CN113302270A - 低pH粉末洗涤剂组合物 - Google Patents
低pH粉末洗涤剂组合物 Download PDFInfo
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- CN113302270A CN113302270A CN201980080256.9A CN201980080256A CN113302270A CN 113302270 A CN113302270 A CN 113302270A CN 201980080256 A CN201980080256 A CN 201980080256A CN 113302270 A CN113302270 A CN 113302270A
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Classifications
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D17/00—Detergent materials or soaps characterised by their shape or physical properties
- C11D17/06—Powder; Flakes; Free-flowing mixtures; Sheets
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/0005—Other compounding ingredients characterised by their effect
- C11D3/0047—Other compounding ingredients characterised by their effect pH regulated compositions
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/386—Preparations containing enzymes, e.g. protease or amylase
- C11D3/38609—Protease or amylase in solid compositions only
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2411—Amylases
- C12N9/2414—Alpha-amylase (3.2.1.1.)
- C12N9/2417—Alpha-amylase (3.2.1.1.) from microbiological source
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
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Abstract
本发明涉及低pH粉末洗涤剂组合物，该组合物包含具有α‑淀粉酶活性并且与SEQ ID NO:1‑19具有至少60％，如65％、如70％、如75％、如80％、如85％、如90％、如95％、如97％序列同一性的至少一种多肽，并且该组合物的pH值低于9.5。
Description
序列表的引用
本申请包含计算机可读形式的序列表。该计算机可读形式通过引用并入本文。
技术领域
本发明涉及粉末洗涤剂组合物，特别涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶的至少一种多肽。
背景技术
α-淀粉酶多年来一直用于衣物洗涤，其中熟知的是α-淀粉酶在去除含淀粉或基于淀粉的污渍中具有有益的效果。
WO 95/26397披露了在30℃-60℃范围内的温度下测量时具有良好洗涤性能的碱性芽孢杆菌淀粉酶。
WO 00/60060和WO 00/60058披露了具有良好洗涤性能的另外的细菌α-淀粉酶。
WO 2016/180748披露了α-淀粉酶变体，在低温下提高了洗涤性能。证明这些变体在液体洗涤剂组合物中具有良好性能。
洗涤剂组合物可以以不同的配制品如液体配制品和粉末配制品提供。
由于粉末配制品中的成分呈低含水量的固体形式，因此粉末配制品中的酶也是固体形式的，并且以此形式至少在某种程度上受到保护而不被蛋白水解降解。粉末洗涤剂配制品通常是高度碱性的，且pH值高于9.0，并且甚至高于10.0的pH值也并不少见。此外，粉末洗涤剂可含有漂白剂。
液体洗涤剂配制品通常含有水，且其中的酶通常是可溶性形式，这意味着与粉末配制品中的酶相比，液体洗涤剂中的酶更容易降解如蛋白水解降解。
粉末洗涤剂和液体洗涤剂配制品之间的显著差异，因此通常在这两种配制品中使用不同的酶，不能因为酶在例如粉末洗涤剂配制品中表现良好得出结论认为该酶在液体洗涤剂配制品中也会表现良好，反之亦然。
发明内容
本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性的至少一种多肽，其中该组合物中α-淀粉酶的存在令人惊讶地发现产生了改善的清洁性能。
本发明提供了粉末洗涤剂组合物，该组合物包含具有α-淀粉酶活性并且与SEQ IDNO:1-19具有至少60％，如65％、如70％、如75％、如80％、如85％、如90％、如95％、如97％序列同一性的至少一种多肽，并且其中该组合物的pH值低于9.5；例如在7.0-9.5的范围内；例如在7.5至9.0的范围内；例如在8.0至9.0的范围内。
本发明还涉及本文所述的组合物在清洁过程如衣物洗涤或餐具洗涤中的用途，并且涉及使用低pH洗涤剂组合物进行清洁的方法。
定义
餐具洗涤组合物：术语“餐具洗涤组合物”是指旨在用于清洁盘子、餐桌用具、罐、锅、餐具的组合物和用于清洁厨房中硬表面区域的所有形式的组合物。本发明不限于任何特定类型的餐具洗涤组合物或任何特定的洗涤剂。
酶去垢力益处：术语“酶洗涤益处”在本文中定义为将酶添加至洗涤剂中与不具有该酶的相同洗涤剂相比的有利效果。可以由酶提供的重要洗涤益处是污渍去除伴随在洗涤和/或清洁之后无可见污垢或可见污垢非常少、防止或减少在洗涤过程中所释放的污垢再沉积(也被称作抗再沉积的效果)、完全或部分地恢复纺织品的白度(也被称作变白的效果)，该纺织品最初是白色的，但是在反复使用和洗涤后获得淡灰或淡黄色外观。不直接与催化污渍去除或防止污垢再沉积相关的纺织品护理益处对于酶洗涤益处而言也是重要的。此类纺织品护理益处的实例是防止或减少染料从一织物转移至另一织物或同一织物的另一部分(也被称作染料转移抑制或抗返染的效果)，从织物表面去除突出或断裂的纤维以减少起球倾向或去除已经存在的球或绒毛(也被称作抗起球的效果)，改善织物柔软性，使织物的颜色澄清以及去除陷在织物或服装的纤维中的微粒状污垢。酶漂白是一种另外的酶洗涤益处，其中通常将催化活性用于催化漂白组分(例如过氧化氢或其他过氧化物)的形成。
改善的洗涤性能：术语“改善的洗涤性能”在此定义为相对于洗涤剂组合物的洗涤性能增加的洗涤性能，例如增加的去污。术语“改善的洗涤性能”包括在衣物中的洗涤性能。
硬表面清洁：本文将术语“硬表面清洁”定义为清洁硬表面，其中硬表面可以包括地板、桌子、墙壁、屋顶等，连同硬物体的表面，如汽车(汽车洗涤)和餐具(餐具洗涤)。餐具洗涤包括但不限于，清洁盘子、杯子、玻璃杯、碗、刀叉餐具(例如匙、刀、叉)、上菜用具、陶瓷、塑料、金属、瓷器、玻璃及丙烯酸酯。
粉末洗涤剂组合物：术语“粉末洗涤剂组合物”在本文中定义为其中全部或大部分成分呈固体干燥形式的洗涤剂组合物。粉末通常由包含一种或多种粉末和或颗粒的混合物组成。术语粉末洗涤剂组合物包括单位剂型，例如已通过将一种或多种粉末组合、压制或附聚成较大的结构制成的、以干燥形式呈现的洗涤块(tab)、洗涤片(tablet)。因此，粉末洗涤剂组合物中的含水量应足够低，以防止组合物粘附或意外附聚成更大的结构。为了简单起见，本说明书和权利要求书中通常称为“粉末”组合物。除非从上下文中另有说明或是明显的，否则本文使用的术语“粉末”应理解为还包括如上所述的固体形式，例如颗粒和洗涤块。
粉末洗涤剂组合物的pH：粉末洗涤剂组合物的pH意指表示粉末洗涤剂组合物在水中的即时可用水溶液在20℃的pH。为了测量粉末洗涤剂组合物的pH，第一步是制备每升水5g粉末洗涤剂组合物的溶液，然后使用众所周知的技术和/或用于pH测量的装置测量溶液中的pH。
在粉末洗涤剂组合物呈单位剂型的形式的情况下，将一单位剂量溶解于15升水中，测量该溶液在20℃的pH，并且该pH值被认为是粉末洗涤剂组合物单位剂型的pH。
序列同一性：两个氨基酸序列之间或两个核苷酸序列之间的关联度通过参数“序列同一性”来描述。出于本发明的目的，使用如在EMBOSS软件包(EMBOSS：欧洲分子生物学开放软件包(EMBOSS:The European Molecular Biology Open Software Suite)，Rice等人,2000,Trends Genet.[遗传学趋势]16:276-277)(优选5.0.0版本或更新版本)的尼德尔程序中所实施的尼德曼-翁施算法(Needleman-Wunsch algorithm)(Needleman和Wunsch,1970,J.Mol.Biol.[分子生物学杂志]48:443-453)来确定两个氨基酸序列之间的序列同一性。所使用的参数是空位开放罚分10、空位延伸罚分0.5和EBLOSUM62(BLOSUM62的EMBOSS版)取代矩阵。将标记为“最长同一性”的Needle的输出(使用非简化选项获得)用作同一性百分比并且计算如下：
(相同的残基x 100)/(比对长度-比对中的空位总数)
出于本发明的目的，使用如在EMBOSS软件包(EMBOSS：欧洲分子生物学开放软件包(EMBOSS:The European Molecular Biology Open Software Suite)，Rice等人,2000,同上)(优选5.0.0版本或更新版本)的尼德尔程序中所实施的尼德曼-翁施算法(Needleman和Wunsch,1970,同上)来确定两个脱氧核糖核苷酸序列之间的序列同一性。所使用的参数是空位开放罚分10、空位延伸罚分0.5和EDNAFULL(NCBI NUC4.4的EMBOSS版)取代矩阵。将标记为“最长同一性”的Needle的输出(使用非简化选项获得)用作同一性百分比并且计算如下：
(相同的脱氧核糖核苷酸x 100)/(比对长度-比对中的空位总数)
纺织品：术语“纺织品”意指任何纺织品材料，所述任何纺织品材料包括纱线、纱线中间体、纤维、非机织材料、天然材料、合成材料、以及任何其他纺织品材料，由这些材料制成的织物和由织物制成的产品(例如，服装和其他制品)。该纺织品或织物可以处于针织品、机织物(woven)、牛仔布(denim)、非机织物、毡、纱线、以及毛巾布的形式。该纺织品可以是基于纤维素的，如天然纤维素制品，包括棉、亚麻/亚麻布、黄麻、苎麻、剑麻或椰壳纤维，或者人造纤维素(例如，来源于木浆)，包括粘胶纤维/人造丝、乙酸纤维素纤维(三胞)、莱赛尔纤维(lyocell)或其共混物。该纺织品或织物也可以不基于纤维素，如天然聚酰胺，包括羊毛、驼毛、羊绒、马海毛、兔毛和蚕丝，或合成聚合物如尼龙、芳族聚酰胺、聚酯、丙烯酸、聚丙烯和氨纶/弹性纤维(spandex/elastane)、或其共混物以及基于纤维素和不基于纤维素的纤维的共混物。共混物的实例是棉和/或人造丝/粘胶纤维与一种或多种伴随材料的共混物，该伴随材料如羊毛、合成纤维(例如聚酰胺纤维、丙烯酸纤维、聚酯纤维、聚氯乙烯纤维、聚氨酯纤维、聚脲纤维、芳族聚酰胺纤维)和/或含纤维素的纤维(例如人造丝/粘胶纤维、苎麻、亚麻/亚麻布、黄麻、乙酸纤维素纤维、莱赛尔纤维)。织物可以是常规的可洗涤衣物，例如有污渍的家用衣物。当使用术语织物或服装时，旨在也包括广义术语纺织品。
变体：术语“变体”意指在一个或多个位置处包含改变(即，取代、插入和/或缺失)的具有α-淀粉酶活性的多肽。取代意指用不同的氨基酸替代占据某一位置的氨基酸；缺失意指去除占据某一位置的氨基酸；而插入意指在邻接并且紧随占据某一位置的氨基酸之后添加氨基酸。
洗涤性能：术语“洗涤性能”被用作酶在例如洗涤或硬表面清洁过程中去除存在于有待清洁的物体上的污渍的能力。洗涤性能的改善可以通过计算本文的所谓的强度值(Int)来量化。术语“洗涤性能”和“餐具洗涤性能”可互换使用。
变体命名惯例
出于本发明的目的，使用在SEQ ID NO:1中披露的多肽来确定另一种α-淀粉酶中的相应氨基酸残基。将另一种α-淀粉酶的氨基酸序列与SEQ ID NO:1中披露的脂肪酶进行比对，并且基于比对，使用如在EMBOSS包(EMBOSS：欧洲分子生物学开放软件套件，Rice等人,2000,Trends Genet.[遗传学趋势]16:276-277)(优选5.0.0版本或更新版本)的尼德尔程序中所实施的尼德曼-翁施算法(Needleman和Wunsch,1970,J.Mol.Biol.[分子生物学杂志]48:443-453)来确定与SEQ ID NO:1中披露的多肽中的任何氨基酸残基相对应的氨基酸位置编号。所使用的参数是空位开放罚分10、空位延伸罚分0.5和EBLOSUM62(BLOSUM62的EMBOSS版)取代矩阵。
可以通过使用若干计算机程序，使用其相应默认参数比对多个多肽序列来确定在另一种α-淀粉酶中的对应氨基酸残基的鉴定，所述计算机程序包括但不限于MUSCLE(通过对数预期的多序列比较；版本3.5或更新版本；Edgar,2004,Nucleic Acids Research[核酸研究]32:1792-1797)，MAFFT(版本6.857或更新版本；Katoh和Kuma,2002,Nucleic AcidsResearch[核酸研究]30:3059-3066；Katoh等人,2005,Nucleic Acids Research[核酸研究]33:511-518；Katoh和Toh,2007,Bioinformatics[生物信息学]23:372-374；Katoh等人,2009,Methods in Molecular Biology[分子生物学方法]537:39-64；Katoh和Toh,2010,Bioinformatics[生物信息学]26:1899-1900)以及采用ClustalW(1.83或更新版本；Thompson等人,1994,Nucleic Acids Research[核酸研究]22:4673-4680)的EMBOSS EMMA。
对于已知结构的蛋白质，若干工具和资源可用于检索并产生结构比对。例如，蛋白质的SCOP超家族已经在结构上进行比对，并且那些比对是可访问且可下载的。可以使用多种算法如距离比对矩阵(Holm和Sander,1998,Proteins[蛋白质]33:88-96)或组合延伸(Shindyalov和Bourne,1998,Protein Engineering[蛋白质工程]11:739-747)比对两种或更多种蛋白质结构，并且这些算法的实施可以另外用于查询具有感兴趣结构的结构数据库，以便发现可能的结构同源物(例如，Holm和Park,2000,Bioinformatics[生物信息学]16:566-567)。
在描述本发明的变体中，为了便于参考，对以下所述的命名法进行了改编。采用了已接受的IUPAC单字母或三字母的氨基酸缩写。
取代：对于氨基酸取代，使用以下命名法：原始氨基酸、位置、取代的氨基酸。相应地，将在位置226处的苏氨酸被丙氨酸取代表示为“Thr226Ala”或“T226A”。多个突变通过加号(“+”)分开，例如“Gly205Arg+Ser411Phe”或“G205R+S411F”代表在位置205和位置411处的甘氨酸(G)和丝氨酸(S)分别被精氨酸(R)和苯丙氨酸(F)取代。
缺失：对于氨基酸缺失，使用以下命名法：原始氨基酸、位置、*。相应地，将在位置195处的甘氨酸的缺失表示为“Gly195*”或“G195*”。多个缺失通过加号(“+”)分开，例如“Gly195*+Ser411*”或“G195*+S411*”。
多重修饰：包含多个修饰的变体由加号(“+”)分开，例如“Arg170Tyr+Gly195Glu”或“R170Y+G195E”代表在位置170和位置195处的精氨酸和甘氨酸分别被酪氨酸和谷氨酸取代。
不同修饰：在一个位置上可能引入不同修饰的情况下，这些不同的改变由逗号分开，例如“Arg170Tyr,Glu”表示在位置170上的精氨酸被酪氨酸或谷氨酸取代。因此，“Tyr167Gly,Ala+Arg170Gly,Ala”表示以下变体：
“Tyr167Gly+Arg170Gly”、“Tyr167Gly+Arg170Ala”、“Tyr167Ala+Arg170Gly”、和“Tyr167Ala+Arg170Ala”。
具体实施方式
本发明在一个方面涉及粉末洗涤剂组合物，该组合物包含具有α-淀粉酶活性并且与SEQ ID NO:1-19具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽，并且其中该组合物的pH值低于9.5。
在一个实施例中，本发明涉及具有不超过约9的pH(如在7.0-9.5的范围内；如在7.5至9.0的范围内；如在8.0至9.0的范围内)的组合物。
从本说明书中显而易见的是，如本文所用的术语“低pH”是指相对于常规粉末洗涤剂、例如用于衣物洗涤或餐具洗涤的那些(如上所述，其通常具有使用中高于9且经常高于10的pH)较低的pH。
如上所述，本文使用的术语“粉末”应理解为指固体干燥形式的组合物。本发明的“粉末”通常由包含一种或多种粉末和/或颗粒的混合物组成，但也包括例如单位剂型、如洗涤块。
在一个实施例中，本发明的组合物的pH低于约9.5，如不超过约9.4、如不超过约9.3、如不超过约9.2、如不超过约9.1、如不超过约9.0、如不超过约8.9、如不超过约8.8、如不超过约8.7、如不超过约8.6、如不超过约8.5、如不超过约8.4、如不超过约8.3、如不超过约8.2、如不超过约8.1、或不超过约8.0。另一方面，组合物通常具有至少约7的pH，例如至少约7.1、至少约7.2、至少约7.3、至少约7.4、至少约7.5、至少约7.6、至少约7.7、至少约7.8、或至少约7.9。在所有情况下，如上所述在5g/l溶液中确定pH。
在一些实施例中，pH可以是例如在从约7.0至不超过约9.0的范围内，例如从约7.2至约8.9、如从约7.4至约8.8、如从约7.6至约8.7、如从约7.8至约8.6。
在一个实施例中，如上所述在5g/l溶液中确定的，pH可以在从约7.0至约8.2的范围内，例如从约7.2至约8.0。
在另一个实施例中，如上所述在5g/l溶液中确定的，pH可以在从约7.8至约8.8的范围内，例如从约8.0至约8.6。
应当注意，尽管如上所述pH通常在5g/l溶液中确定，但预期对于单位剂型、例如洗涤块，可以通过在20℃将一单位(例如一个洗涤块)溶解于15l的去离子水中并测量该溶液的pH来确定pH。
本发明的粉末洗涤剂组合物具有低pH值。如本领域技术人员将理解的，pH值通常在水溶液中测量而不是在干燥粉末中测量，因此应该理解，干燥组合物如本披露中的粉末洗涤剂组合物的pH值意指所讨论的组合物的水溶液的pH值。因此，根据本发明，通过制备粉末洗涤剂组合物在水中的5g/L溶液以及在洗涤剂组合物完全溶解于水中后测量pH值(使用本领域中已知的用于pH测量的技术)，确定粉末洗涤剂组合物的pH。
应当注意，尽管如上所述pH通常在5g/l溶液中确定，但预期对于单位剂型、例如洗涤块，可以通过在20℃将一单位(例如一个洗涤块)溶解于15l的去离子水中并测量该溶液的pH来确定pH。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:1-19具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:1-19具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少两种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:1-19具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少三种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:1-19具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少四种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:1-19具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少五种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:1-19具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种或多种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:1具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:2具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:3具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:4具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:5具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:6具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:7具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:8具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:9具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:10具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:11具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:12具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:13具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:14具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:15具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:16具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:17具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:18具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:19具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的至少一种多肽。
在一个方面，本发明涉及低pH粉末洗涤剂组合物，其包含具有α-淀粉酶活性并且与SEQ ID NO:1-19具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％序列同一性的多肽的变体。
在一个实施例中，本发明涉及具有α-淀粉酶活性的多肽的变体，其包含在对应于SEQ ID NO:11的多肽的位置G304、W140、W189、D134、E260、F262、W284、W347、W439、W469、G476、和G477的两个或更多个(几个)位置处的改变，其中每个改变独立地是取代、缺失或插入，并且其中该变体与SEQ ID NO:11的多肽具有至少60％、至少65％、至少70％、至少75％、至少80％、或至少87％、但少于100％的序列同一性，并且其中该变体具有α-淀粉酶活性。
在一个实施例中，具有α-淀粉酶活性的多肽是SEQ ID NO:11的变体，包括选自由以下组成的组的取代：
W140Y+N195F+V206Y+Y243F+E260G+G477E，
W140Y+N195F+V206Y+Y243F+E260T+W284D，
W140Y+N195F+V206Y+Y243F+W284D，
G109A+W140Y+N195F+V206Y+Y243F+E260G，
W140Y+N195F+V206Y+Y243F+E260G，
N195F+V206Y+Y243F+E260K+W284D，
D134E+G476E，
W140Y+N195F+V206Y+Y243F+E260G+G476E，
W140Y+W189G+N195F+V206Y+Y243F+E260G，
W140Y+N195F+V206Y+Y243F+E260G+S303G，
W140Y+W189T+N195F+V206Y+Y243F+E260G，
W140Y+N195F+V206Y+Y243F+E260G+W284D，
Y100I+W140Y+N195F+V206Y+Y243F+E260G，
W140Y+N195F+V206Y+Y243F+E260G+G337N，
W140Y+N195F+V206Y+Y243F+E260G+W439R
G109A+W140Y+E194D+N195F+V206Y+Y243F+E260G
G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E
T51I+Y100I+G109A+W140Y+N195F+V206Y+Y243F+E260G
T51I+G109A+W140Y+N195F+V206Y+Y243F+E260G+W439R
T51I+S52Q+N54K+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E
W140Y+N195F+V206Y+Y243F+E260G+G304R+G476K
W140Y+N195F+V206Y+Y243F+E260G+W284R+G477K
W140Y+N195F+V206Y+Y243F+E260G+W284F+G477R，以及
N195F+V206Y+Y243F+E260G+W284D。
优选地，该变体还包括在对应于SEQ ID NO:11的位置G182*+D183*或D183*+G184*的位置处的删除。
待用于本发明的组合物中的变体的优选实例包括如下变体，这些变体包含从选自以下的组中选择的改变或由其组成：
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G477E，
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260T+W284D，
D183*+G184*+W140Y+N195F+V206Y+Y243F+W284D，
D183*+G184*+G109A+W140Y+N195F+V206Y+Y243F+E260G，
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G，
D183*+G184*+N195F+V206Y+Y243F+E260K+W284D，
D183*+G184*+D134E+G476E，
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G476E，
D183*+G184*+W140Y+W189G+N195F+V206Y+Y243F+E260G，
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+S303G，
D183*+G184*+W140Y+W189T+N195F+V206Y+Y243F+E260G，
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284D，
D183*+G184*+Y100I+W140Y+N195F+V206Y+Y243F+E260G，
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G337N，
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W439R
D183*+G184*+G109A+W140Y+E194D+N195F+V206Y+Y243F+E260G
D183*+G184*+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E
D183*+G184*+T51I+Y100I+G109A+W140Y+N195F+V206Y+Y243F+E260G
D183*+G184*+T51I+G109A+W140Y+N195F+V206Y+Y243F+E260G+W439R
D183*+G184*+T51I+S52Q+N54K+G109A+W140Y+N195F+V206Y+Y243F+E260G+G476E
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G304R+G476K
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284R+G477K
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+W284F+G477R，以及
D183*+G184*+N195F+V206Y+Y243F+E260G+W284D，
H1*+G109A+W140Y+D183*+G184*+N195F+I206Y+Y243F+E260G+N280S+G304R+E391A+G476K，
H1*+G109A+W140Y+D183*+G184*+N195F+I206Y+Y243F+E260G+W284H+G304R+E391A+G476K，
H1*+G109A+W140Y+D183*+G184*+N195F+I206Y+Y243F+E260G+N280S+G304R+K320A+M323N+E391A+G476K，
H1*+G7A+G109A+W140Y+D183*+G184*+N195F+I206Y+Y243F+E260G+N280S+G304R+E391A+G476K，以及
H1*+G7A+G109A+W140Y+D183*+G184*+N195F+I206Y+Y243F+E260G+N280S+W284H+G304R+M323N+E391A+G476K。
在一个实施例中，在本发明的粉末洗涤剂组合物中具有α-淀粉酶活性的SEQ IDNO:11的变体，该变体包括如下改多肽包含变或由其组成：
D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G304R+G476K，以及
H1*+G7A+G109A+W140Y+D183*+G184*+N195F+I206Y+Y243F+E260G+N280S+G304R+E391A+G476K。
在一个实施例中，本发明涉及在SEQ ID NO:1的多肽的氨基酸序列内的一个或多个(例如，几个)位置处包含突变的、具有α-淀粉酶活性的多肽的变体，其中所述变体氨基酸序列包含氨基酸位置202和/或186处的突变，其中根据SEQ ID NO:2进行编号；并且与SEQID NO:1或2具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％、至少95％但少于100％的序列同一性。
在一个实施例中，本发明涉及具有SEQ ID NO:1的氨基酸序列的多肽的变体，其中该多肽具有α-淀粉酶活性，其中所述变体氨基酸序列包含在对应于位置202和/或186的氨基酸位置处的突变，其中根据SEQ ID NO:2进行编号，其条件是位置186处的突变不是A186G，并且其中所述变体与SEQ ID NO:1或2具有至少60％的序列同一性。
在一个实施例中，本发明涉及氨基酸序列的多肽的变体，其包含在氨基酸位置202处(其中根据SEQ ID NO:2进行编号)的取代，例如L202M。在一个实施例中，多肽是由取代L202M组成的变体氨基酸序列，其中根据SEQ ID NO:2进行编号。
在另一个实施例中，本发明涉及多肽的变体，其不包含在位置202处但包含在至少位置186处(其不是取代A186D)以及潜在地其他位置处的突变，其中根据SEQ ID NO:2进行编号。
在另一个实施例中，本发明涉及氨基酸序列的多肽的变体，其包含在氨基酸位置186处(其中根据SEQ ID NO:2进行编号)的取代，例如A186D。
在一个实施例中，本发明涉及SEQ ID NO:1的多肽的变体，其在对应于位置51、186、202、246和334的一个或多个位置处具有突变(例如取代、缺失、和/或插入)，其中根据SEQ ID NO:2中所示的氨基酸序列进行编号。因此，本发明的变体在对应于位置202和/或186的至少一个位置处包含突变，其中根据SEQ ID NO:2进行编号并且可以进一步包含在位置51、246和334中的一个或多个处的突变，并且其中在变体在位置202处包含突变的情况下，则它还可以在位置186处(以及潜在地还在任何其他位置)包含突变，反之亦然。即，当变体在位置186处包含突变时，则它还可以在位置202处(以及还潜在地在任何其他位置)包含突变。应当理解，当根据本发明的变体在其他位置包含突变时，其不一定在位置202和位置186处包含突变。
在一个特定实施例中，具有α-淀粉酶活性的多肽与SEQ ID NO:1或2的氨基酸序列具有至少60％、至少65％、至少70％、至少75％、至少80％、或至少87％但少于100％至少80％，如至少85％、如至少90％、如至少92％、如至少93％、如至少94％、如至少95％、如至少96％、如至少97％、如至少98％、如至少99％但少于100％的序列同一性。
因此，相对于SEQ ID NO:2的氨基酸序列，该多肽可以包含以下取代中的一个或多个：A51T、A186D、L202M、T246(I/L/V)和S334T。
本领域技术人员将会理解，本发明多肽的不同实例将与SEQ ID NO:1的序列具有不同程度的氨基酸序列同一性。因此，多肽可以包含氨基酸序列或由其组成，该氨基酸序列与SEQ ID NO:1具有至少85％序列同一性，例如与SEQ ID NO:1具有至少90％、至少95％、至少96％、至少97％、至少98％、或至少99％的序列同一性。在一个实施例中，相对于SEQ IDNO:1的氨基酸序列，在该多肽内的变体的数量是在1个和20个之间，例如，在1个和10个突变之间或在1个和5个突变之间，如1个、2个、3个、4个、5个、6个、7个、8个、9个或10个突变。
在具有α-淀粉酶活性的多肽的一个具体的子集中，该多肽由具有选自下组的突变的SEQ ID NO:1的氨基酸序列、或其具有α淀粉酶活性的片段组成，该组由以下组成：A51T+A186D、A51T+L202M、A51T+T246I、A51T+S334T、A186D+L202M、A186D+T246I、A186D+S334T、L202M+T246I、L202M+S334T、T246I+S334T、A51T+A186D+L202M、A51T+A186D+T246I、A51T+A186D+S334T、A51T+L202M+T246I、A51T+L202M+S334T、A51T+T246I+S334T、A186D+L202M+T246I、A186D+L202M+S334T、A186D+T246I+S334T、L202M+T246I+S334T、A51T+A186D+L202M+T246I、A51T+A186D+L202M+S334T、A51T+A186D+T246I+S334T、A51T+L202M+T246I+S334T、A186D+L202M+T246I+S334T、和A51T+A186D+L202M+T246I+S334T。
例如，该多肽可以包含具有选自下组的突变的SEQ ID NO:1的氨基酸序列、或由其组成，该组由以下组成：
(a)L202M；
(b)A186D；
(c)L202M+T246I+S334T；
(d)L202M+T246L+S334T；
(e)A51T+L202M+T246I+S334T；
(f)L202M+T246V；以及
(g)L202M+T246V+S334T；
其中根据SEQ ID NO:2中所示的氨基酸序列进行氨基酸位置的编号。
在一个实施例中，α-淀粉酶是SEQ ID NO:19的母体α-淀粉酶的变体α-淀粉酶，其中该变体选自由以下组成的组：
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+R87S+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+T40G+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+T51K+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72H+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37H+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37M+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37V+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37S+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37Y+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37R+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+F113W+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+F113S+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+F113N+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+R116Q+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+R116V+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+R116K+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q125P+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+S381G+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+M246T+K391A+G476K，
H1*+A37L+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+R116A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+R116H+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q125A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q172R+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+D377S+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+D377A+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+S381A+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+G346P+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+I214H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+I214S+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+A420Q+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+A420S+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+A420K+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+A420L+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+K391Y+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+P473A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+P473G+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+T444D+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+T444Y+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+S280W+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+S280L+K391A+G476K，
H1*+N54S+V56T+K72S+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+T40K+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+G346T+K391A+G476K，
H1*+G50A+N54S+V56T+G109A++Q172N+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K+K484S，
H1*K+G50A+N54S+V56T+G109A+W167F+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+L173V+A174S+G182*+D183*+N195F+A204T+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+W140Y+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+G346T+K391A+G476K+G477A，
H1*+N54S+V56T+G109AG255A++A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+G50A+N54S+V56T+G109A+F113L+R116L+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+W167F+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+W167F+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+R320V+K391A+G476Y，
H1*+N54S+V56T+G109A+R116M+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q172Y+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+I214L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+L217V+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+L217H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+P211D+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+K391Q+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+T444H+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+T444V+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+T444R+G476K，
H1*+G50A+N54S+V56T+G109A+L173V+A174S+G182*+D183*+N195F+A204V+V206L+K391A+G476K，
H1*+G7K+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+R320A+K391A+G476K,
H1*+N54S+V56T+G109A+Q172D+A174S+G182*+D183*+N195F+V206L+G346T+K391A+G476K+G477A，
H1*+N54S+V56T+G109A+Q118S+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+R320A+K391A+G476K，
H1*+N54S+V56T+G109A+T134E+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+Y382M+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+T134E+A174S+G182*+D183*+N195F+V206L+G255A+K391A+G476K，
H1*+N54S+V56T+K72R+M105F+G109A+A174S+G182*+D183*+N195F+V206L+G255A+K391A+G476K，
H1*+N54S+V56T+M105F+G109A+T134E+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+R320A+S323N+D377H+K391A+G476K，
H1*+T51E+N54S+V56T+G109A+A174H+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+L173A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+Q299V+K391A+G476K，
H1*+N54S+V56T+G109A+L173G+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+T355L+K391A+G476K，
H1*+G50A+N54S+V56T+G109A+A174S+G182*+D183*+N195F+A204T+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+A174S+G182*+D183*+N195F+V206L+R320A+S323N+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+M246L+K391A+G476K，
H1*+N54S+V56T+K72Q+G109A+W167H+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+T134E+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+T134E+A174S+G182*+D183*+N195F+V206L+D377H+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+T134E+A174S+G182*+D183*+G184T+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q118R+T134E+A174S+G182*+D183*+N195F+V206L+G255A+K391A+G476K，
H1*+N54S+V56T+G109A+Q118R+A174S+G182*+D183*+N195F+V206L+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+T134E+A174S+G182*+D183*+G184T+N195F+V206L+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+Q172D+A174Q+G182*+D183*+N195F+V206L+K391A+G476K+G477A，
H1*+N54S+V56T+K72R+G109A+A174S+G182*+D183*+G184T+N195F+V206L+G255A+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+G184T+N195F+V206L+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+G255A+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+T134E+A174S+G182*+D183*+G184T+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+V238T+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+V291A+D377H+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+T134E+A174S+G182*+D183*+G184T+N195F+V206L+G255A+K391A+G476K，
H1*+N54S+V56T+G109A+L173H+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72N+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72N+G109A+W167L+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q118R+T134E+A174S+G182*+D183*+N195F+V206L+G255A+D377H+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+A174S+G182*+D183*+N195F+G184T+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+Q118R+A174S+G182*+D183*+N195F+V206L+G255A+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+G182*+D183*+N195F+V206L+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+D377H+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+D377H+K391A，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+D377S+K391A+G476K，
H1*+N54S+V56T+G109A+R116E+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+T134E+A174S+G182*+D183*+N195F+V206L+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+R116S+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+R116I+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+R116G+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+T51A+N54S+V56T+G109A+Q172S+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+T51E+N54S+V56T+G109A+Q172T+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+T51S+N54S+V56T+G109A+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+T51E+N54S+V56T+G109A+Q172L+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+T51E+N54S+V56T+G109A+Q172R+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q172K+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+Q172S+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72H+G109A+Q172T+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72Q+G109A+Q172R+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+Q172T+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72M+G109A+Q172S+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72Q+G109A+Q172M+A174S+G182*+D183*+N195F+V206L+K391A+G476K
H1*+N54S+V56T+K72S+G109A+A174H+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72Q+G109A+A174D+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+A174D+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72Q+G109A+A174K+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174G+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+A174Q+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72Q+G109A+A174G+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72S+G109A+A174N+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+A174H+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+G50A+N54S+V56T+G109A+A174E+G182*+D183*+N195F+A204T+V206L+S323N+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+A204G+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+Y382I+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+I214W+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+A204S+V206L+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+T334S+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+A263G+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+M286F+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+L235V+D377H+K391A+G476K，
H1*+N54S+V56T+M105L+G109A+A174S+G182*+D183*+N195F+V206L+D377H+K391A+G476K，
H1*+N54S+V56T+M105I+G109A+A174S+G182*+D183*+N195F+V206L+D377H+K391A+G476K，
H1*+N54S+V56T+M105V+G109A+A174S+G182*+D183*+N195F+V206L+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+M246I+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+L250I+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+L250V+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391Y+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391V+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391M+G476K，
H1*+N54S+V56T+G109A+T165G+A174S+G182*+D183*+N195F+V206L+D377H+K391A+G476K，
H1*+N54S+V56T+G109S+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174V+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+M246F+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+M246V+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+M246S+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391R+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391H+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391W+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391I+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+L250T+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+L250A+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+L250F+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+L250M+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+Q172H+L173A+A174P+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+G184T+N195F+V206L+G255A+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+S376T+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+S376V+K391A+G476K，
H1*+N54S+V56T+G109A+Q172E+L173I+A174N+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q172D+L173A+A174T+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+Y267M+D377H+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+V238G+K391A+G476K，
H1*+N54S+V56T+M105V+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+V264T+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+V264I+K391A+G476K，
H1*+N54S+V56T+G109A+W167H+Q172N+L173A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q172N+L173T+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q172E+L173M+A174H+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+W167Y+Q172E+L173V+A174H+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+L173A+A174T+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+V291A+F328L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+M246L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+L250V+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+L250F+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+L250M+K391A+G476K，
H1*+N54S+V56T+G109A+T165G+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+T51A+N54S+V56T+G109A+Q172R+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391L+G476K，
H1*+N54S+V56T+G109A+F113Q+R116H+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109S+F113Q+R116Q+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q172D+A174S+G182*+D183*+N195F+A204G+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+K72R+G109A+T134E+A174S+G182*+D183*+N195F+A204G+V206L+G255A+K391A+P473R+G476K，
H1*+N54S+V56T+K72R+G109A+W167F+A174S+G182*+D183*+N195F+A204G+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+A174S+G182*+D183*+G184T+N195F+A204G+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+G109A+W167F+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+G109A+W167F+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+P473G+G476K，
H1*+N54S+V56T+G109A+W167F+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+K72R+G109A+W167F+Q172R+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+R116H+W167F+Q172R+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+M105F+G109A+A174S+G182*+D183*+N195F+V206L+R320A+S323N+K391A+C474V+G476K，
H1*+N54S+V56T+K72R+G109A+A174S+G182*+D183*+N195F+V206L+G255A+K391A+C474V+G476K，
H1*+N54S+V56T+G109A+T134E+A174S+G182*+D183*+N195F+V206L+K391A+C474V+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+D377H+K391A+C474V+G476K，
H1*+N54S+V56T+K72R+G109A+A174S+G182*+D183*+N195F+V206L+K391A+C474V+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+A265G+K391A+T444Q+G476K+G477A，
H1*+A37V+N54S+V56T+G109A+W167F+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+G346T+G477A+G476K，
H1*+N54S+V56T+G109A+R116Q+Q172D+A174S+G182*+D183*+N195F+V206L+G346T+K391A+T444Q+G477A+G476K，
H1*+N54S+V56T+G109A+R116Q+W167F+Q172N+A174S+G182*+D183*+N195F+V206L+G346T+K391A+T444Q+G477A+G476K，
H1*+N54S+V56T+G109S+R116Q+W167F+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109S+W167F+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+F113Q+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+G476K
H1*+N54S+V56T+G109A+F113Q+R116H+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+G476K，
H1*+N54S+V56T+G109A+F113Q+A174S+G182*+D183*+N195F+V206L+K391A+C474V+G476K，
H1*+N54S+V56T+G109A+Q172N+A174S+G182*+D183*+N195F+V206L+V264I+K391A+C474V+G476K，
H1*+N54S+V56T+G109A+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+C474V+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+S280Q+H321Y+K391A+C474V+G476K，
H1*+N54S+V56T+K72R+G109A+T134E+A174S+G182*+D183*+N195F+V206L+G255A+K391A+C474V+G476K，
H1*+N54S+V56T+G109A+F113Q+R116H+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+P473R+G476K，
H1*+N54S+V56T+G109A+F113Q+R116Q+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+G476K，
H1*+N54S+V56T+G109A+R116H+W167F+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+F113Q+R116H+T165G+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+G476K，
H1*+N54S+V56T+G109A+F113Q+R116Q+T165G+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+G476K，
H1*+N54S+V56T+G109A+F113Q+R116H+W167F+Q172R+A174S+G182*+D183*+N195F+V206L+F289I+K391A+G476K，
H1*+N54S+V56T+G109A+F113Q+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+G476K，
H1*+N54S+V56T+G109A+F113Q+R116H+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+F113Q+R116Q+Q172M+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+G476K，
H1*+N54S+V56T+G109A+F113Q+R116H+Q172M+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+R116H+Q172M+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+R116Q+A174S+G182*+D183*+N195F+V206L+A265G+K391A+T444Q+P473R+G476K，
H1*+N54S+V56T+G109A+R116Q+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+P473R+G476K，
H1*+N54S+V56T+K72R+G109A+T134E+T165G+A174S+G182*+D183*+N195F+V206L+G255A+K391A+G476K，
H1*+N54S+V56T+G109A+F113Q+R116H+W167F+Q172R+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+R116Q+W167F+Q172R+A174S+G182*+D183*+N195F+V206L+A265G+K391A+P473R+G476K，
H1*+N54S+V56T+G109A+F113Q+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+T51V+N54S+V56T+G109A+Q172D+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+S280Q+H321Y+K391A+G476K，
H1*+N54S+V56T+G109A+F113Q+R116H+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+F113Q+R116H+Q172G+L173V+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+P473R+G476K，
H1*+N54S+V56T+G109A+R116Q+W167F+Q172N+L173V+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+G476K，
H1*+N54S+V56T+G109A+G133Q+W167F+Q172N+A174S+G182*+D183*+N195F+V206L+Q395P+K391A+G476K，
H1*+N54S+V56T+G109A+F113Q+R116H+W167F+Q172N+L173V+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37V+N54S+V56T+K72R+G109A+R116H+W167F+Q172R+A174S+G182*+D183*+G184T+N195F+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+K72R+G109A+F113Q+R116H+W167F+Q172G+L173V+A174S+G182*+D183*+G184T+N195F+V206L+K391A+T444Q+P473R+G476K，
H1*+N54S+V56T+G109S+R116Q+W167F+Q172R+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109S+R116Q+W167F+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+G109S+F113Q+W167F+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37V+N54S+V56T+G109S+F113Q+W167F+Q172R+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+R116H+T134E+W167F+Q172G+L173V+A174S+G182*+D183*+N195F+V206L+G255A+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+R116H+T134E+W167F+Q172G+L173V+A174S+G182*+D183*+N195F+V206L+G255A+K391A+Q395P+T444Q+P473R+G476K，
H1*+N54S+V56T+K72R+G109A+F113Q+R116Q+W167F+Q172G+A174S+G182*+D183*+G184T+N195F+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+K72R+G109A+F113Q+R116Q+T134E+W167F+Q172R+A174S+G182*+D183*+N195F+V206L+G255A+K391A+T444Q+P473R+G476K，
H1*+N54S+V56T+K72R+G109A+F113Q+R116H+T134E+W167F+Q172R+A174S+G182*+D183*+N195F+V206L+G255A+K391A+Q395P+G476K，
H1*+N54S+V56T+K72R+G109A+T134E+W167F+Q172N+A174S+G182*+D183*+N195F+V206L+G255A+A265G+K391A+T444Q+P473R+G476K，
H1*+N54S+V56T+K72R+G109A+G133Q+W167F+Q172R+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+F113Q+R116Q+W167F+Q172G+A174S+G182*+D183*+N195F+A204G+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+F113Q+R116Q+W167F+Q172N+L173V+A174S+G182*+D183*+G184T+N195F+V206L+K391A+T444Q+P473R+G476K，
H1*+A37H+N54S+V56T+G109A+F113Q+W167F+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+F113Q+W167F+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+T459N+G476K，
H1*+N54S+V56T+G109A+F113Q+R116Q+A174S+G182*+D183*+N195F+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+A60V+G109A+R116Q+W167F+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+A60V+G109A+R116H+Q172R+L173V+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37V+N54S+V56T+A60V+G109A+R116Q+T165G+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37H+N54S+V56T+A60V+G109A+R116Q+T165G+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+A60V+G109A+R116Q+W167F+Q172G+L173V+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+A60V+G109A+R116Q+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37H+N54S+V56T+A60V+G109A+R116Q+W167F+Q172R+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37H+N54S+V56T+A60V+G109A+R116Q+W167F+Q172N+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+A60V+G109A+R116Q+T165G+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+P473R+G476K，
H1*+N54S+V56T+A60V+G109A+R116Q+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+P473R+G476K，
H1*+N54S+V56T+A60V+G109A+R116Q+W167F+Q172N+L173V+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+A60V+G109A+R116Q+W167F+Q172R+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+R116A+W167F+Q172R+A174S+G182*+D183*+N195F+A204G+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+G109A+R116A+A174S+G182*+D183*+N195F+A204G+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+G109A+R116A+T165G+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+P473R+G476K，
H1*+N54S+V56T+G109A+G182*+D183*+N195F+K391A+G476K，
H1*+N54S+V56T+G109A+G182*+D183*+N195F+V206L，
N54S+V56T+G109A+G182*+D183*+N195F，
N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+G476K，
H1*+N54S+V56T+A174S+G182*+D183*+N195F+V206L+G273R+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L，
N54S+V56T+G109A+A174S+G182*+D183*+N195F+K391A+G476K，
H1*+N54S+V56T+G109A+F113Q+Q172M+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+F113Q+T134E+A174S+G182*+D183*+N195F+V206L+G255A+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+F113Q+T134E+A174S+G182*+D183*+N195F+V206L+G255A+K391A+P473R+G476K，
H1*+N54S+V56T+K72R+G109A+R116Q+A174S+G182*+D183*+G184T+N195F+V206L+K391A+G476K，
H1*+A37V+N54S+V56T+G109A+R116A+T165G+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37V+N54S+V56T+G109A+R116Q+T165G+A174S+G182*+D183*+N195F+V206L+M246F+K391A+G476K，
H1*+V56A+G109A+A174S+G182*+D183*+N195F+V206L+G476K，
H1*+N54S+V56T+K72R+G109A+F113Q+R116H+A174S+G182*+D183*+G184T+N195F+V206L+K391A+T444Q+G476K，
H1*+N54S+V56T+G109S+F113Q+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+G476K，
H1*+N54S+V56T+G109A+F113Q+R116H+W167F+Q172G+L173V+A174S+G182*+D183*+N195F+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+G109A+F113Q+R116Q+Q172N+A174S+G182*+D183*+N195F+V206L+A265G+K391A+P473R+G476K，
H1*+N54S+V56T+G109A+F113Q+W167F+Q172N+A174S+G182*+D183*+N195F+A204G+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+G109A+F113Q+R116H+W167F+Q172R+A174S+G182*+D183*+N195F+A204G+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+G109A+R116A+A174S+G182*+D183*+N195F+V206L+K391A+T444Q+P473R+G476K，
H1*+N54S+V56T+G109A+R116H+Q172R+A174S+G182*+D183*+N195F+A204G+V206L+K391A+P473R+G476K，
H1*+A37V+N54S+V56T+G109A+R116Q+T165G+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37H+N54S+V56T+G109A+R116H+T165G+A174S+G182*+D183*+N195F+V206L+M246F+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+K391A+G476K，
H1*+N54S+V56T+G109A+G133S+A174S+G182*+D183*+N195F+K391A+G476H，
H1*+N54S+V56T+G109A+R116A+Q172N+L173V+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+A37V+N54S+V56T+G109A+R116H+T165G+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+P45A+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+C474V+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+P364A+K391A+C474V+G476K，
H1*+N54S+V56T+G109A+Q118R+T134E+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+G184T+N195F+V206L+K391A+G476K，
H1*+G50A+T51A+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+
G476K,
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+V264F+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+Y382L+K391A+G476K，
H1*+G7K+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K,
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+G255A+Q256A+K391A+G476K，
H1*+N54S+V56T+G109A+W167Y+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+W167H+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+T165V+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174N+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174Q+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N16H+V17L+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+Y382F+K391A+G476K，
H1*+Q32S+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+Q385L+K391A+G476K，
H1*+N54S+V56T+R87S+G109A+R171H+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+R116H+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174*+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+K72R+G109A+F113Q+R116Q+W167F+Q172G+A174S+G182*+D183*+G184T+N195F+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+K72R+G109A+F113Q+R116Q+W167F+Q172G+A174S+G182*+D183*+G184T+N195F+V206L+P473R+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172G+A174*+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+F113Q+R116Q+W167F+Q172G+A174S+G182*+D183*+G184T+N195F+K391A+P473R+G476K，
H1*+N54S+V56T+G109A+R116W+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206F+K391A+G476K，
H1*+V56T+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+N260G+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+G184T+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+Y295N+K391A+G476K，
H1*+N54S+V56T+G109A+W167F+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+A186N+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+A288V+K391A+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+G255A+K391A+G476K，
H1*+N54S+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+W48F+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+V291A+K391A+G476K，
H1*+N54S+V56T+G109A+F113Q+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+R320A+S323N+K391A+G476K，
H1*+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+K72R+G109A+F113Q+R116H+W167F+Q172G+A174S+G182*+D183*+G184T+N195F+V206L+K391A+P473R+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+N270T+K391A+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+N260G+K391A+G476K，
H1*+N54S+V56T+G109A+F113Q+R116H+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q172G+A174S+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+A186D+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+A225V+K391A+G476K，
H1*+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+K391A+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+G255S+K391A+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174*+G182*+D183*+N195F+V206L+K269Q+K391A+G476K，
H1*+N54S+V56T+G109A+A174S+G182*+D183*+N195F+V206L+G255S+K391A+G476K，
H1*+N54S+V56T+G109A+Q169E+Q172K+A174S+G182*+D183*+N195F+V206L+K391A+G476K；其中根据SEQ ID NO:19进行编号。
在一个实施例中，本发明涉及具有α-淀粉酶活性的多肽，该多肽包含A和B结构域，该A和B结构域获得自包含SEQ ID NO:3的氨基酸序列的α-淀粉酶，其A和B结构域在此还被披露为SEQ ID NO:4。在本发明的一个实施例中，形成A和B结构域的氨基酸序列与SEQ IDNO:4的氨基酸序列具有至少60％的同一性，如至少65％、如至少70％、如至少75％的同一性，例如至少78％、至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或100％的序列同一性。
在一个实施例中，本发明涉及包含C结构域供体的具有α-淀粉酶活性的多肽，该C结构域供体获得自包含SEQ ID NO:8的氨基酸序列的α-淀粉酶(从其确定C结构域对应于氨基酸398-483(本文还披露为SEQ ID NO:8))。在本发明的一个实施例中，形成C结构域的氨基酸序列与SEQ ID NO:8的氨基酸序列具有至少60％的同一性，如至少65％、如至少70％、如至少75％的同一性，例如至少78％、至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或100％的序列同一性。
在本发明的一个实施例中，具有α-淀粉酶活性的多肽包含A和B结构域以及C结构域，其中形成A和B结构域的氨基酸序列与SEQ ID NO:4的氨基酸序列具有至少60％序列同一性，并且形成C结构域的氨基酸序列与SEQ ID NO:8的氨基酸序列具有至少60％序列同一性。
在本发明的一个实施例中，形成A和B结构域的氨基酸序列与SEQ ID NO:4的氨基酸序列具有至少60％同一性，如至少65％、如至少70％、如75％的同一性，例如至少78％、至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或100％的序列同一性，并且形成C结构域的氨基酸序列与SEQ ID NO:8具有至少60％同一性。
在本发明的一个实施例中，形成A和B结构域的氨基酸序列与SEQ ID NO:4的氨基酸序列具有至少75％同一性，例如至少78％、至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或100％的序列同一性，并且形成C结构域的氨基酸序列与SEQ ID NO:8具有至少80％同一性。
在本发明的一个实施例中，形成A和B结构域的氨基酸序列与SEQ ID NO:4的氨基酸序列具有至少75％同一性，例如至少78％、至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或100％的序列同一性，并且形成C结构域的氨基酸序列与SEQ ID NO:8具有至少85％同一性。
在本发明的一个实施例中，形成A和B结构域的氨基酸序列与SEQ ID NO:4的氨基酸序列具有至少75％同一性，例如至少78％、至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或100％的序列同一性，并且形成C结构域的氨基酸序列与SEQ ID NO:8具有至少90％同一性。
在本发明的一个实施例中，形成A和B结构域的氨基酸序列与SEQ ID NO:4的氨基酸序列具有至少75％同一性，例如至少78％、至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或100％的序列同一性，并且形成C结构域的氨基酸序列与SEQ ID NO:8具有至少95％同一性。
在本发明的另一个实施例中，对应于SEQ ID NO:3中的181和182的氨基酸是缺失的。在本发明的又另一个实施例中，对应于SEQ ID NO:3中的183和184的氨基酸是缺失的。在另外的实施例中，对应于SEQ ID NO:3中的182与183或181与183或182与184是缺失的。因而，本发明还涉及一种融合多肽，该融合多肽包含SEQ ID NO:3的A和B结构域以及来自SEQID NO:8的α-淀粉酶的C结构域，并且另外具有对应于SEQ ID NO:1中的183和184的氨基酸的缺失。本发明另外涉及一种融合多肽，该融合多肽在此披露为SEQ ID NO:9。
在另外的优选实施例中，本发明的多肽可以进一步包括在对应于SEQ ID NO:8氨基酸序列的位置6、22、23和29的一个或多个位置处在C结构域中的氨基酸取代。
在某些实施例中，本发明的多肽包含在对应于SEQ ID NO:8氨基酸序列的位置6、22、23和29中任何两个位置处在C结构域中的氨基酸取代。在另一个实施例中，本发明的多肽包含在对应于SEQ ID NO:8的氨基酸序列的位置6、22、23和29中任何三个位置处的氨基酸取代。在优选实施例中，本发明的多肽包含在对应于SEQ ID NO:8的氨基酸序列的位置6、22、23和29的每个位置处的氨基酸取代。在位置6处的氨基酸取代可以是I6A、I6C、I6D、I6E、I6F、I6G、I6H、I6K、I6L、I6M、I6N、I6P、I6Q、I6R、I6S、I6T、I6V、I6W或I6Y中任一项。优选地，是I6L。在位置22处的氨基酸取代可以是A22C、A22D、A22E、A22F、A22G、A22H、A22I、A22K、A22L、A22M、A22N、A22P、A22Q、A22R、A22S、A22T、A22V、A22W或A22Y中任一项。优选地，是A22H。在位置23处的氨基酸取代可以是A23C、A23D、A23E、A23F、A23G、A23H、A23I、A23K、A23L、A23M、A23N、A23P、A23Q、A23R、A23S、A23T、A23V、A23W或A23Y中任一项。优选地，是A23P。在位置29处的氨基酸取代可以是A29C、A29D、A29E、A29F、A29G、A29H、A29I、A29K、A29L、A29M、A29N、A29P、A29Q、A29R、A29S、A29T、A29V、A29W或A29Y中任一项。优选地，是A29T。
在本发明的另一个实施例中，多肽包含A和B结构域，该A和B结构域与具有SEQ IDNO:4的氨基酸序列的A和B结构域具有至少75％序列同一性；并且还包括C结构域，该C结构域与具有SEQ ID NO:8的氨基酸序列的C结构域具有至少70％、至少75％、至少80％、至少85％、至少90％、至少91％、至少92％、至少93％、至少94％、至少95％、至少96％、至少97％、至少98％、至少99％或100％序列同一性。在优选实施例中，组成C结构域的氨基酸序列包括对应于SEQ ID NO:8的氨基酸序列的I6L、A22H、A23P和A29T的取代中的每个。
在优选实施例中，本发明的多肽包含A和B结构域以及C结构域，其中形成A和B结构域的氨基酸序列与SEQ ID NO:4的氨基酸序列具有至少75％的同一性并且包括对应于SEQID NO:4的位置183和184的氨基酸的缺失，并且形成C结构域的氨基酸序列与SEQ ID NO:8的氨基酸序列具有至少75％的同一性。在一个实施例中，多肽包含对应于SEQ ID NO:8的氨基酸序列的I6L、A22H、A23P和A29T的取代。
在另一个优选实施例中，本发明的多肽包含A和B结构域以及C结构域，其中形成A和B结构域的氨基酸序列与SEQ ID NO:4的氨基酸序列具有至少75％的同一性并且包括对应于SEQ ID NO:4的位置181和182的氨基酸的缺失，并且形成C结构域的氨基酸序列与SEQID NO:8的氨基酸序列具有至少75％的同一性。
在另一个实施例中，本发明的A和B结构域的氨基酸序列包括SEQ ID NO:4的序列，并且C结构域的氨基酸序列包括SEQ ID NO:8的序列。在又另一个实施例中，本发明的A和B结构域的氨基酸序列由SEQ ID NO:4的序列组成，并且C结构域的氨基酸序列由SEQ ID NO:8的序列组成。
还如以上在所有以上提及的实施例中的优选实施例中所提及的，对应于SEQ IDNO:4的181+182或181+183或182+184或182+183或183+184的氨基酸是缺失的。优选的，缺失的是氨基酸181+182或183+184。最优选的是对应于SEQ ID NO:4的183+184的氨基酸缺失。
另外，还如以上所提及的，对应于SEQ ID NO:8的氨基酸6、22、23和29的氨基酸中的一个或多个可以被取代。在以上提及的实施例的一个实施例中，对应于SEQ ID NO:8的位置6中的氨基酸的氨基酸被亮氨酸所取代。对应于SEQ ID NO:8的位置22中的氨基酸的氨基酸可以被组氨酸所取代。对应于SEQ ID NO:8的位置23中的氨基酸的氨基酸可以被脯氨酸所取代。对应于SEQ ID NO:8的位置29中的氨基酸的氨基酸可以被苏氨酸所取代。
α-淀粉酶由于它们在低温下的良好性能最初产生和使用，但诸位发明人令人惊讶地发现这些α-淀粉酶变体在低pH粉末洗涤剂组合物中也具有良好性能。
本发明不限于用于确定α-淀粉酶活性的特定方法，但可以使用用于确定α-淀粉酶活性的任何公认的方法。用于确定α-淀粉酶活性的一种优选方法是基于PNP-G7底物的切割。PNP-G7是4,6-亚乙基(G7)-对硝基苯基(G1)-α,D-麦芽庚糖苷的缩写，为一种在α-葡糖苷酶存在下的嵌段寡糖。切割之后，α-葡糖苷酶进一步消化水解底物以释放游离PNP分子，该分子具有黄色并且因而可通过可见分光光度法在λ＝405nm(400-420nm.)测量。含有PNP-G7底物和α-葡糖苷酶的试剂盒由罗氏/日立公司(Roche/Hitachi)制造(目录号11876473)。
除了本文具体披露的氨基酸改变之外，本发明组合物中的α-淀粉酶变体可包括在一个或多个其他位置处的另外的改变。这些另外的改变可以具有微小性质，即，不会显著地影响蛋白质的折叠和/或活性的保守氨基酸取代或插入；典型地为1-30个氨基酸的小缺失；小的氨基末端或羧基末端延伸，如氨基末端的甲硫氨酸残基；多达20-25个残基的小接头肽；或小的延伸，其通过改变净电荷或另一功能(例如聚组氨酸段、抗原表位或结合结构域)来促进纯化。
保守取代的实例是在下组之内：碱性氨基酸(精氨酸、赖氨酸和组氨酸)、酸性氨基酸(谷氨酸和天冬氨酸)、极性氨基酸(谷氨酰胺和天冬酰胺)、疏水性氨基酸(亮氨酸、异亮氨酸和缬氨酸)、芳香族氨基酸(苯丙氨酸、色氨酸和酪氨酸)以及小氨基酸(甘氨酸、丙氨酸、丝氨酸、苏氨酸和甲硫氨酸)。通常不会改变比活性的氨基酸取代是本领域已知的并且例如由H.Neurath和R.L.Hill,1979,于The Proteins[蛋白质],Academic Press[学术出版社],纽约中描述。常见的保守取代基团包括但不限于：G＝A＝S；I＝V＝L＝M；D＝E；Y＝F；以及N＝Q(其中例如“G＝A＝S”表示这三个氨基酸可以彼此取代)。
可替代地，这些氨基酸改变具有这样的性质：改变多肽的物理化学特性。例如，氨基酸改变可以提高多肽的热稳定性、改变底物特异性、改变最适pH，等等。
可以根据本领域已知的程序，如定点诱变或丙氨酸扫描诱变(Cunningham和Wells,1989,Science[科学]244:1081-1085)来鉴定多肽中的必需氨基酸。在后一种技术中，在分子中的每个残基处引入单丙氨酸突变，并测试所得的突变分子的α-淀粉酶活性以鉴定对分子活性关键的氨基酸残基。还参见，Hilton等人,1996,J.Biol.Chem.[生物化学杂志]271:4699-4708。酶或其他生物学相互作用的活性位点还可以通过对结构的物理分析来确定，如通过诸如下述技术来确定：核磁共振、晶体学、电子衍射或光亲和标记，连同对推定的接触位点氨基酸进行突变。参见例如，de Vos等人,1992,Science[科学]255:306-312；Smith等人,1992,J.Mol.Biol.[分子生物学杂志]224:899-904；Wlodaver等人,1992,FEBSLett.[欧洲生化学会联合会快报]309:59-64。还可以从与相关多肽的比对来推断必需氨基酸的身份。
本发明还涉及粉末洗涤剂组合物，其进一步包含至少一种或多种洗涤剂组分。
在一个实施例中，组合物另外包含至少一种洗涤剂组分。
洗涤剂组合物的洗涤剂组分的选择在普通技术人员技术内并且包括常规成分，包括以下列出的示例性、非限制性组分。对于织物护理，组分的选择可以包括以下考虑：待清洁的织物的类型、污垢的类型和/或程度、进行清洁时的温度、以及洗涤剂产品的配制。
在具体实施例中，洗涤剂组合物包含具有α-淀粉酶的多肽和一种或多种非天然存在的洗涤剂组分，如表面活性剂、水溶助剂、助洗剂、共助洗剂、螯合剂(chelator)或螯合试剂(chelating agent)、漂白系统或漂白组分、聚合物、织物调色剂、织物调理剂、增泡剂、抑泡剂、分散剂、染料转移抑制剂、荧光增白剂、香料、光学增亮剂、杀细菌剂、杀真菌剂、污垢悬浮剂、污垢释放聚合物、抗再沉积剂、酶抑制剂或稳定剂、酶活化剂、抗氧化剂以及增溶剂。洗涤剂组合物通常包括至少表面活性剂和助洗剂。
在具体实施例中，洗涤剂组合物包含具有α-淀粉酶的多肽和一种或多种天然存在的洗涤剂组分，如表面活性剂、水溶助剂、助洗剂、共助洗剂、螯合剂或螯合试剂、漂白系统或漂白组分、聚合物、织物调色剂、织物调理剂、增泡剂、抑泡剂、分散剂、染料转移抑制剂、荧光增白剂、香料、光学增亮剂、杀细菌剂、杀真菌剂、污垢悬浮剂、污垢释放聚合物、抗再沉积剂、酶抑制剂或稳定剂、酶活化剂、抗氧化剂以及增溶剂。洗涤剂组合物通常包括至少表面活性剂和助洗剂。
在一个实施例中，组合物还包含选自下组的一种或多种另外的酶，该组由以下组成：蛋白酶、脂肪酶、角质酶、α-淀粉酶、糖酶、纤维素酶、果胶酶、甘露聚糖酶、β-淀粉酶、支链淀粉酶、过水解酶、磷脂酶阿拉伯糖酶、半乳聚糖酶、木聚糖酶、果胶裂解酶、半乳糖醛酸酶、半纤维素酶、木葡聚糖酶、核酸酶、卵磷脂酶、氧化酶及其混合物。
洗涤剂组合物可以是例如呈常规的或压制的粉末、颗粒、均匀的片剂、或具有两层或更多层的片剂的形式。
本发明还涉及本发明的组合物在清洁过程，如衣物洗涤或硬表面清洁如餐具洗涤中的用途。
在一个实施例中，本发明涉及ADW(自动餐具洗涤)组合物，该组合物包含与一种或多种另外的ADW组合物组分组合的所披露的α-淀粉酶变体。另外的组分的选择处于技术人员的能力范围内并且包括常规的成分，包括下文所述的示例性非限制性组分。
在一个实施例中，本发明涉及衣物洗涤剂组合物，其包含与一种或多种另外的衣物洗涤剂组合物组分组合的所披露的α-淀粉酶变体。另外的组分的选择处于技术人员的能力范围内并且包括常规的成分，包括下文所述的示例性非限制性组分。
在一个实施例中，可以将具有α-淀粉酶活性的多肽以对应于以下的量添加至洗涤剂组合物中：0.001-200mg酶蛋白/升洗涤液，优选0.005-50mg酶蛋白/升洗涤液，特别是0.001-10mg酶蛋白/升洗涤液。
用于衣物洗涤的颗粒状组合物可以例如包括按该组合物的重量计0.001％-20％、如0.01％-10％、如0.05％-5％的酶蛋白。
自动餐具洗涤(ADW)组合物例如可以包括按该组合物的重量计0.001％-30％，例如0.01％-20％、如0.1％-15％、如0.5％-10％的酶蛋白。
该洗涤剂组合物可以配制在用于衣物洗涤的颗粒洗涤剂中。例如，这类洗涤剂可包含：
a)每克组合物至少0.001mgα-淀粉酶
b)阴离子表面活性剂，优选地5wt％至50wt％
c)非离子表面活性剂，优选地1wt％至8wt％
d)助洗剂，优选地5wt％至40wt％，如碳酸盐、沸石、磷酸盐助洗剂、钙掩蔽助洗剂或络合剂。
尽管根据具体的功能性对以下提及的组分由通用标题进行分类，但是这并不被解释为限制，因为如将被本领域技术人员所理解，组分可以包含另外的功能性。
表面活性剂
洗涤剂组合物可以包含一种或多种表面活性剂，它们可以是阴离子型和/或阳离子型和/或非离子型和/或半极性型和/或兼性离子型、或其混合物。在特定的实施例中，洗涤剂组合物包括一种或多种非离子表面活性剂和一种或多种阴离子表面活性剂的混合物。该一种或多种表面活性剂典型地以按重量计从约0.1％至60％(如约1％至约40％、或约3％至约20％、或约3％至约10％)的水平存在。基于所希望的清洁应用来选择所述一种或多种表面活性剂，并且所述表面活性剂包括本领域中已知的任何一种或多种常规表面活性剂。可以利用本领域中已知的用于在洗涤剂中使用的任何表面活性剂。表面活性剂降低洗涤剂中的表面张力，这使得清洁的污渍被提起并分散，并且然后洗去。
当包括于其中时，洗涤剂通常将含有按重量计从约1％至约40％，如从约5％至约30％，包含从约5％至约15％或从约20％至约25％的阴离子表面活性剂。阴离子表面活性剂的非限制性实例包括硫酸盐和磺酸盐，特别是直链烷基苯磺酸盐(LAS)、LAS的异构体、支链烷基苯磺酸盐(BABS)、苯基链烷磺酸盐、α-烯烃磺酸盐(AOS)、烯烃磺酸盐、链烯烃磺酸盐、链烷-2,3-二基双(硫酸盐)、羟基链烷磺酸盐以及二磺酸盐、烷基硫酸盐(AS)(例如十二烷基硫酸钠(SDS))、脂肪醇硫酸盐(FAS)、伯醇硫酸盐(PAS)、醇醚硫酸盐(AES或AEOS或FES，也被称为醇乙氧基硫酸盐或脂肪醇醚硫酸盐)、仲链烷磺酸盐(SAS)、石蜡烃磺酸盐(PS)、酯磺酸盐、磺化的脂肪酸甘油酯、α-磺酸基脂肪酸甲酯(α-SFMe或SES)(包括磺酸甲酯(MES))、烷基琥珀酸或烯基琥珀酸、十二烯基/十四烯基琥珀酸(DTSA)、氨基酸的脂肪酸衍生物、磺酸基琥珀酸或皂的二酯和单酯、及其组合。
当包括于其中时，洗涤剂通常将含有按重量计从约0％至约10％的阳离子表面活性剂。阳离子型表面活性剂的非限制性实例包括烷基二甲基乙醇季胺(ADMEAQ)、十六烷基三甲基溴化铵(CTAB)、二甲基二硬脂酰氯化铵(DSDMAC)、以及烷基苄基二甲基铵、烷基季铵化合物、烷氧基化季铵(AQA)化合物、及其组合。
当包括于其中时，洗涤剂通常将含有按重量计从约0.2％至约40％的非离子表面活性剂，例如从约0.5％至约30％、尤其是从约1％至约20％、从约3％至约10％、如从约3％至约5％或从约8％至约12％。非离子型表面活性剂的非限制性实例包括醇乙氧基化物(AE或AEO)、醇丙氧基化物、丙氧基化的脂肪醇(PFA)、烷氧基化的脂肪酸烷基酯(例如乙氧基化的和/或丙氧基化的脂肪酸烷基酯)、烷基酚乙氧基化物(APE)、壬基酚乙氧基化物(NPE)、烷基多糖苷(APG)、烷氧基化胺、脂肪酸单乙醇酰胺(FAM)、脂肪酸二乙醇酰胺(FADA)、乙氧基化的脂肪酸单乙醇酰胺(EFAM)、丙氧基化的脂肪酸单乙醇酰胺(PFAM)、多羟基烷基脂肪酸酰胺、或葡糖胺的N-酰基N-烷基衍生物(葡糖酰胺(GA)、或脂肪酸葡糖酰胺(FAGA))、连同在SPAN和TWEEN商品名下可获得的产品、及其组合。
当包括在其中时，洗涤剂通常将含有按重量计从约0％至约10％的半极性表面活性剂。半极性型表面活性剂的非限制性实例包括氧化胺(AO)(例如烷基二甲基氧化胺)、N-(椰油基烷基)-N,N-二甲基氧化胺和N-(牛油-烷基)-N,N-双(2-羟乙基)氧化胺、脂肪酸链烷醇酰胺和乙氧基化的脂肪酸链烷醇酰胺及其组合。
当包括在其中时，洗涤剂通常将含有按重量计从约0％至约10％的两性离子表面活性剂。兼性离子型表面活性剂的非限制性实例包括甜菜碱、烷基二甲基甜菜碱、磺基甜菜碱、及其组合。
助洗剂和共助洗剂
洗涤剂组合物可以含有按重量计约0-65％(如约5％至约45％)的洗涤剂助洗剂或共助洗剂、或其混合物。在餐具洗涤洗涤剂中，助洗剂的水平典型地是40％-65％，特别是50％-65％。助洗剂和螯合剂例如通过从液体中除去金属离子来软化洗涤水。助洗剂和/或共助洗剂可以特别是与Ca和Mg形成水溶性络合物的螯合试剂。可以利用本领域中已知的用于在衣物洗涤剂中使用的任何助洗剂和/或共助洗剂。助洗剂的非限制性实例包括沸石、二磷酸盐(焦磷酸盐)、三磷酸盐例如三磷酸钠(STP或STPP)、碳酸盐例如碳酸钠、可溶性硅酸盐例如硅酸钠、层状硅酸盐(例如来自赫斯特公司(Hoechst)的SKS-6)、乙醇胺例如2-氨基乙-1-醇(MEA)、二乙醇胺(DEA，也称为亚氨基二乙醇)、三乙醇胺(TEA，也称为2,2’,2”-次氨基三乙醇)、以及羧甲基菊粉(CMI)及其组合。
在优选的实施例中，洗涤剂组合物不含磷酸盐。
洗涤剂组合物还可以含有按重量计0％-20％，例如约5％至约10％的洗涤剂共助洗剂或其混合物。洗涤剂组合物可以包括单独的共助洗剂，或与助洗剂(例如沸石助洗剂)组合。共助洗剂的非限制性实例包括聚丙烯酸酯的均聚物或其共聚物，如聚(丙烯酸)(PAA)或共聚(丙烯酸/马来酸)(PAA/PMA)。另外的非限制性实例包括柠檬酸盐、螯合剂(如氨基羧酸盐、氨基多羧酸盐和膦酸盐)、以及烷基琥珀酸或烯基琥珀酸。另外的具体实例包括2,2’,2”-次氨基三乙酸(NTA)、乙二胺四乙酸(EDTA)、二亚乙基三胺五乙酸(DTPA)、亚氨基二丁二酸(iminodisuccinic acid)(IDS)、乙二胺-N,N’-二丁二酸(EDDS)、甲基甘氨酸二乙酸(MGDA)、谷氨酸-N,N-二乙酸(GLDA)、1-羟基乙烷-1,1-二膦酸(HEDP)、乙二胺四-(亚甲基膦酸)(EDTMPA)、二亚乙基三胺五(亚甲基膦酸)(DTPMPA或DTMPA)、N-(2-羟乙基)亚氨基二乙酸(EDG)、天冬氨酸-N-单乙酸(ASMA)、天冬氨酸-N,N-二乙酸(ASDA)、天冬氨酸-N-单丙酸(ASMP)、亚氨基二丁二酸(iminodisuccinic acid)(IDA)、N-(2-磺甲基)-天冬氨酸(SMAS)、N-(2-磺乙基)-天冬氨酸(SEAS)、N-(2-磺甲基)-谷氨酸(SMGL)、N-(2-磺乙基)-谷氨酸(SEGL)、N-甲基亚氨基二乙酸(MIDA)、α-丙氨酸-N,N-二乙酸(α-ALDA)、丝氨酸-N,N-二乙酸(SEDA)、异丝氨酸-N,N-二乙酸(ISDA)、苯丙氨酸-N,N-二乙酸(PHDA)、邻氨基苯甲酸-N,N-二乙酸(ANDA)、磺胺酸-N,N-二乙酸(SLDA)、牛磺酸-N,N-二乙酸(TUDA)以及磺甲基-N,N-二乙酸(SMDA)、N-(2-羟乙基)-亚乙基二胺-N,N’,N’-三乙酸盐(HEDTA)、二乙醇甘氨酸(DEG)、二亚乙基三胺五(亚甲基膦酸)(DTPMP)、氨基三(亚甲基膦酸)(ATMP)及其组合和盐。另外的示例性助洗剂和/或共助洗剂描述于例如WO 2009/102854和US 5,977,053中。
本发明的α-淀粉酶变体也可以配制在餐具洗涤组合物中，优选是自动餐具洗涤组合物(ADW)，该组合物包含：
a)至少0.001mg的根据本发明的活性α-淀粉酶，和
b)10wt％-50wt％的助洗剂，优选地选自柠檬酸、甲基甘氨酸-N,N-二乙酸(MGDA)和/或谷氨酸-N,N-二乙酸(GLDA)及其混合物，和
c)至少一种漂白组分。
漂白系统
洗涤剂可以含有按重量计0-50％，如约0.1％至约25％的漂白系统。漂白系统去除经常因氧化所致的褪色，并且许多漂白剂还具有强的杀菌特性，并且用于消毒和灭菌。可以利用本领域中已知的用于在衣物洗涤剂中使用的任何漂白系统。合适的漂白系统组分包括漂白催化剂、光漂白剂、漂白活化剂、过氧化氢源，例如过碳酸钠和过硼酸钠，预成型过酸及其混合物。合适的预成型过酸包括但不限于：过氧羧酸及盐、过碳酸及盐、过亚氨酸(perimidic acids)及盐、过氧单硫酸及盐(例如过硫酸氢钾(Oxone(R))、及其混合物。漂白系统的非限制性实例包括与过酸形成漂白活化剂组合的基于过氧化物的漂白系统，其可以包含例如无机盐，包括碱金属盐例如过硼酸盐的钠盐(通常为单或四水合物)、过碳酸盐、过硫酸盐、过磷酸盐、过硅酸盐。
术语漂白活化剂在本文意指与过氧化物漂白剂(像过氧化氢)反应以形成过酸的化合物。以此方式形成的过酸构成活化的漂白剂。在此将使用的适合漂白活化剂包括属于酯酰胺、酰亚胺或酸酐类别的那些。合适的实例是四乙酰基乙二胺(TAED)、4-[(3,5,5-三甲基己酰)氧基]苯磺酸钠(ISONOBS)、二过氧月桂酸、4-(十二酰基氧基)苯磺酸盐(LOBS)、4-(癸酰基氧基)苯磺酸盐、4-(癸酰基氧基)苯甲酸盐(DOBS)、4-(壬酰基氧基)-苯磺酸盐(NOBS)和/或披露于WO 98/17767中的那些。目的漂白活化剂的具体家族披露于EP624154中并且该家族中特别优选的是乙酰柠檬酸三乙酯(ATC)。ATC或短链甘油三酯如三乙酸甘油酯具有环境友好的优点，因为它最终降解成柠檬酸和醇。此外，乙酰柠檬酸三乙酯和三醋精在存储时在产品中具有良好的水解稳定性，并且是有效的漂白活化剂。最后，ATC为衣物洗涤添加剂提供良好的助洗能力。可替代地，漂白系统可以包含例如酰胺、酰亚胺或砜类型的过氧酸。漂白系统还可以包含过酸，例如6-(苯二甲酰亚氨基)过己酸(PAP)。漂白系统还可以包括漂白催化剂或促进剂。
可以用于本发明组合物中的漂白催化剂的一些非限制性实例包括草酸锰、乙酸锰、锰胶原、钴-胺催化剂和锰三氮杂环壬烷(MnTACN)催化剂；特别优选的是锰与1,4,7-三甲基-1,4,7-三氮杂环壬烷(Me3-TACN)或1,2,4,7-四甲基-1,4,7-三氮杂环壬烷(Me4-TACN)的络合物，特别是Me3-TACN，如双核锰络合物[(Me3-TACN)Mn(O)3Mn(Me3-TACN)](PF6)2、和[2,2',2”-次氮基三(乙烷-1,2-二基氮烷基亚基-κN-甲基亚基)三酚并-κ3O]锰(III)。这些漂白催化剂还可以是其他金属化合物，如铁或钴络合物。
在一些实施例中，漂白组分可以是选自下组的有机催化剂，该组由具有下式的有机催化剂组成：
(iii)及其混合物；其中每个R1独立地是包含从9至24个碳的支链烷基基团或包含从11至24个碳的直链烷基基团，优选地，每个R1独立地是包含从9至18个碳的支链烷基基团或包含从11至18个碳的直链烷基基团，更优选地，每个R1独立地选自下组，所述组由以下组成：2-丙基庚基、2-丁基辛基、2-戊基壬基、2-己基癸基、正-十二烷基、正-十四烷基、正-十六烷基、正-十八烷基、异-壬基、异-癸基、异-十三基和异-十五烷基。其他示例性漂白系统描述于例如WO 2007/087258、WO 2007/087244、WO 2007/087259以及WO 2007/087242中。合适的光漂白剂可以例如是磺化的酞菁锌。
优选地，除了漂白催化剂，特别是有机漂白催化剂以外，漂白组分还包含过酸源。过酸源可以选自(a)预形成的过酸；(b)过碳酸盐、过硼酸盐或过硫酸盐(过氧化氢源)，优选与漂白活化剂组合；和(c)过水解酶以及酯，用于在纺织品或硬表面处理步骤中在水的存在下原位形成过酸。
水溶助剂
水溶助剂是溶解水溶液中的疏水化合物(或相反地，非极性环境中的极性物质)的化合物。通常，水溶助剂具有亲水和疏水两种特征(所谓的两亲特性，如由表面活性剂已知的)；然而水溶助剂的分子结构一般并不有利于自发自聚集，参见例如Hodgdon和Kaler的综述,2007,Current Opinion in Colloid&Interface Science[胶体&界面科学新见]12:121-128。水溶助剂并不显示临界浓度，高于该浓度就会发生如对表面活性剂而言所发现的自聚集以及脂质形成胶束、薄层或其他明确定义的中间相。相反，许多水溶助剂显示连续类型的聚集过程，在该过程中聚集物的大小随着浓度增加而增长。然而，许多水溶助剂改变了含有极性和非极性特征的物质的系统(包括水、油、表面活性剂、和聚合物的混合物)的相行为、稳定性、和胶体特性。通常地在从药学、个人护理、食品到技术应用的各个产业中使用水溶助剂。水溶助剂在洗涤剂组合物中的使用允许例如更浓的表面活性剂配制品(如在通过去除水而压制液体洗涤剂的过程中)而不引起不希望的现象，例如相分离或高黏度。
所述洗涤剂可以含有按重量计0％-5％，例如约0.5％至约5％，或约3％至约5％的水溶助剂。可以利用本领域中已知的用于在洗涤剂中使用的任何水溶助剂。水溶助剂的非限制性实例包括苯磺酸钠、对甲苯磺酸钠(STS)、二甲苯磺酸钠(SXS)、枯烯磺酸钠(SCS)、伞花烃磺酸钠、氧化胺、醇和聚乙二醇醚、羟基萘甲酸钠、羟基萘磺酸钠、乙基己基硫酸钠及其组合。
聚合物
洗涤剂可以含有按重量计0-10％(如0.5％-5％、2％-5％、0.5％-2％或0.2％-1％)的聚合物。可以利用本领域中已知的用于在洗涤剂中使用的任何聚合物。该聚合物可以作为如上文提到的共助洗剂起作用，或可以提供抗再沉积、纤维保护、污垢释放、染料转移抑制、油脂清洁和/或防沫特性。一些聚合物可以具有多于一种的上文提到的特性和/或多于一种的下文提到的基序。示例性聚合物包括(羧甲基)纤维素(CMC)、聚(乙烯醇)(PVA)、聚(乙烯吡咯烷酮)(PVP)、聚(乙二醇)或聚(环氧乙烷)(PEG)、乙氧基化的聚(亚乙基亚胺)、羧甲基菊粉(CMI)、和聚羧化物，例如PAA、PAA/PMA、聚-天冬氨酸、和甲基丙烯酸月桂酯/丙烯酸共聚物、疏水修饰CMC(HM-CMC)和硅酮、对苯二甲酸和低聚乙二醇的共聚物、聚(对苯二甲酸乙二酯)和聚(氧乙烯对苯二甲酸乙二酯)的共聚物(PET-POET)、PVP、聚(乙烯基咪唑)(PVI)、聚(乙烯吡啶-N-氧化物)(PVPO或PVPNO)以及聚乙烯吡咯烷酮-乙烯基咪唑(PVPVI)。另外的示例性聚合物包括磺化的聚羧酸酯、聚环氧乙烷和聚环氧丙烷(PEO-PPO)以及乙氧基硫酸二季铵盐。其他示例性聚合物披露于例如WO 2006/130575中。还考虑了以上提到的聚合物的盐。
织物调色剂
本发明的洗涤剂组合物还可以包括织物调色剂，例如染料或色素，当配制在洗涤剂组合物中时，当该织物与一种洗涤液接触时织物调色剂可以沉积在织物上，该洗涤液包括该洗涤剂组合物，并且因此通过可见光的吸收/反射改变该织物的色彩。荧光增白剂发射至少一些可见光。相比之下，当织物调色剂吸收至少部分可见光谱时，它们改变表面的色彩。适合的织物调色剂包括染料和染料-粘土轭合物，并且还可以包括颜料。合适的染料包括小分子染料和聚合物染料。合适的小分子染料包括选自由落入颜色索引(Colour Index)(C.I.)分类的以下染料组成的组的小分子染料：直接蓝、直接红、直接紫、酸性蓝、酸性红、酸性紫、碱性蓝、碱性紫和碱性红、或其混合物，例如如WO 2005/003274、WO 2005/003275、WO 2005/003276以及EP 1876226(通过引用并入本文)中所描述的。该洗涤剂组合物优选包括从约0.00003wt％至约0.2wt％、从约0.00008wt％至约0.05wt％、或甚至从约0.0001wt％至约0.04wt％的织物调色剂。该组合物可以包括从0.0001wt％至0.2wt％的织物调色剂，当该组合物处于单位剂量袋的形式时，这可以是尤其优选的。合适的调色剂还披露于例如WO2007/087257和WO 2007/087243中。
另外的酶
洗涤剂组分或洗涤剂组合物可包含一种或多种酶，如蛋白酶、脂肪酶、角质酶、α-淀粉酶、糖酶、纤维素酶、果胶酶、甘露聚糖酶、β-淀粉酶、支链淀粉酶、过水解酶、磷脂酶阿拉伯糖酶、半乳聚糖酶、木聚糖酶、果胶裂解酶、半乳糖醛酸酶、半纤维素酶、木葡聚糖酶、核酸酶、卵磷脂酶、氧化酶及其混合物。
选择的一种或多种酶的特性应当与所选择的洗涤剂相容(即，最适pH，与其他酶成分或非酶成分的相容性等)。
纤维素酶
合适的纤维素酶包括细菌或真菌来源的酶的单组分和混合物。也可以考虑化学修饰的或蛋白质工程改造的突变体。纤维素酶可以例如是单组分或单组分内切-1,4-β-葡聚糖酶(也称为内切葡聚糖酶)的混合物。
合适的纤维素酶包括来自芽孢杆菌属、假单胞菌属、腐质霉属、毁丝霉属、镰孢属、梭孢壳属、木霉属和枝顶孢属的那些。示例性纤维素酶包括真菌纤维素酶，该真菌纤维素酶来自特异腐质霉(US 4,435,307)或来自木霉属，例如里氏木霉或绿色木霉。其他合适的纤维素酶来自梭孢壳属，例如在WO 96/29397中描述的土生梭孢壳霉(Thielaviaterrestris)或在US 5,648,263、US 5,691,178、US 5,776,757、WO 89/09259和WO 91/17244中披露的由嗜热毁丝霉和尖孢镰孢产生的真菌纤维素酶。如WO 02/099091和JP2000210081中所述，来自芽孢杆菌属的纤维素酶也是相关的。合适的纤维素酶是具有护理益处的碱性或中性纤维素酶。纤维素酶的实例在EP 0 495 257、EP 0 531 372、WO 96/11262、WO 96/29397、WO 98/08940中描述。其他实例是例如描述于WO 94/07998、EP 0 531315、US 5,457,046、US 5,686,593、US 5,763,254、WO 95/24471、WO 98/12307中的那些纤维素酶变体。
其他纤维素酶是具有以下序列的内切-β-1,4-葡聚糖酶，所述序列与WO 2002/099091的SEQ ID NO:2的位置1至位置773的氨基酸序列具有至少97％同一性，或具有以下序列的家族44木葡聚糖酶，所述序列与WO 2001/062903的SEQ ID NO:2的位置40-559具有至少60％同一性。
可商购的纤维素酶包括
Classic、
(诺维信公司(Novozymes A/S))、
PuradaxHA和Puradax EG(可从杰尼卡国际公司(Genencor International Inc.)获得)和KAC-500(B)TM(花王公司(Kao Corporation))。
甘露聚糖酶
合适的甘露聚糖酶包括细菌或真菌来源的那些。包括化学或遗传修饰的突变体。甘露聚糖酶可以是家族5或26的碱性甘露聚糖酶。它可以是来自芽孢杆菌属或腐质霉属的野生型，特别是粘琼脂芽孢杆菌、地衣芽孢杆菌、嗜碱芽孢杆菌(B.halodurans)、克劳氏芽孢杆菌或特异腐质霉。合适的甘露聚糖酶描述于WO 1999/064619中。可商购的甘露聚糖酶是Mannaway(诺维信公司)。
过氧化物酶/氧化酶
合适的过氧化物酶/氧化酶包括植物、细菌或真菌来源的那些。包括化学修饰的突变体或蛋白质工程化的突变体。有用的过氧化物酶的实例包括来自鬼伞属，例如来自灰盖鬼伞的过氧化物酶，及其变体，如在WO 93/24618、WO 95/10602、以及WO 98/15257中描述的那些。可商购的过氧化物酶包括GuardzymeTM(诺维信公司)。
合适的过氧化物酶优选包括包含由国际生物化学与分子生物学联合会(IUBMB)命名委员会(Nomenclature Committee of the International Union of Biochemistryand Molecular Biology)陈述的酶分类EC 1.11.1.7，或源自其中的展示出过氧化物酶活性的任何片段的过氧化物酶。
合适的过氧化物酶包括植物、细菌或真菌来源的那些。包括化学修饰的突变体或蛋白质工程化的突变体。有用的过氧化物酶的实例包括来自拟鬼伞属，例如来自灰盖拟鬼伞(C.cinerea)的过氧化物酶(EP 179,486)，及其变体，如在WO 93/24618、WO 95/10602以及WO 98/15257中所描述的那些。
合适的过氧化物酶还包括卤代过氧化物酶，例如氯过氧化物酶、溴过氧化物酶以及展现出氯过氧化物酶或溴过氧化物酶活性的化合物。根据其对卤素离子的特异性将卤代过氧化物酶进行分类。氯过氧化物酶(E.C.1.11.1.10)催化从氯离子形成次氯酸盐。卤代过氧化物酶可以是氯过氧化物酶。优选地，卤代过氧化物酶是钒卤代过氧化物酶，即含钒酸盐的卤代过氧化物酶。在优选方法中，将含钒酸盐的卤代过氧化物酶与氯根离子来源组合。
已从许多不同真菌，特别是从暗色丝孢菌(dematiaceous hyphomycete)真菌组中分离出了卤代过氧化物酶，例如卡尔黑霉属(Caldariomyces)(例如，煤卡尔黑霉(C.fumago))、链格孢属、弯孢属(例如，疣枝弯孢(C.verruculosa)和不等弯孢(C.inaequalis))、内脐蠕孢属、细基格孢属以及葡萄孢属。
还已从细菌，例如假单胞菌属(例如吡咯假单胞菌(P.pyrrocinia))和链霉菌属(例如，金色链霉菌(S.aureofaciens))中分离出了卤代过氧化物酶。
卤代过氧化物酶源自弯孢属物种(Curvularia sp.)，特别是疣枝弯孢(Curvularia verruculosa)或不等弯孢(Curvularia inaequalis)，如描述于WO 95/27046中的不等弯孢CBS 102.42或来自描述于WO 97/04102中的疣枝弯孢CBS 147.63或疣枝弯孢CBS 444.70；或来自描述于WO 01/79459中的Drechslera hartlebii、如描述于WO 01/79458中的盐沼小树状霉(Dendryphiella salina)、如描述于WO 01/79461中的Phaeotrichoconis crotalarie、或如描述于WO 01/79460中的棉丝菌属物种(Geniculosporium sp.)。
适合的氧化酶具体包括由酶分类EC 1.10.3.2所包含的任何漆酶或源自其的展现出漆酶活性的任何片段、或展现出类似活性的化合物，例如儿茶酚氧化酶(EC 1.10.3.1)、邻氨基苯酚氧化酶(EC 1.10.3.4)或胆红素氧化酶(EC 1.3.3.5)。
优选的漆酶是微生物来源的酶。这些酶可以来源于植物、细菌或真菌(包括丝状真菌和酵母)。
来自真菌的合适实例包括可来源于以下的菌株的漆酶：曲霉属，脉孢菌属(例如，粗糙脉孢菌)，柄孢壳菌属，葡萄孢属，金钱菌属(Collybia)，层孔菌属(Fomes)，香菇属，侧耳属，栓菌属(例如，长绒毛栓菌和变色栓菌)，丝核菌属(例如，立枯丝核菌(R.solani))，拟鬼伞属(例如，灰盖拟鬼伞、毛头拟鬼伞(C.comatus)、弗瑞氏拟鬼伞(C.friesii)及C.plicatilis)，小脆柄菇属(Psathyrella)(例如，白黄小脆柄菇(P.condelleana))，斑褶菇属(例如，蝶形斑褶菇(P.papilionaceus))，毁丝霉属(例如，嗜热毁丝霉)，Schytalidium(例如，S.thermophilum)，多孔菌属(例如，P.pinsitus)，射脉菌属(例如，射脉侧菌(P.radiata))(WO 92/01046)或革盖菌属(例如，毛革盖菌(C.hirsutus))(JP 2238885)。
来自细菌的合适的实例包括可来源于芽孢杆菌属的菌株的漆酶。
优选的是来源于拟鬼伞属或毁丝霉属的漆酶；特别是来源于灰盖拟鬼伞的漆酶，如披露于WO 97/08325中；或来源于嗜热毁丝霉，如披露于WO95/33836中。
蛋白酶
合适的蛋白酶可以是任何来源的，但优选地是细菌或真菌来源的，任选地呈蛋白质工程化的或化学修饰的突变体的形式。蛋白酶可以是碱性蛋白酶，例如丝氨酸蛋白酶或金属蛋白酶。丝氨酸蛋白酶可以例如是S1家族的(如胰蛋白酶)或S8家族的(如枯草杆菌蛋白酶(subtilisin))。金属蛋白酶可以例如是嗜热菌蛋白酶，例如来自M4家族的嗜热菌蛋白酶，或另一种金属蛋白酶，如来自M5、M7或M35家族的那些。
术语“枯草杆菌酶(subtilase)”是指根据Siezen等人,Protein Eng.[蛋白质工程]4(1991)719-737和Siezen等人,Protein Sci.[蛋白质科学]6(1997)501-523的丝氨酸蛋白酶的亚组。丝氨酸蛋白酶是特征为在活性位点具有与底物形成共价加合物的丝氨酸的蛋白酶的亚组。枯草杆菌酶可以被划分为六个亚类：枯草杆菌蛋白酶家族、嗜热蛋白酶家族、蛋白酶K家族、羊毛硫氨酸抗生素肽酶家族、Kexin家族和Pyrolysin家族。
尽管适于洗涤剂用途的蛋白酶可以从多种生物(包括例如曲霉属等真菌)获得，但洗涤剂蛋白酶通常已从细菌(特别是从芽孢杆菌属)获得。衍生出枯草杆菌酶的芽孢杆菌属物种的实例包括迟缓芽孢杆菌(Bacillus lentus)、嗜碱芽孢杆菌(Bacillusalkalophilus)、枯草芽孢杆菌(Bacillus subtilis)、解淀粉芽孢杆菌(Bacillusamyloliquefaciens)、地衣芽孢杆菌(Bacillus licheniformis)、短小芽孢杆菌(Bacilluspumilus)和吉氏芽孢杆菌(Bacillus gibsonii)。特定的枯草杆菌蛋白酶包括枯草杆菌蛋白酶lentus、枯草杆菌蛋白酶Novo、枯草杆菌蛋白酶Carlsberg、枯草杆菌蛋白酶BPN’、枯草杆菌蛋白酶309、枯草杆菌蛋白酶147和枯草杆菌蛋白酶168、以及例如蛋白酶PD138(在WO93/18140中描述)。其他有用的蛋白酶是例如在WO 01/16285和WO 02/16547中描述的那些。
胰蛋白酶样蛋白酶的实例包括镰孢属(Fusarium)蛋白酶(在WO 94/25583和WO2005/040372中描述)，以及来源于纤维单胞菌(Cellumonas)的糜蛋白酶(在WO 2005/052161和WO 2005/052146中描述)。
金属蛋白酶的实例包括在WO 2007/044993中描述的中性金属蛋白酶，如来源于解淀粉芽孢杆菌的那些，以及例如在WO 2015/158723和WO 2016/075078中描述的金属蛋白酶。
有用的蛋白酶的实例是在WO 89/06279、WO 92/19729、WO 96/34946、WO 98/20115、WO 98/20116、WO 99/11768、WO 01/44452、WO 03/006602、WO 2004/003186、WO2004/041979、WO 2007/006305、WO 2011/036263、WO 2014/207227、WO 2016/087617和WO2016/174234中描述的蛋白酶变体。优选的蛋白酶变体可以例如包含选自下组的一个或多个突变，该组由以下组成：S3T、V4I、S9R、S9E、A15T、S24G、S24R、K27R、N42R、S55P、G59E、G59D、N60D、N60E、V66A、N74D、S85R、A96S、S97G、S97D、S97A、S97SD、S99E、S99D、S99G、S99M、S99N、S99R、S99H、S101A、V102I、V102Y、V102N、S104A、G116V、G116R、H118D、H118N、A120S、S126L、P127Q、S128A、S154D、A156E、G157D、G157P、S158E、Y161A、R164S、Q176E、N179E、S182E、Q185N、A188P、G189E、V193M、N198D、V199I、Q200L、Y203W、S206G、L211Q、L211D、N212D、N212S、M216S、A226V、K229L、Q230H、Q239R、N246K、S253D、N255W、N255D、N255E、L256E、L256D T268A和R269H，其中位置编号对应于WO 2016/001449的SEQ ID NO 1中示出的迟缓芽孢杆菌蛋白酶的位置。具有这些突变中的一个或多个的蛋白酶变体优选地是在WO2016/001449的SEQ ID NO:1中示出的迟缓芽孢杆菌蛋白酶(
也称为枯草杆菌蛋白酶309)的变体或在WO 2016/001449的SEQ ID NO:2中示出的解淀粉芽孢杆菌蛋白酶(BPN’)的变体。此类蛋白酶变体与WO 2016/001449的SEQ ID NO:1或SEQ ID NO:2优选地具有至少80％序列同一性。
另一种目的蛋白酶是来自迟缓芽孢杆菌DSM 5483的碱性蛋白酶(如例如在WO 91/02792中所述)及其变体(这些变体例如在WO 92/21760、WO 95/23221、EP 1921147、EP1921148和WO 2016/096711中描述)。
可替代地，蛋白酶可以是具有WO 2004/067737的SEQ ID NO:1的TY145蛋白酶的变体，例如在以下一个或多个位置(对应于WO 2004/067737的SEQ ID NO:1的位置27、109、111、171、173、174、175、180、182、184、198、199和297)处包含取代的变体，其中所述蛋白酶变体与WO 2004/067737的SEQ ID NO:1具有至少75％但少于100％的序列同一性。目的TY145变体描述于例如WO 2015/014790、WO 2015/014803、WO 2015/014804、WO 2016/097350、WO 2016/097352、WO 2016/097357和WO 2016/097354中。
优选的蛋白酶的实例包括：
(a)包含选自下组的两个或更多个取代的WO 2016/001449的SEQ ID NO:1的变体，该组由以下组成：S9E、N43R、N76D、Q206L、Y209W、S259D和L262E，例如具有取代S9E、N43R、N76D、V205I、Q206L、Y209W、S259D、N261W和L262E，或具有取代S9E、N43R、N76D、N185E、S188E、Q191N、A194P、Q206L、Y209W、S259D和L262E的变体，其中位置编号基于WO 2016/001449的SEQ ID NO:2的编号；
(b)WO 2016/001449的SEQ ID NO:1的多肽的具有突变S99SE的变体，其中位置编号基于WO 2016/001449的SEQ ID NO:2的编号；
(c)WO 2016/001449的SEQ ID NO:1的多肽的具有突变S99AD的变体，其中位置编号基于WO 2016/001449的SEQ ID NO:2的编号；
(d)具有取代Y167A+R170S+A194P的WO 2016/001449的SEQ ID NO:1的多肽的变体，其中位置编号基于WO 2016/001449的SEQ ID NO:2的编号；
(e)具有取代S9R+A15T+V68A+N218D+Q245R的WO 2016/001449的SEQ ID NO:1的多肽的变体，其中位置编号基于WO 2016/001449的SEQ ID NO:2的编号；
(f)具有取代S9R+A15T+G61E+V68A+A194P+V205I+Q245R+N261D的WO 2016/001449的SEQ ID NO:1的多肽的变体，其中位置编号基于WO 2016/001449的SEQ ID NO:2的编号；
(g)具有取代S99D+S101R/E+S103A+V104I+G160S的WO 2016/001449的SEQ ID NO:1的多肽的变体；例如，具有取代S3T+V4I+S99D+S101E+S103A+V104I+G160S+V205I的WO2016/001449的SEQ ID NO:1的变体，其中位置编号基于WO 2016/001449的SEQ ID NO:2的编号；
(h)具有取代S24G+S53G+S78N+S101N+G128A/S+Y217Q的WO 2016/001449的SEQ IDNO:2的多肽的变体，其中位置编号基于WO 2016/001449的SEQ ID NO:2的编号；
(i)以登录号BER84782披露于GENESEQP中的多肽，其对应于WO 2017/210295中的SEQ ID NO:302；
(j)具有取代S99D+S101E+S103A+V104I+S156D+G160S+L262E的WO 2016/001449的SEQ ID NO:1的多肽的变体，其中位置编号基于WO 2016/001449的SEQ ID NO:2的编号；
(k)具有取代S9R+A15T+G61E+V68A+N76D+S99G+N218D+Q245R的WO 2016/001449的SEQ ID NO:1的多肽的变体，其中位置编号基于WO 2016/001449的SEQ ID NO:2的编号；
(l)具有取代V68A+S106A的WO 2016/001449的SEQ ID NO:1的多肽的变体，其中位置编号基于WO 2016/001449的SEQ ID NO:2的编号；以及
(m)具有取代S27K+N109K+S111E+S171E+S173P+G174K+S175P+F180Y+G182A+L184F+Q198E+N199+T297P的WO 2004/067737的SEQ ID NO:1的多肽的变体，其中位置编号基于WO2004/067737的SEQ ID NO:1的编号。
在以下一个或多个位置(对应于WO 2004/067737的SEQ ID NO:1的位置171、173、175、179或180)处包含取代的蛋白酶变体，其中所述蛋白酶变体与WO 2004/067737的SEQID NO:1具有至少75％但少于100％的序列同一性。
在一个实施例中，蛋白酶是包含突变S99AD的、SEQ ID NO:20的多肽的变体，其中位置编号对应于SEQ ID NO:21的多肽的位置，例如是与SEQ ID NO:20具有至少80％、至少85％、至少90％、至少95％、至少96％、至少97％或至少98％序列同一性的变体。在一个实施例中，蛋白酶包含具有突变S99AD的、SEQ ID NO:20的多肽或由其组成。
在一个实施例中，蛋白酶是包含突变S99SE的、SEQ ID NO:20的多肽的变体，其中位置编号对应于SEQ ID NO:21的多肽的位置，例如是与SEQ ID NO:21具有至少80％、至少85％、至少90％、至少95％、至少96％、至少97％或至少98％序列同一性的变体。在一个实施例中，蛋白酶包含具有突变S99SE的、SEQ ID NO:20的多肽或由其组成。
在一个实施例中，蛋白酶是包含突变Y167A+R170S+A194P的、SEQ ID NO:20的多肽的变体，其中位置编号对应于SEQ ID NO:21的多肽的位置，例如是与SEQ ID NO:20具有至少80％、至少85％、至少90％、至少95％、至少96％、至少97％或至少98％序列同一性的变体。在一个实施例中，蛋白酶包含具有突变Y167A+R170S+A194P的、SEQ ID NO:21的多肽或由其组成。
在一个实施例中，蛋白酶是包含突变S9E+N43R+N76D+V205I+Q206L+Y209W+S259D+N261W+L262E的、SEQ ID NO:20的多肽的变体，其中位置编号对应于SEQ ID NO:21的多肽的位置，例如是与SEQ ID NO:20具有至少80％、至少85％、至少90％或至少95％序列同一性的变体。在一个实施例中，蛋白酶包含具有突变S9E+N43R+N76D+V205I+Q206L+Y209W+S259D+N261W+L262E的、SEQ ID NO:20的多肽或由其组成。
在一个实施例中，蛋白酶是包含突变S3T+V4I+S99D+S101R+S103A+V104I+G160S+V199M+V205I+L217D的、SEQ ID NO:20的多肽的变体，其中位置编号对应于SEQ ID NO:21的多肽的位置，例如是与SEQ ID NO:20具有至少80％、至少85％、至少90％或至少95％序列同一性的变体。在一个实施例中，蛋白酶包含具有突变S3T+V4I+S99D+S101R+S103A+V104I+G160S+V199M+V205I+L217D的、SEQ ID NO:20的多肽或由其组成。
在一个实施例中，蛋白酶是包含突变S3T+V4I+S99D+S101E+S103A+V104I+G160S+V205I的、SEQ ID NO:20的多肽的变体，其中位置编号对应于SEQ ID NO:21的多肽的位置，例如是与SEQ ID NO:20具有至少80％、至少85％、至少90％或至少95％序列同一性的变体。在一个实施例中，蛋白酶包含具有突变S3T+V4I+S99D+S101E+S103A+V104I+G160S+V205I的、SEQ ID NO:20的多肽或由其组成。
在一个实施例中，蛋白酶是包含突变S99D+S101E+S103A+V104I+G160S的、SEQ IDNO:20的多肽的变体，其中位置编号对应于SEQ ID NO:21的多肽的位置，例如是与SEQ IDNO:20具有至少80％、至少85％、至少90％、至少95％或至少96％序列同一性的变体。在一个实施例中，蛋白酶包含具有突变S99D+S101E+S103A+V104I+G160S的、SEQ ID NO:20的多肽或由其组成。
在一个实施例中，蛋白酶是包含突变S99D+S101E+S103A+V104I+S156D+G160S+L262E的、SEQ ID NO:20的多肽的变体，其中位置编号对应于SEQ ID NO:21的多肽的位置，例如是与SEQ ID NO:20具有至少80％、至少85％、至少90％或至少95％序列同一性的变体。在一个实施例中，蛋白酶包含具有突变S99D+S101E+S103A+V104I+S156D+G160S+L262E的、SEQ ID NO:20的多肽或由其组成。
在一个实施例中，蛋白酶是包含突变S87N+S101G+V104N的、SEQ ID NO:20的多肽的变体，其中位置编号对应于SEQ ID NO:21的多肽的位置，例如是与SEQ ID NO:20具有至少80％、至少85％、至少90％、至少95％、至少96％、至少97％或至少98％序列同一性的变体。在一个实施例中，蛋白酶包含具有突变S87N+S101G+V104N的、SEQ ID NO:20的多肽或由其组成。
在一个实施例中，蛋白酶包含SEQ ID NO:21的多肽或由其组成。
在一个实施例中，蛋白酶是包含突变Y217L的、SEQ ID NO:21的多肽的变体，例如是与SEQ ID NO:21具有至少80％、至少85％、至少90％、至少95％、至少96％、至少97％或至少98％序列同一性的变体。在一个实施例中，蛋白酶包含具有突变Y217L的、SEQ ID NO:21的多肽或由其组成。
在一个实施例中，蛋白酶是包含突变S24G+S53G+S78N+S101N+G128S+Y217Q的、SEQID NO:21的多肽的变体，例如是与SEQ ID NO:21具有至少80％、至少85％、至少90％、至少95％或至少96％序列同一性的变体。在一个实施例中，蛋白酶包含具有突变S24G+S53G+S78N+S101N+G128S+Y217Q的、SEQ ID NO:21的多肽或由其组成。
在一个实施例中，蛋白酶是包含突变S24G+S53G+S78N+S101N+G128A+Y217Q的、SEQID NO:21的多肽的变体，例如是与SEQ ID NO:21具有至少80％、至少85％、至少90％、至少95％或至少96％序列同一性的变体。在一个实施例中，蛋白酶包含具有突变S24G+S53G+S78N+S101N+G128A+Y217Q的、SEQ ID NO:21的多肽或由其组成。
在一个实施例中，蛋白酶包含SEQ ID NO:22的多肽或由其组成。
在一个实施例中，蛋白酶是SEQ ID NO:22的多肽的变体，与SEQ ID NO:22具有至少80％、至少85％、至少90％或至少95％的序列同一性。蛋白酶可以是例如包含选自下组的一个或多个突变的SEQ ID NO:22的多肽的变体，该组由以下组成：S27K、N109K、S111E、S171E、S173P、G174K、S175P、F180Y、G182A、L184F、Q198E、N199K和T297P，例如，包含所述突变中的1、2、3、4、5、6、7、8、9、10、11、12个或全部。
在一个实施例中，蛋白酶是包含突变S27K+N109K+S111E+S171E+S173P+G174K+S175P+F180Y+G182A+L184F+Q198E+N199K+T297P的、SEQ ID NO:22的多肽的变体，例如是与SEQ ID NO:22具有至少80％、至少85％、至少90％或至少95％序列同一性的变体。在一个实施例中，蛋白酶包含具有突变S27K+N109K+S111E+S171E+S173P+G174K+S175P+F180Y+G182A+L184F+Q198E+N199K+T297P的、SEQ ID NO:22的多肽或由其组成。
合适的可商购的蛋白酶包括以下列商标名出售的那些：
DuralaseTM、DurazymTM、
Ultra、
Ultra、PrimaseTM、
Ultra、
Ultra、
Blaze
100T、Blaze
125T、Blaze
150T、Blaze
200T、
Uno、
In和
Excel(诺维信公司(Novozymes A/S))，以下列商标名出售的那些：MaxataseTM、MaxacalTM、
Ox、
OxP、
FN2TM、FN3TM、FN4exTM、
ExcellenzTM P1000、ExcellenzTM P1250、EraserTM、
P100、Purafect Prime、Preferenz P110TM、Effectenz P1000TM、
Effectenz P1050TM、
Ox、Effectenz TM P2000、PurafastTM、
OpticleanTM和
(丹斯尼克公司/杜邦公司(Danisco/DuPont))、BLAP(在US 5352604的图29中显示的序列)及其变体(汉高公司(Henkel AG))、以及来自花王株式会社(Kao)的KAP(嗜碱芽孢杆菌枯草杆菌蛋白酶)。
脂肪酶和角质酶
合适的脂肪酶和角质酶包括细菌或真菌来源的那些。包括化学修饰的突变体酶或蛋白质工程化的突变体酶。实例包括来自嗜热真菌属的脂肪酶，例如，如描述于EP 258068和EP 305216中的来自疏绵状嗜热丝孢菌(早先命名为疏棉状腐质霉)；来自腐质霉属的角质酶，例如特异腐质霉(WO96/13580)；来自假单胞菌属的菌株的脂肪酶(这些中的一些现在改名为伯克霍尔德菌属)，例如产碱假单胞菌或类产碱假单胞菌(EP 218272)、洋葱假单胞菌(EP 331376)、假单胞菌属菌株SD705(WO 95/06720和WO 96/27002)、威斯康星假单胞菌(P.wisconsinensis)(WO 96/12012)；GDSL-型链霉菌属脂肪酶(WO 10/065455)；来自稻瘟病菌的角质酶(WO 10/107560)；来自门多萨假单胞菌的角质酶(US 5,389,536)；来自褐色嗜热裂孢菌(Thermobifida fusca)的脂肪酶(WO 11/084412)；嗜热脂肪土芽孢杆菌脂肪酶(WO 11/084417)；来自枯草芽孢杆菌的脂肪酶(WO 11/084599)；以及来自灰色链霉菌(WO11/150157)和始旋链霉菌(S.pristinaespiralis)的脂肪酶(WO 12/137147)。
其他实例是脂肪酶变体，例如EP 407225、WO 92/05249、WO 94/01541、WO 94/25578、WO 95/14783、WO 95/30744、WO 95/35381、WO 95/22615、WO 96/00292、WO 97/04079、WO 97/07202、WO 00/34450、WO 00/60063、WO 01/92502、WO 07/87508以及WO 09/109500中所述的那些。
优选的商业化脂肪酶产品包括LipolaseTM、LipexTM；LipolexTM和LipocleanTM(诺维信公司)，Lumafast(来自杰能科公司(Genencor))以及Lipomax(来自吉斯特布罗卡德斯公司(Gist-Brocades))。
仍其他实例是有时称为酰基转移酶或过水解酶的脂肪酶，例如与南极假丝酵母(Candida antarctica)脂肪酶A具有同源性的酰基转移酶(WO 10/111143)、来自耻垢分枝杆菌(Mycobacterium smegmatis)的酰基转移酶(WO 05/56782)、来自CE 7家族的过水解酶(WO 09/67279)以及耻垢分枝杆菌过水解酶的变体(特别是来自亨斯迈纺织品染化有限公司(Huntsman Textile Effects Pte Ltd)的商业产品Gentle Power Bleach中所用的S54V变体)(WO 10/100028)。
淀粉酶
组合物可包含一种或多种另外的α-淀粉酶。
合适的淀粉酶可以是α-淀粉酶或葡糖淀粉酶并且可以具有细菌或真菌起源。包括化学修饰的突变体或蛋白质工程化的突变体。淀粉酶包括例如从芽孢杆菌属、例如地衣芽孢杆菌的特定菌株(更详细地描述于GB 1,296,839中)获得的α-淀粉酶。
适合的淀粉酶包括具有WO 95/10603中的SEQ ID NO:2的淀粉酶或其与SEQ IDNO:3具有90％序列同一性的变体。优选的变体描述于WO 94/02597、WO 94/18314、WO 97/43424中以及WO 99/19467的SEQ ID NO:4中，例如在以下位置的一个或多个中具有取代的变体：15、23、105、106、124、128、133、154、156、178、179、181、188、190、197、201、202、207、208、209、211、243、264、304、305、391、408和444。
不同的合适的淀粉酶包括具有WO 02/10355中的SEQ ID NO 6的淀粉酶或其与SEQID NO:6具有90％序列同一性的变体。SEQ ID NO:6的优选的变体是在位置181和182中具有缺失并且在位置193中具有取代的那些。
其他合适的淀粉酶是包含示于WO 2006/066594的SEQ ID NO:6中的来源于解淀粉芽孢杆菌的α-淀粉酶的残基1-33和示于WO 2006/066594的SEQ ID NO:4中的地衣芽孢杆菌α-淀粉酶的残基36-483的杂合α-淀粉酶或其具有90％序列同一性的变体。此杂合α-淀粉酶的优选的变体是在以下位置的一个或多个中具有取代、缺失或插入的那些：G48、T49、G107、H156、A181、N190、M197、I201、A209和Q264。包含示于WO 2006/066594的SEQ ID NO:6中的来源于解淀粉芽孢杆菌的α-淀粉酶的残基1-33和SEQ ID NO:4的残基36-483的杂合α-淀粉酶的最优选的变体是具有以下取代的那些：
M197T；
H156Y+A181T+N190F+A209V+Q264S；或者
G48A+T49I+G107A+H156Y+A181T+N190F+I201F+A209V+Q264S。
其他适合的淀粉酶是具有WO 99/19467中的SEQ ID NO:6序列的淀粉酶或与SEQID NO:6具有90％序列同一性的其变体。SEQ ID NO:6的优选的变体是在以下位置中的一个或多个中具有取代、缺失或插入的那些：R181、G182、H183、G184、N195、I206、E212、E216和K269。特别优选的淀粉酶是在位置R181和G182、或位置H183和G184中具有缺失的那些。
可以使用的另外的淀粉酶是具有WO 96/23873的SEQ ID NO:1、SEQ ID NO:3、SEQID NO:2或SEQ ID NO:7的那些或其与SEQ ID NO:1、SEQ ID NO:2、SEQ ID NO:3或SEQ IDNO:7具有90％序列同一性的变体。SEQ ID NO:1、SEQ ID NO:2、SEQ ID NO:3或SEQ ID NO:7的优选的变体是在以下位置中的一个或多个中具有取代、缺失或插入的那些：140、181、182、183、184、195、206、212、243、260、269、304和476，使用WO 96/23873的SEQ ID 2进行编号。更优选的变体是在选自181、182、183和184的两个位置(例如181和182、182和183、或位置183和184)中具有缺失的那些。SEQ ID NO:1、SEQ ID NO:2或SEQ ID NO:7的最优选的淀粉酶变体是在位置183和184中具有缺失并且在位置140、195、206、243、260、304和476中的一个或多个中具有取代的那些。
可以使用的其他淀粉酶是具有WO 2008/153815的SEQ ID NO:2、WO01/66712中的SEQ ID NO:10的淀粉酶或其与WO 2008/153815的SEQ ID NO:2具有90％序列同一性或与WO01/66712中的SEQ ID NO:10具有90％序列同一性的变体。WO 01/66712中的SEQ ID NO:10的优选的变体是在以下位置的一个或多个中具有取代、缺失或插入的那些：176、177、178、179、190、201、207、211和264。
另外的合适的淀粉酶是具有WO 2009/061380的SEQ ID NO:2的淀粉酶或其与SEQID NO:2具有90％序列同一性的变体。SEQ ID NO:2的优选的变体是在以下位置中的一个或多个中具有C-末端截短和/或取代、缺失或插入的那些：Q87、Q98、S125、N128、T131、T165、K178、R180、S181、T182、G183、M201、F202、N225、S243、N272、N282、Y305、R309、D319、Q320、Q359、K444和G475。SEQ ID NO:2的更优选的变体是在以下位置的一个或多个中具有取代的那些：Q87E,R、Q98R、S125A、N128C、T131I、T165I、K178L、T182G、M201L、F202Y、N225E,R、N272E,R、S243Q,A,E,D、Y305R、R309A、Q320R、Q359E、K444E以及G475K，和/或在位置R180和/或S181或T182和/或G183处具有缺失的那些。SEQ ID NO:2的最优选的淀粉酶变体是具有以下取代的那些：
N128C+K178L+T182G+Y305R+G475K；
N128C+K178L+T182G+F202Y+Y305R+D319T+G475K；
S125A+N128C+K178L+T182G+Y305R+G475K；或者
S125A+N128C+T131I+T165I+K178L+T182G+Y305R+G475K，
其中所述变体是C-末端截短的并且任选地进一步包含在位置243处的取代和/或在位置180和/或位置181处的缺失。
另外的合适的淀粉酶是具有WO 2013/184577的SEQ ID NO:1的淀粉酶或其与SEQID NO:1具有90％序列同一性的变体。SEQ ID NO:1的优选的变体是在以下位置的一个或多个中具有取代、缺失或插入的那些：K176、R178、G179、T180、G181、E187、N192、M199、I203、S241、R458、T459、D460、G476和G477。SEQ ID NO:1的更优选的变体是在以下位置的一个或多个中具有取代的那些：K176L、E187P、N192FYH、M199L、I203YF、S241QADN、R458N、T459S、D460T、G476K、以及G477K中具有取代，和/或在位置R178和/或S179或T180和/或G181中具有缺失的那些。SEQ ID NO:1的最优选的淀粉酶变体包含以下取代：
E187P+I203Y+G476K
E187P+I203Y+R458N+T459S+D460T+G476K
并且任选地进一步包含在位置241处的取代和/或在位置178和/或位置179处的缺失。
另外的合适的淀粉酶是具有WO 2010/104675的SEQ ID NO:1的淀粉酶或其与SEQID NO:1具有90％序列同一性的变体。SEQ ID NO:1的优选的变体是在以下位置的一个或多个中具有取代、缺失或插入的那些：N21、D97、V128、K177、R179、S180、I181、G182、M200、L204、E242、G477和G478。
SEQ ID NO:1的更优选的变体是在以下位置的一个或多个中具有取代的那些：N21D、D97N、V128I、K177L、M200L、L204YF、E242QA、G477K和G478K，和/或在位置R179和/或S180或I181和/或G182中具有缺失的那些。SEQ ID NO:1的最优选的淀粉酶变体包含取代N21D+D97N+V128I，并且任选地进一步包含在位置200处的取代和/或在位置180和/或位置181处的缺失。
其他合适的淀粉酶是具有WO 01/66712中的SEQ ID NO:12的α-淀粉酶或与SEQ IDNO:12具有至少90％序列同一性的变体。优选的淀粉酶变体是在WO 01/66712中的SEQ IDNO:12的以下位置的一个或多个中具有取代、缺失或插入的那些：R28、R118、N174；R181、G182、D183、G184、G186、W189、N195、M202、Y298、N299、K302、S303、N306、R310、N314；R320、H324、E345、Y396、R400、W439、R444、N445、K446、Q449、R458、N471、N484。特别优选的淀粉酶包括具有D183和G184的缺失并且具有取代R118K、N195F、R320K及R458K的变体，以及在选自下组的一个或多个位置处另外具有取代的变体：M9、G149、G182、G186、M202、T257、Y295、N299、M323、E345和A339，最优选的是另外在所有这些位置中具有取代的变体。
其他实例是例如描述于WO 2011/098531、WO 2013/001078及WO 2013/001087中的那些淀粉酶变体。可商购的淀粉酶包括DuramylTM、TermamylTM、FungamylTM、Stainzyme TM、Stainzyme PlusTM、NatalaseTM、Liquozyme X、BANTM、
和
Prime(来自诺维信公司)，以及RapidaseTM、PurastarTM/EffectenzTM、Powerase、Preferenz S1000、Preferenz S100、Preferenz S110和Preferenz S210(来自杰能科国际有限公司/杜邦公司)。
一种优选的淀粉酶是WO 2016/180748中具有SEQ ID NO:13的淀粉酶的变体，具有改变H1*+N54S+V56T+K72R+G109A+F113Q+R116Q+W167F+Q172G+A174S+G182*+D183*+G184T+N195F+V206L+K391A+P473R+G476K。
另一种优选的淀粉酶是WO 2013/001078中具有SEQ ID NO:1的淀粉酶的变体，具有改变D183*+G184*+W140Y+N195F+V206Y+Y243F+E260G+G304R+G476K。
另一种优选的淀粉酶是WO 2018/141707中具有SEQ ID NO:1的淀粉酶的变体，具有改变H1*+G7A+G109A+W140Y+G182*+D183*+N195F+V206Y+Y243F+E260G+N280S+G304R+E391A+G476K。
另一种优选的淀粉酶是WO 2017/191160中具有SEQ ID NO:1的淀粉酶的变体，具有改变L202M+T246V。
核酸酶
合适的核酸酶包括脱氧核糖核酸酶(DNA酶)和核糖核酸酶(RNA酶)，它们是分别催化DNA或RNA主链中磷酸二酯键的水解切割从而降解DNA和RNA的任何酶。根据活性的位点有两种主要的分类。外切核酸酶从末端消化核酸。内切核酸酶作用于靶分子中间的区域。核酸酶优选为DNA酶，其优选可得自微生物，优选细菌；特别地，可得自芽孢杆菌属的物种的DNA酶是优选的；特别地，可获自食物芽孢杆菌(Bacillus cibi)、枯草芽孢杆菌或地衣芽孢杆菌的DNA酶是优选的。这些DNA酶的实例在WO 2011/098579、WO 2014/087011和WO 2017/060475中描述。
辅料
还可以利用本领域中已知的用于在衣物洗涤剂中使用的任何洗涤剂组分。其他任选的洗涤剂组分包括防腐蚀剂、防缩剂、抗污垢再沉积剂、抗皱剂、杀细菌剂、粘合剂、腐蚀抑制剂、崩解剂(disintegrant)/崩解试剂(disintegration agent)、染料、酶稳定剂(包括硼酸、硼酸盐、CMC和/或多元醇，例如丙二醇)、织物调理剂(包括粘土)、填充剂/加工助剂、荧光增白剂/光学增亮剂、增泡剂、泡沫(泡)调节剂、香料、污垢悬浮剂、软化剂、抑泡剂、晦暗抑制剂以及芯吸剂，单独或组合使用。可以利用本领域中已知的用于在衣物洗涤剂中使用的任何成分。此类成分的选择完全在技术人员的技术范围内。
分散剂：本发明的洗涤剂组合物还可以包含分散剂。特别地，粉末洗涤剂可以包含分散剂。合适的水溶性有机材料包括均聚合或共聚合的酸或其盐，其中聚羧酸包含被不多于两个碳原子彼此分开的至少两个羧基。合适的分散剂例如描述于Powdered Detergents[粉状洗涤剂],Surfactant science series[表面活性剂科学系列],第71卷,马塞尔德克尔公司,1997中。
染料转移抑制剂：本发明的洗涤剂组合物还可以包括一种或多种染料转移抑制剂。合适的聚合物染料转移抑制剂包括但不限于聚乙烯吡咯烷酮聚合物、多胺N-氧化物聚合物、N-乙烯吡咯烷酮与N-乙烯基咪唑的共聚物、聚乙烯噁唑烷酮以及聚乙烯咪唑或其混合物。当在主题组合物中存在时，染料转移抑制剂可以按该组合物的重量计以从约0.0001％至约10％、从约0.01％至约5％或甚至从约0.1％至约3％的水平存在。
荧光增白剂：本发明的洗涤剂组合物将优选地还包含另外的组分，这些组分可以给正在清洁的物品着色，例如荧光增白剂或光学增亮剂。当存在时，该增亮剂优选地以约0.01％至约05％的水平存在。在本发明的组合物中可以使用合适用于在衣物洗涤剂组合物中使用的任何荧光增白剂。最常用的荧光增白剂是属于以下类别的那些：二氨芪-磺酸衍生物、二芳基吡唑啉衍生物和二苯基-联苯乙烯衍生物。荧光增白剂的二氨芪-磺酸衍生物型的实例包括以下的钠盐：4,4'-双-(2-二乙醇氨基-4-苯胺基-s-三嗪-6-基氨基)芪-2,2'-二磺酸盐；4,4'-双-(2,4-二苯胺基-s-三嗪-6-基氨基)芪-2.2'-二磺酸盐；4,4'-双-(2-苯胺基-4(N-甲基-N-2-羟基-乙基氨基)-s-三嗪-6-基氨基)芪-2,2'-二磺酸盐，4,4'-双-(4-苯基-2,1,3-三唑-2-基)芪-2,2'-二磺酸盐；4,4'-双-(2-苯胺基-4(1-甲基-2-羟基-乙氨基)-s-三嗪-6-基氨基)芪-2,2'-二磺酸盐和2-(二苯乙烯基-4"-萘-1.,2':4,5)-1,2,3-三唑-2"-磺酸盐。优选的荧光增白剂是可从汽巴–嘉基股份有限公司(Ciba-Geigy AG)(巴塞尔，瑞士)获得的天来宝(Tinopal)DMS和天来宝CBS。天来宝DMS是4,4'-双-(2-吗啉基-4苯胺基-s-三嗪-6-基氨基)芪二磺酸盐的二钠盐。天来宝CBS是2,2'-双-(苯基-苯乙烯基)二磺酸盐的二钠盐。还优选的是荧光增白剂是可商购的Parawhite KX，由印度孟买的派拉蒙矿物与化学品公司(Paramount Minerals and Chemicals)供应。合适用于本发明中使用的其他荧光剂包括1-3-二芳基吡唑啉和7-氨烷基香豆素。适当的荧光增亮剂水平包括从约0.01wt％、从0.05wt％、从约0.1wt％或甚至从约0.2wt％的较低水平至0.5wt％或甚至0.75wt％的较高水平。
污垢释放聚合物：本发明的洗涤剂组合物还可以包括一种或多种污垢释放聚合物，这些聚合物帮助从织物(如棉和基于聚酯的织物)去除污垢，特别是从基于聚酯的织物去除疏水性污垢。污垢释放聚合物可以例如是非离子型或阴离子型对苯二甲酸基的聚合物、聚乙烯基己内酰胺和相关共聚物、乙烯基接枝共聚物、聚酯聚酰胺，参见例如PowderedDetergents[粉末洗涤剂],Surfactant science series[表面活性剂科学系列],第71卷,第7章,马塞尔德克尔公司。另一种类型的污垢释放聚合物是包含核芯结构和附接至该核芯结构的多个烷氧基化基团的两亲性烷氧基化油污清洁聚合物。核芯结构可以包含聚烷基亚胺结构或聚烷醇胺结构，如在WO 2009/087523中详细描述的(通过引用并入本文)。此外，任意接合的共聚物是适合的污垢释放聚合物。适合的接合的共聚物更详细地描述于WO 2007/138054、WO 2006/108856以及WO 2006/113314中(将其通过引用特此结合)。其他污垢释放聚合物是取代的多糖结构，尤其是取代的纤维素结构，如修饰的纤维素衍生物，如EP1867808或WO 03/040279中所描述的那些(将二者都通过引用并入本文)。合适的纤维素聚合物包括纤维素、纤维素醚、纤维素酯、纤维素酰胺及其混合物。合适的纤维素聚合物包括阴离子修饰的纤维素、非离子修饰的纤维素、阳离子修饰的纤维素、兼性离子修饰的纤维素及其混合物。合适的纤维素聚合物包括甲基纤维素、羧甲基纤维素、乙基纤维素、羟乙基纤维素、羟丙基甲基纤维素、酯羧甲基纤维素及其混合物。
抗再沉积剂：本发明的洗涤剂组合物还可以包括一种或多种抗再沉积剂，如羧甲基纤维素(CMC)、聚乙烯醇(PVA)、聚乙烯吡咯烷酮(PVP)、聚氧乙烯和/或聚乙二醇(PEG)、丙烯酸的均聚物、丙烯酸和马来酸的共聚物、和乙氧基化的聚乙亚胺。以上在污垢释放聚合物下所描述的基于纤维素的聚合物还可以作为抗再沉积剂起作用。
其他合适的辅料包括但不限于防缩剂、抗皱剂、杀细菌剂、粘合剂、载体、染料、酶稳定剂、织物柔软剂、填料、泡沫调节剂、水溶助剂、香料、色素、抑泡剂、溶剂以及用于液体洗涤剂的结构剂和/或结构弹性剂。
洗涤剂产品的配制
一种或多种洗涤剂酶，即α-淀粉酶和任选地一种或多种另外的酶可以通过添加含有一种或多种酶的单独的添加剂，或通过添加包含这些酶的组合添加剂而被包含于洗涤剂组合物中。可以配制包含一种或多种酶的洗涤剂添加剂，例如，配制为颗粒，特别是无粉尘颗粒。
本发明的洗涤剂组合物可以呈任何方便的形式，例如，常规的或压制的粉末、颗粒、均匀的片剂、具有两层或更多层的片剂。粉末组合物(例如粉末、颗粒或片剂)也可以形成在多个隔室小袋或荚中的复合组合物(例如隔室)的一部分。
小袋(荚)可以被配置为单个或多个隔室，并且可以是适用于保存该组合物的任何形式、任何形状、和任何材料，不允许该组合物在与水接触之前从该小袋中释放出。袋由水溶性膜制成，它包含了一个内部体积。该内体积可以分成小袋的区室。优选的膜是聚合材料，优选地被成型为膜或薄片的聚合物。优选的聚合物、共聚物或其衍生物选自聚丙烯酸酯、和水溶性丙烯酸酯共聚物、甲基纤维素、羧甲基纤维素、糊精钠、乙基纤维素、羟乙基纤维素、羟丙基甲基纤维素、麦芽糊精、聚甲基丙烯酸酯，最优选地是聚乙烯醇共聚物以及羟丙基甲基纤维素(HPMC)。优选地，聚合物在膜例如PVA中的水平是至少约60％。优选的平均分子量典型地将是约20,000至约150,000。膜还可以是共混物组合物，该共混物组合物包括可水解降解并且水可溶的聚合物共混物，例如聚乳酸和聚乙烯醇(已知在商标名称(Tradereference)M8630下，如由美国印第安纳州盖里(Gary,Indiana,US)的克里斯克拉夫特工业产品公司(Chris Craft In.Prod.)销售)加增塑剂，像甘油、乙二醇、丙二醇、山梨醇及其混合物。小袋可以包含例如由水溶性膜分隔的固体衣物洗涤剂组合物或部分组分和/或液体清洁组合物或部分组分。用于液体组分的室在构成上可以与包含固体的室不同。参见，例如US 2009/0011970。
可以由水可溶的小袋中或片的不同层中的隔室来将洗涤剂成分物理地彼此分离，从而避免了部件之间的负储存相互作用。在洗涤溶液中，每个室的不同溶解曲线还可以引起选择的组分的延迟溶解。
颗粒洗涤剂配制品
颗粒(粉末)洗涤剂中通常使用呈颗粒形式的酶，包含含酶的核芯和任选的一个或多个包衣。用于制备核芯的各种方法在本领域中是众所周知的，并且包括例如a)喷雾干燥含液体酶的溶液，b)产生分层产物，其中酶被包衣为围绕预形成的惰性核芯颗粒的层，例如使用流化床装置进行，c)将酶吸收到预形成的核芯的表面上和/或其中，d)挤出含酶的糊剂，e)将含酶的粉末悬浮在熔融蜡中并雾化以产生粒状产品，f)通过将含酶的液体添加到制粒组分的干粉组合物中来进行混合制粒，g)通过研磨或粉碎较大的颗粒、小丸等对含酶的核芯进行粒度减小，以及h)流化床制粒。可以干燥含酶的核芯(例如使用流化床干燥器或用于在进料或酶工业中干燥颗粒的其他已知方法)，导致通常为0.1-10％w/w水的含水量。
含酶的核芯可任选地提供涂层以改善储存稳定性和/或减少粉尘形成。通常用于洗涤剂的酶颗粒的一种涂层是盐涂层，通常是无机盐涂层，其可以是例如用流体床以盐的溶液施加的。可以使用的其他包衣材料是例如聚乙二醇(PEG)、甲基羟基-丙基纤维素(MHPC)和聚乙烯醇(PVA)。颗粒可含有多于一个涂层，例如盐涂层，然后是诸如PEG、MHPC或PVA的材料的另外的涂层。
有关酶颗粒及其制备的更多信息，参见WO 2013/007594以及例如：WO 2009/092699、EP 1705241、EP 1382668、WO 2007/001262、US 6,472,364、WO 2004/074419和WO2009/102854。
微生物
洗涤剂组分以及洗涤剂组合物还可以包含一种或多种微生物，如一种或多种真菌、酵母、或细菌。
在实施例中，一种或多种微生物是脱水(例如通过冻干)的细菌或酵母，例如乳酸杆菌菌株。
在另一个实施例中，微生物是一种或多种微生物孢子/芽孢(与营养细胞相对)，例如细菌芽孢；或真菌孢子、分生孢子、菌丝。优选地，该一种或多种孢子/芽孢是芽孢杆菌芽孢；甚至更优选地，该一种或多种孢子/芽孢是枯草芽孢杆菌、地衣芽孢杆菌、解淀粉芽孢杆菌、或巨大芽孢杆菌的芽孢。
微生物可以按与酶相同的方式包含在洗涤剂组合物中。
酶配制品以及洗涤剂配制品可包含一种或多种微生物或微生物体。通常，在酶/洗涤剂配制品中可以使用任何合适的量/浓度的任何一种或多种微生物。微生物可以作为唯一的生物活性成分使用，但它们也可以与上述一种或多种酶结合使用。
添加一种或多种微生物的目的可以是如在WO 2012/112718中所述的减少恶臭。其他目的可包括原位产生合意的生物化合物，或者用一种或多种微生物接种/占据位置，以竞争地预防其他非希望的微生物形式占据相同位置(竞争排斥)。
术语“微生物”通常意指通过显微镜可见的小的生物。微生物通常以单细胞或细胞集落的形式存在。一些微生物可能是多细胞的。微生物包括原核生物(例如，细菌和古生菌)和真核生物(例如，一些真菌、藻类、原生动物)。细菌的实例可以是革兰氏阳性细菌或革兰氏阴性细菌。细菌的实例形式包括营养细胞和芽孢。真菌的实例可以是酵母、霉菌和蕈类。真菌的实例形式包括菌丝和孢子。在本文中，病毒可以被认为是微生物。
微生物可以是重组或非重组的。在一些实例中，微生物可以产生可用于包含在洗涤剂组合物中的各种物质(例如，酶)。来自微生物的提取物或提取物的级分可用于洗涤剂中。在洗涤剂中也可以使用培养微生物的培养基或来自培养基的提取物或分离物。在微生物的一些具体实例中，由微生物，其提取物、培养基和级分产生的物质可以特别排除在洗涤剂之外。在一些实例中，微生物或由微生物产生或提取的物质可以激活、增强、保存、延长洗涤剂活性或洗涤剂所含的组分的活性等。
通常，可以使用本领域已知的方法培养微生物。然后可以以各种方式处理或配制微生物。在一些实例中，微生物可以是干的(例如，冻干的)。在一些实例中，微生物可以是被包封的(例如，喷雾干燥)。许多其他处理或配制也是可能的。这些处理或制备有利于随时间和/或在洗涤剂组分存在下保留微生物活力。然而，在一些实例中，洗涤剂中的微生物可能是无活力的。可以在使用之前或在使用洗涤剂时将经处理/配制的微生物添加到洗涤剂中。
在一个实施例中，微生物是芽孢杆菌属物种，例如选自下组的至少一种芽孢杆菌属物种，该组由以下组成：枯草芽孢杆菌、解淀粉芽孢杆菌、地衣芽孢杆菌、萎缩芽孢杆菌、短小芽孢杆菌、巨大芽孢杆菌或其组合。在优选的实施例中，上述芽孢杆菌属物种均处于芽孢形式，这显著提高了储存稳定性。
用途
本发明还涉及用于在纺织品和织物的洗涤例如家用衣物洗涤和工业衣物洗涤中使用洗涤剂组合物的方法。
本发明还涉及根据本发明的洗涤剂组合物在清洁过程如衣物洗涤(包括工业清洁)、ADW和硬表面清洁中的用途。对于清洁而言重要的污物和污渍由许多不同物质构成，并且已经开发全都具有不同底物特异性的一系列不同的酶用于在涉及衣物和硬表面清洁(例如餐具洗涤)两者中使用。这些酶被认为提供酶洗涤益处，因为与不具有酶的同一过程相比，它们在其应用的清洁过程中特异性地改善污渍去除。本领域已知的去污酶包括以下酶，例如蛋白酶、淀粉酶、脂肪酶、角质酶、纤维素酶、内切葡聚糖酶、木葡聚糖酶、果胶酶、果胶裂解酶、黄原胶酶、过氧化物酶、卤代过氧酶、过氧化氢酶以及甘露聚糖酶。
在另一方面，本发明涉及洗衣方法，该方法可用于家用洗衣以及工业洗衣。此外，本发明涉及用于洗涤纺织品(例如织物、服装(garment)、衣服(cloth)等)的方法，其中该方法包括用洗涤溶液处理该纺织品，该洗涤溶液包含本发明的洗涤剂组合物。可以使用家用或工业洗衣机进行洗衣或可以使用本发明的洗涤剂组合物手动地进行洗衣。
另一方面，本发明涉及餐具洗涤(包括ADW或硬表面清洁)方法，该方法可用于家用清洁以及工业清洁。此外，本发明涉及用于餐具洗涤或硬表面清洁的方法，其中该方法包括用含有本发明的洗涤剂组合物的洗涤溶液处理餐具或硬表面。例如，可以使用本发明的洗涤剂组合物用家用洗碗机或用手进行餐具洗涤或硬表面清洁。
本发明的洗涤剂组合物可以配制为(例如)手洗或机洗衣物洗涤剂组合物，包括适用于预处理有污迹的织物的洗衣添加剂组合物，和漂洗添加的织物软化剂组合物，或配制为用于一般家用硬表面清洁操作的洗涤剂组合物，或配制用于手洗或机洗餐具洗涤操作。
该清洁过程或纺织品护理过程可以例如是衣物洗涤过程、餐具洗涤过程或硬表面(例如浴室瓷砖、地板、桌面、排水管、水槽以及脸盆)清洁。衣物洗涤过程可以例如是家用衣物洗涤，但是也可以是工业衣物洗涤。此外，本发明涉及用于洗涤织物和/或衣物的方法，其中该方法包括用含有本发明的洗涤剂组合物的洗涤溶液处理织物。例如，可以在机器洗涤或者在手动洗涤中进行清洁过程或纺织品保养过程。洗液可以是例如含有洗涤剂组合物的水性洗液。
另一方面，本发明涉及包含5-100g粉末洗涤剂的洗涤剂组合物，该组合物包含具有α-淀粉酶活性的至少一种多肽，并且还包含一种或多种洗涤剂组分；本发明还涉及该组合物在清洁过程如衣物洗涤或餐具洗涤中的用途，其中该组合物的pH不超过约9.0。在这一方面，组合物可以例如包含8-80g，例如10-60g的粉末洗涤剂。在一个实施例中，这一方面的洗涤剂组合物是压制的组合物，例如呈高度压制的粉末或洗涤块的形式，包括例如10-50g、如10-40g、如10-30g或10-20g的粉末洗涤剂。
该方面还涉及一种清洁方法，特别是用于清洁织物或纺织品，或用于餐具洗涤，该方法包括在适于清洁所述织物/纺织品或餐具的条件下使本方面的洗涤剂组合物与织物/纺织品或餐具接触。
根据该方面的组合物及其用途和清洁方法中的α-淀粉酶可以是上文进一步描述的任何α-淀粉酶。
洗涤方法
本发明提供一种使用本发明的包含α-淀粉酶的洗涤剂组合物的清洁方法，特别是用于清洁织物或纺织品，或用于餐具洗涤。
清洁方法包括在适于清洁物体的条件下使物体与包含α-淀粉酶的洗涤剂组合物接触。在优选实施例中，洗涤剂组合物用于衣物或餐具洗涤过程中。
另一个实施例涉及用于从织物或纺织品去除污渍的方法，该方法包括在适合于清洁该物体的条件下将该织物或纺织品与本发明的组合物接触。
另一个实施例涉及用于从餐具去除污渍的方法，该方法包括在适合于清洁该物体的条件下将该餐具与本发明的组合物接触。
可以在溶液中按以下浓度使用组合物：从约100ppm，优选地500ppm至约15,000ppm。水温范围典型地是从约5℃至约95℃，包括约10℃、约15℃、约20℃、约25℃、约30℃、约35℃、约40℃、约45℃、约50℃、约55℃、约60℃、约65℃、约70℃、约75℃、约80℃、约85℃和约90℃。水与织物比率典型地是从约1:1至约30:1。
可以使用常规稳定剂和蛋白酶抑制剂来稳定本发明的洗涤剂组合物的一种或多种酶，例如多元醇(如丙二醇或甘油)、糖或糖醇、不同的盐(例如NaCl、KCl)、乳酸、甲酸、硼酸、或硼酸衍生物(例如，芳族硼酸酯、或苯基硼酸衍生物(如4-甲酰苯基硼酸))、或肽醛(如二肽醛、三肽醛或四肽醛或醛类似物)(或者具有形式B1-B0-R，其中R是H、CH3、CX3、CHX2、或CH2X(X＝卤素)，B0是单个氨基酸残基(优选地具有一个任选取代的脂肪族或芳香族侧链)；并且B1由一个或多个氨基酸残基组成(优选一个、两个或三个)，任选地包含一个N-末端保护基团，或者如描述于WO 2009/118375、WO 98/13459中的)或者蛋白类型的蛋白酶抑制剂，例如RASI、BASI、WASI(水稻、大麦和小麦的双功能α-淀粉酶/枯草杆菌蛋白酶抑制剂)或CI2或SSI。该组合物可以例如如WO 92/19709、WO 92/19708和US 6,472,364中所述进行配制。在一些实施例中，本文采用的这些酶由存在于为这些酶提供此类离子的成品组合物中的锌(II)、钙(II)和/或镁(II)离子的水溶性来源，连同其他金属离子(例如，钡(II)、钪(II)、铁(II)、锰(II)、铝(III)、锡(II)、钴(II)、铜(II)、镍(II)以及氧钒(IV))稳定化。
通过以下实例进一步描述本发明，这些实例不应当理解为对本发明的范围进行限制。
实例
材料与方法
PNP-G7测定
用于衣物洗涤的自动机械应力测定(AMSA)
为了评估在衣物洗涤中的洗涤性能，使用自动机械应力测定(AMSA)进行洗涤实验。使用AMSA，可以检査大量小体积酶洗涤剂溶液的洗涤性能。AMSA板具有许多用于测试溶液的槽和盖子，盖子针对所有槽开口强力挤压衣物洗涤样品(待洗涤的纺织品)。在洗涤时间期间，板、测试溶液、纺织品和盖子剧烈摇动以使测试溶液与纺织品接触并以规则的、周期性摆动方式施加机械应力。关于进一步描述，参见WO 02/42740，尤其是第23-24页的“Special method embodiments”[特殊方法实施例]段落。
在以下指定的实验条件下进行衣物洗涤实验：
模式洗涤剂和测试材料如下：
测试了以下α-淀粉酶：
测试材料获得自测试材料BV中心(Center for Testmaterials BV)，邮政信箱120，3133KT弗拉尔丁恩，荷兰。
通过将CaCl2、MgCl2、以及NaHCO3(Ca2+:Mg2+:NaHCO3＝4:1:7.5)添加到测试系统中将水硬度调节至15°dH。在洗涤之后，将纺织品用自来水冲洗并干燥。
以洗涤过的纺织品的色彩亮度测量洗涤性能。也可以将亮度表示为当用白光照射样品时从样品反射的光的强度。当样品受污时，反射光的强度低于干净样品。换言之，更清洁的样品将反射更多的光线，并将具有更高的强度。因此，反射光的强度可以用来衡量洗涤性能。
颜色测量使用专业平台式扫描仪(Kodak iQsmart,Kodak,Midtager 29,DK-2605
Denmark)来进行，该扫描仪用于获取经洗涤的纺织品的图像。
为了从扫描的图像中提取光强度的值，将来自图像的24-位像素值转化为红色、绿色以及蓝色(RGB)的值。通过将RGB值作为向量进行加和并且然后取所得向量的长度来计算强度值(Int)：
实例1：不同淀粉酶在不同模式洗涤剂中的洗涤性能
如上所述，在AMSA的不同模式洗涤剂中研究了α-淀粉酶编号1的洗涤性能。在20℃确定的强度值在表1中示出，并且在40℃确定的强度值在表2中示出。
表1.在不同洗衣模式洗涤剂中，在20℃，相对于不含淀粉酶的洗涤剂确定淀粉酶
变体的Δ强度值
表2.在不同洗衣模式洗涤剂中，在40℃，相对于不含淀粉酶的洗涤剂确定具有α-
淀粉酶的多肽的Δ强度值
从表1和2中清楚地是，在20℃和40℃，α-淀粉酶编号1在低pH粉末洗涤剂洗衣粉模式洗涤剂1、2和3中均显示出显著的洗涤性能，而在高pH粉末洗涤剂洗衣粉模式洗涤剂4中均没有可检测到的显著性能。
实例2.洗涤性能，与现有技术α-淀粉酶相比。
在20℃和40℃，在低pH粉末洗涤剂洗衣粉模式洗涤剂1、2和3中和在高pH粉末洗涤剂洗衣粉模式洗涤剂4中，将具有α-淀粉酶的多肽的洗涤性能(两次测量的平均值)与具有α-淀粉酶的参比多肽进行了比较。将结果归一化，使现有技术α-淀粉酶的结果设定为1.0
在20℃和40℃进行的AMSA结果显示在表3和表4中。
表3：在20℃的相对洗涤性能
表4：在40℃的相对洗涤性能
该数据清楚地表明，在20℃和40℃，淀粉酶变体在低pH粉末洗涤剂中显示出优于现有技术的α-淀粉酶的洗涤性能，而在高pH粉末洗涤剂中观察到性能急剧下降。
序列表
<110> 诺维信公司（Novozymes A/S）
<120> 低pH粉末洗涤剂组合物
<130> 14757-WO-PCT
<160> 22
<170> PatentIn 3.5版
<210> 1
<211> 483
<212> PRT
<213> 芽孢杆菌属
<400> 1
His His Asn Gly Thr Asn Gly Thr Leu Met Gln Tyr Phe Glu Trp Tyr
1 5 10 15
Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Arg Ser Asp Ala Ser
20 25 30
Asn Leu Lys Asp Lys Gly Ile Ser Ala Val Trp Ile Pro Pro Ala Trp
35 40 45
Lys Gly Ala Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Ile Arg Thr Lys Tyr Gly
65 70 75 80
Thr Arg Asn Gln Leu Gln Ala Ala Val Asn Ala Leu Lys Ser Asn Gly
85 90 95
Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp
100 105 110
Ala Thr Glu Met Val Lys Ala Val Glu Val Asn Pro Asn Asn Arg Asn
115 120 125
Gln Glu Val Ser Gly Glu Tyr Thr Ile Glu Ala Trp Thr Lys Phe Asp
130 135 140
Phe Pro Gly Arg Ala Asn Thr His Ser Asn Phe Lys Trp Arg Trp Tyr
145 150 155 160
His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Lys Leu Asn Asn Arg
165 170 175
Ile Tyr Lys Phe Arg Thr Lys Ala Trp Asp Trp Glu Val Asp Thr Glu
180 185 190
Phe Gly Asn Tyr Asp Tyr Leu Leu Tyr Ala Asp Ile Asp Met Asp His
195 200 205
Pro Glu Val Val Asn Glu Leu Arg Asn Trp Gly Val Trp Tyr Thr Asn
210 215 220
Thr Leu Gly Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His Ile Lys
225 230 235 240
Tyr Ser Phe Thr Arg Asp Trp Ile Asn His Val Arg Ser Ala Ile Gly
245 250 255
Lys Asn Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu Gly Ala
260 265 270
Ile Glu Asn Tyr Leu Asn Lys Thr Asn Trp Asn His Ser Val Phe Asp
275 280 285
Val Pro Leu His Phe Asn Leu Tyr Tyr Ala Ser Lys Ser Gly Gly Asn
290 295 300
Tyr Asp Met Arg Gln Ile Phe Asn Gly Thr Val Val Gln Lys His Pro
305 310 315 320
Thr His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro Glu Glu
325 330 335
Ser Leu Glu Ser Phe Val Arg Glu Trp Phe Lys Pro Leu Ala Tyr Ala
340 345 350
Leu Thr Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr Gly Asp
355 360 365
Tyr Tyr Gly Ile Pro Thr His Gly Val Pro Ala Met Lys Ser Lys Ile
370 375 380
Asp Pro Ile Leu Glu Ala Arg Gln Lys Tyr Ala Tyr Gly Arg Gln Asn
385 390 395 400
Asp Tyr Leu Asp His His Asn Ile Ile Gly Trp Thr Arg Glu Gly Asn
405 410 415
Thr Ala His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp Gly Ala
420 425 430
Gly Gly Asn Lys Trp Met Phe Val Gly Arg Asn Lys Ala Gly Gln Val
435 440 445
Trp Thr Asp Ile Thr Gly Asn Lys Ala Gly Thr Val Thr Ile Asn Ala
450 455 460
Asp Gly Trp Gly Asn Phe Ser Val Asn Gly Gly Ser Val Ser Ile Trp
465 470 475 480
Val Asn Lys
<210> 2
<211> 485
<212> PRT
<213> 芽孢杆菌属
<400> 2
His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr
1 5 10 15
Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Arg Ser Asp Ala Ser
20 25 30
Asn Leu Lys Asp Lys Gly Ile Ser Ala Val Trp Ile Pro Pro Ala Trp
35 40 45
Lys Gly Ala Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Ile Arg Thr Lys Tyr Gly
65 70 75 80
Thr Arg Asn Gln Leu Gln Ala Ala Val Asn Ala Leu Lys Ser Asn Gly
85 90 95
Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp
100 105 110
Ala Thr Glu Met Val Arg Ala Val Glu Val Asn Pro Asn Asn Arg Asn
115 120 125
Gln Glu Val Ser Gly Glu Tyr Thr Ile Glu Ala Trp Thr Lys Phe Asp
130 135 140
Phe Pro Gly Arg Gly Asn Thr His Ser Asn Phe Lys Trp Arg Trp Tyr
145 150 155 160
His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Lys Leu Asn Asn Arg
165 170 175
Ile Tyr Lys Phe Arg Gly Asp Gly Lys Gly Trp Asp Trp Glu Val Asp
180 185 190
Thr Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Ile Asp Met
195 200 205
Asp His Pro Glu Val Val Asn Glu Leu Arg Asn Trp Gly Val Trp Tyr
210 215 220
Thr Asn Thr Leu Gly Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His
225 230 235 240
Ile Lys Tyr Ser Phe Thr Arg Asp Trp Ile Asn His Val Arg Ser Ala
245 250 255
Thr Gly Lys Asn Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu
260 265 270
Gly Ala Ile Glu Asn Tyr Leu Asn Lys Thr Asn Trp Asn His Ser Val
275 280 285
Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Lys Ser Gly
290 295 300
Gly Asn Tyr Asp Met Arg Gln Ile Phe Asn Gly Thr Val Val Gln Arg
305 310 315 320
His Pro Met His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro
325 330 335
Glu Glu Ala Leu Glu Ser Phe Val Glu Glu Trp Phe Lys Pro Leu Ala
340 345 350
Tyr Ala Leu Thr Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr
355 360 365
Gly Asp Tyr Tyr Gly Ile Pro Thr His Gly Val Pro Ala Met Lys Ser
370 375 380
Lys Ile Asp Pro Ile Leu Glu Ala Arg Gln Lys Tyr Ala Tyr Gly Arg
385 390 395 400
Gln Asn Asp Tyr Leu Asp His His Asn Ile Ile Gly Trp Thr Arg Glu
405 410 415
Gly Asn Thr Ala His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp
420 425 430
Gly Ala Gly Gly Asn Lys Trp Met Phe Val Gly Arg Asn Lys Ala Gly
435 440 445
Gln Val Trp Thr Asp Ile Thr Gly Asn Arg Ala Gly Thr Val Thr Ile
450 455 460
Asn Ala Asp Gly Trp Gly Asn Phe Ser Val Asn Gly Gly Ser Val Ser
465 470 475 480
Ile Trp Val Asn Lys
485
<210> 3
<211> 485
<212> PRT
<213> 芽孢杆菌属物种 707
<400> 3
His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr
1 5 10 15
Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Asn Ser Asp Ala Ser
20 25 30
Asn Leu Lys Ser Lys Gly Ile Thr Ala Val Trp Ile Pro Pro Ala Trp
35 40 45
Lys Gly Ala Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly
65 70 75 80
Thr Arg Ser Gln Leu Gln Ala Ala Val Thr Ser Leu Lys Asn Asn Gly
85 90 95
Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp
100 105 110
Ala Thr Glu Met Val Arg Ala Val Glu Val Asn Pro Asn Asn Arg Asn
115 120 125
Gln Glu Val Thr Gly Glu Tyr Thr Ile Glu Ala Trp Thr Arg Phe Asp
130 135 140
Phe Pro Gly Arg Gly Asn Thr His Ser Ser Phe Lys Trp Arg Trp Tyr
145 150 155 160
His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Arg Leu Asn Asn Arg
165 170 175
Ile Tyr Lys Phe Arg Gly His Gly Lys Ala Trp Asp Trp Glu Val Asp
180 185 190
Thr Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Ile Asp Met
195 200 205
Asp His Pro Glu Val Val Asn Glu Leu Arg Asn Trp Gly Val Trp Tyr
210 215 220
Thr Asn Thr Leu Gly Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His
225 230 235 240
Ile Lys Tyr Ser Phe Thr Arg Asp Trp Ile Asn His Val Arg Ser Ala
245 250 255
Thr Gly Lys Asn Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu
260 265 270
Gly Ala Ile Glu Asn Tyr Leu Gln Lys Thr Asn Trp Asn His Ser Val
275 280 285
Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Lys Ser Gly
290 295 300
Gly Asn Tyr Asp Met Arg Asn Ile Phe Asn Gly Thr Val Val Gln Arg
305 310 315 320
His Pro Ser His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro
325 330 335
Glu Glu Ala Leu Glu Ser Phe Val Glu Glu Trp Phe Lys Pro Leu Ala
340 345 350
Tyr Ala Leu Thr Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr
355 360 365
Gly Asp Tyr Tyr Gly Ile Pro Thr His Gly Val Pro Ala Met Arg Ser
370 375 380
Lys Ile Asp Pro Ile Leu Glu Ala Arg Gln Lys Tyr Ala Tyr Gly Lys
385 390 395 400
Gln Asn Asp Tyr Leu Asp His His Asn Ile Ile Gly Trp Thr Arg Glu
405 410 415
Gly Asn Thr Ala His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp
420 425 430
Gly Ala Gly Gly Ser Lys Trp Met Phe Val Gly Arg Asn Lys Ala Gly
435 440 445
Gln Val Trp Ser Asp Ile Thr Gly Asn Arg Thr Gly Thr Val Thr Ile
450 455 460
Asn Ala Asp Gly Trp Gly Asn Phe Ser Val Asn Gly Gly Ser Val Ser
465 470 475 480
Ile Trp Val Asn Lys
485
<210> 4
<211> 399
<212> PRT
<213> 合成构建体
<400> 4
His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr
1 5 10 15
Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Asn Ser Asp Ala Ser
20 25 30
Asn Leu Lys Ser Lys Gly Ile Thr Ala Val Trp Ile Pro Pro Ala Trp
35 40 45
Lys Gly Ala Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly
65 70 75 80
Thr Arg Ser Gln Leu Gln Ala Ala Val Thr Ser Leu Lys Asn Asn Gly
85 90 95
Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp
100 105 110
Ala Thr Glu Met Val Arg Ala Val Glu Val Asn Pro Asn Asn Arg Asn
115 120 125
Gln Glu Val Thr Gly Glu Tyr Thr Ile Glu Ala Trp Thr Arg Phe Asp
130 135 140
Phe Pro Gly Arg Gly Asn Thr His Ser Ser Phe Lys Trp Arg Trp Tyr
145 150 155 160
His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Arg Leu Asn Asn Arg
165 170 175
Ile Tyr Lys Phe Arg Gly His Gly Lys Ala Trp Asp Trp Glu Val Asp
180 185 190
Thr Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Ile Asp Met
195 200 205
Asp His Pro Glu Val Val Asn Glu Leu Arg Asn Trp Gly Val Trp Tyr
210 215 220
Thr Asn Thr Leu Gly Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His
225 230 235 240
Ile Lys Tyr Ser Phe Thr Arg Asp Trp Ile Asn His Val Arg Ser Ala
245 250 255
Thr Gly Lys Asn Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu
260 265 270
Gly Ala Ile Glu Asn Tyr Leu Gln Lys Thr Asn Trp Asn His Ser Val
275 280 285
Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Lys Ser Gly
290 295 300
Gly Asn Tyr Asp Met Arg Asn Ile Phe Asn Gly Thr Val Val Gln Arg
305 310 315 320
His Pro Ser His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro
325 330 335
Glu Glu Ala Leu Glu Ser Phe Val Glu Glu Trp Phe Lys Pro Leu Ala
340 345 350
Tyr Ala Leu Thr Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr
355 360 365
Gly Asp Tyr Tyr Gly Ile Pro Thr His Gly Val Pro Ala Met Arg Ser
370 375 380
Lys Ile Asp Pro Ile Leu Glu Ala Arg Gln Lys Tyr Ala Tyr Gly
385 390 395
<210> 5
<211> 86
<212> PRT
<213> 合成构建体
<400> 5
Lys Gln Asn Asp Tyr Leu Asp His His Asn Ile Ile Gly Trp Thr Arg
1 5 10 15
Glu Gly Asn Thr Ala His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser
20 25 30
Asp Gly Ala Gly Gly Ser Lys Trp Met Phe Val Gly Arg Asn Lys Ala
35 40 45
Gly Gln Val Trp Ser Asp Ile Thr Gly Asn Arg Thr Gly Thr Val Thr
50 55 60
Ile Asn Ala Asp Gly Trp Gly Asn Phe Ser Val Asn Gly Gly Ser Val
65 70 75 80
Ser Ile Trp Val Asn Lys
85
<210> 6
<211> 483
<212> PRT
<213> 解淀粉芽孢杆菌
<400> 6
Val Asn Gly Thr Leu Met Gln Tyr Phe Glu Trp Tyr Thr Pro Asn Asp
1 5 10 15
Gly Gln His Trp Lys Arg Leu Gln Asn Asp Ala Glu His Leu Ser Asp
20 25 30
Ile Gly Ile Thr Ala Val Trp Ile Pro Pro Ala Tyr Lys Gly Leu Ser
35 40 45
Gln Ser Asp Asn Gly Tyr Gly Pro Tyr Asp Leu Tyr Asp Leu Gly Glu
50 55 60
Phe Gln Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly Thr Lys Ser Glu
65 70 75 80
Leu Gln Asp Ala Ile Gly Ser Leu His Ser Arg Asn Val Gln Val Tyr
85 90 95
Gly Asp Val Val Leu Asn His Lys Ala Gly Ala Asp Ala Thr Glu Asp
100 105 110
Val Thr Ala Val Glu Val Asn Pro Ala Asn Arg Asn Gln Glu Thr Ser
115 120 125
Glu Glu Tyr Gln Ile Lys Ala Trp Thr Asp Phe Arg Phe Pro Gly Arg
130 135 140
Gly Asn Thr Tyr Ser Asp Phe Lys Trp His Trp Tyr His Phe Asp Gly
145 150 155 160
Ala Asp Trp Asp Glu Ser Arg Lys Ile Ser Arg Ile Phe Lys Phe Arg
165 170 175
Gly Glu Gly Lys Ala Trp Asp Trp Glu Val Ser Ser Glu Asn Gly Asn
180 185 190
Tyr Asp Tyr Leu Met Tyr Ala Asp Val Asp Tyr Asp His Pro Asp Val
195 200 205
Val Ala Glu Thr Lys Lys Trp Gly Ile Trp Tyr Ala Asn Glu Leu Ser
210 215 220
Leu Asp Gly Phe Arg Ile Asp Ala Ala Lys His Ile Lys Phe Ser Phe
225 230 235 240
Leu Arg Asp Trp Val Gln Ala Val Arg Gln Ala Thr Gly Lys Glu Met
245 250 255
Phe Thr Val Ala Glu Tyr Trp Gln Asn Asn Ala Gly Lys Leu Glu Asn
260 265 270
Tyr Leu Asn Lys Thr Ser Phe Asn Gln Ser Val Phe Asp Val Pro Leu
275 280 285
His Phe Asn Leu Gln Ala Ala Ser Ser Gln Gly Gly Gly Tyr Asp Met
290 295 300
Arg Arg Leu Leu Asp Gly Thr Val Val Ser Arg His Pro Glu Lys Ala
305 310 315 320
Val Thr Phe Val Glu Asn His Asp Thr Gln Pro Gly Gln Ser Leu Glu
325 330 335
Ser Thr Val Gln Thr Trp Phe Lys Pro Leu Ala Tyr Ala Phe Ile Leu
340 345 350
Thr Arg Glu Ser Gly Tyr Pro Gln Val Phe Tyr Gly Asp Met Tyr Gly
355 360 365
Thr Lys Gly Thr Ser Pro Lys Glu Ile Pro Ser Leu Lys Asp Asn Ile
370 375 380
Glu Pro Ile Leu Lys Ala Arg Lys Glu Tyr Ala Tyr Gly Pro Gln His
385 390 395 400
Asp Tyr Ile Asp His Pro Asp Val Ile Gly Trp Thr Arg Glu Gly Asp
405 410 415
Ser Ser Ala Ala Lys Ser Gly Leu Ala Ala Leu Ile Thr Asp Gly Pro
420 425 430
Gly Gly Ser Lys Arg Met Tyr Ala Gly Leu Lys Asn Ala Gly Glu Thr
435 440 445
Trp Tyr Asp Ile Thr Gly Asn Arg Ser Asp Thr Val Lys Ile Gly Ser
450 455 460
Asp Gly Trp Gly Glu Phe His Val Asn Asp Gly Ser Val Ser Ile Tyr
465 470 475 480
Val Gln Lys
<210> 7
<211> 397
<212> PRT
<213> 合成构建体
<400> 7
Val Asn Gly Thr Leu Met Gln Tyr Phe Glu Trp Tyr Thr Pro Asn Asp
1 5 10 15
Gly Gln His Trp Lys Arg Leu Gln Asn Asp Ala Glu His Leu Ser Asp
20 25 30
Ile Gly Ile Thr Ala Val Trp Ile Pro Pro Ala Tyr Lys Gly Leu Ser
35 40 45
Gln Ser Asp Asn Gly Tyr Gly Pro Tyr Asp Leu Tyr Asp Leu Gly Glu
50 55 60
Phe Gln Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly Thr Lys Ser Glu
65 70 75 80
Leu Gln Asp Ala Ile Gly Ser Leu His Ser Arg Asn Val Gln Val Tyr
85 90 95
Gly Asp Val Val Leu Asn His Lys Ala Gly Ala Asp Ala Thr Glu Asp
100 105 110
Val Thr Ala Val Glu Val Asn Pro Ala Asn Arg Asn Gln Glu Thr Ser
115 120 125
Glu Glu Tyr Gln Ile Lys Ala Trp Thr Asp Phe Arg Phe Pro Gly Arg
130 135 140
Gly Asn Thr Tyr Ser Asp Phe Lys Trp His Trp Tyr His Phe Asp Gly
145 150 155 160
Ala Asp Trp Asp Glu Ser Arg Lys Ile Ser Arg Ile Phe Lys Phe Arg
165 170 175
Gly Glu Gly Lys Ala Trp Asp Trp Glu Val Ser Ser Glu Asn Gly Asn
180 185 190
Tyr Asp Tyr Leu Met Tyr Ala Asp Val Asp Tyr Asp His Pro Asp Val
195 200 205
Val Ala Glu Thr Lys Lys Trp Gly Ile Trp Tyr Ala Asn Glu Leu Ser
210 215 220
Leu Asp Gly Phe Arg Ile Asp Ala Ala Lys His Ile Lys Phe Ser Phe
225 230 235 240
Leu Arg Asp Trp Val Gln Ala Val Arg Gln Ala Thr Gly Lys Glu Met
245 250 255
Phe Thr Val Ala Glu Tyr Trp Gln Asn Asn Ala Gly Lys Leu Glu Asn
260 265 270
Tyr Leu Asn Lys Thr Ser Phe Asn Gln Ser Val Phe Asp Val Pro Leu
275 280 285
His Phe Asn Leu Gln Ala Ala Ser Ser Gln Gly Gly Gly Tyr Asp Met
290 295 300
Arg Arg Leu Leu Asp Gly Thr Val Val Ser Arg His Pro Glu Lys Ala
305 310 315 320
Val Thr Phe Val Glu Asn His Asp Thr Gln Pro Gly Gln Ser Leu Glu
325 330 335
Ser Thr Val Gln Thr Trp Phe Lys Pro Leu Ala Tyr Ala Phe Ile Leu
340 345 350
Thr Arg Glu Ser Gly Tyr Pro Gln Val Phe Tyr Gly Asp Met Tyr Gly
355 360 365
Thr Lys Gly Thr Ser Pro Lys Glu Ile Pro Ser Leu Lys Asp Asn Ile
370 375 380
Glu Pro Ile Leu Lys Ala Arg Lys Glu Tyr Ala Tyr Gly
385 390 395
<210> 8
<211> 85
<212> PRT
<213> 合成构建体
<400> 8
Pro Gln His Asp Tyr Ile Asp His Pro Asp Val Ile Gly Trp Thr Arg
1 5 10 15
Glu Gly Asp Ser Ser Ala Ala Lys Ser Gly Leu Ala Ala Leu Ile Thr
20 25 30
Asp Gly Pro Gly Gly Ser Lys Arg Met Tyr Ala Gly Leu Lys Asn Ala
35 40 45
Gly Thr Trp Tyr Asp Ile Thr Gly Asn Arg Ser Asp Thr Val Lys Ile
50 55 60
Gly Ser Asp Gly Trp Gly Glu Phe His Val Asn Asp Gly Ser Val Ser
65 70 75 80
Ile Tyr Val Gln Lys
85
<210> 9
<211> 483
<212> PRT
<213> 合成构建体
<400> 9
His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr
1 5 10 15
Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Asn Ser Asp Ala Ser
20 25 30
Asn Leu Lys Ser Lys Gly Ile Thr Ala Val Trp Ile Pro Pro Ala Trp
35 40 45
Lys Gly Ala Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly
65 70 75 80
Thr Arg Ser Gln Leu Gln Ala Ala Val Thr Ser Leu Lys Asn Asn Gly
85 90 95
Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp
100 105 110
Ala Thr Glu Met Val Arg Ala Val Glu Val Asn Pro Asn Asn Arg Asn
115 120 125
Gln Glu Val Thr Gly Glu Tyr Thr Ile Glu Ala Trp Thr Arg Phe Asp
130 135 140
Phe Pro Gly Arg Gly Asn Thr His Ser Ser Phe Lys Trp Arg Trp Tyr
145 150 155 160
His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Arg Leu Asn Asn Arg
165 170 175
Ile Tyr Lys Phe Arg Gly Lys Ala Trp Asp Trp Glu Val Asp Thr Glu
180 185 190
Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Ile Asp Met Asp His
195 200 205
Pro Glu Val Val Asn Glu Leu Arg Asn Trp Gly Val Trp Tyr Thr Asn
210 215 220
Thr Leu Gly Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His Ile Lys
225 230 235 240
Tyr Ser Phe Thr Arg Asp Trp Ile Asn His Val Arg Ser Ala Thr Gly
245 250 255
Lys Asn Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu Gly Ala
260 265 270
Ile Glu Asn Tyr Leu Gln Lys Thr Asn Trp Asn His Ser Val Phe Asp
275 280 285
Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Lys Ser Gly Gly Asn
290 295 300
Tyr Asp Met Arg Asn Ile Phe Asn Gly Thr Val Val Gln Arg His Pro
305 310 315 320
Ser His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro Glu Glu
325 330 335
Ala Leu Glu Ser Phe Val Glu Glu Trp Phe Lys Pro Leu Ala Tyr Ala
340 345 350
Leu Thr Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr Gly Asp
355 360 365
Tyr Tyr Gly Ile Pro Thr His Gly Val Pro Ala Met Arg Ser Lys Ile
370 375 380
Asp Pro Ile Leu Glu Ala Arg Gln Lys Tyr Ala Tyr Gly Pro Gln His
385 390 395 400
Asp Tyr Ile Asp His Pro Asp Val Ile Gly Trp Thr Arg Glu Gly Asp
405 410 415
Ser Ser Ala Ala Lys Ser Gly Leu Ala Ala Leu Ile Thr Asp Gly Pro
420 425 430
Gly Gly Ser Lys Arg Met Tyr Ala Gly Leu Lys Asn Ala Gly Glu Thr
435 440 445
Trp Tyr Asp Ile Thr Gly Asn Arg Ser Asp Thr Val Lys Ile Gly Ser
450 455 460
Asp Gly Trp Gly Glu Phe His Val Asn Asp Gly Ser Val Ser Ile Tyr
465 470 475 480
Val Gln Lys
<210> 10
<211> 514
<212> PRT
<213> 芽孢杆菌属
<400> 10
Met Lys Gln Gln Lys Arg Leu Tyr Ala Arg Leu Leu Thr Leu Leu Phe
1 5 10 15
Ala Leu Ile Phe Leu Leu Pro His Ser Ala Ala Ala Ala His His Asn
20 25 30
Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp His Leu Pro Asn
35 40 45
Asp Gly Asn His Trp Asn Arg Leu Arg Asp Asp Ala Ser Asn Leu Arg
50 55 60
Asn Arg Gly Ile Thr Ala Ile Trp Ile Pro Pro Ala Trp Lys Gly Thr
65 70 75 80
Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr Asp Leu Gly
85 90 95
Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly Thr Arg Ser
100 105 110
Gln Leu Glu Ser Ala Ile His Ala Leu Lys Asn Asn Gly Val Gln Val
115 120 125
Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp Ala Thr Glu
130 135 140
Asn Val Leu Ala Val Glu Val Asn Pro Asn Asn Arg Asn Gln Glu Ile
145 150 155 160
Ser Gly Asp Tyr Thr Ile Glu Ala Trp Thr Lys Phe Asp Phe Pro Gly
165 170 175
Arg Gly Asn Thr Tyr Ser Asp Phe Lys Trp Arg Trp Tyr His Phe Asp
180 185 190
Gly Val Asp Trp Asp Gln Ser Arg Gln Phe Gln Asn Arg Ile Tyr Lys
195 200 205
Phe Arg Gly Asp Gly Lys Ala Trp Asp Trp Glu Val Asp Ser Glu Asn
210 215 220
Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Val Asp Met Asp His Pro
225 230 235 240
Glu Val Val Asn Glu Leu Arg Arg Trp Gly Glu Trp Tyr Thr Asn Thr
245 250 255
Leu Asn Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His Ile Lys Tyr
260 265 270
Ser Phe Thr Arg Asp Trp Leu Thr His Val Arg Asn Ala Thr Gly Lys
275 280 285
Glu Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu Gly Ala Leu
290 295 300
Glu Asn Tyr Leu Asn Lys Thr Asn Trp Asn His Ser Val Phe Asp Val
305 310 315 320
Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Asn Ser Gly Gly Asn Tyr
325 330 335
Asp Met Ala Lys Leu Leu Asn Gly Thr Val Val Gln Lys His Pro Met
340 345 350
His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro Gly Glu Ser
355 360 365
Leu Glu Ser Phe Val Gln Glu Trp Phe Lys Pro Leu Ala Tyr Ala Leu
370 375 380
Ile Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr Gly Asp Tyr
385 390 395 400
Tyr Gly Ile Pro Thr His Ser Val Pro Ala Met Lys Ala Lys Ile Asp
405 410 415
Pro Ile Leu Glu Ala Arg Gln Asn Phe Ala Tyr Gly Thr Gln His Asp
420 425 430
Tyr Phe Asp His His Asn Ile Ile Gly Trp Thr Arg Glu Gly Asn Thr
435 440 445
Thr His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp Gly Pro Gly
450 455 460
Gly Glu Lys Trp Met Tyr Val Gly Gln Asp Lys Ala Gly Gln Val Trp
465 470 475 480
His Asp Ile Thr Gly Asn Lys Pro Gly Thr Val Thr Ile Asn Ala Asp
485 490 495
Gly Trp Ala Asn Phe Ser Val Asn Gly Gly Ser Val Ser Ile Trp Val
500 505 510
Lys Arg
<210> 11
<211> 485
<212> PRT
<213> 芽孢杆菌属
<400> 11
His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp His
1 5 10 15
Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Arg Asp Asp Ala Ser
20 25 30
Asn Leu Arg Asn Arg Gly Ile Thr Ala Ile Trp Ile Pro Pro Ala Trp
35 40 45
Lys Gly Thr Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly
65 70 75 80
Thr Arg Ser Gln Leu Glu Ser Ala Ile His Ala Leu Lys Asn Asn Gly
85 90 95
Val Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp
100 105 110
Ala Thr Glu Asn Val Leu Ala Val Glu Val Asn Pro Asn Asn Arg Asn
115 120 125
Gln Glu Ile Ser Gly Asp Tyr Thr Ile Glu Ala Trp Thr Lys Phe Asp
130 135 140
Phe Pro Gly Arg Gly Asn Thr Tyr Ser Asp Phe Lys Trp Arg Trp Tyr
145 150 155 160
His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Gln Phe Gln Asn Arg
165 170 175
Ile Tyr Lys Phe Arg Gly Asp Gly Lys Ala Trp Asp Trp Glu Val Asp
180 185 190
Ser Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Val Asp Met
195 200 205
Asp His Pro Glu Val Val Asn Glu Leu Arg Arg Trp Gly Glu Trp Tyr
210 215 220
Thr Asn Thr Leu Asn Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His
225 230 235 240
Ile Lys Tyr Ser Phe Thr Arg Asp Trp Leu Thr His Val Arg Asn Ala
245 250 255
Thr Gly Lys Glu Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu
260 265 270
Gly Ala Leu Glu Asn Tyr Leu Asn Lys Thr Asn Trp Asn His Ser Val
275 280 285
Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Asn Ser Gly
290 295 300
Gly Asn Tyr Asp Met Ala Lys Leu Leu Asn Gly Thr Val Val Gln Lys
305 310 315 320
His Pro Met His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro
325 330 335
Gly Glu Ser Leu Glu Ser Phe Val Gln Glu Trp Phe Lys Pro Leu Ala
340 345 350
Tyr Ala Leu Ile Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr
355 360 365
Gly Asp Tyr Tyr Gly Ile Pro Thr His Ser Val Pro Ala Met Lys Ala
370 375 380
Lys Ile Asp Pro Ile Leu Glu Ala Arg Gln Asn Phe Ala Tyr Gly Thr
385 390 395 400
Gln His Asp Tyr Phe Asp His His Asn Ile Ile Gly Trp Thr Arg Glu
405 410 415
Gly Asn Thr Thr His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp
420 425 430
Gly Pro Gly Gly Glu Lys Trp Met Tyr Val Gly Gln Asp Lys Ala Gly
435 440 445
Gln Val Trp His Asp Ile Thr Gly Asn Lys Pro Gly Thr Val Thr Ile
450 455 460
Asn Ala Asp Gly Trp Ala Asn Phe Ser Val Asn Gly Gly Ser Val Ser
465 470 475 480
Ile Trp Val Lys Arg
485
<210> 12
<211> 483
<212> PRT
<213> 芽孢杆菌属
<400> 12
His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp His
1 5 10 15
Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Arg Asp Asp Ala Ser
20 25 30
Asn Leu Arg Asn Arg Gly Ile Thr Ala Ile Trp Ile Pro Pro Ala Trp
35 40 45
Lys Gly Thr Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly
65 70 75 80
Thr Arg Ser Gln Leu Glu Ser Ala Ile His Ala Leu Lys Asn Asn Gly
85 90 95
Val Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp
100 105 110
Ala Thr Glu Asn Val Leu Ala Val Glu Val Asn Pro Asn Asn Arg Asn
115 120 125
Gln Glu Ile Ser Gly Asp Tyr Thr Ile Glu Ala Trp Thr Lys Phe Asp
130 135 140
Phe Pro Gly Arg Gly Asn Thr Tyr Ser Asp Phe Lys Trp Arg Trp Tyr
145 150 155 160
His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Gln Phe Gln Asn Arg
165 170 175
Ile Tyr Lys Phe Arg Gly Lys Ala Trp Asp Trp Glu Val Asp Ser Glu
180 185 190
Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Val Asp Met Asp His
195 200 205
Pro Glu Val Val Asn Glu Leu Arg Arg Trp Gly Glu Trp Tyr Thr Asn
210 215 220
Thr Leu Asn Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His Ile Lys
225 230 235 240
Tyr Ser Phe Thr Arg Asp Trp Leu Thr His Val Arg Asn Ala Thr Gly
245 250 255
Lys Glu Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu Gly Ala
260 265 270
Leu Glu Asn Tyr Leu Asn Lys Thr Asn Trp Asn His Ser Val Phe Asp
275 280 285
Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Asn Ser Gly Gly Asn
290 295 300
Tyr Asp Met Ala Lys Leu Leu Asn Gly Thr Val Val Gln Lys His Pro
305 310 315 320
Met His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro Gly Glu
325 330 335
Ser Leu Glu Ser Phe Val Gln Glu Trp Phe Lys Pro Leu Ala Tyr Ala
340 345 350
Leu Ile Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr Gly Asp
355 360 365
Tyr Tyr Gly Ile Pro Thr His Ser Val Pro Ala Met Lys Ala Lys Ile
370 375 380
Asp Pro Ile Leu Glu Ala Arg Gln Asn Phe Ala Tyr Gly Thr Gln His
385 390 395 400
Asp Tyr Phe Asp His His Asn Ile Ile Gly Trp Thr Arg Glu Gly Asn
405 410 415
Thr Thr His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp Gly Pro
420 425 430
Gly Gly Glu Lys Trp Met Tyr Val Gly Gln Asp Lys Ala Gly Gln Val
435 440 445
Trp His Asp Ile Thr Gly Asn Lys Pro Gly Thr Val Thr Ile Asn Ala
450 455 460
Asp Gly Trp Ala Asn Phe Ser Val Asn Gly Gly Ser Val Ser Ile Trp
465 470 475 480
Val Lys Arg
<210> 13
<211> 513
<212> PRT
<213> 脂环酸芽孢杆菌属-FL
<400> 13
Met Lys Ile Arg Asn Gly Trp Lys Lys Thr Leu Thr Leu Leu Phe Ala
1 5 10 15
Leu Ile Phe Leu Leu Pro His Ser Ala Ala Ala Thr Phe Ala Gly Asp
20 25 30
Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr Leu Pro Asn Asp Gly
35 40 45
Thr Leu Trp Thr Lys Met Gly Ser Asp Ala Ser His Leu Lys Ser Ile
50 55 60
Gly Ile Thr Gly Val Trp Phe Pro Pro Ala Tyr Lys Gly Gln Ser Gln
65 70 75 80
Ser Asp Val Gly Tyr Gly Val Tyr Asp Met Tyr Asp Leu Gly Glu Phe
85 90 95
Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly Thr Lys Ala Gln Leu
100 105 110
Gln Ser Ala Ile Thr Ser Leu His Asn Asn Gly Ile Gln Ala Tyr Gly
115 120 125
Asp Val Val Leu Asn His Arg Met Gly Ala Asp Ala Thr Glu Thr Ile
130 135 140
Ser Ala Val Glu Val Asn Pro Ser Asn Arg Asn Gln Val Thr Ser Gly
145 150 155 160
Ala Tyr Asn Ile Ser Ala Trp Thr Asp Phe Glu Phe Pro Gly Arg Gly
165 170 175
Asn Thr Tyr Ser Ser Phe Lys Trp His Ser Tyr Tyr Phe Asp Gly Val
180 185 190
Asp Trp Asp Gln Ser Arg Gln Leu Ser Gly Lys Ile Tyr Gln Ile Gln
195 200 205
Gly Thr Gly Lys Ala Trp Asp Trp Glu Val Asp Ser Glu Asn Gly Asn
210 215 220
Tyr Asp Tyr Leu Met Gly Ala Asp Ile Asp Tyr Asp His Pro Asp Val
225 230 235 240
Gln Thr Glu Val Lys Asn Trp Gly Lys Trp Phe Val Asn Thr Leu Asn
245 250 255
Leu Asp Gly Val Arg Leu Asp Ala Val Lys His Ile Lys Phe Asp Tyr
260 265 270
Met Ser Ser Trp Leu Ser Ser Val Lys Ser Thr Thr Gly Lys Ser Asn
275 280 285
Leu Phe Ala Val Gly Glu Tyr Trp Asn Thr Ser Leu Gly Ala Leu Glu
290 295 300
Asn Tyr Glu Asn Lys Thr Asn Trp Ser Met Ser Leu Phe Asp Val Pro
305 310 315 320
Leu His Met Asn Phe Gln Ala Ala Ala Asn Gly Gly Gly Tyr Tyr Asp
325 330 335
Met Arg Asn Leu Leu Asn Asn Thr Met Met Lys Asn His Pro Ile Gln
340 345 350
Ala Val Thr Phe Val Asp Asn His Asp Thr Glu Pro Gly Gln Ala Leu
355 360 365
Gln Ser Trp Val Ser Asp Trp Phe Lys Pro Leu Ala Tyr Ala Thr Ile
370 375 380
Leu Thr Arg Gln Glu Gly Tyr Pro Cys Val Phe Tyr Gly Asp Tyr Tyr
385 390 395 400
Gly Ile Pro Ser Gln Ser Val Ser Ala Lys Ser Thr Trp Leu Asp Lys
405 410 415
Gln Leu Ser Ala Arg Lys Ser Tyr Ala Tyr Gly Thr Gln His Asp Tyr
420 425 430
Leu Asp Asn Gln Asp Val Ile Gly Trp Thr Arg Glu Gly Asp Ser Ala
435 440 445
His Ala Gly Ser Gly Leu Ala Thr Val Met Ser Asp Gly Pro Gly Gly
450 455 460
Ser Lys Thr Met Tyr Val Gly Thr Ala His Ala Gly Gln Val Phe Lys
465 470 475 480
Asp Ile Thr Gly Asn Arg Thr Asp Thr Val Thr Ile Asn Ser Ala Gly
485 490 495
Asn Gly Thr Phe Pro Cys Asn Gly Gly Ser Val Ser Ile Trp Val Lys
500 505 510
Gln
<210> 14
<211> 486
<212> PRT
<213> 脂环酸芽孢杆菌属物种
<400> 14
Thr Phe Ala Gly Asp Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr
1 5 10 15
Leu Pro Asn Asp Gly Thr Leu Trp Thr Lys Met Gly Ser Asp Ala Ser
20 25 30
His Leu Lys Ser Ile Gly Ile Thr Gly Val Trp Phe Pro Pro Ala Tyr
35 40 45
Lys Gly Gln Ser Gln Ser Asp Val Gly Tyr Gly Val Tyr Asp Met Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly
65 70 75 80
Thr Lys Ala Gln Leu Gln Ser Ala Ile Thr Ser Leu His Asn Asn Gly
85 90 95
Ile Gln Ala Tyr Gly Asp Val Val Leu Asn His Arg Met Gly Ala Asp
100 105 110
Ala Thr Glu Thr Ile Ser Ala Val Glu Val Asn Pro Ser Asn Arg Asn
115 120 125
Gln Val Thr Ser Gly Ala Tyr Asn Ile Ser Ala Trp Thr Asp Phe Glu
130 135 140
Phe Pro Gly Arg Gly Asn Thr Tyr Ser Ser Phe Lys Trp His Ser Tyr
145 150 155 160
Tyr Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Gln Leu Ser Gly Lys
165 170 175
Ile Tyr Gln Ile Gln Gly Thr Gly Lys Ala Trp Asp Trp Glu Val Asp
180 185 190
Ser Glu Asn Gly Asn Tyr Asp Tyr Leu Met Gly Ala Asp Ile Asp Tyr
195 200 205
Asp His Pro Asp Val Gln Thr Glu Val Lys Asn Trp Gly Lys Trp Phe
210 215 220
Val Asn Thr Leu Asn Leu Asp Gly Val Arg Leu Asp Ala Val Lys His
225 230 235 240
Ile Lys Phe Asp Tyr Met Ser Ser Trp Leu Ser Ser Val Lys Ser Thr
245 250 255
Thr Gly Lys Ser Asn Leu Phe Ala Val Gly Glu Tyr Trp Asn Thr Ser
260 265 270
Leu Gly Ala Leu Glu Asn Tyr Glu Asn Lys Thr Asn Trp Ser Met Ser
275 280 285
Leu Phe Asp Val Pro Leu His Met Asn Phe Gln Ala Ala Ala Asn Gly
290 295 300
Gly Gly Tyr Tyr Asp Met Arg Asn Leu Leu Asn Asn Thr Met Met Lys
305 310 315 320
Asn His Pro Ile Gln Ala Val Thr Phe Val Asp Asn His Asp Thr Glu
325 330 335
Pro Gly Gln Ala Leu Gln Ser Trp Val Ser Asp Trp Phe Lys Pro Leu
340 345 350
Ala Tyr Ala Thr Ile Leu Thr Arg Gln Glu Gly Tyr Pro Cys Val Phe
355 360 365
Tyr Gly Asp Tyr Tyr Gly Ile Pro Ser Gln Ser Val Ser Ala Lys Ser
370 375 380
Thr Trp Leu Asp Lys Gln Leu Ser Ala Arg Lys Ser Tyr Ala Tyr Gly
385 390 395 400
Thr Gln His Asp Tyr Leu Asp Asn Gln Asp Val Ile Gly Trp Thr Arg
405 410 415
Glu Gly Asp Ser Ala His Ala Gly Ser Gly Leu Ala Thr Val Met Ser
420 425 430
Asp Gly Pro Gly Gly Ser Lys Thr Met Tyr Val Gly Thr Ala His Ala
435 440 445
Gly Gln Val Phe Lys Asp Ile Thr Gly Asn Arg Thr Asp Thr Val Thr
450 455 460
Ile Asn Ser Ala Gly Asn Gly Thr Phe Pro Cys Asn Gly Gly Ser Val
465 470 475 480
Ser Ile Trp Val Lys Gln
485
<210> 15
<211> 485
<212> PRT
<213> 芽孢杆菌属
<400> 15
His His Asp Gly Thr Asn Gly Thr Ile Met Gln Tyr Phe Glu Trp Asn
1 5 10 15
Val Pro Asn Asp Gly Gln His Trp Asn Arg Leu His Asn Asn Ala Gln
20 25 30
Asn Leu Lys Asn Ala Gly Ile Thr Ala Ile Trp Ile Pro Pro Ala Trp
35 40 45
Lys Gly Thr Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly
65 70 75 80
Thr Lys Ala Glu Leu Glu Arg Ala Ile Arg Ser Leu Lys Ala Asn Gly
85 90 95
Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp
100 105 110
Phe Thr Glu Arg Val Gln Ala Val Glu Val Asn Pro Gln Asn Arg Asn
115 120 125
Gln Glu Val Ser Gly Thr Tyr Gln Ile Glu Ala Trp Thr Gly Phe Asn
130 135 140
Phe Pro Gly Arg Gly Asn Gln His Ser Ser Phe Lys Trp Arg Trp Tyr
145 150 155 160
His Phe Asp Gly Thr Asp Trp Asp Gln Ser Arg Gln Leu Ala Asn Arg
165 170 175
Ile Tyr Lys Phe Arg Gly Asp Gly Lys Ala Trp Asp Trp Glu Val Asp
180 185 190
Thr Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Val Asp Met
195 200 205
Asp His Pro Glu Val Ile Asn Glu Leu Asn Arg Trp Gly Val Trp Tyr
210 215 220
Ala Asn Thr Leu Asn Leu Asp Gly Phe Arg Leu Asp Ala Val Lys His
225 230 235 240
Ile Lys Phe Ser Phe Met Arg Asp Trp Leu Gly His Val Arg Gly Gln
245 250 255
Thr Gly Lys Asn Leu Phe Ala Val Ala Glu Tyr Trp Lys Asn Asp Leu
260 265 270
Gly Ala Leu Glu Asn Tyr Leu Ser Lys Thr Asn Trp Thr Met Ser Ala
275 280 285
Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Gln Ala Ser Asn Ser Ser
290 295 300
Gly Asn Tyr Asp Met Arg Asn Leu Leu Asn Gly Thr Leu Val Gln Arg
305 310 315 320
His Pro Ser His Ala Val Thr Phe Val Asp Asn His Asp Thr Gln Pro
325 330 335
Gly Glu Ala Leu Glu Ser Phe Val Gln Gly Trp Phe Lys Pro Leu Ala
340 345 350
Tyr Ala Thr Ile Leu Thr Arg Glu Gln Gly Tyr Pro Gln Val Phe Tyr
355 360 365
Gly Asp Tyr Tyr Gly Ile Pro Ser Asp Gly Val Pro Ser Tyr Arg Gln
370 375 380
Gln Ile Asp Pro Leu Leu Lys Ala Arg Gln Gln Tyr Ala Tyr Gly Arg
385 390 395 400
Gln His Asp Tyr Phe Asp His Trp Asp Val Ile Gly Trp Thr Arg Glu
405 410 415
Gly Asn Ala Ser His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp
420 425 430
Gly Pro Gly Gly Ser Lys Trp Met Tyr Val Gly Arg Gln Lys Ala Gly
435 440 445
Glu Val Trp His Asp Met Thr Gly Asn Arg Ser Gly Thr Val Thr Ile
450 455 460
Asn Gln Asp Gly Trp Gly His Phe Phe Val Asn Gly Gly Ser Val Ser
465 470 475 480
Val Trp Val Lys Arg
485
<210> 16
<211> 483
<212> PRT
<213> 芽孢杆菌属
<400> 16
His His Asp Gly Thr Asn Gly Thr Ile Met Gln Tyr Phe Glu Trp Asn
1 5 10 15
Val Pro Asn Asp Gly Gln His Trp Asn Arg Leu His Asn Asn Ala Gln
20 25 30
Asn Leu Lys Asn Ala Gly Ile Thr Ala Ile Trp Ile Pro Pro Ala Trp
35 40 45
Lys Gly Thr Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly
65 70 75 80
Thr Lys Ala Glu Leu Glu Arg Ala Ile Arg Ser Leu Lys Ala Asn Gly
85 90 95
Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp
100 105 110
Phe Thr Glu Arg Val Gln Ala Val Glu Val Asn Pro Gln Asn Arg Asn
115 120 125
Gln Glu Val Ser Gly Thr Tyr Gln Ile Glu Ala Trp Thr Gly Phe Asn
130 135 140
Phe Pro Gly Arg Gly Asn Gln His Ser Ser Phe Lys Trp Arg Trp Tyr
145 150 155 160
His Phe Asp Gly Thr Asp Trp Asp Gln Ser Arg Gln Leu Ala Asn Arg
165 170 175
Ile Tyr Lys Phe Arg Gly Lys Ala Trp Asp Trp Glu Val Asp Thr Glu
180 185 190
Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Val Asp Met Asp His
195 200 205
Pro Glu Val Ile Asn Glu Leu Asn Arg Trp Gly Val Trp Tyr Ala Asn
210 215 220
Thr Leu Asn Leu Asp Gly Phe Arg Leu Asp Ala Val Lys His Ile Lys
225 230 235 240
Phe Ser Phe Met Arg Asp Trp Leu Gly His Val Arg Gly Gln Thr Gly
245 250 255
Lys Asn Leu Phe Ala Val Ala Glu Tyr Trp Lys Asn Asp Leu Gly Ala
260 265 270
Leu Glu Asn Tyr Leu Ser Lys Thr Asn Trp Thr Met Ser Ala Phe Asp
275 280 285
Val Pro Leu His Tyr Asn Leu Tyr Gln Ala Ser Asn Ser Ser Gly Asn
290 295 300
Tyr Asp Met Arg Asn Leu Leu Asn Gly Thr Leu Val Gln Arg His Pro
305 310 315 320
Ser His Ala Val Thr Phe Val Asp Asn His Asp Thr Gln Pro Gly Glu
325 330 335
Ala Leu Glu Ser Phe Val Gln Gly Trp Phe Lys Pro Leu Ala Tyr Ala
340 345 350
Thr Ile Leu Thr Arg Glu Gln Gly Tyr Pro Gln Val Phe Tyr Gly Asp
355 360 365
Tyr Tyr Gly Ile Pro Ser Asp Gly Val Pro Ser Tyr Arg Gln Gln Ile
370 375 380
Asp Pro Leu Leu Lys Ala Arg Gln Gln Tyr Ala Tyr Gly Arg Gln His
385 390 395 400
Asp Tyr Phe Asp His Trp Asp Val Ile Gly Trp Thr Arg Glu Gly Asn
405 410 415
Ala Ser His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp Gly Pro
420 425 430
Gly Gly Ser Lys Trp Met Tyr Val Gly Arg Gln Lys Ala Gly Glu Val
435 440 445
Trp His Asp Met Thr Gly Asn Arg Ser Gly Thr Val Thr Ile Asn Gln
450 455 460
Asp Gly Trp Gly His Phe Phe Val Asn Gly Gly Ser Val Ser Val Trp
465 470 475 480
Val Lys Arg
<210> 17
<211> 399
<212> PRT
<213> 芽孢杆菌属
<400> 17
His His Asp Gly Thr Asn Gly Thr Ile Met Gln Tyr Phe Glu Trp Asn
1 5 10 15
Val Pro Asn Asp Gly Gln His Trp Asn Arg Leu His Asn Asn Ala Gln
20 25 30
Asn Leu Lys Asn Ala Gly Ile Thr Ala Ile Trp Ile Pro Pro Ala Trp
35 40 45
Lys Gly Thr Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly
65 70 75 80
Thr Lys Ala Glu Leu Glu Arg Ala Ile Arg Ser Leu Lys Ala Asn Gly
85 90 95
Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp
100 105 110
Phe Thr Glu Arg Val Gln Ala Val Glu Val Asn Pro Gln Asn Arg Asn
115 120 125
Gln Glu Val Ser Gly Thr Tyr Gln Ile Glu Ala Trp Thr Gly Phe Asn
130 135 140
Phe Pro Gly Arg Gly Asn Gln His Ser Ser Phe Lys Trp Arg Trp Tyr
145 150 155 160
His Phe Asp Gly Thr Asp Trp Asp Gln Ser Arg Gln Leu Ala Asn Arg
165 170 175
Ile Tyr Lys Phe Arg Gly Asp Gly Lys Ala Trp Asp Trp Glu Val Asp
180 185 190
Thr Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Val Asp Met
195 200 205
Asp His Pro Glu Val Ile Asn Glu Leu Asn Arg Trp Gly Val Trp Tyr
210 215 220
Ala Asn Thr Leu Asn Leu Asp Gly Phe Arg Leu Asp Ala Val Lys His
225 230 235 240
Ile Lys Phe Ser Phe Met Arg Asp Trp Leu Gly His Val Arg Gly Gln
245 250 255
Thr Gly Lys Asn Leu Phe Ala Val Ala Glu Tyr Trp Lys Asn Asp Leu
260 265 270
Gly Ala Leu Glu Asn Tyr Leu Ser Lys Thr Asn Trp Thr Met Ser Ala
275 280 285
Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Gln Ala Ser Asn Ser Ser
290 295 300
Gly Asn Tyr Asp Met Arg Asn Leu Leu Asn Gly Thr Leu Val Gln Arg
305 310 315 320
His Pro Ser His Ala Val Thr Phe Val Asp Asn His Asp Thr Gln Pro
325 330 335
Gly Glu Ala Leu Glu Ser Phe Val Gln Gly Trp Phe Lys Pro Leu Ala
340 345 350
Tyr Ala Thr Ile Leu Thr Arg Glu Gln Gly Tyr Pro Gln Val Phe Tyr
355 360 365
Gly Asp Tyr Tyr Gly Ile Pro Ser Asp Gly Val Pro Ser Tyr Arg Gln
370 375 380
Gln Ile Asp Pro Leu Leu Lys Ala Arg Gln Gln Tyr Ala Tyr Gly
385 390 395
<210> 18
<211> 86
<212> PRT
<213> 脂环酸芽孢杆菌属
<400> 18
Thr Gln His Asp Tyr Leu Asp Asn Gln Asp Val Ile Gly Trp Thr Arg
1 5 10 15
Glu Gly Asp Ser Ala His Ala Gly Ser Gly Leu Ala Thr Val Met Ser
20 25 30
Asp Gly Pro Gly Gly Ser Lys Thr Met Tyr Val Gly Thr Ala His Ala
35 40 45
Gly Gln Val Phe Lys Asp Ile Thr Gly Asn Arg Thr Asp Thr Val Thr
50 55 60
Ile Asn Ser Ala Gly Asn Gly Thr Phe Pro Cys Asn Gly Gly Ser Val
65 70 75 80
Ser Ile Trp Val Lys Gln
85
<210> 19
<211> 485
<212> PRT
<213> 合成构建体
<400> 19
His His Asp Gly Thr Asn Gly Thr Ile Met Gln Tyr Phe Glu Trp Asn
1 5 10 15
Val Pro Asn Asp Gly Gln His Trp Asn Arg Leu His Asn Asn Ala Gln
20 25 30
Asn Leu Lys Asn Ala Gly Ile Thr Ala Ile Trp Ile Pro Pro Ala Trp
35 40 45
Lys Gly Thr Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr
50 55 60
Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly
65 70 75 80
Thr Lys Ala Glu Leu Glu Arg Ala Ile Arg Ser Leu Lys Ala Asn Gly
85 90 95
Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp
100 105 110
Phe Thr Glu Arg Val Gln Ala Val Glu Val Asn Pro Gln Asn Arg Asn
115 120 125
Gln Glu Val Ser Gly Thr Tyr Gln Ile Glu Ala Trp Thr Gly Phe Asn
130 135 140
Phe Pro Gly Arg Gly Asn Gln His Ser Ser Phe Lys Trp Arg Trp Tyr
145 150 155 160
His Phe Asp Gly Thr Asp Trp Asp Gln Ser Arg Gln Leu Ala Asn Arg
165 170 175
Ile Tyr Lys Phe Arg Gly Asp Gly Lys Ala Trp Asp Trp Glu Val Asp
180 185 190
Thr Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Val Asp Met
195 200 205
Asp His Pro Glu Val Ile Asn Glu Leu Asn Arg Trp Gly Val Trp Tyr
210 215 220
Ala Asn Thr Leu Asn Leu Asp Gly Phe Arg Leu Asp Ala Val Lys His
225 230 235 240
Ile Lys Phe Ser Phe Met Arg Asp Trp Leu Gly His Val Arg Gly Gln
245 250 255
Thr Gly Lys Asn Leu Phe Ala Val Ala Glu Tyr Trp Lys Asn Asp Leu
260 265 270
Gly Ala Leu Glu Asn Tyr Leu Ser Lys Thr Asn Trp Thr Met Ser Ala
275 280 285
Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Gln Ala Ser Asn Ser Ser
290 295 300
Gly Asn Tyr Asp Met Arg Asn Leu Leu Asn Gly Thr Leu Val Gln Arg
305 310 315 320
His Pro Ser His Ala Val Thr Phe Val Asp Asn His Asp Thr Gln Pro
325 330 335
Gly Glu Ala Leu Glu Ser Phe Val Gln Gly Trp Phe Lys Pro Leu Ala
340 345 350
Tyr Ala Thr Ile Leu Thr Arg Glu Gln Gly Tyr Pro Gln Val Phe Tyr
355 360 365
Gly Asp Tyr Tyr Gly Ile Pro Ser Asp Gly Val Pro Ser Tyr Arg Gln
370 375 380
Gln Ile Asp Pro Leu Leu Lys Ala Arg Gln Gln Tyr Ala Tyr Gly Thr
385 390 395 400
Gln His Asp Tyr Leu Asp Asn Gln Asp Val Ile Gly Trp Thr Arg Glu
405 410 415
Gly Asp Ser Ala His Ala Gly Ser Gly Leu Ala Thr Val Met Ser Asp
420 425 430
Gly Pro Gly Gly Ser Lys Thr Met Tyr Val Gly Thr Ala His Ala Gly
435 440 445
Gln Val Phe Lys Asp Ile Thr Gly Asn Arg Thr Asp Thr Val Thr Ile
450 455 460
Asn Ser Ala Gly Asn Gly Thr Phe Pro Cys Asn Gly Gly Ser Val Ser
465 470 475 480
Ile Trp Val Lys Gln
485
<210> 20
<211> 269
<212> PRT
<213> 迟缓芽孢杆菌
<400> 20
Ala Gln Ser Val Pro Trp Gly Ile Ser Arg Val Gln Ala Pro Ala Ala
1 5 10 15
His Asn Arg Gly Leu Thr Gly Ser Gly Val Lys Val Ala Val Leu Asp
20 25 30
Thr Gly Ile Ser Thr His Pro Asp Leu Asn Ile Arg Gly Gly Ala Ser
35 40 45
Phe Val Pro Gly Glu Pro Ser Thr Gln Asp Gly Asn Gly His Gly Thr
50 55 60
His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu
65 70 75 80
Gly Val Ala Pro Ser Ala Glu Leu Tyr Ala Val Lys Val Leu Gly Ala
85 90 95
Ser Gly Ser Gly Ser Val Ser Ser Ile Ala Gln Gly Leu Glu Trp Ala
100 105 110
Gly Asn Asn Gly Met His Val Ala Asn Leu Ser Leu Gly Ser Pro Ser
115 120 125
Pro Ser Ala Thr Leu Glu Gln Ala Val Asn Ser Ala Thr Ser Arg Gly
130 135 140
Val Leu Val Val Ala Ala Ser Gly Asn Ser Gly Ala Gly Ser Ile Ser
145 150 155 160
Tyr Pro Ala Arg Tyr Ala Asn Ala Met Ala Val Gly Ala Thr Asp Gln
165 170 175
Asn Asn Asn Arg Ala Ser Phe Ser Gln Tyr Gly Ala Gly Leu Asp Ile
180 185 190
Val Ala Pro Gly Val Asn Val Gln Ser Thr Tyr Pro Gly Ser Thr Tyr
195 200 205
Ala Ser Leu Asn Gly Thr Ser Met Ala Thr Pro His Val Ala Gly Ala
210 215 220
Ala Ala Leu Val Lys Gln Lys Asn Pro Ser Trp Ser Asn Val Gln Ile
225 230 235 240
Arg Asn His Leu Lys Asn Thr Ala Thr Ser Leu Gly Ser Thr Asn Leu
245 250 255
Tyr Gly Ser Gly Leu Val Asn Ala Glu Ala Ala Thr Arg
260 265
<210> 21
<211> 275
<212> PRT
<213> 解淀粉芽孢杆菌
<400> 21
Ala Gln Ser Val Pro Tyr Gly Val Ser Gln Ile Lys Ala Pro Ala Leu
1 5 10 15
His Ser Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp
20 25 30
Ser Gly Ile Asp Ser Ser His Pro Asp Leu Lys Val Ala Gly Gly Ala
35 40 45
Ser Met Val Pro Ser Glu Thr Asn Pro Phe Gln Asp Asn Asn Ser His
50 55 60
Gly Thr His Val Ala Gly Thr Val Ala Ala Leu Asn Asn Ser Ile Gly
65 70 75 80
Val Leu Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu
85 90 95
Gly Ala Asp Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu
100 105 110
Trp Ala Ile Ala Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly
115 120 125
Pro Ser Gly Ser Ala Ala Leu Lys Ala Ala Val Asp Lys Ala Val Ala
130 135 140
Ser Gly Val Val Val Val Ala Ala Ala Gly Asn Glu Gly Thr Ser Gly
145 150 155 160
Ser Ser Ser Thr Val Gly Tyr Pro Gly Lys Tyr Pro Ser Val Ile Ala
165 170 175
Val Gly Ala Val Asp Ser Ser Asn Gln Arg Ala Ser Phe Ser Ser Val
180 185 190
Gly Pro Glu Leu Asp Val Met Ala Pro Gly Val Ser Ile Gln Ser Thr
195 200 205
Leu Pro Gly Asn Lys Tyr Gly Ala Tyr Asn Gly Thr Ser Met Ala Ser
210 215 220
Pro His Val Ala Gly Ala Ala Ala Leu Ile Leu Ser Lys His Pro Asn
225 230 235 240
Trp Thr Asn Thr Gln Val Arg Ser Ser Leu Glu Asn Thr Thr Thr Lys
245 250 255
Leu Gly Asp Ser Phe Tyr Tyr Gly Lys Gly Leu Ile Asn Val Gln Ala
260 265 270
Ala Ala Gln
275
<210> 22
<211> 311
<212> PRT
<213> 芽孢杆菌属
<400> 22
Ala Val Pro Ser Thr Gln Thr Pro Trp Gly Ile Lys Ser Ile Tyr Asn
1 5 10 15
Asp Gln Ser Ile Thr Lys Thr Thr Gly Gly Ser Gly Ile Lys Val Ala
20 25 30
Val Leu Asp Thr Gly Val Tyr Thr Ser His Leu Asp Leu Ala Gly Ser
35 40 45
Ala Glu Gln Cys Lys Asp Phe Thr Gln Ser Asn Pro Leu Val Asp Gly
50 55 60
Ser Cys Thr Asp Arg Gln Gly His Gly Thr His Val Ala Gly Thr Val
65 70 75 80
Leu Ala His Gly Gly Ser Asn Gly Gln Gly Val Tyr Gly Val Ala Pro
85 90 95
Gln Ala Lys Leu Trp Ala Tyr Lys Val Leu Gly Asp Asn Gly Ser Gly
100 105 110
Tyr Ser Asp Asp Ile Ala Ala Ala Ile Arg His Val Ala Asp Glu Ala
115 120 125
Ser Arg Thr Gly Ser Lys Val Val Ile Asn Met Ser Leu Gly Ser Ser
130 135 140
Ala Lys Asp Ser Leu Ile Ala Ser Ala Val Asp Tyr Ala Tyr Gly Lys
145 150 155 160
Gly Val Leu Ile Val Ala Ala Ala Gly Asn Ser Gly Ser Gly Ser Asn
165 170 175
Thr Ile Gly Phe Pro Gly Gly Leu Val Asn Ala Val Ala Val Ala Ala
180 185 190
Leu Glu Asn Val Gln Gln Asn Gly Thr Tyr Arg Val Ala Asp Phe Ser
195 200 205
Ser Arg Gly Asn Pro Ala Thr Ala Gly Asp Tyr Ile Ile Gln Glu Arg
210 215 220
Asp Ile Glu Val Ser Ala Pro Gly Ala Ser Val Glu Ser Thr Trp Tyr
225 230 235 240
Thr Gly Gly Tyr Asn Thr Ile Ser Gly Thr Ser Met Ala Thr Pro His
245 250 255
Val Ala Gly Leu Ala Ala Lys Ile Trp Ser Ala Asn Thr Ser Leu Ser
260 265 270
His Ser Gln Leu Arg Thr Glu Leu Gln Asn Arg Ala Lys Val Tyr Asp
275 280 285
Ile Lys Gly Gly Ile Gly Ala Gly Thr Gly Asp Asp Tyr Ala Ser Gly
290 295 300
Phe Gly Tyr Pro Arg Val Lys
305 310
Claims (15)
1.一种粉末洗涤剂组合物，该组合物包含具有α-淀粉酶活性并且与SEQ ID NO:1-19具有至少60％，如65％、如70％、如75％、如80％、如85％、如90％、如95％、如97％序列同一性的至少一种多肽，其中该组合物的pH值低于9.5，其中pH是在20℃、在该组合物在去离子水中的5g/l溶液中确定的。
2.根据权利要求1所述的组合物，其中该pH是在7.0-9.5的范围内；如在7.5至9.0的范围内；如在8.0至9.0的范围内。
3.根据前述权利要求中任一项所述的组合物，其中该具有α-淀粉酶活性的多肽与SEQID NO:1-19具有至少60％、至少65％、至少70％、至少75％、至少80％、至少85％、至少90％或至少95％但少于100％的序列同一性。
4.根据前述权利要求中任一项所述的组合物，其中该组合物还包含一种或多种洗涤剂组分。
5.根据前述权利要求中任一项所述的组合物，其中洗涤剂组分选自下组，该组由以下组成：表面活性剂、水溶助剂、助洗剂、共助洗剂、螯合剂或螯合试剂、漂白系统或漂白组分、聚合物、织物调色剂、织物调理剂、增泡剂、抑泡剂、分散剂、染料转移抑制剂、荧光增白剂、香料、光学增亮剂、杀细菌剂、杀真菌剂、污垢悬浮剂、污垢释放聚合物、抗再沉积剂、酶抑制剂或稳定剂、酶活化剂、抗氧化剂以及增溶剂。
6.根据前述权利要求中任一项所述的组合物，其中该助洗剂选自下组，该组由以下组成：磷酸盐、柠檬酸钠助洗剂、碳酸钠、硅酸钠、钠和沸石。
7.根据前述权利要求中任一项所述的组合物，其中该表面活性剂是阴离子的和/或非离子的。
8.根据权利要求7所述的组合物，其中该阴离子表面活性剂选自：直链烷基苯磺酸(LAS)的LAS异构体、醇醚硫酸盐(AEO、AEOS)和月桂基醚硫酸钠和月桂醇聚醚硫酸钠(SLES)。
9.根据权利要求7所述的组合物，其中该非离子表面活性剂选自：醇乙氧基化物(AE或AEO)、醇丙氧基化物、醇丙氧基化物、丙氧基化的脂肪醇(PFA)、烷氧基化的脂肪酸烷基酯(例如乙氧基化的和/或丙氧基化的脂肪酸烷基酯)、烷基酚乙氧基化物(APE)、壬基酚乙氧基化物(NPE)、烷基多糖苷(APG)、烷氧基化胺、脂肪酸单乙醇酰胺(FAM)、脂肪酸二乙醇酰胺(FADA)、乙氧基化的脂肪酸单乙醇酰胺(EFAM)、丙氧基化的脂肪酸单乙醇酰胺(PFAM)、多羟基烷基脂肪酸酰胺、葡糖胺的N-酰基N-烷基衍生物(葡糖酰胺(GA)、或脂肪酸葡糖酰胺(FAGA))及其组合。
10.根据前述权利要求中任一项所述的组合物，该组合物还包含选自下组的一种或多种另外的酶，该组由以下组成：蛋白酶、脂肪酶、角质酶、α-淀粉酶、糖酶、纤维素酶、果胶酶、甘露聚糖酶、β-淀粉酶、支链淀粉酶、过水解酶、磷脂酶阿拉伯糖酶、半乳聚糖酶、木聚糖酶、果胶裂解酶、半乳糖醛酸酶、半纤维素酶、木葡聚糖酶、核酸酶、卵磷脂酶、氧化酶及其混合物。
11.根据前述权利要求中任一项所述的组合物，该组合物是衣物洗涤或餐具洗涤组合物。
12.根据权利要求11所述的组合物，其中该餐具洗涤组合物是自动餐具洗涤(ADW)洗涤剂组合物、皂条或粉末餐具洗涤组合物，例如ADW单位剂量洗涤剂组合物以及例如手动餐具洗涤(HDW)洗涤剂组合物。
13.根据权利要求11中任一项所述的组合物，其中该组合物是单位剂型，如洗涤块或小袋。
14.根据权利要求1-13中任一项所述的组合物在清洁过程如衣物洗涤或包括餐具洗涤和工业清洁的硬表面清洁中的用途。
15.一种清洁方法，特别是用于清洁织物或纺织品，或用于餐具洗涤，该方法包括在适于清洁织物/纺织品或餐具的条件下使织物/纺织品或餐具与根据权利要求1-12中任一项所述的洗涤剂组合物接触。
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| EP18209710.5 | 2018-12-03 | ||
| EP18209710 | 2018-12-03 | ||
| EP18209711 | 2018-12-03 | ||
| EP18209714.7 | 2018-12-03 | ||
| EP18209711.3 | 2018-12-03 | ||
| EP18209714 | 2018-12-03 | ||
| EP19192257 | 2019-08-19 | ||
| EP19192257.4 | 2019-08-19 | ||
| PCT/EP2019/083304 WO2020114965A1 (en) | 2018-12-03 | 2019-12-02 | LOW pH POWDER DETERGENT COMPOSITION |
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| EP (1) | EP3891264A1 (zh) |
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Also Published As
| Publication number | Publication date |
|---|---|
| EP3891264A1 (en) | 2021-10-13 |
| US20220017844A1 (en) | 2022-01-20 |
| WO2020114965A1 (en) | 2020-06-11 |
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