CN101646760B - 稳定的酶溶液和制造方法 - Google Patents
稳定的酶溶液和制造方法 Download PDFInfo
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- CN101646760B CN101646760B CN200880010193.1A CN200880010193A CN101646760B CN 101646760 B CN101646760 B CN 101646760B CN 200880010193 A CN200880010193 A CN 200880010193A CN 101646760 B CN101646760 B CN 101646760B
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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
- 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
- C11D3/38663—Stabilised liquid enzyme 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
- C11D3/02—Inorganic compounds ; Elemental compounds
- C11D3/04—Water-soluble compounds
- C11D3/046—Salts
-
- 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
- C11D3/02—Inorganic compounds ; Elemental compounds
- C11D3/04—Water-soluble compounds
- C11D3/046—Salts
- C11D3/048—Nitrates or nitrites
-
- 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
- C11D3/166—Organic compounds containing borium
-
- 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
- C11D3/20—Organic compounds containing oxygen
- C11D3/2075—Carboxylic acids-salts thereof
Abstract
本发明涉及在包含于液体洗涤剂组合物中的酶的储存过程中的稳定化。
Description
发明领域
本文公开内容涉及包含或包括酶、抑制剂和抑制剂加强剂(inhibitorbooster)的液体组合物。
背景
在含酶液体如含酶液体洗涤剂中的储存稳定性问题是公知的。这点在含蛋白酶的液体洗涤剂中尤是如此。
现有技术已经广泛地致力于改进储存稳定性,例如通过加入蛋白酶抑制剂来进行。
已知硼酸和代硼酸(boronic acid)可逆地抑制蛋白水解酶。关于代硼酸对一种丝氨酸蛋白酶枯草溶菌素(subtilisin)的抑制的讨论在Molecular &Cellular Biochemistry 51,1983,第5-32页中提供。
代硼酸具有非常不同的作为枯草溶菌素抑制剂的能力。仅含烷基如甲基、丁基或2-环己基乙基的代硼酸是弱抑制剂,其中甲基硼酸是最弱的抑制剂,而带有芳香基如苯基、4-甲氧苯基或3,5-二氯苯基的代硼酸是良好的抑制剂,其中3,5-二氯苯基硼酸是尤其有效的一种(参见Keller等,Biochem.Biophys.Res.Com.176,1991,第401-405页)。
还已知在相对于硼的3-位具有取代的芳基硼酸是可逆性蛋白酶抑制剂。在WO 92/19707中,乙酰氨基苯基硼酸(acetamidophenyl boronic acid)作为蛋白水解酶抑制剂得到描述。
另外EP 0 832 174描述了接近苯基硼酸(adjacent to the phenyl boronicacid)在对位用>C=O取代的苯基硼酸衍生物在液体中具有良好的作为酶稳定剂的能力。
在配制、制造和包装包括敏感酶的液体酶组合物中仍有改进的空间,以提供在运输和储存期间不丧失(loose)酶活性的洗涤剂组合物。
发明内容
本发明的目的是提供具有改进的酶稳定性的液体酶组合物。本发明另一个目的是提供用于制造所述液体酶组合物的方法。
已经发现向包括酶和抑制剂如苯基硼酸或苯基硼酸衍生物的液体酶组合物加入抑制剂加强剂如可溶性盐使抑制剂效果显著改进,并由此相对于酶活性改进酶的储存稳定性。
本发明提供一种包括酶组分、苯基硼酸组分或其衍生物和水溶性盐组分的液体酶组合物。
本发明还涉及所述液体酶组合物的制造及其用途。
本发明的目的已经通过提供一种包括酶组分、苯基硼酸组分或其衍生物和溶解的盐组分的液体组合物而获得。在多个实施方案中,酶组分是蛋白酶如丝氨酸蛋白酶。盐组分可以包括阳离子如Cu、Ca、Mg、Zn、Na、K、NH4和它们的组合。在多个实施方案中,盐组分可以包括选自下组的阳离子:Mg、Zn、NH4和它们的组合。在一些实施方案中,盐组分包括阴离子,其包含氯离子、硫酸根、硝酸根、磷酸根、碳酸根、甲酸根和它们的组合。另外,盐组分可以包括阴离子如氯离子、硫酸根、硝酸根和它们的组合。
在特定的实施方案中,阳离子选自由Cu、Ca、Mg、Zn、Na、K、NH4组成的组且阴离子选自由氯离子、硫酸根、硝酸根、磷酸根、碳酸根和甲酸根组成的组。
在一些实施方案中,液体组合物的pH为7-10.5,而在一些实施方案中,液体组合物的pH为8-9.5。
在一些实施方案中,盐组分以占总组合物重量的0.1%-20%的量存在。
本文公开内容的目的还通过提供洗涤剂组合物,如洗衣用洗涤剂组合物或洗碟用组合物而实现。
本发明的目的还可以通过提供一种用于制造液体组合物的方法来实现,所述方法包括如下步骤:提供液体;向a)的液体加入水溶性盐;与b)同时或在b)之后,在a)中加入酶和苯基硼酸或其衍生物;和混合所述液体组合物。在多个实施方案中,所述方法也可以包括将pH调节至7-9.5,或8-9的步骤。
本发明的目的还通过使用根据本文公开内容的组合物清洁物体来实现。
还通过使用盐组分来加强或增强液体酶组合物中苯基硼酸或其衍生物的抑制剂效果来实现本发明的目的。
定义
如本文所使用,术语“%RH”是指空气的相对湿度。100%RH是在固定的温度下水分饱和的空气,且因此%RH反映了空气的百分比湿分饱和度(percent moisture saturation)。
关于化合物或物质的术语“恒定湿度”(在本发明的上下文中有时简写为CH)是指大气的%RH与同所述化合物的固相接触的化合物饱和水溶液平衡,其全部限制在给定温度下的封闭空间内。此定义是根据“Handbook of chemistryandphysics”CRC Press,Inc.,Cleveland,USA,第58版,p E46,1977-1978页。因此就某种化合物而言CH20℃=50%是指在20℃下50%湿度的空气将与该化合物的饱和水溶液平衡。因此,术语恒定湿度是化合物吸湿性质的量度。
关于化合物的术语“pH”在本发明的上下文中应理解为该化合物的10%w/w水溶液的pH。
优选实施方案详述
本文公开内容涉及包括一种或多种酶组分、一种或多种抑制剂和一种或多种抑制剂加强剂的液体酶组合物。已经发现如果抑制剂是代硼酸或其衍生物,那么盐在液体酶组合物中发挥抑制剂加强剂的作用。
不希望受到本文公开内容的任何理论的限制,认为洗涤剂组合物中的苯基硼酸衍生物的抑制剂作用受到碱性pH和高含水量(水活度(water activity))的组合的负面影响。在碱性pH下,代硼酸经由它的抗碱反应(antibase-reaction)而变为带电荷的以增加水溶性。另外,这降低分子对光解位点的亲和力,所述分子有抑制光解位点的倾向。平衡(1)向不受抑制的蛋白酶一侧(右)移动:
EZ是蛋白酶,I是抑制剂,而EZ[I]是失活的复合物。
通过降低抑制剂在洗涤剂基质中的溶解度,平衡(1)将向受抑制的蛋白酶复合物一侧(左)移动——降低抑制剂将从溶液中析出的可能性。
苯环是高疏水性的,因此认为向洗涤剂组合物加入一种或多种盐组分将使其不利于苯环存在于溶液中,而更有可能与蛋白酶的活性位点相互作用。
还认为加强作用可能附带蛋白酶中的一些小的结构变化,其促进抑制剂更好地匹配入活性位点中。
抑制剂组分
根据本文公开内容一种或多种抑制剂存在于组合物中。在多个实施方案中,本发明的酶抑制剂是代硼酸和/或其衍生物。
在本发明的特定实施方案中,抑制剂是苯基硼酸和/或其衍生物。
本发明涵盖包括代硼酸或其衍生物的液体酶组合物。在特定的实施方案中,本发明涵盖包含苯基硼酸或其衍生物的液体酶组合物。
在本发明的特定实施方案中,抑制剂是萘硼酸衍生物。
所述抑制剂组分以足以提供有益效果的量存在。在多个实施方案中,加入的抑制剂组分的量是总液体组合物的0.1%-20%(w/w),在一些实施方案中,其量是总组合物的0.5%-8%(w/w),并且在一些实施方案中,其量是总组合物的1%-5%(w/w)。
在本发明的特定实施方案中,抑制剂的量是总液体组合物的1%(w/w)以上。在本发明更特定的实施方案中,抑制剂组分的量是总液体组合物的1.5%(w/w)以上。在本发明最特定的实施方案中,抑制剂的量是总液体组合物的2%(w/w)以上。
在本发明的特定实施方案中,以总组合物的至少0.1%(w/w)的量向酶液体组合物加入抑制剂。在本发明更特定的实施方案中,以总组合物的至少0.5%(w/w)的量向液体酶组合物加入抑制剂。在甚至更特定的实施方案中,以总组合物的至少1%(w/w)的量向液体酶组合物加入抑制剂。在本发明最特定的实施方案中,以总组合物的至少1.5%(w/w)的量向液体酶组合物加入抑制剂组分。
在本发明的特定实施方案中,加入酶液体组合物中的抑制剂的量是低于总组合物的20%(w/w)的量。在本发明更特定的实施方案中,加入酶液体组合物的抑制剂的量是低于总组合物的15%(w/w)的量。在本发明甚至更特定的实施方案中,加入酶液体组合物的抑制剂的量低于总组合物的10%(w/w)。在本发明最特定的实施方案中,加入酶液体组合物的抑制剂的量低于总组合物的5%(w/w)。
用于根据本文公开内容的用途的合适苯基硼酸衍生物的非限定性实例具有下式:
其中R选自下组:氢、羟基、C1-C6烷基、取代的C1-C6烷基、C1-C6烯基和取代的C1-C6烯基。
在本文公开内容的一个实施方案中,液体组合物包括酶组分和上文公开的式子的苯基硼酸衍生的酶抑制剂,其中R是C1-C6烷基,具体而言其中R是CH3、CH3CH2或CH3CH2CH2,或其中R是氢。在本文公开内容的一个实施方案中,酶的抑制剂是4-甲酰-苯基-硼酸(4-FPBA)。
在多个实施方案中,抑制剂的合适的非限定性实例包括选自下组的化合物:
噻吩-2硼酸、噻吩-3硼酸、乙酰胺苯基硼酸、苯并呋喃-2硼酸、萘-1硼酸、萘-2硼酸、2-FPBA、3-FBPA、4-FPBA、1-噻蒽硼酸、4-二苯并呋喃硼酸、5-甲基噻吩-2硼酸、硫茚硼酸(thionaphthene boronic acid)、呋喃-2硼酸、呋喃-3硼酸、4,4联苯-二硼酸(4,4biphenyl-diborinic acid)、6-羟基-2-萘(6-hydroxy-2-naphtalene)、4-(甲硫基)苯基硼酸、4(三甲基-甲硅烷基)苯基硼酸、3-溴代噻吩硼酸、4-甲基噻吩硼酸、2-萘基硼酸、5-溴噻吩硼酸(5-bromothiphene boronic acid)、5-氯代噻吩硼酸、二甲基噻吩硼酸、2-溴代苯基硼酸、3-氯代苯基硼酸、3-甲氧基-2-噻吩、对-甲基-苯乙基硼酸、2-噻蒽硼酸、二苯并噻吩硼酸、4-羧基苯基硼酸、9-蒽基硼酸、3,5二氯苯基硼酸、二苯基硼酸酐、邻-氯代苯基硼酸、对-氯代苯基硼酸、间-溴代苯基硼酸、对-溴代苯基硼酸、对-氟代苯基硼酸、对-甲苯基硼酸、邻-甲苯基硼酸、辛基硼酸、1,3,5三甲基苯基硼酸、3-氯代-4-氟代苯基硼酸、3-氨基苯基硼酸、3,5-二-(三氟甲基)苯基硼酸、2,4二氯代苯基硼酸、4-甲氧基苯基硼酸和他们的组合。
适合作为抑制剂的合适代硼酸衍生物的其它非限定性实例在US4,963,655、US 5,159,060、WO 95/12655、WO 95/29223、WO 92/19707、WO94/04653、WO 94/04654、US 5442100、US 5488157和US 5472628中描述(通过提述将它们整体并入本文)。
在一个实施方案中,组合物包含酶、抑制剂组分(其中所述组分是代硼酸或其衍生物)和抑制剂加强剂组分。
抑制剂加强剂组分
根据本文公开内容一种或多种抑制剂加强剂存在于组合物中。抑制剂加强剂可以以足以提供有益效果的量存在,例如,抑制剂加强剂可以以有效量存在。
在一个实施方案中,抑制剂加强剂是水溶性的。在本文公开内容的上下文中,抑制剂加强剂在20℃可以具有100克水中至少1克的溶解度,如在20℃在100克水中至少2克的溶解度。在本文公开内容的一些实施方案中,抑制剂加强剂是溶解的形式。在一个抑制剂加强剂是盐的实施方案中,将所述盐溶解,并且因此是离子形式。在一些实施方案中,仅部分盐溶解,而剩余的为固体形式。
抑制剂加强剂能够增加或增强抑制剂组分对酶组分的效果。在多个实施方案中,抑制剂加强剂可以是一种或多种可溶性盐。
合适的可溶性盐的非限定性实例可以是无机盐或有机盐,和它们的组合。合适的阳离子的非限定性实例是铵或金属离子和碱金属或碱土金属离子,如钠、钾、镁、钙、锌或铝,和它们的组合。阴离子的非限定性实例包括氯离子、碘离子、硫酸根、亚硫酸根、亚硫酸氢根、硫代硫酸根、膦酸根、磷酸根、磷酸二氢根、磷酸氢根、次磷酸根、焦磷酸二氢根、硝酸根、氯离子、碳酸根、碳酸氢根、正(偏)硅酸根,简单有机酸(少于10个碳原子,例如6个或更少的碳原子),如柠檬酸根、苹果酸根、马来酸根、丙二酸根、琥珀酸根、乳酸根、甲酸根、乙酸根、丁酸根、丙酸根、苯甲酸根、酒石酸根、抗坏血酸根或葡糖酸根,和它们的组合。具体而言,可以使用碱金属和碱土金属的硫酸盐、亚硫酸盐、磷酸盐、膦酸盐、硝酸盐、氯化物或碳酸盐,或简单有机酸的盐如柠檬酸盐、丙二酸盐或乙酸盐,和它们的组合。具体非限定性实例包括NaH2PO4、Na2HPO4、Na3PO4、(NH4)H2PO4、K2HPO4、KH2PO4、Na2SO4、K2SO4、KHSO4、ZnSO4、MgSO4、CuSO4、Mg(NO3)2、(NH4)2SO4、四硼酸钠(sodium borate)、乙酸镁、柠檬酸钠和它们的组合物。
盐也可以是水合盐,即带有结晶的结合水的晶体盐水合物,如WO99/32595中所述。水合盐的实例包括七水合硫酸镁(MgSO4(7H2O))、七水合硫酸锌(ZnSO4(7H2O))、七水合磷酸氢钠(Na2HPO4(7H2O))、六水合硝酸镁(Mg(NO3)2(6H2O))、十水合硼酸钠、二水合柠檬酸钠和四水合乙酸镁。
在本发明的特定实施方案中,盐选自下组:MgCl2、MgSO4、Mg(NO3)2、ZnCl2、ZnSO4、ZN(NO3)2、NH4Cl、NH4NO3、(NH4)2SO4、CaCl2、NaCl、KCl、Na2SO4、NaNO3、NaH2PO4、C2H3NaO2、NaHCO3和甲酸钠。在本发明的另一特定实施方案中,盐选自下组:MgCl2、MgSO4、Mg(NO3))2、ZnCl2、ZnSO4、Zn(NO3)2、NH4Cl、NH4NO3、(NH4)2SO4、CaCl2、KCl、Na2SO4、NaNO3、NaH2PO4、C2H3NaO2、NaHCO3和甲酸钠。
在本发明的特定实施方案中,盐选自下组:MgCl2、MgSO4、Mg(NO3))2、ZnCl2、ZnSO4、Zn(NO3)2、NH4Cl、NH4NO3、(NH4)2SO4、KCl、Na2SO4、NaNO3、NaH2PO4、C2H3NaO2和甲酸钠。
在本发明的另一特定实施方案中,盐选自下组:MgCl2、MgSO4、Mg(NO3))2、ZnCl2、ZnSO4、Zn(NO3)2、NH4Cl、NH4NO3、(NH4)2SO4、NaNO3和NaH2PO4。
在本发明的另一特定实施方案中,盐选自下组:MgCl2、MgSO4、Mg(NO3))2、NH4Cl、NH4NO3、(NH4)2SO4、NaNO3和NaH2PO4。
在本发明的另一特定实施方案中,盐选自下组:MgCl2、MgSO4、Mg(NO3))2、NH4Cl、NH4NO3和(NH4)2SO4。
在本发明的特定实施方案中,阳离子选自Mg、Zn、Na、K或NH4。在本发明更特定的实施方案中,阳离子选自Mg或NH4。
在本发明的特定实施方案中,阴离子选自氯离子、硫酸根和硝酸根。
可以以液体或固体形式向液体洗涤剂加入抑制剂加强剂。如果抑制剂加强剂以液体形式加入,那么具体而言作为水成液(aqueous liquid)加入。
在一个实施方案中,组合物不包含磷酸二氢钠或三水合乙酸钠。
在多个实施方案中,用于根据本文公开内容的用途的组合物以有效量含有一种或多种抑制剂加强剂以改进稳定性和/或延长的储存期限。本文使用的“有效量”是指按照本文公开内容的抑制剂加强剂组分的量足以对根据本文公开内容的液体酶组合物的稳定性或储存期限引起特定的正面效益。正面效益可以是本质上用于化妆(cosmetic in nature),或与活性相关,或是二者的组合。例如,在一些实施方案中,在应激条件下当与不含抑制剂加强剂的类似组合物相比时,酶的残余活性可为2倍、3倍、4倍、5倍、6倍、7倍、8倍、9倍、10倍长。本文使用的应激条件包括,但不限于,在40℃的高温下储存四周。在多个实施方案中,正面效益是通过将液体酶组合物与抑制剂组分和抑制剂加强剂组分的组合接触从而改进液体酶组合物的稳定性和/或储存期限来实现的。
例如,在一些实施方案中,在应激条件下酶的残余活性可以高于10%、20%、30%、40%、50%、60%、70%,其中应激条件包括在40℃的高温下储存四周。
应用的特定抑制剂加强剂组分浓度通常依赖于应用该组合物的目的。例如,所述浓度可能依赖于所用酶的类型和溶液中稳定性和/或储存问题的严重性而不同。在多个实施方案中,将一种或多种抑制剂加强剂应用于液体酶组合物,从而使抑制剂加强剂浓度为占总组合物重量的0.1%-20%的量。在多个实施方案中,一种或多种抑制剂加强剂以占总组合物重量约0.5%-10%的量存在。
在多个抑制剂加强剂是一种或多种盐的实施方案中,向洗涤剂加入的盐的量在特定的实施方案中是总洗涤剂组合物重量的0.1%-20%。在另外的特定实施方案中,向洗涤剂加入的盐的量按重量计是0.5%-10%。在另一特定的实施方案中,向洗涤剂加入的盐的量按重量计是0.8-5%。在更进一步的特定实施方案中,向洗涤剂加入的盐的量按重量计是1-3%。
在特定的实施方案中,洗涤剂中存在的阳离子的量按重量计是0.005%-10%。在另一特定的实施方案中,洗涤剂中存在的阳离子的量按重量计是0.05%-4%。在另外的特定实施方案中,洗涤剂中存在的阳离子的量按重量计是0.1%-2%。
在一个实施方案中,组合物包含酶、抑制剂组分和抑制剂加强剂组分,其中所述抑制剂加强剂是一种或多种盐。
酶
能够按照本发明稳定化的酶在本发明的上下文中称为“洗涤酶(detersiveenzyme)”,其用于本文是指任何在洗涤循环过程中发挥它们的作用的酶,例如,在洗涤应用中具有清洁、织物护理、抗再沉积和去污作用,并且所述酶是为了这样的目的而加入的。
根据本发明,液体组合物含有至少一种酶。所述酶可以是任何可以通过商业方法获得的酶,具体而言是选自下组的酶:蛋白酶、淀粉酶、脂肪酶、纤维素酶、裂合酶、氧化还原酶和它们的任意组合。也包括来自同一类别的多种酶(例如多种蛋白酶)的混合物。
根据本发明,包含蛋白酶的液体组合物是优选的。在特定的实施方案中,优选的是包含两种或更多种酶的液体组合物,其中第一种酶是蛋白酶,并且第二种酶选自下组:淀粉酶、脂肪酶、纤维素酶、裂合酶和氧化还原酶。在更特定的实施方案中,第二种酶是脂肪酶。
应该理解的是,酶变体(例如,通过重组技术产生的)包括在术语“酶”的含义内。这样的酶变体的实例公开在例如EP 251,446(Genencor)、WO 91/00345(Novo Nordisk)、EP 525,610(Solvay)和WO 94/02618(Gist-Brocades NV)中。
可以根据来自NC-IUBMB,1992的酶命名手册(handbook EnzymeNomenclature from NC-IUBMB,1992)对酶进行分类,也可参见因特网上的ENZYME网站:http://www.expasy.ch/enzyme/。ENZYME是有关酶的命名的信息库。其主要基于国际生物化学和分子生物学联合会命名委员会(Nomenclature Committee of the Intemational Union of Biochemistry andMolecular Biology)(IUB-MB),Academic Press,Inc.,1992的建议,并且其描述了已为之提供了EC(酶学委员会)号的经表征的酶的各种类型(BairochA.TheENZYME database,2000,Nucleic Acids Res 28:304-305)。这种IUB-MB酶命名法是基于它们的底物特异性,有时基于它们的分子机制;这样的分类法不反映这些酶的结构特征。
几年前已经提出了另一种基于氨基酸序列相似性将某些糖苷水解酶,如内切葡聚糖酶、木聚糖酶、半乳聚糖酶、甘露聚糖酶、葡聚糖酶和α-半乳糖苷酶,归类到家族中。目前它们属于90个不同的家族:参见CAZy(ModO)因特网网站(Coutinho,P.M.& Henrissat,B.(1999)Carbohydrate-ActiveEnzymes服务器,在URL:http://afmb.cnrs-mrs.fr/~cazy/CAZY/index.html上(相应的论文:Coutinho,P.M.& Henrissat,B.(1999)Carbohydrate-active enzymes:an integrated database approach.In“Recent Advances in CarbohydrateBioengineering”,H.J.Gilbert,G.Davies,B.Henrissat和B.Svensson编辑,TheRoyal Society of Chemistry,Cambridge,第3-12页;Coutinho,P.M.& Henrissat,B.(1999)The modular structure of cellulases and other carbohydrate-activeenzymes:an integrated database approach.In“Genetics,Biochemistry andEcology of Cellulose Degradation”.,K.Ohmiya,K.Hayashi,K.Sakka,Y.Kobayashi,S.Karita和T.Kimura编辑,Uni Publishers Co.,Tokyo,第15-23页)。
液体酶添加剂优选包含蛋白酶,如丝氨酸蛋白酶。
蛋白酶:合适的蛋白酶包括动物、植物或微生物来源的那些蛋白酶。微生物来源是优选的。包括化学修饰的或遗传修饰的突变体。蛋白酶可以是丝氨酸蛋白酶,优选是碱性微生物蛋白酶或胰蛋白酶-样蛋白酶。碱性蛋白酶的实例是枯草溶菌素,特别是源自芽孢杆菌属(Bacillus)的那些枯草溶菌素,例如枯草溶菌素Novo、枯草溶菌素Carlsberg、枯草溶菌素309、枯草溶菌素147和枯草溶菌素168(在WO 89/06279中描述)。胰蛋白酶-样蛋白酶的实例是胰蛋白酶(例如,猪或牛来源的)和WO 89/06270中描述的镰孢属(Fusarium)蛋白酶。在本发明的特定实施方案中,蛋白酶是丝氨酸蛋白酶。丝氨酸蛋白酶或丝氨酸内肽酶(较新名称)是一类肽酶,其特征在于酶活性中心内存在丝氨酸残基。
丝氨酸蛋白酶:丝氨酸蛋白酶是催化肽键水解的酶,并且在所述酶的活性部位存在必要的丝氨酸残基(White,Handler和Smith,1973″Principles ofBiochemistry,″第五版,McGraw-Hill Book Company,NY,第271-272页)。
细菌丝氨酸蛋白酶分子量在20,000-45,000道尔顿的范围。它们受到二异丙基氟磷酸的抑制。它们水解简单的末端酯,并且在活性上类似于真核生物的胰凝乳蛋白酶,其也是一种丝氨酸蛋白酶。更狭义的术语,碱性蛋白酶,包括一个亚组,反映了一些丝氨酸蛋白酶从pH 9.0-11.0的高最适pH(综述参见Priest(1977)Bacteriological Rev.41711-753)。
枯草蛋白酶(subtilase):Siezen等(1991),Protein Eng.,4719-737提出了暂时定名为枯草蛋白酶的丝氨酸蛋白酶的亚组。它们通过对先前称作枯草溶菌素-样(subtilisin-like)蛋白酶的丝氨酸蛋白酶的超过40个氨基酸序列的同源性分析来定义。先前将枯草溶菌素定义为由革兰氏阳性细菌或真菌产生的丝氨酸蛋白酶,而根据Siezen等枯草溶菌素目前是枯草蛋白酶的亚组。已鉴定出多种枯草溶菌素,并且已经测定了许多枯草溶菌素的氨基酸序列。这些包括超过六种来自芽孢杆菌属菌株的枯草溶菌素,即,枯草溶菌素168、枯草溶菌素BPN′、枯草溶菌素Carlsberg、枯草溶菌素Y、枯草溶菌素amylosacchariticus和肠系膜肽酶(mesentericopeptidase)(Kurihara等(1972)J.Biol.Chem.2475629-5631;Wells等(1983)Nucleic Acids Res.117911-7925;Stahl和Ferrari(1984)J.Bacteriol.159811-819,Jacobs等(1985)Nucl.AcidsRes.138913-8926;Nedkov等(1985)Biol.Chem.Hoppe-Seyler 366421-430,Svendsen等(1986)FEBS Lett.196228-232),一种来自放线菌目的枯草溶菌素,来自普通高温放线菌(Thermoactinomyces vulgaris)的耐热蛋白酶(thermitase)(Meloun等(1985)FEBS Lett.198195-200),和一种真菌枯草溶菌素,来自白麦轴霉(Tritirachium album)的蛋白酶K(proteinase K)(Jany和Mayer(1985)Biol.Chem.Hoppe-Seyler 366584-492)。为了进一步参考在下文再现了Siezen等的表I。
枯草溶菌素在物理和化学上是充分表征的。除了对这些酶的初级结构(氨基酸序列)的认识,已经测定了枯草溶菌素的超过50种高解析度X射线结构,其描绘了底物结合、过渡态、产物、至少三种不同的蛋白酶抑制剂,并且定义了天然变异的结构影响(structural consequence)(Kraut(1977)Ann.Rev.Biochem.46331-358)。
枯草蛋白酶的一个亚组,I-S1,包含“经典”枯草溶菌素,如枯草溶菌素168,枯草溶菌素BPN′,枯草溶菌素Carlsberg(ALCALASE,NovozymesA/S)和枯草溶菌素DY。
枯草蛋白酶的另一个亚组I-S2,是由Siezen等(见上文)识别的。将亚组I-S2蛋白酶描述为高碱性枯草溶菌素并且包括如下的酶,如枯草溶菌素PB92(MAXACAL,Gist-Brocades NV)、枯草溶菌素309(SAVINASE(洒维奈斯),Novozymes A/S)、枯草溶菌素147(ESPERASE,Novozymes A/S)和碱性弹性蛋白酶YaB。
枯草蛋白酶基因的随机和定点突变都源于对酶的物理和化学性质的认识和贡献出的涉及枯草蛋白酶催化活性、底物特异性、三级结构等的信息(Wells等(1987)Proc.Natl.Acad.Sci.U.S.A.84;1219-1223;Wells等(1986)Phil.Trans.R.Soc.Lond.A.317415-423;Hwang和Warshel(1987)Biochem.262669-2673;Rao等,(1987)Nature 328551-554。
涵盖这个领域的最近的公开文献是与切割底物中特定靶序列(第24和64位)的变体的设计有关的Carter等(1989)Proteins 6 240-248;讨论多个之前公开的结果的Graycar等(1992)Annals of the New York Academy of Sciences 67271-79;和同样综述了之前的结果的Takagi(1993)Int.J.Biochem.25 307-312。
可以通过商业方法获得的蛋白酶(肽酶)的实例包括KannaseTM、EverlaseTM、EsperaseTM、AlcalaseTM、NeutraseTM、DurazymTM、SavinaseTM、OvozymeTM、PyraseTM、Pancreatic Trypsin NOVO(PTN)、Bio-FeedTM Pro和Clear-LensTM Pro(全部可从Novozymes A/S,Bagsvaerd,Denmark获得)。其它优选的蛋白酶包括在WO 01/58275和WO 01/58276中描述的那些蛋白酶。
其它可以通过商业方法获得的蛋白酶包括RonozymeTM Pro、MaxataseTM、MaxacalTM、MaxapemTM、OpticleanTM、PropeaseTM、PurafectTM和Purafect OxTM(可从Genencor International Inc.,Gist-Brocades,BASF或DSM Nutritional Products获得)。
脂肪酶:合适的脂肪酶包括细菌或真菌来源的那些。包括化学修饰或遗传修饰的突变体。
有用的脂肪酶的实例包括疏棉状腐质霉(Humicola lanuginosa)脂肪酶,例如,如EP 258 068和EP 305 216中描述的;曼赫根毛霉(Rhizomucor miehei)脂肪酶,例如,如EP 238 023中所述;念珠菌属(Candida)脂肪酶,例如南极念珠菌(C.antarctica)脂肪酶,例如,EP 214 761中描述的南极念珠菌脂肪酶A或B;假单胞菌属(Pseudomonas)脂肪酶如类产碱假单胞菌(P.pseudoalcaligenes)和产碱假单胞菌(P.alcaligenes)脂肪酶,例如,如EP 218 272中所述,洋葱假单胞菌(P.cepacia)脂肪酶,例如,如EP 331 376中所述,施氏假单胞菌(P.stutzeri)脂肪酶,例如,如BP 1,372,034中所公开,荧光假单胞菌(P.fluorescens)脂肪酶;芽孢杆菌属(Bacillus)脂肪酶,例如,枯草芽孢杆菌(B.subtilis)脂肪酶(Dartois等,(1993),Biochemica et Biophysica acta 1131,253-260),嗜热脂肪芽孢杆菌(B.stearothermophilus)脂肪酶(JP 64/744992)和短小芽孢杆菌(B.pumilus)脂肪酶(WO 91/16422)。
另外,许多克隆的脂肪酶可能是有用的,包括Yamaguchi等,(1991),Gene103,61-67)描述的沙门柏干酪青霉(Penicillium camenbertii)脂肪酶;念珠地丝菌(白地霉)(Geotricum candidum)脂肪酶(Schimada,Y.等,(1989),J.Biochem.106,383-388),和多种根霉属(Rhizopus)脂肪酶如德氏根霉(R.delemar)脂肪酶(Hass,M.J等,(1991),Gene 109,117-113),雪白根霉(R.niveus)脂肪酶(Kugimiya等,(1992),Biosci.Biotech.Bio-chem.56,716-719)和米根霉(R.oryzae)脂肪酶。
脂肪分解酶的其它类型例如角质酶也可能是有用的,例如,如WO88/09367中所述的源自门多萨假单胞菌(Pseudomonas mendocina)的角质酶,或源自豌豆腐皮镰孢(Fusarium solani pisi)的角质酶(例如WO 90/09446中所述)。
可以通过商业方法获得的脂肪酶的实例包括LipexTM、LipoprimeTM、LipopanTM、LipolaseTM、LipolaseTM Ultra、LipozymeTM、PalataseTM、ResinaseTM、NovozymTM 435和LecitaseTM(全部可从Novozymes A/S获得)。
其它可以通过商业方法获得的脂肪酶包括LumafastTM(来自GenencorInternational Inc.的门多萨假单胞菌脂肪酶);LipomaxTM(来自Gist-Brocades/Genencor Int.Inc.的类产碱假单胞菌脂肪酶;和来自Solvay enzymes的芽孢杆菌属菌种脂肪酶。另外的脂肪酶可从其它供应商例如Lipase P″Amano″(Amano Pharmaceutical Co.Ltd.)获得。
淀粉酶:合适的淀粉酶(α和/或B)包括细菌或真菌来源的那些。包括化学修饰的或遗传修饰的突变体。例如,淀粉酶包括获得自地衣芽孢杆菌(B.licheniformis)的特殊菌株的α-淀粉酶,其在英国专利说明书(British PatentSpecification)No.1,296,839中更详细地描述。可以通过商业方法获得的淀粉酶是DuramylTM、TermamylTM、FungamylTM和BANTM(可从Novozymes A/S获得)和RapidaseTM和Maxamyl PTM(可从Gist-Brocades获得)。
纤维素酶:合适的纤维素酶包括细菌或真菌来源的那些。包括化学修饰的或遗传修饰的突变体。合适的纤维素酶在US 4,435,307中公开,其公开了从特异腐质霉(Humicola insolens)产生的真菌纤维素酶。特别合适的纤维素酶是具有颜色保护效益(color care benefits)的纤维素酶。这样的纤维素酶的实例是在欧洲专利申请No.0 495 257中描述的纤维素酶。
氧化还原酶:适合于在液体组合物中使用的任何氧化还原酶,例如,过氧化物酶或氧化酶如漆酶,可以用于本文。在本文中合适的过氧化物酶包括植物、细菌或真菌来源的那些。包括化学修饰的或遗传修饰的突变体。合适的过氧化物酶的实例是源自鬼伞属(Coprinus)菌株,例如,灰盖鬼伞(C.cinerius)或长根鬼伞(C.macrorhizus)的那些,或源自芽孢杆菌属菌株,例如,短小芽孢杆菌的那些,特别是根据WO 91/05858的过氧化物酶。在本文中合适的漆酶包括细菌或真菌来源的那些。包括化学修饰的或遗传修饰的突变体。合适的漆酶的实例是可以从栓菌属菌株,例如,长绒毛栓菌(T.villosa)或变色栓菌(T.versicolor),或可以从鬼伞属菌株,例如,灰盖鬼伞,或可以从毁丝霉属(Myceliophthora)菌株,例如嗜热毁丝霉(M.thermophila)获得的那些。
可以存在于本发明液体中的酶的类型包括氧化还原酶(EC 1.-.-.-),转移酶(EC 2.-.-.-),水解酶(EC 3.-.-.-),裂合酶(EC 4.-.-.-),异构酶(EC 5.-.-.-)和连接酶(EC 6.-.-.-)。
在本发明的上下文中优选的氧化还原酶是过氧化物酶(EC 1.11.1),漆酶(EC 1.10.3.2)和葡萄糖氧化酶(EC 1.1.3.4)。可以通过商业方法获得的氧化还原酶(EC 1.-.-.-)的实例是GluzymeTM(可从Novozymes A/S获得的酶)。另外的氧化还原酶可从其它供应商获得。优选的转移酶是任何以下亚类中的转移酶:
a转移一碳基团的转移酶(EC 2.1);
b转移醛或酮残基的转移酶(EC 2.2);酰基转移酶(EC 2.3);
c糖基转移酶(EC 2.4);
d转移除甲基之外的烷基或芳基的转移酶(EC 2.5);和
e转移含氮基团的转移酶(EC 2.6)。
在本发明的上下文中最优选的转移酶类型是转谷氨酰胺酶(蛋白质-谷氨酰胺γ-谷氨酰转移酶;EC 2.3.2.13)。
合适的转谷氨酰胺酶的其它实例在WO 96/06931(Novo Nordisk A/S)中描述。
在本发明的上下文中优选的水解酶是:羧酸酯水解酶(EC 3.1.1.-)如脂肪酶(EC 3.1.1.3);肌醇六磷酸酶(EC 3.1.3.-),例如3-肌醇六磷酸酶(EC 3.1.3.8)和6-肌醇六磷酸酶(EC 3.1.3.26);糖苷酶(EC 3.2,其属于本文称作“糖酶”的组),如α-淀粉酶(EC 3.2.1.1);肽酶(EC 3.4,也称为蛋白酶);和其它羰基水解酶。可以通过商业方法获得的肌醇六磷酸酶的实例包括Bio-FeedTMPhytase(Novozymes),RonozymeTM P(DSM Nutritional Products),NatuphosTM(BASF),FinaseTM(AB Enzymes)和PhyzymeTM产品系列(Danisco)。其它优选的肌醇六磷酸酶包括在WO 98/28408、WO 00/43503和WO 03/066847中描述的那些。
在本文上下文中,术语“糖酶”不仅用于指能够断裂糖链(例如淀粉或纤维素)特别是五元和六元环结构的酶(即糖苷酶,EC 3.2),还指能够异构化糖的酶,例如将六元环结构如D-葡萄糖异构为五元环结构如D-果糖的酶。
相关的糖酶包括以下(EC号在圆括号内):
α-淀粉酶(EC 3.2.1.1)、β-淀粉酶(EC 3.2.1.2)、葡聚糖1,4-α-葡糖苷酶(EC3.2.1.3)、内切-1,4-β-葡聚糖酶(纤维素酶,EC 3.2.1.4)、内切-1,3(4)-β-葡聚糖酶(EC 3.2.1.6)、内切-1,4-β-木聚糖酶(EC 3.2.1.8)、葡聚糖酶(dextranase)(EC3.2.1.11)、壳多糖酶(EC 3.2.1.14)、多聚半乳糖醛酸酶(EC 3.2.1.15)、溶菌酶(EC 3.2.1.17)、β-葡糖苷酶(EC 3.2.1.21)、α-半乳糖苷酶(EC 3.2.1.22)、β-半乳糖苷酶(EC 3.2.1.23)、淀粉-1,6-葡糖苷酶(EC 3.2.1.33)、木聚糖1,4-β-木糖苷酶(EC 3.2.1.37)、葡聚糖内切-1,3-β-D-葡糖苷酶(EC 3.2.1.39)、α-糊精内切-1,6-α-葡糖苷酶(EC3.2.1.41)、蔗糖α-葡糖苷酶(EC 3.2.1.48)、葡聚糖内切-1,3-α-葡糖苷酶(EC 3.2.1.59)、葡聚糖1,4-β-葡糖苷酶(EC 3.2.1.74)、葡聚糖内切-1,6-β-葡糖苷酶(EC 3.2.1.75)、半乳聚糖酶(EC 3.2.1.89)、阿拉伯聚糖内切-1,5-α-L-阿拉伯糖苷酶(EC 3.2.1.99)、乳糖酶(EC 3.2.1.108)、壳聚糖酶(chitosanase)(EC 3.2.1.132)和木糖异构酶(EC 5.3.1.5)。
可以通过商业方法获得的糖酶的实例包括Alpha-GalTM、Bio-FeedTMAlpha、Bio-FeedTM Beta、Bio-FeedTM Plus、Bio-FeedTM Wheat、Bio-FeedTM Z、NovozymeTM 188、CarezymeTM、CelluclastTM、CellusoftTM、CelluzymeTM、CeremylTM、CitrozymTM、DenimaxTM、DezymeTM、DextrozymeTM、DuramylTM、EnergexTM、FinizymTM、FungamylTM、GamanaseTM、GlucanexTM、LactozymTM、LiquezymeTM、MaltogenaseTM、NatalaseTM、PentopanTM、PectinexTM、PromozymeTM、PulpzymeTM、NovamylTM、TermamylTM、AMGTM(Amyloglucosidase Novo)、MaltogenaseTM、SweetzymeTM和AquazymTM(全部可从Novozymes A/S获得)。其它糖酶可从其它供应商获得,例如RoxazymeTM和RonozymeTM产品系列(DSM Nutritional Products),AvizymeTM、PorzymeTM和GrindazymeTM产品系列(Danisco,Fimnfeeds),以及NatugrainTM(BASF)、PurastarTM和PurastarTM OxAm(Genencor)。
其它可以通过商业方法获得的酶包括MannawayTM、PectawayTM、StainzymeTM和RenozymeTM。
液体洗涤剂
根据本发明,液体洗涤剂组合物将在酶、抑制剂和抑制剂加强剂之外还包括一种或多种表面活性剂。例如,所述洗涤剂组合物可以是洗衣用洗涤剂组合物或洗碟用洗涤剂组合物。
洗涤剂将通常含有0-50%的阴离子表面活性剂如直链烷基苯磺酸酯(盐)(LAS)、α-烯烃磺酸酯(盐)(AOS)、烷基硫酸酯(盐)(脂肪醇硫酸酯(盐))(AS)、醇乙氧基硫酸酯(盐)(alcohol ethoxysulfate)(AEOS或AES)、仲烷基磺酸酯(盐)(secondary alkanesulfonate)(SAS)、α-磺基脂肪酸甲酯、烷基-或烯基琥珀酸或肥皂(soap)。洗涤剂也可以含有0-40%的非离子表面活性剂,例如醇乙氧基化物(AEO或AE)、醇丙氧基化物、羧化醇乙氧基化物(carboxylated alcoholethoxylates)、壬基苯酚乙氧基化物、烷基聚糖苷(alkylpolyglycoside)、烷基二甲基氧化胺(alkyldimethylamineoxide)、乙氧基化脂肪酸单乙醇酰胺(ethoxylatedfatty acid monoethanolamide)、脂肪酸单乙醇酰胺或多羟基烷基脂肪酸酰胺(例如如WO 92/06154中所述)。
通常洗涤剂含有1-65%的助洗剂(detergent builder),但是一些洗碟用洗涤剂可以含有甚至高达90%的助洗剂,或络合剂如沸石,二磷酸盐,三磷酸盐,膦酸盐,柠檬酸盐,氮川三醋酸(NTA),乙二胺四乙酸(EDTA),二亚乙基三胺五乙酸(DTMPA),烷基-或烯基琥珀酸,可溶性硅酸盐或层状硅酸盐(例如来自Hoechst的SKS-6)。
助洗剂可以细分为含磷型和不含磷型。含磷的无机碱性助洗剂的实例包括水溶性盐,特别是碱金属焦磷酸盐、正磷酸盐、聚磷酸盐和膦酸盐。不含磷的无机助剂的实例包括水溶性碱金属碳酸盐、硼酸盐和硅酸盐以及层状二硅酸盐和各种类型的水不溶性晶状或无定形铝硅酸盐(alumino silicate),其中沸石是最为人所知的代表。
合适的有机助剂的非限定性实例包括琥珀酸、丙二酸、脂肪酸丙二酸、脂肪酸磺酸、羧甲氧基琥珀酸、聚乙酸、羧酸、聚羧酸、氨基聚羧酸和聚乙酰羧酸的碱金属盐、铵盐或取代的铵盐。洗涤剂也可以是无助洗的(unbuilt),即基本上不含助洗剂。
洗涤剂可以包含或包括一种或多种聚合物。非限定性实例是羧甲基纤维素(CMC)、聚(乙烯基吡咯烷酮)(PVP)、聚乙二醇(PEG)、聚(乙烯醇)(PVA)、聚羧化物(polycarboxylates)如聚丙烯酸酯(polyacrylates)、聚马来酸酯、马来酸/丙烯酸共聚物和甲基丙烯酸月桂酯/丙烯酸共聚物。
洗涤剂组合物可以含有氯/溴型或氧型漂白剂。漂白剂可以是涂覆的或包囊的。无机氯/溴型漂白剂的实例是锂、钠或钙的次氯酸盐或次溴酸盐以及氯化磷酸三钠(chlorinatedtrisodium phosphate)。漂白体系也可以包含H2O2源如过硼酸盐或过碳酸盐,其可以与形成过酸的漂白剂活化剂如四乙酰乙二胺(TAED)或壬酰氧苯磺酸盐(NOBS)组合。
有机氯/溴型漂白剂的实例是杂环N-溴代和N-氯代亚胺,如三氯异氰尿酸、三溴异氰尿酸、二溴异氰尿酸和二氯异氰尿酸,和它们与水溶阳离子如钾和钠的盐。乙内酰脲(海因)化合物(hydantoin compound)也是合适的。漂白体系也可以包含例如酰胺、酰亚胺或砜型的过氧酸(peroxyacid)。
在洗碟用洗涤剂中,氧漂白剂是优选的,例如以无机过酸盐(persalt)的形式,优选与漂白剂前体一起或作为过氧酸化合物。合适的过氧漂白化合物的典型实例是碱金属的过硼酸盐(四水合物和一水合物),碱金属的过碳酸盐,过硅酸盐(persilicate)和过磷酸盐(perphosphate)。优选的活化物质是TAED或NOBS。
本发明的洗涤剂组合物的酶可以额外地使用常规稳定剂稳定化,所述稳定剂例如多元醇如丙二醇或甘油,糖或糖醇,或乳酸。
洗涤剂也可以含有其他常规洗涤剂成分如,例如,织物整理剂(fabricconditioner)包括粘土、反絮凝物质(deflocculant material)、泡沫促进剂/泡沫抑制剂(在洗碟用洗涤剂泡沫抑制剂中)、抑泡剂、防腐蚀剂、悬污剂、抗污物再沉积剂、染料、脱水剂、杀菌剂、光亮剂(optical brightener)或香料。
pH(按使用浓度在水溶液中测量)将通常为中性或碱性,例如在7-11的范围内。在本发明的特定实施方案中,pH为7-9.5。在本发明更特定的实施方案中,pH为8-9。已经发现了对于某些洗涤剂,如果洗涤剂的pH为8-9,那么洗涤剂的效果特别好。
以下非限定性实施例进一步阐明了按照本文公开内容的组合物、方法和处理。应该注意的是本公开内容不限于所述实施例中示例的特定细节。
实施例
实施例1
储存稳定性试验
洗涤剂基底:
55g阴离子表面活性剂Na-LAS
105g阴离子表面活性剂Surfac LC70
25g非离子表面活性剂Neodol 25-3
30g非离子表面活性剂Neodol 25-7
40g NaCO3
33g SXS(二甲苯磺酸钠40重量%水溶液)
17g一水合柠檬酸盐
10g STS(甲苯磺酸钠)
10g乙醇
pH调节至pH 9(NaOH)
加水至1000g
pH 9
将所述洗涤剂基底用水按1∶1.5稀释(diluted 1∶1.5 water)。
加入的盐的量为按重量计占稀释的洗涤剂基底3%的盐。
蛋白酶以0.173KNPU-S/g、395u/g比活性的量加入。
4-FPBA以0.17mg/g经稀释洗涤剂基底+盐的量加入。
选择的储存条件是在40℃储存4周。
测试的盐 | 残余活性(40℃4周) | 阳离子 | 阴离子 |
氯化镁 | 79% | Mg | Cl |
硝酸镁 | 55% | Mg | NO3 |
氯化铵 | 49% | NH4 | Cl |
硫酸铵 | 43% | NH4 | SO4 |
硝酸铵 | 41% | NH4 | NO3 |
硫酸镁 | 37% | Mg | SO4 |
氯化钾 | 34% | K | Cl |
氯化钠 | 32% | Na | Cl |
甲酸钠 | 29% | Na | CHO2 |
氯化钙 | 22% | Ca | Cl |
硫酸钠 | 22% | Na | SO4 |
硝酸钠 | 20% | Na | NO3 |
乙酸钠 | 20% | Na | C2H3O2 |
氯化铝 | 16% | Al | Cl |
碳酸钠 | 15% | Na | CO3 |
磷酸钠 | 13% | Na | PO4 |
无盐 | 6% | … | … |
柠檬酸钠 | 1% | Na | C6H5O7 |
能够得出结论,大多数盐对包含苯基硼酸衍生物的洗涤剂基底的稳定性具有正面影响。最有希望的阳离子应为镁和铝。
实施例2
储存稳定性试验
洗涤剂基底:
55g阴离子表面活性剂Na-LAS
105g阴离子表面活性剂Surfac LC70
25g非离子表面活性剂Neodol 25-3
30g非离子表面活性剂Neodol 25-7
40g NaCO3
33g SXS(二甲苯磺酸钠40重量%水溶液)
17g一水合柠檬酸盐
10g STS(甲苯磺酸钠)
10g乙醇
pH调节至pH 9(NaOH)
加水至1000g
将所述洗涤剂基底用水按1∶1.5稀释。
加入的盐的量为按重量计占洗涤剂3%的盐。
蛋白酶以0.173KNPU-S/g、395u/g比活性的量加入。
4-FPBA以0.17mg/g洗涤剂+盐的量加入。
选择的储存条件是在40℃储存4周。
测试的盐 | 残余活性(在40℃2周) | 阳离子 | 阴离子 |
氯化锌 | 102% | Zn | Cl |
硫酸锌 | 88% | Zn | SO4 |
无盐 | 33% | … | … |
两种锌盐都显示了对稳定性的显著改进。
应理解的是,可以对本文公开的实施方案进行各种修改。因此,不应将上述描述理解为限制,他们仅作为对实施方案的示例说明。本领域那些技术人员将会预见到本文所附权利要求范围和宗旨内的其他修改。
Claims (14)
1.一种液体洗涤剂组合物,其包含酶组分、苯基硼酸组分或其衍生物和溶解的盐组分,其中所述盐组分包含一种或多种阴离子,该一种或多种阴离子选自氯离子、硫酸根、硝酸根、乙酸根或它们的组合,且加入的盐组分的量按重量计是该组合物的0.5%-10%,其中所述酶组分是蛋白酶,所述苯基硼酸或其衍生物是
其中R选自下组:氢、羟基、C1-C6烷基、取代的C1-C6烷基、C1-C6烯基和取代的C1-C6烯基。
2.权利要求1的液体洗涤剂组合物,其中所述蛋白酶是丝氨酸蛋白酶。
3.权利要求1或2的液体洗涤剂组合物,其中所述盐组分包含一种或多种阳离子,其中所述一种或多种阳离子包含Ca、Mg、Zn、Na、K或NH4,和它们的组合。
4.权利要求1或2的液体洗涤剂组合物,其中所述盐组分包含一种或多种选自下组的阳离子:Ca、Mg、Zn、Na、K、NH4和它们的组合。
5.权利要求1或2的液体洗涤剂组合物,其中所述盐组分包含一种或多种选自下组的阳离子:Zn、Mg、NH4和它们的组合。
6.权利要求1或2的液体洗涤剂组合物,其中所述液体组合物的pH是7-10.5。
7.权利要求1或2的液体洗涤剂组合物,其中所述液体组合物的pH是8-9.5。
8.权利要求1或2的液体洗涤剂组合物,其中所述液体组合物是洗衣用洗涤剂组合物。
9.权利要求1或2的液体洗涤剂组合物,其中所述液体组合物是洗碟用组合物。
10.一种用于制造权利要求1-9中任一项的液体洗涤剂组合物的方法,其包括如下步骤:
a)提供液体;
b)向a)的液体加入溶解的盐组分;
c)与b)同时或在b)之后将酶和苯基硼酸或其衍生物加入a)中;和
d)混合液体组合物,
其中,所述苯基硼酸或其衍生物是:
其中R选自下组:氢、羟基、C1-C6烷基、取代的C1-C6烷基、C1-C6烯基和取代的C1-C6烯基。
11.权利要求10的方法,进一步包括将pH调节至7-9.5的步骤。
12.权利要求10的方法,进一步包括将pH调节至8-9的步骤。
13.权利要求1-9的组合物用于清洁物体的用途。
14.溶解的盐用于加强苯基硼酸或其衍生物在含蛋白酶液体洗涤剂组合物中的抑制剂效果的用途,其中所述盐包含一种或多种阴离子,该一种或多种阴离子选自氯离子、硫酸根、硝酸根、乙酸根或它们的组合,且加入的盐的量按重量计是该组合物的0.5%-10%,此外,所述苯基硼酸或其衍生物是
其中R选自下组:氢、羟基、C1-C6烷基、取代的C1-C6烷基、C1-C6烯基和取代的C1-C6烯基。
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DE102010038497A1 (de) | 2010-07-27 | 2012-02-02 | Henkel Ag & Co. Kgaa | Stabilisierte flüssige enzymhaltige Tensidzubereitung |
FI123942B (fi) | 2010-10-29 | 2013-12-31 | Ab Enzymes Oy | Sieniperäisen seriiniproteaasin variantteja |
DK177132B1 (da) | 2010-11-05 | 2012-01-30 | Respekt Danmark As | Maskinopvask |
FI123425B (fi) | 2011-03-31 | 2013-04-30 | Ab Enzymes Oy | Proteaasientsyymi ja sen käytöt |
US8921299B2 (en) | 2011-07-25 | 2014-12-30 | The Procter & Gamble Company | Detergents having acceptable color |
EP2716644B1 (en) | 2012-10-03 | 2017-04-05 | The Procter and Gamble Company | A stable enzyme stabilizer premix |
CN106471110A (zh) * | 2014-07-03 | 2017-03-01 | 诺维信公司 | 改进的非蛋白酶类酶稳定化 |
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CN107043762A (zh) * | 2017-04-21 | 2017-08-15 | 江苏福隆生物技术有限公司 | 一种提高辣根过氧化物酶保存稳定性的保存稳定剂及其保存方法 |
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