CN103052704A - 用于提高清洁的鼠李糖脂和酶的组合物 - Google Patents
用于提高清洁的鼠李糖脂和酶的组合物 Download PDFInfo
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- CN103052704A CN103052704A CN2011800357973A CN201180035797A CN103052704A CN 103052704 A CN103052704 A CN 103052704A CN 2011800357973 A CN2011800357973 A CN 2011800357973A CN 201180035797 A CN201180035797 A CN 201180035797A CN 103052704 A CN103052704 A CN 103052704A
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- rhamnolipid
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- lipase
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- C—CHEMISTRY; METALLURGY
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- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
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- C—CHEMISTRY; METALLURGY
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- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
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Abstract
包含鼠李糖脂和脂肪酶的洗涤剂组合物,其中由单-鼠李糖脂构成的鼠李糖脂的重量百分比超过50wt%,优选超过80wt%,更优选超过90wt%。
Description
技术领域
本发明涉及包含单-鼠李糖脂和酶的清洁组合物。
背景
鼠李糖脂是一类糖脂。它们由鼠李糖结合β-羟基脂肪酸构成。鼠李糖是一种糖。脂肪酸遍布于动物和植物中。脂肪酸末端的羧基端连接鼠李糖。鼠李糖脂是只有三种常见元素碳、氢和氧的化合物。它们是晶体酸。鼠李糖脂可以通过细菌铜绿假单胞菌(Pseudomonas aeruginosa)的菌株来产生。存在两个主要的鼠李糖脂组:单-鼠李糖脂和二-鼠李糖脂。
单-鼠李糖脂具有单个鼠李糖糖环。通过铜绿假单胞菌产生的典型单-鼠李糖脂是L-鼠李糖酰基-β-羟基癸酰基-β-羟基癸酸酯(RhaC10C10)。可以称为Rha-C10-C10,其式为C26H48O9。单-鼠李糖脂具有单个鼠李糖糖环。IUPAC名称为3-[3-[(2R,3R,4R,5R,6S)-3,4,5-三羟基-6-甲基氧杂环己-2-基]氧基癸酰氧基]癸酸。
二-鼠李糖脂具有两个鼠李糖糖环。典型的二-鼠李糖脂是L-鼠李糖酰基-L-鼠李糖酰基-β-羟基癸酰基-β-羟基癸酸酯(Rha2C10C10)。可以称为Rha-Rha-C10-C10,其式为C32H58O13。IUPAC名称为3-[3-[4,5-二羟基-6-甲基-3-(3,4,5-三羟基-6-甲基氧杂环己-2-基)氧基氧杂环己-2-基]氧基癸酰基氧基]癸酸。
实践中,多种具有不同烷基链长组合的其他次要成分(取决于碳源和细菌菌株)与上述更常见的鼠李糖脂结合存在。可以通过生产方法来控制单-鼠李糖脂和二-鼠李糖脂的比例。一些细菌只产生单-鼠李糖脂,参见US5767090实施例1,一些酶可以将单-鼠李糖脂转化成二-鼠李糖脂。
在各种出版物中,单-鼠李糖脂标注为Rha-,其可以缩写为Rh或RL2。相似地,二-鼠李糖脂标注为Rha-Rha或Rh-Rh-或RL1。出于历史原因,“鼠李糖脂2”是单-鼠李糖脂,而“鼠李糖脂1”是二-鼠李糖脂。这导致一些文献中在“RL1”和“RL2”使用上含糊不清。在本发明的整个专利申请文件中,我们使用术语单-和二-鼠李糖脂,以避免这种可能的混淆。然而,如果使用缩写,则R1是单-鼠李糖脂,而R2是双-鼠李糖脂。关于现有技术中术语混乱的更多信息,请参见US4814272的介绍。
检测到通过以下细菌产生的以下鼠李糖脂:(C12:1、C14:1表示具有双键的脂肪酰基链)。
通过铜绿假单胞菌产生的鼠李糖脂(单-鼠李糖脂):
Rha-C8-C10,Rha-C10-C8,Rha-C10-C10,Rha-C10-C12,Rha-C10-C12:1,Rha-C12-C10,Rha-C12:1-C10
通过铜绿假单胞菌产生的鼠李糖脂(二-鼠李糖脂):
Rha-Rha-C8-C10,Rha-Rha-C8-C12:1,Rha-Rha-C10-C8,Rha-Rha-C10-C10,Rha-Rha-C10-C12:1,Rha-Rha-C10-C12,Rha-Rha-C12-C10,Rha-Rha-C12:1-C12,Rha-Rha-C10-C14:1
通过铜绿假单胞菌产生的鼠李糖脂(未经确认是单-还是二-鼠李糖脂):
C8-C8,C8-C10,C10-C8,C8-C12:1,C12:1-C8,C10-C10,C12-C10,C12:1-C10,C12-C12,C12:1-C12,C14-C10,C14:1-C10,C14-C14。
通过绿针假单胞菌(P.chlororaphis)产生的鼠李糖脂(只有单-鼠李糖脂):Rha-C10-C8,Rha-C10-C10,Rha-C12-C10,Rha-C12:1-C10,Rha-C12-C12,Rha-C12:1-C12,Rha-C14-C10,Rha-C14:1-C10。
通过类鼻疽伯克霍尔德菌(Burkholdera pseudomallei)产生的鼠李糖脂(只有二-鼠李糖脂):
Rha-Rha-C14-C14。
通过植物伯克霍尔德菌(Burkholdera (假单胞菌(Pseudomonas))plantarii)产生的鼠李糖脂(只有二-鼠李糖脂):
Rha-Rha-C14-C14。
因为以各种化学式制备了鼠李糖脂,每一种都具有不同的HLB,因此已知可以制备或混合鼠李糖脂来具有各种起泡性能。鼠李糖脂是同时具有亲水端和亲脂端的阴离子型表面活性剂。当它们的浓度升高至一定水平时,已知鼠李糖脂以胶束在液体内部结合在一起。
已表明具有两个较短脂肪酸的鼠李糖脂在降低表面张力中以及作为乳化剂的活性更大。那些少见的具有单个脂肪酸链的鼠李糖脂没有这么有效。
细菌铜绿假单胞菌是在土壤中、水中和植物上天然发现的。在代谢上,铜绿假单胞菌是化能异养的,通常是需氧的,利用多种有机化合物作为碳源和氮源。
在美国典型培养物保藏中心(ATCC)的档案中,存在超过100株的铜绿假单胞菌。还存在许多只有商业鼠李糖脂制造商可以获得的菌株。此外,可能存在上千个被全世界各种研究机构分离出的菌株。已进行一些工作将它们分组。每个菌株具有不同的特征,包括产生多少鼠李糖脂、产生哪种类型的鼠李糖脂、它代谢成何物和它生长的条件。只有少数菌株已经得到了详尽的研究。
通过评估和选择,可以分离铜绿假单胞菌的菌株从而以较高浓度和更高效率制备鼠李糖脂。还可以选择菌株来产生较少的副产物以及代谢不同的给料或污染物。这种生产很大程度上受到细菌生长环境的影响。
各种文献已经提出在洗涤剂组合物中使用鼠李糖脂。US2004/0152613A1(Ecover)以及EP1445302描述了几种包括具有鼠李糖脂表面活性剂和合成的常规表面活性剂的混合物的组合物。在实施例5中,通过将表面活性剂体系加入包含沸石助剂但不含酶的常规洗衣粉基料中来对其进行测试。
US5417879 A1(Unilever)提出了将胶束糖脂和层状表面活性剂混合的组合物,该表面活性剂可以是糖脂类,也可不是。提出以0.5至50g/l来使用组合物。使用鼠李糖脂的实施例没有使用任何酶。在第12栏第24至25行,提出可能将生物表面活性剂结合未公开含量的酶。为了获得酶与鼠李糖脂的组合物,需要从该文献进行几种选择。
US2006106120描述了微生物、生物表面活性剂和塑料降解酶的混合物,用于人工材料的生物修复。生物表面活性剂可以是鼠李糖脂(第62段)。酶可以是脂肪酶(第64段)。没有给出优选的任何鼠李糖脂的成分。没有举例说明鼠李糖脂,也没有举例说明脂肪酶,并且没有具体要求保护鼠李糖脂。
US2004072713A(Unilever)公开了一种用于酶促织物清洁方法的制品,所述制品含有一种或多种类型的能够分泌用于所述织物清洁方法中的酶的无害微生物。除了酶以外,如果微生物还能够产生其他有助于清洁方法的化学物质(例如,生物表面活性剂,例如EP924221中所述的脂多糖),则尤其有用。这些生物表面活性剂不是鼠李糖脂。所产生的生物表面活性剂的水平实际上非常低,并且确定不会超过0.5g/l。据称微生物能够产生和分泌有用的洗衣酶,如氧化还原酶、糖酶、蛋白酶、脂肪酶、转移酶和糖苷酶。
在“Lipase and biosurfactant production for utilisation in bioremediationof vegetable oils and hydrocarbon(用于植物油和烃的生物去除中的脂酶和生物表面活性剂的生产)”,Martins VG等(2008)Quimica Nova第31期第8卷,1942-1947和“Isolation and characterisation of a lipiddegrading bacterium and its application to lipid containing wastewatertreatment(脂质降解细菌的分离和表征及其在含脂质废水处理中的应用)”,Matsumiya Y.等(2007)Journal of Bioscience and Bioengineering第103期,第4卷,325-330中公开了微生物同时产生酶和生物表面活性剂的能力。
US2006080785A(Nero)描述了通过将具有生物表面活性剂和酶的组合物施用于地毯而进行地毯清洁;并用圆套清洁材料。源自海带的酶没有得到进一步的详细说明。提及未知组成的鼠李糖脂,但没有举例说明。
发明概述
本发明提供了一种具有新的单鼠李糖脂与二鼠李糖脂比率并结合脂肪酶的洗涤剂组合物。存在的单-鼠李糖脂的含量高于存在的二-鼠李糖脂(如果有的话)的含量。优选地,组合物中的至少80wt%,更优选至少90wt%,或甚至100wt%的鼠李糖脂是单-鼠李糖脂。
脂肪酶优选源自真菌或细菌源。对于细菌源,我们包括了由已经从细菌中克隆的基因在其他微生物(如酵母)中的表达物。
鼠李糖脂优选以0.5至40wt%的量存在。脂肪酶优选以0.0001至5wt%的量存在。
洗涤剂组合物优选是未加助剂的(unbuilt)。即,不存在沸石、磷酸盐或硅酸盐助剂。
洗涤剂组合物优选是液体洗涤剂组合物,如果存在柠檬酸助剂,其最大水平限于2wt%。组合物作为洗衣洗涤剂尤其有用,并且可以有利地用于在低于15℉的低水硬度的水中清洗。由此在预先软化的水中清洗衣物和组合物的方法与本发明的组合物一起使用特别有利。
将组合物用于除去衣物、尤其是棉布上的脂肪污垢。因为在现代洗衣洗涤剂中所包括的许多非常复杂的污垢去除和污垢释放技术在聚酯布上效果最好,因此对于从棉布去除污垢的关注渐增。因此,将本发明的洗涤剂体系结合聚酯污垢释放聚合物是有利的。
在本发明的洗涤剂组合物中使用单-鼠李糖脂和脂肪酶产生增强的清洁益处,并且可能与合成的阴离子型表面活性剂(例如C12-14烷基苯磺酸盐合成阴离子型表面活性剂)协同作用。这种表面活性剂通常用于洗衣洗涤剂组合物中,并且通常与非离子型表面活性剂(如US5417879中使用的乙氧基化非离子型表面活性剂)一起使用。出于环境原因,希望从组合物中消除这种非离子型表面活性剂。具有所要求的单鼠李糖脂与二鼠李糖脂比率的鼠李糖脂提供了该非离子型表面活性剂成分的合适替代物,尤其是当用于除去脂肪污垢时,并且特别是当用于从棉布除去污垢时。该组合物适于低清洗温度和快速清洗时间,这有利于节约能量和时间。
优选的脂肪污垢是牛肉脂肪。
发明详述
大部分生物表面活性剂通过细菌对可再生给料的作用产生,并且日益变成目前合成表面活性剂可持续替代品的越来越可行的选择。在目前商业化的现有生物表面活性剂资源中,通过Pseudomonas Aeg降解油和脂肪形成的鼠李糖脂,当以通过细菌分解方法产生的成分浓度使用时,清洁效果不好。然而,当将所表达的鼠李糖脂的单鼠李糖脂和二鼠李糖脂成分被提取出来,并且将单-鼠李糖脂与脂肪酶一起使用时,获得了较好的清洁结果。此外,通过生产掺混物并且与合成的阴离子型表面活性剂混合,可以获得进一步增强的去污力。
洗涤剂组合物可以包含通常在洗衣液中发现的其他成分。尤其是聚酯类的污垢释放聚合物、水溶助长剂、遮光剂、着色剂、香料、其他酶、其他表面活性剂、诸如香料或护理添加剂的成分的微胶囊、软化剂、用于抵抗污垢再次沉积的聚合物、漂白剂、漂白活化剂和漂白催化剂、抗氧化剂、pH控制剂和缓冲剂、增稠剂、用于流变修正的外部结构剂、视觉指示物,其含有或不含包埋在其中的功能性成分,和本领域技术人员已知的其他成分。组合物优选是液体的,并且有利地包装在多剂量瓶子或单位剂量可溶性袋子中。
酶
用于本发明组合物中的合适的脂肪酶包括细菌、真菌或酵母来源的那些。包括化学修饰或蛋白质工程突变体。有用的脂肪酶的实例包括来自腐质霉属(Humicola)(Thermomyces同义)的脂肪酶,例如EP258068和EP305216中所述的来自柔毛腐质霉(H.lanuginosa)(T.lanuginosus),或如WO96/13580中所述的来自特异腐质霉(H.insolens);假丝单胞菌属脂肪酶,例如,来自产碱假单胞菌(P.alcaligenes)或类产碱假单胞菌(P.pseudoalcaligenes)(EP218272)、洋葱假单胞菌(P.cepacia)(EP331376)、斯氏假单胞菌(GB1372034)、荧光假单胞菌(P.fluorescens)、假单胞菌属菌株SD705(WO95/06720和WO96/27002)、威斯康星假单胞菌(P.wisconsinensis)(WO96/12012);芽孢杆菌属脂肪酶,例如,来自枯草芽孢杆菌(B.subtilis)(Dartois等(1993),Biochemicaet Biophysica Acta,1131,253-360)、嗜热芽孢杆菌(B.stearothermophilus)(JP64/74992)或短小芽孢杆菌(B.pumilus)(WO91/16422)。
其他实例有脂肪酶变体,如WO 92/05249、WO 94/01541、EP 407225、EP 260 105、WO 95/35381、WO 96/00292、WO 95/30744、WO94/25578、WO 95/14783、WO 95/22615、WO 97/04079、WO 97/07202、US2008004186、US2006205628、US5869438、US6017866、US2002110854、US6939702、US2009221034、US200802425、US2004053360、US2005281912、US2006075518、US2005059130、US20041542180、US2003199069、WO98106215和WO08088489中所述的那些。
合适脂肪酶的另外实例在以下文献中描述和提及,但不限于此:Juardo等,J Surfact Deterg(2007)10:61-70;Horchani等,J MolecularCatalysis:Enzymatic 56(2009)237-245;Aloulou等,Biochimica etBiophysica acta 1771(2007)1446-1456;Mogensen等,Biochemistry(2005)44:1719-1730;Nicanuzia dos Prazeres等,Brazilian J of Microbiology(2006)37:505-509;Fernandez-Lorente 等,Biotechnology andBioengineering,97:第2卷242-250;Gilbert(1993)Enzyme Microb.Technol.第15卷634-645;Bora和Kalita(2008)J of Chemical Technologyand Biotechnology 83:688-693;Liu等(2009)Biochemical EngineeringJournal 46:265-270;Thirunavukarasu等(2008)Process Biochemistry 43:701-706;Saisubramanian等(2008)Appl Biochem Biotechnol 150:139-156;Joshi和Vinay(2007)Res J Biotech第2卷第2期50-56。酵母脂肪酶,如来自假丝酵母属(Candida)和隐球菌属(Cryptococcus)的那些,是合适的。
优选的可购得的脂肪酶包括LipolaseTM和Lipolase UltraTM、LipexTM、Novozym 525L(Novozymes A/S)。
组合物可以包含归类为EC 3.1.1.74的角质酶。本发明中使用的角质酶可以是任何来源的。优选地,角质酶是微生物来源的,特别是细菌、真菌或酵母来源的。
角质酶(酯酶)是能够降解角质的酶。在一个优选的实施方案中,角质酶源自曲霉属(Aspergillus)的菌株,特别是米曲霉(Aspergillusoryzae);链格孢属(Alternaria)的菌株,特别是芸苔生链格孢(Alternariabrassiciola);镰刀霉属(Fusarium)的菌株,特别是腐皮镰刀霉(Fusariumsolani)、Fusarium solani pisi,Fusarium roseum culmorum或Fusariumroseum sambucium;长孺孢属(Helminthosporum)的菌株,特别是小麦长孺孢(Helminthosporum sativum);腐质霉属(Humicola)的菌株,特别是特意腐质霉(Humicola insolens);假单胞菌属的菌株,特别是门多萨假单胞菌(Pseudomonas mendocina)或恶臭假单胞菌(Pseudomonasputida);丝核菌属(Rhizoctonia)的菌株,特别是立枯丝核菌(Rhizoctoniasolani);链霉菌属(Streptomyces)的菌株,特别是疮痂链霉菌(Streptomycesscabies);或细基格孢属(Ulocladium)的菌株,特别是群生细基格孢(Ulocladium consortiale)。在一个最优选的实施方案中,角质酶源自特异腐质霉的菌株,特别是菌株特异腐质霉DSM 1800。特异腐质霉角质酶在WO96/13580中有描述。角质酶可以是变体,如WO00/34550和WO01/92502中公开的变体中的一种,所述文献在此通过引证的方式并入本文。优选的角质酶变体包括WO01/92502的实施例2中所列的变体,它们在此通过引证的方式具体并入本文。
其他合适的酯酶是US2002012959,WO09085743,WO09002480,US2002137177,US2003024009,US2010151542,US2003032161,US2002007518和US2007167344中描述的那些。这还包括转移酶酶类。
优选的市售角质酶包括NOVOZYMTM 51032(可从丹麦NovozymesA/S获得)。
组合物还可以包含归类为EC 3.1.1.4和/或EC 3.1.1.32的磷脂酶。如本文所用,术语磷脂酶是一种对磷脂具有活性的酶。磷脂,如卵磷脂或磷脂酰胆碱,由用两个脂肪酸在外侧(sn-1)和中间(sn-2)位置酯化并且用磷酸在第三个位置酯化的甘油构成;磷酸进而可被酯化成氨基-醇。磷脂酶是参与磷脂水解的酶。可以区分几种类型的磷脂酶活性,包括磷脂酶A1和A2,其水解一个脂肪酰基(各自在sn-1和sn-2位置)以形成溶血磷脂;和溶血磷脂酶(或磷脂酶B),其可以水解溶血磷脂中剩余的脂肪酰基。磷脂酶C和磷脂酶D(磷酸二酯酶)各自释放二酰基甘油或磷脂酸。
术语磷脂酶包括具有磷脂酶活性的酶,例如,磷脂酶A(A1或A2)、磷脂酶B活性、磷脂酶C活性或磷脂酶D活性。本文所用的与本发明的酶有关的术语“磷脂酶A”意在涵盖具有磷脂酶A1和/或磷脂酶A2活性的酶。磷脂酶活性可以通过还具有其他活性的酶来提供,如,例如,具有磷脂酶活性的脂肪酶。磷脂酶活性,例如,可以来自具有磷脂酶副活性的脂酶。在本发明的其他实施方式中,磷脂酶酶活性通过基本上只具有磷脂酶活性且其中磷脂酶酶活性不是副活性的酶来提供。
磷脂酶可以是任何来源的,例如,动物来源的(例如哺乳动物),例如,来自胰腺(例如牛或猪胰腺),或蛇毒或蜂毒。优选,磷脂酶可以是微生物来源的,例如,来自丝状真菌、酵母或细菌,如以下的属或种,曲霉属(Aspergillus),例如,黑曲霉(A.niger);网柄菌属(Dictyostelium),例如,盘基网柄菌(D.discoideum);毛霉属(Mucor),例如,爪哇毛霉(M.javanicus)、高大毛霉(M.mucedo)、细孢毛霉(M.subtilissimus);脉孢菌属(Neurospora),例如,粗糙脉孢菌(N.crassa);根毛霉属(Rhizomucor),例如,微小根毛霉(R.pusillus);根霉属(Rhizopus),例如,少根根霉(R.arrhizus)、日本根霉(R.japonicus)、葡枝根霉(R.stolonifer);核盘菌属(Sclerotinia),例如,大豆核盘菌(S.libertiana);毛藓菌属(Trichophyton),例如,红色毛藓菌(T.rubrum);维氏核盘菌属(Whetzelinia),例如,W.sclerotiorum;芽孢杆菌属(Bacillus),例如,巨大芽孢杆菌(B.megaterium)、枯草芽孢杆菌(B.subtilis);柠檬酸杆菌属(Citrobacter),例如,弗氏柠檬酸杆菌(C.freundii);肠杆菌属(Enterobacter),例如,产气肠杆菌(E.aerogenes)、阴沟肠杆菌(E.cloacae);爱德华菌属(Edwardsiella),迟缓爱德华菌(E.tarda);欧文氏菌属(Erwinia),例如,草生欧文氏菌(E.herbicola);埃希氏菌属(Eecherichia),例如,大肠埃希氏菌(E.coli);克雷伯氏菌属(Klebsiella),例如,肺炎克雷伯氏菌(K.pneumoniae);变形杆菌属(Proteus),例如,普通变形杆菌(P.vulgaris);普罗威登斯菌属(Providencia),例如,斯氏普罗维登斯菌(P.stuartii);沙门氏菌属(Salmonella),例如,鼠伤寒沙门氏菌(S.typhimurium);沙雷氏菌属(Serratia),例如,液化沙雷氏菌(S.liquefasciens)、粘质沙雷氏菌(S.marcescens);志贺氏菌属(Shigella),例如,福氏志贺氏菌(S.flexneri);链霉菌属(Streptomyces),例如,紫红链霉菌(S.violeceoruber);耶尔森氏菌属(Yersinia),例如,小肠结肠炎耶尔森氏菌(Y.Enterocolitica)。例如,磷脂酶可以是真菌的,例如,来自核菌纲(Pyrenomycetes),如镰刀霉(Fusarium)属,如黄色镰刀霉(F.culmorum)、异孢镰刀霉(F.heterosporum)、腐皮镰刀霉(F.solani)的菌株,或尖孢镰刀霉(F.oxysporum)的菌株。磷脂酶还可以来自曲霉属内的丝状真菌菌株,如泡盛曲霉(Aspergillus awamori)、臭曲霉(Aspergillus foetidus)、日本曲霉(Aspergillus japonicus)、黑曲霉(Aspergillus niger)或米曲霉(Aspergillus oryzae)的菌株。
优选的磷脂酶源自腐质霉属(Humicola)的菌株,尤其是柔毛腐质霉(Humicola lanuginosa)。磷脂酶可以是变体,如WO00/32758中公开的变体中的一种,该文献通过引证的方式并入本文。优选的磷脂酶变体包括WO00/32758的实施例5中所列的变体,它们通过引证的方式具体并入本文。在另一个优选的实施方案中,磷脂酶是WO04/111216中所述的一种,尤其是实施例1的表格中所列的变体。
在另一个优选的实施方案中,磷脂酶源自镰刀霉属(Fusarium),尤其是尖孢镰刀霉(Fusarium oxysporum)的菌株。磷脂酶可以是WO98/026057中相关的一种,呈现于源自尖孢镰刀霉(Fusariumoxysporum)DSM 2672的SEQ ID NO:2中,或其变体。
在本发明的一个优选实施方案中,磷脂酶是磷脂酶A1(EC.3.1.1.32)。在本发明的另一个优选实施方案中,磷脂酶是磷脂酶A2(EC.3.1.1.4).
市售磷脂酶的实例包括LECITASETM和LECITASETM ULTRA,YIELSMAX或LIPOPAN F(可从Novozymes A/S丹麦获得)。
组合物可以进一步包含增强组合物去污力的其他酶,如软化剂、淀粉酶(例如来自丹麦Novo Nordisk A/S的Fungamyl(R))、脂肪酶(例如,来自丹麦Novo Nordisk A/S的Novocor(R)AD)、纤维素酶(例如,Celluzyme(R)、Carezyme(R)和/或Celluclast(R),全部来自Novo NordiskA/S,丹麦)、木聚糖酶(例如,来自丹麦Novo Nordisk A/S的Biofeed(R)PLUS或Shearzyme(TM))、β-葡聚糖酶(例如,来自丹麦Novo NordiskA/S的Viscozyme(R)或Ultraflo(TM))、果胶酶(例如,来自丹麦NovoNordisk A/S的Pectinex(TM))、过氧化物酶(例如,来自丹麦Novo NordiskA/S的Guardzyme(TM))、漆酶(例如,获自Myceliophthora或Polyporus)、用于过氧化物酶/漆酶的增强剂(例如,PPT或甲基丁香酸甲基丁香酸酯或其衍生物)和/或缓冲剂(例如,柠檬酸)。
现在将参照以下非限制性实施例来进一步描述本发明。
实施例
实施例1
检测了各种脂肪酶和鼠李糖脂的组合物,以测定它们除去有色牛肉污垢的能力。
通过将4mg蛋白质/升浓度的脂肪酶与所需浓度的洗涤剂表面活性剂一起分散在pH调节至8和水硬度为12°FH的磷酸盐缓冲盐水(PBS)中来制备清洗溶液。在37℃下将10ml清洗溶液在25ml塑料小瓶中在轨道培养器中以200rpm搅拌30分钟而混合。然后加入染有苏丹红着色的牛肉脂肪污渍的棉布样品(大约1cm2)并且将小瓶返回振荡培养器中。在定时的间隔下取出样品、在冷水中漂洗并且在37℃下干燥。使用Macbeth Colour Eye监测残余的颜色,并且与未处理的染有污渍的布相比。
细菌酶是“Lipomax”,是Gist-Brocades(M.M.M.J.Cox,H.B.M.Lenting,L.J.S.M.Mulleners和J.M.van der Laan)的WO94/25578中描述的产碱假单胞菌脂酶(Pseudomonas alcaligenes)的细菌产生的脂酶变体M21L。
真菌酶是“Lipolase”,如EP025068中所述源自柔毛腐质霉(Humicolalanguginosa),并且可以从NovoZymes A/S获得。
表面活性剂的详细内容如下:
RL=鼠李糖脂:一种细菌来源的生物表面活性剂。作为RBR425(25%AM)从Jeneil购得。分析了该材料的组成,并且在以下的表3中给出。
除了市售供应的鼠李糖脂,通过将其分离成R1和R2富集级分,来制备另外两种样品。将这些级分也结合脂肪酶来使用。表1和2中给出了1小时和4小时的清洁数据。
表1-1小时
鼠李糖脂 | 无酶 | 细菌酶 | 真菌酶 |
0.5g/l RL | 1.15 | 8.88 | 1.04 |
0.5g/l R1 | 9.85 | 11.31 | 12.25 |
0.5g/l R2 | 0.80 | 8.87 | 1.05 |
表2-4小时
鼠李糖脂 | 无酶 | 细菌酶 | 真菌酶 |
0.5g/l RL | 1.18 | 10.68 | 1.89 |
0.5g/l R1 | 14.52 | 12.43 | 14.19 |
0.5g/l R2 | 1.14 | 11.42 | 2.85 |
表3-通过HPLC/MS对JBR425的分析
鼠李糖脂同源物 | m/z | % |
二-C10-C8 | 621 | 1.6 |
二-C8-C10 | 621 | 1.3 |
二-C10-C10 | 649 | 67.4 |
二-C10-C12:1 | 675 | 0.78 |
二-C12:1-C10 | 675 | 0.016 |
二-C10-C12 | 677 | 3.18 |
二-C12-C10 | 677 | 1.12 |
单-C10-C8 | 475 | 0.63 |
单-C8-C10 | 475 | 0.47 |
单C10-C10 | 503 | 21.6 |
单-C10-C12:1 | 529 | 0.69 |
单-C12:1-C10 | 529 | 0.014 |
单C10-C12 | 531 | 1.12 |
单-C12-C10 | 531 | 0.023 |
为进行我们在表3中的这种鼠李糖脂的分析,使用12M HCl将已知量的JBR425酸化至pH 3,并且置于冰箱中过夜。然后使用2∶1的氯仿和乙醇混合物将上清液萃取三次。然后通过旋转蒸发来除去溶剂,然后将分离的鼠李糖脂混合物重新溶解于甲醇中。
使用连接有Ion Trap Electrospray ionization Mass Spectrometer(离子阱电喷雾电离质谱仪)的HPLC对混合物进行分离和表征。电离模式是使用50-1200Da扫描范围的负模式。用于分离的柱子是Phenomenex lunaC18 250×4.6mm 5μm柱。用移动相即水(移动相A)和乙腈(移动相B)在30分钟内从60∶40(A∶B)改变至30∶70(A∶B)的梯度而进行分离。然后将系统保持5分钟,随后恢复至起始条件,所有流速为0.5ml/min。注射体积为10μl。
Claims (16)
1.包含鼠李糖脂和脂肪酶的洗涤剂组合物,其中由单-鼠李糖脂构成的鼠李糖脂的重量百分比超过50wt%,优选超过80wt%,更优选超过90wt%。
2.根据权利要求1的组合物,其中100wt%的所述鼠李糖脂是单-鼠李糖脂。
3.根据前述权利要求的任一项的组合物,其中所述鼠李糖脂以0.5-40wt%的含量存在。
4.根据前述权利要求的任一项的组合物,其中所述脂肪酶以0.0001-5wt%的含量存在。
5.根据前述权利要求的任一项的组合物,进一步包含至少5wt%的合成阴离子型表面活性剂。
6.如前述权利要求的任一项所要求的组合物,其中所述合成阴离子型表面活性剂包含C12-14直链烷基苯磺酸盐。
7.根据前述权利要求的任一项的组合物,其具有小于2%的洗涤剂助剂。
8.根据前述权利要求的任一项的组合物,其具有的合成阴离子型表面活性剂多于鼠李糖脂。
9.根据前述权利要求的任一项的组合物,其是洗衣液,包含10至40%总的表面活性剂。
10.根据权利要求9的洗衣液,包含少于或等于2%的柠檬酸。
11.根据前述权利要求的任一项的洗衣洗涤剂,进一步包含聚酯类的污垢释放聚合物。
12.根据权利要求1至11任一项的组合物的用途,用于在具有低于15℉的低水硬度的水中的清洗。
13.根据权利要求1至11任一项的组合物的用途,用于从衣物上去除脂肪污垢。
14.根据权利要求13的用途,其中所述脂肪污垢从棉布上去除。
15.根据权利要求13或14的用途,其中所述脂肪污垢是牛肉脂肪。
16.用于清洗基体的方法,其包括将基体浸入水中,将根据权利要求1-11的组合物加入水中来形成清洗液,以及清洗所述基体的步骤,其特征在于清洗周期时间少于60分钟,优选少于30分钟,并且水温低于35℃。
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CN107427438A (zh) * | 2015-02-27 | 2017-12-01 | 赢创德固赛有限公司 | 包含鼠李醣脂和硅氧烷的组合物 |
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CN108473915A (zh) * | 2015-08-28 | 2018-08-31 | 荷兰联合利华有限公司 | 具有脂肪酶和生物表面活性剂的洗涤组合物 |
CN106591013A (zh) * | 2016-11-30 | 2017-04-26 | 大连百奥泰科技有限公司 | 一种生物洗涤剂组合物 |
Also Published As
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ES2561553T3 (es) | 2016-02-26 |
WO2012010406A1 (en) | 2012-01-26 |
BR112013000108A2 (pt) | 2017-06-13 |
EP2596087A1 (en) | 2013-05-29 |
BR112013000108B1 (pt) | 2021-05-11 |
CN107955721A (zh) | 2018-04-24 |
EP2596087B1 (en) | 2015-12-16 |
ZA201300377B (en) | 2014-03-26 |
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