101A-L111V-P129E,S101A-S103A-V104L-G159E-A232V-Q245R-N248D-H249R,S101A-S103S-V104L-G159E-A232V-Q245R-N248D-H249R,S101A-V104L-S128N-A158E-R186H,S101G-S103A-V104I-P129E-L148I-S188D-A232V-Q245R-N248D,S101G-S103A-V104I-S128N-P129E-A158E-A232V-N238R-Q245R-N248D,S101G-S103A-V104I-S128N-P129E-S188D-A232V-Q245R-N248D-H249R,S101G-S103S-V104L-G159E-A232V-Q245R-N248D-H249R,S101 S-S103A-V104L-G159E-A232V-Q245R-N248D-H249R,S128N-P129E-R186H,T022A-A158E-R186H-H249R-E271F,T022A-S024K-S101G-S103A-V104I-S128N-A158E-G159E-A232V-Q245R-N248D,T022A-S024R-S101G-S103A-V104I-S128N-P129E-A158E-S188D-A232V-Q245R-N248D-H249R,T022A-S024R-S103A-V104I-S128I-G159D-S188D-A232V-N248D,T022A-S101G-S103A-V104I-L148I-S188D-A232V-Q245R-N248D,T022A-S101G-S103A-V104I-S128N-G159E-A232V-Q245R-N248D,T022A-S101G-S103A-V104I-S128N-P129E-A158E-A232V-N238R-Q245R-N248D,T022A-S128N-A158E-R186H,T022A-S128N-R186H-S188D-,T022A-V104L-A158E-H249R,T022A-V104L-R186H-S188D-H249R,T022L-G023A-K027R-S101D-V104L-S216F,T022L-S078N-N116L-P129E-S256N,T022L-S078N-S128D-T213A,T033S-G118R-P129E-A194E-P239G,T033S-S105T-S188D-S216F,V026F-V104L-S256N-A272F,T022A-N043R-S103A-V104I-S128I-G159D-S188D-A232V-Q245R-N248D,T022A-S024R-N043R-S103A-V104I-G118R-G159D-S188D-L217D-A232V-N248D,T022A-S101D-S103A-V104I-G118R-G159D-S188D-L217D-A232V-Q245R-N248D-E271F,T022A-S103A-V104I-G118R-P129E-G159D-S188D-A232V-Q245R-N248D-E271F,T022A-S103A-V104I-G159D-S188D-L217D-A232V-Q245R-N248D-E271F,T022A-S103A-V104I-S128I-P129E-G...

【专利技术属性】
技术研发人员：V·Y·阿列克谢耶夫，N·S·阿明，K·奥古斯汀，L·G·卡斯康佩雷拉，K·D·科利尔，D·A·埃斯特尔，J·T·小凯利斯，A·J·保罗斯，P·F·苏特，G·S·沃德，J·姚，
申请(专利权)人：丹尼斯科美国公司，宝洁公司，
类型：发明
国别省市：美国;US