CN1242040A - 防止石洗中回染的方法 - Google Patents
防止石洗中回染的方法 Download PDFInfo
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- CN1242040A CN1242040A CN96197599A CN96197599A CN1242040A CN 1242040 A CN1242040 A CN 1242040A CN 96197599 A CN96197599 A CN 96197599A CN 96197599 A CN96197599 A CN 96197599A CN 1242040 A CN1242040 A CN 1242040A
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Abstract
本发明涉及一种在染色纤维素织物表面形成局部颜色密度变化的方法,和用于该方法的组合物。
Description
本发明涉及一种在染色纤维素织物表面形成局部颜色密度变化的方法，和用于该方法的组合物。
在用染色纤维素织物制衣的过程中，如用靛蓝染色的斜纹粗绵布denim制牛仔服时，常常处理这种粗绵布使其具有“石洗”外观(在棉布表面的局部颜色磨损)。这可以通过将denim在含机械磨损剂如浮石、磨损性纤维素酶或它们的混合物的水介质中搅拌而获得。这种处理优选在近中性pH下进行，因而优选使用在此pH范围内具有高活性的纤维素酶。过去，一般通过培养天然微生物产生纤维素酶制剂，这种制剂都含有许多不同纤维素酶成分的混合物。使用这样的混合纤维素酶制剂的一种方法公开在US 4832864(Ecolab所有)中。
重组DNA技术的迅速发展使得可能得到高收率的单一成分酶，因而使用单一成分纤维素酶的方法已引起人们更大的兴趣。例如，WO91/17243和WO95/09225(Novo Nordisk)描述了一种使用单一成分内切聚糖酶的方法，所述酶称为EG V，分子量为～43kD，来自腐质霉Humicola insolens菌株DSM 1800，在近中性pH有最佳活性。WO94/21801(Genencor)描述了一种称为EG III的单一成分纤维素酶在“石洗”中的应用，该酶来自木霉T.longibrachiatum，据报道最佳pH范围为5.5-6.0，在碱性pH下保持显著的活性。WO 95/16782(Genencor International)提示来自木霉属的其他单一成分纤维素酶在“石洗”中的应用，但这些纤维素酶具酸性，在中性pH下基本上没有活性。
在已知的“石洗”方法中一个普遍问题是回染，即通过磨损已脱下的染料沉集在织物或衣服的一些部位造成所希望的颜色密度变化被淡化或使衣服的浅色部分被染色。
我们意外地发现，加入一定类型的纤维素酶(以下称为第一成分)可减少回染。这种纤维素酶本身没有显著的磨损效果。
因此，本发明提供一种在染色纤维素织物表面形成局部颜色密度变化的方法，包括将织物在pH在6.5-9范围内并含有下列成分的水介质中搅拌：
第一成分，它是(a)能够水解纤维三糖和/或对硝基苯基-β-1，4-纤维二糖苷的第5族纤维素酶，或(b)第7族纤维素酶，
和第二成分，它是(a)机械磨损剂或(b)具有磨损活性的纤维素酶，
其中每种纤维素酶在pH7下显示其最大活性的至少30％。
本发明的另一方面提供用于所述方法的组合物，其包括上述第一和第二成分。
在本说明书和权利要求中，适用下列定义：
术语“纤维素酶”指对纤维素水解有贡献的酶，如纤维二糖水解酶(酶的命名E.C.3.2.1.91)、内切葡聚糖酶(以下简称为“EG”，E.C.3.2.1.4)或β-葡糖苷酶(E.C.3.2.1.21)。
根据氨基酸序列的相似性和Henrissat.B等在《生物化学杂志》(Biochem.J.)(1991)，280，309-16页和Henrissat，B等在《生物化学杂志》，293，781-788页中描述的分类系统，将纤维素酶分为不同的族。
用于本发明的纤维素酶优选是单一成分，即用于本发明的水介质应该除指出的酶外不含其他纤维素酶成分。单一成分的酶可用重组DNA技术廉价获得，即克隆编码该单一成分的DNA序列，然后用该DNA序列转化适当的宿主细胞并在该宿主中表达该成分。
因而，编码有用纤维素酶的DNA序列可通过下述一般方法分离：
-从本说明书以后提到的微生物之一在适当的载体中克隆一个DNA文库，
-用所述载体转化适当的酵母宿主细胞，
-在适当的条件下培养宿主细胞以表达由该DNA文库中某克隆编码的目的酶，
-通过确定由这种克隆产生的酶的纤维素酶活性来筛选阳性克隆，及
-从这种克隆中分离酶的编码DNA。
WO94/14953(Novo Nordisk)进一步公开了该通用方法，其内容引入本文作为参考。
编码有用纤维素酶的DNA序列例如可通过筛选微生物的cDNA文库并挑选表达适当活性(即纤维素酶活性)的克隆而分离得到。
编码同类酶的DNA序列(即类似DNA序列)可从其他微生物获得。例如，这种DNA序列可通过相似方法筛选另一真菌的cDNA文库而获得，例如曲霉属的菌株，特别是棘孢曲霉或黑曲霉的菌株，木霉属的菌株，特别是T.resei、绿色木霉、T.longibrachiatum、T.harzianum或康宁木霉的菌株，或Neocallimatix属、Piromyces属、青霉属、蘑菇属或Phanerochaete属的菌株。
或者，编码有用纤维素酶的DNA也可以按熟知的程序、利用在已知DNA序列上制备的合成寡核苷酸探针，方便地从适当的DNA来源分离得到，如从上述任何生物体分离。
然后可将DNA序列插入重组表达载体中。这可以是可方便地进行重组DNA操作用的任何载体，载体的选择常取决于其所要转入的宿主细胞。例如，载体可以是自主复制的载体，即作为染色体外单位存在的载体，其复制不依赖于染色体的复制，例如为质粒。或者，载体可以是当引入宿主细胞时被整合在宿主细胞基因组中并随着所整合的染色体一起复制的载体。
编码纤维素酶的DNA序列在载体中应与适当的启动子和终止序列有效地连接。启动子可以是在所选的宿主细胞中显示转录活性的任何DNA序列，可来自与宿主细胞同源或异源的蛋白编码基因。用于将纤维素酶编码DNA序列与启动子和终目子分别连接的方法，和用于将它们插入适当载体的方法都是本领域技术人员熟知的(参见例如，Sambrook等，分子克隆实验室指南(MolecularCloning.A Laboratory Manual，纽约冷泉港，1989)。
用该DNA序列转化的宿主细胞优选是真核细胞，特别是真菌细胞如酵母或丝状真菌细胞。特别是，属于曲霉属或木霉属种类的细胞，最优选米曲霉或黑曲霉细胞。转化真菌细胞的方法涉及，按本身已知的方法进行原生质体形成和原生质体转化，然后再生细胞壁。使用曲霉作为宿主微生物在EP 238023(Novo Nordisk A/S)中有描述，其内容引入这里作为参考。宿主细胞还可以是醇母细胞，如酵母属菌株，特别是酿酒酵母、克鲁弗酵母或葡萄汁酵母，裂殖酵母属菌株，如粟酒裂殖酵母，汉逊酵母属菌株，毕赤酵母属菌株、Yarrowia属菌株如Yarrowia lipolytica或克鲁维酵母属菌株如乳酸克鲁维酵母。
在本说明书中，术语“同源”或“同源序列”以一个或多个氨基酸残基而分别区别于下文序列表中每一序列的氨基酸序列。同源序列可以是通过改变序列表中所示序列而造成的序列，序列改变例如为在氨基酸序列的一个或多个不同位点取代一个或多个氨基酸残基，在酶的任一个或两个末端或在氨基酸序列中的一个或多个位点缺失一个或多个氨基酸残基，或在氨基酸序列中一个或多个位点插入一个或多个氨基酸残基。
但如对本领域技术人员显见的那样，氨基酸的改变优选是轻微的，即不显著影响蛋白折叠或活性的保守性氨基酸取代，小的缺失，常为1至约30个氨基酸；小的氨基或羧基端延伸，如氨基端的甲硫氨酸残基，最长约20-25个残基的小接头肽，或用于纯化的小延伸，如多组氨酸尾，抗原性表位或结合域。一般性描述见Ford等，蛋白表达和纯化(Protein Expression and Purification)2：95-107，1991。保守性取代的例子有碱性氨基酸内(如精氨酸、赖氨酸、组氨酸)、酸性氨基酸内(如谷氨酸和天冬氨酸)、极性氨基酸内(如谷氨酰胺和天冬酰胺)、疏水氨基酸内(如亮氨酸、异亮氨酸、缬氨酸)、芳香氨基酸内(如苯丙氨酸、色氨酸、酪氨酸)和小氨基酸内(如甘氨酸、丙氨酸、丝氨酸、苏氨酸、甲硫氨酸)。
这种取代可以在分子功能关键区之外进行并仍产生活性多肽，这也是本领域技术人员显见的。对本发明DNA构建物编码的多肽活性关键的(从而优选不进行取代的)氨基酸，可用本领域已知的方法确定，如定点诱变或丙氨酸扫描诱变(Cunningham和Wells，科学(Science)244，1081-1085，1989)。在后一技术中，在每分子中的每一个残基上引入突变，然后测检这些突变分子的生物活性(即纤维素酶活性)，以确定对分子活性至关重要的氨基酸残基。底物-酶作用位点也可通过晶体结构分析确定，晶体结构是用诸如核磁共振、晶体学或光亲和标记技术测定的。例如见de Vos等，科学，255：306-312，1992；Smith等，分子生物学杂志(J.Mol.Biol.)224：899-904，1992；Wlodaver等，FEBS Lett 309：59-64，1992。
氨基酸序列的改变可适当地通过改变酶的编码DNA来进行，如按已熟知的操作进行定点诱变或随机诱变或这些技术的组合。或者，同源序列可以是来自与下文序列表中每一氨基酸序列相应的纤维素酶不同来源的酶序列。因此，“同源”可以指例如在一定条件下(如在5×SSC中预浸，在20％甲酰胺、5×Denhardt溶液、pH6.8的50mM磷酸钠、和50mg超声变性牛胸腺DNA溶液中～40℃预杂变l小时，然后在补有100mM ATP的相同溶液～40℃杂变18小时)与具有原氨基酸序列的纤维素酶的编码DNA所杂交的相同探针杂交的DNA编码的多肽。这种同源序列一般与下文序列表中所示的每种氨基酸序列分别具有至少50％的同源程度(按一致性计)，例如至少60％、65％、70％、75％、80％、85％、90％或甚至95％。
上述同源性是按两个序列间一致性程度确定的，显示一种序列衍生于另一序列。这种同源性可适当地借助于本领域已知的计算机程序测定，如GCG程序包中提供的GAP(Needleman，S.B.和Wunsch，C.D.，分子生物学杂志，48：443-453，1970)。染色的纤维素织物
本发明方法可应用于欲在其表面形成局部颜色密度变化的任何类型的染色纤维素织物。具有特别商业意义的例子是斜纹粗棉布denim，特别是用于制蓝牛仔服等的靛蓝染色的denim。
可以以未缝制的织物或从这种织物制得的缝好的衣物形式处理织物。用本发明方法处理新的洁净织物或衣物特别重要。成分1
第一成分是一种在pH7显示其至少30％最佳活性的第5或7族纤维素酶。其存在量为防止回染的有效量，一般为0.05～5mg/l(按纯酶蛋白计)，特别是0.l-0.5/l；一般相当于10-1000ECU/I的活性，特别是100-1000ECU/l的活性；或者0.5-100ECU/g织物的活性。第5族纤维素酶
用于本发明的第5族纤维素酶能够水解纤维三糖和/或对硝基苯基-β-1，4-纤维二糖苷(PNP-Cel)；该纤维素酶可对纤维三直接作用，将其水解成纤维二糖而无葡萄糖形成。纤维素酶水解PNP-Cel的能力可用下述测定方法测定，如果该测定方法给出大于0.1μmol PNP/分钟/ECU的结果，则认为该纤维素酶符合该条件。
第5族纤维素酶优选不含有任何纤维素结合域。它可以是来自细菌菌株如芽孢杆菌属或梭菌属的碱性纤维素酶(如内切葡聚糖酶)。
一种这样的第5族纤维素酶是来自芽孢杆菌株KSM-64(FERM BP-2886)的内切葡聚糖酶。在JP-A4-190793(Kao)和Sumitomo等，生物科学生物技术生物化学(Biosci.Biotech.Biochem.)，56(6)，872-877(1992)中描述了这种纤维素酶和其氨基酸序列。
另一种第5族纤维素酶是来自KSM-635株(FERM BP-1485)的内切葡聚糖酶。在JP-A1-281090(Kao)、US4945 053和Y.Ozaki等，普通微生物学杂志(Journal of GeneralMicrobiology)，1990，136卷，1973-7979页中描述了这种纤维素酶和其氨基酸序列。在上述测定方法中其对PNP-Cel的活性为0.18μmol PNP/分钟/ECU。
第三个第5族纤维素酶是来自1139株的内切葡聚糖酶。在Fukumori F.等，普通微生物学杂志，132：2329-2335(1986)和JP-A 62-232386(Riken)中描述了该纤维素酶和其氨基酸序列。
第四种第5族纤维素酶是来自迟缓芽孢杆菌NCIMB 40250的内切葡聚糖酶Endo 3A，描述在WO 91/107324(Novo Nordisk)。后来发现其中描述的氨基酸序列不正确，改正的序列示于SEQ IDNo：1中。该纤维素酶对PNP-Cel的活性为0.44μmol PNP/分钟/ECU。
第五种第5族纤维素酶是来自芽孢杆菌株NCIMB 40482的纤维素酶，其表观分子量为约45kD，描述在WO 94/01532(NovoNordisk)中。其对PNP-Cel的活性为0.22μmol PNP/分钟/ECU。
第六种第5族纤维素酶是来自解纤维梭菌的内切葡聚糖酶A，描述在E.Faure等，基因(Gene)，84(1)，39-46(1989)和Fierobe H-P等，细菌学杂志(J.Bacteriol.)173(24)，7956-7962(1991)。第7族纤维素酶
用于本发明的第7族纤维素酶可来自真菌菌株，如在下述测定方法中测定的那样，一般能够直接水解纤维三糖成为纤维二糖和葡萄糖，还能够水解PNP-Cel。
这种第7族纤维素酶可衍生自腐质霉菌株，优选H.insolens。一个例子是来自H.insolens菌株DSM 1800的内切葡聚糖酶EGI，描述在WO 91/17244(Novo Nordisk)中。成熟的该纤维素酶具有该文图14中21-435位所示的415个氨基酸的序列，比活性为200 ECU/mg(基于纯酶蛋白)。该纤维素酶还可在C-末端截去多达18个氨基酸，含有至少397个氨基酸。例如，可将其截至402、406、408或412个氨基酸。另一例子是称为内切葡聚糖酶EG I*的其变体，描述在WO 95/24471(Novo Nordisk)中，具有其中图3所示的402个氨基酸的序列。
或者，第7族纤维素酶可来自毁丝霉属的菌株，优选M.thermophila，最优选菌株CBS 117.65。一个实例是WO 95/24471中描述的内切葡聚糖酶，其含有其中图6中所示序列的21-420位氨基酸和任选的1-20位和/或421-456位氨基酸。
另一情况是，第7族纤维素酶来自镰孢属菌株，优选尖镰孢。一个实例是WO 91/17244(Novo Nordisk)和Sheppard，P.O.等，基因150：163-167，1994中描述的来自尖镰孢的内切葡聚糖酶。在后来的文献中给出了正确的氨基酸序列。该纤维素酶的比活性为350 ECU/mg。成分2
第二成分是机械磨损剂和/或磨损性纤维素酶。本发明的优选实施方案中联合使用机械磨损剂和磨损性纤维素酶作为第二成分。
机械磨损剂的例子有浮右、热扩张的珍珠岩和磨损元件(如磨球)。
磨损性纤维素酶是起磨损或颜色消除作用的纤维素酶，如EP220013(Novo Nordisk A/S)所述，其在pH7显示其最佳活性的至少30％。它可以是有纤维素结合域的第12或45族纤维素酶。
用于本发明的第45族纤维素酶可来自腐质霉属菌株，优选H.insolens。一个实例为称为EGV的内切葡聚糖酶，其来自H.insolens菌株DSM 1800，分子量为～43kD。WO 91/17243(NovoNordisk)中描述了该纤维素酶和其氨基酸序列。其比活性为430ECU/mg。
用于本发明的第12族纤维素酶可来自木霉属菌株，优选T.longibrachiatum。一个实例是WO 94/21801(Genencor)中描述的内切葡聚糖酶EG III，其中显示了其氨基酸序列。
第二成分优选以形成局部颜色密度变化的磨损有效量存在。如果第二成分是一种磨损性纤维素酶，其一般存在量为0.05-5mg/l(按纯酶蛋白计)，特别是0.1-0.5mg/l；一般相当于10-1000ECU/l的活性，特别是100-1000 ECU/I；或0.1-100ECU/g织物，特别是0.5-10 ECU/g的活性。操作条件
本发明方法可以在常用于石洗的洗衣机(如washer-extractor)中在常规条件下进行。典型的条件是：温度40-60℃，织物：液体比1∶3到1∶20，15分钟到2小时。也可以使用常规添加剂，如缓冲剂、表面活性剂(阴离子性和/或非离子性)和/或聚合物(如PVP、聚丙烯酸酯和聚丙烯酰胺)。纤维素酶活性物的测定方法
纤维素酶内切活性通过在一振动粘度计中测定CMC(羧甲基纤维素)粘度的下降来测定。1 ECU(内切纤维素酶单位)是40℃下在0.1M磷酸缓冲液(pH7.5)中与1ml 3.40g/L CMC溶液(商品名Aqualon 7LFD)一起保温时，引起粘度降低10倍的酶量。对PNP-Cel水解的测定方法
纤维素酶水解对硝基苯基-β-1，4-纤维二糖苷(PNP-Cel)的能力通过稳态动力学测定，即在405nm直接测定产物对硝基苯酚(PNP)的黄色。测定条件为37℃，pH7.5(0.1M磷酸缓冲液)。水解速率(每分钟的PNP微摩尔数)与纤维素酶活性(ECU)比照，结果用μmol PNP/分钟/ECU表示。
实施例1
按如下用各种纤维素酶组合处理靛蓝染色的denim和白棉布片：pH 7(自来水配制的磷酸缓冲液)温度 55℃设备 Launderometer(多个150ml的容器)成分1 来自H.insolens DSM 1800的EG I 0-2.3 ECU/l，如下所示成分2 来自H.insolens DSM 1800的EG V 0或
0.27 ECU/lDenim 5g/容器白棉布 2片/容器时间 2小时
处理后，收集棉绒并测定，作为纤维素酶磨损作用的表达形式。处理后测定白布片的反射(680mm处的DR，相对于无纤维素酶的实验)，作为回染的表达。结果：
| 成分1ECU/ml | 成分2ECU/ml | 磨损(mg绵绒) | 回染降低(DR) | |
| 参照 | 0 | 0 | - | 0 |
| 0 | 0.27 | 33 | -3.8 | |
| 本发明 | 0.115 | 0.27 | 42 | -0.7 |
| 0.23 | 0.27 | 36 | 3.0 | |
| 1.15 | 0.27 | 38 | 8.2 | |
| 2.3 | 0.27 | 62 | 10.4 |
用成分2纤维素酶得到了良好的磨损。加入成分1纤维素酶显著降低了回染。实施例2
在与实施例1中相同的条件下测试了下列纤维素酶：
成分1：来自H.insolens DSM 1800的EG I或EG I*0、0.67
或1.33ECU/ml。
成分2：来自H.insolens DSM l800的EG V 0.7 ECU/ml
通过测定每次处理后的棉绒量评价磨损效果。每个实验中都观察到了良好的磨损效果，加入EG I或EG I*略有提高。
从680nm处滤液吸光度的提高和白色织物在420mm处反射的提高确定回染的抑制。结果表明，用EG I和EG I*得到了基本相同的回染抑制。实施例3
第一步，12片(5×5cm)蓝色denim与80ml磷酸缓冲液(pH7.0)和0.8ml非离子性表面活性剂于50℃搅拌30分钟，然后过滤，得到蓝色液体。
第二步，5片白绵布与200ml蓝液在50℃下0-100 ECU/l纤维素酶存在下保温30分钟。试验纤维素酶是截短至406、408和412氨基酸的来自H.insolens DSM 1800的EG I的混合物。
漂洗和干燥后，从由Dr.Lange Micro Color Data Station测定的白布片f亮度(L*)的提高确定回染的抑制。结果(5片的平均值)
| ECU/L | 平均亮度(L*) |
| 0 | 83.42 |
| 1 | 84.02 |
| 10 | 86.16 |
| 100 | 92.58 |
结果显示EG I对降低蓝denim到白棉布的回染是有效的。实施例4
按与实施3中相同的方式测定了本发明的碱性芽孢杆菌第5族纤维素酶。结果如下：
| ECU/L | 亮度(L*) |
| 0 | 82.52 |
| 90 | 83.82 |
结果证明该纤维素酶对降低回染也有效。实施例5
将4片(5×5cm)脱浆蓝denim和8片(5×5cm)白色丝光棉布在含有本发明的第5或7族纤维素酶及第45族纤维素酶(每种纤维素酶200 ECU/L)的400ml 50mM磷酸缓冲液(pH7.0)中搅拌。30分钟后，在流动自来水中漂洗织物并风干。
第5族纤维素酶是碱性芽孢杆菌纤维素酶。第7族纤维素酶是来自H.insolens截短至408氨基酸的EG I。第45族纤维素酶是来自H.insolens DSM 1800的EG V。
测定两种织物的亮度(L*)和上清液在680nm的吸光值。结果：
| 纤维素酶家族 | 亮度(L*) | A680 | ||
| 第1成分 | 第2成分 | 脱浆棉布 | Denim | |
| 无 | 第45族 | 84.8 | 22.83 | 0.132 |
| 第7族 | 第45族 | 86.3 | 23.55 | 0.212 |
| 第5族 | 第45族 | 86.2 | 23.50 | 0.238 |
脱浆棉布的结果显示亮度增加，即通过加入本发明的第二成分纤维素酶降低了回染。
结果还表明蓝色denim的亮度增加，即回染降低。肉眼检查表明用本发明方法处理的denim具有更显著的局部颜色密度变化。
上清液的吸光度数据表明处理后有更多的颜料留在液体中。
序列表(1)一般信息：
(i)申请人：
(A)名称：Novo Nordisk A/S
(B)街道：Novo Alle
(C)城市：Bagsvaerd
(E)国家：丹麦
(F)邮编：DK-2880
(G)电话：+45-4444-8888
(H)传真：+45-4449-3256
(ii)发明名称：防止石洗中回染的方法
(iii)序列数：1
(iv)计算机可读形式：
(A)媒介类型：软盘
(B)计算机：IBM PC兼容
(C)操作系统：PC-DOS/MS-DOS
(D)软件：PatentIn Release#1.0，#1.30版(EPO)(2)SEQ ID No：1的信息：(i)序列特征：
(A)长度：551个氨基酸
(B)类型：氨基酸
(C)链型：
(E)拓扑结构：线性(ii)原始来源：
(A)生物体：迟缓芽孢杆菌
(B)菌株：NCIMB 40250(xi)序列描述：SEQ ID NO：1Ala Pro Ala Val Pro Phe Gly Gln Leu Lys Val Gln Gly Asn Gln Leu1 5 10 15Val Gly Gln Ser Gly Gln Ala Val Gln Leu Val Gly Met Ser Ser His
20 25 30Gly Leu Gln Trp Tyr Gly Asn Phe Val Asn Lys Ser Ser Leu Gln Trp
35 40 45Met Arg Asp Asn Trp Gly Ile Asn Val Phe Arg Ala Ala Met Tyr Thr
50 55 60Ala Glu Asp Gly Tyr Ile Thr Asp Pro Ser Val Lys Asn Lys Val Lys65 70 75 80Glu Ala Val Gln Ala Ser Ile Asp Leu Gly Leu Tyr Val Ile Ile Asp
85 90 95Trp His Ile Leu Ser Asp Gly Asn Pro Asn Thr Tyr Lys Ala Gln Ser
100 105 110Lys Ala Phe Phe Gln Glu Met Ala Thr Leu Tyr Gly Asn Thr Pro Asn
115 120 125Val Ile Tyr Glu Ile Ala Asn Glu Pro Asn Gly Asn Val Ser Trp Ala
130 135 140Asp Val Lys Ser Tyr Ala Glu Glu Val Ile Thr Ala Ile Arg Ala Ile145 150 155 160Asp Pro Asp Gly Val Val Ile Val Gly Ser Pro Thr Trp Ser Gln Asp
165 170 175Ile His Leu Ala Ala Asp Asn Pro Val Ser His Ser Asn Val Met Tyr
180 185 190Ala Leu His Phe Tyr Ser Gly Thr His Gly Gln Phe Leu Arg Asp Arg
195 200 205Ile Thr Tyr Ala Met Asn Lys Gly Ala Ala Ile Phe Val Thr Glu Trp
210 215 220Gly Thr Ser Asp Ala Ser Gly Asn Gly Gly Pro Tyr Phe Pro Gln Ser225 230 235 240Lys Glu Trp Ile Asp Phe Leu Asn Ala Arg Lys Ile Ser Trp Val Asn
245 250 255Trp Ser Leu Ala Asp Lys Val Glu Thr Ser Ala Ala Leu Met Pro Gly
260 265 270Ala Ser Pro Thr Gly Gly Trp Thr Asp Ala Gln Leu Ser Glu Ser Gly
275 280 285Lys Trp Val Arg Asp Gln Ile Arg Gln Ala Thr Gly Gly Gly Ser Gly
290 295 300Asn Pro Thr Ala Pro Ala Ala Pro Thr Asn Leu Ser Ala Thr Ala Gly305 310 315 320Asn Ala Gln Val Ser Leu Thr Trp Asn Ala Val Ser Gly Ala Thr Ser
325 330 335Tyr Thr Val Lys Arg Ala Thr Thr Ser Gly Gly Pro Tyr Thr Asn Val
340 345 350Ala Thr Gly Val Thr Ala Thr Ser Tyr Thr Asn Thr Gly Leu Thr Asn
355 360 365Gly Thr Thr Tyr Tyr Tyr Val Val Ser Ala Ser Asn Ser Ala Gly Ser
370 375 380Ser Ala Asn Ser Ala Gln Ala Ser Ala Thr Pro Ala Ser Gly Gly Ala385 390 395 400Ser Thr Gly Asn Leu Val Val Gln Tyr Lys Val Gly Asp Thr Ser Ala
405 410 415Thr Asp Asn Gln Met Lys Pro Ser Phe Asn Ile Lys Asn Asn Gly Thr
420 425 430Thr Pro Val Asn Leu Ser Gly Leu Lys Leu Arg Tyr Tyr Phe Thr Lys
435 440 445Asp Gly Thr Ala Asp Met Ser Ala Ser Phe Asp Trp Ala Gln Ile Gly
450 455 460Ala Ser Asn Val Ser Ala Ala Phe Ala Asn Phe Thr Gly Ser Asn Thr465 470 475 480Asp Thr Tyr Val Glu Leu Ser Phe Ser Ala Gly Ser Gly Ser Ile Pro
485 490 495Ala Gly Gly Gln Thr Gly Asp Ile Gln Leu Arg Met Tyr Lys Thr Asp
500 505 510Trp Ser Asn Phe Asn Glu Ala Asn Asp Tyr Ser Tyr Asp Gly Ala Lys
515 520 525Thr Ala Tyr Ala Asp Trp Asn Arg Val Thr Leu His Gln Asn Gly Thr
530 535 540Leu Val Trp Gly Thr Thr Pro545 550
Claims (13)
1.一种在染色纤维素织物表面形成局部颜色密度变化的方法，包括将织物在pH在6.5-9范围内并含有下列成分的水介质中搅拌：
第一成分，它是
(a)能够水解纤维三糖和/或对硝基苯基-β-1，4-纤维二糖苷的第5族纤维素酶，或
(b)第7族纤维素酶，
和第二成分，它是
(a)机械磨损剂或
(b)具有磨损活性的纤维素酶；
其中每种纤维素酶在pH7下显示其最大活性的至少30％。
2.权利要求1的方法，其中织物是靛蓝染色的denim。
3.权利要求1或2的方法，其中第一成分是来自细菌菌株、优选芽孢杆菌属菌株的无纤维素结合域的第5族纤维素酶。
4.权利要求3的方法，其中第5族纤维素酶来自选自芽孢杆菌KSM-64、1139、KSM-635、NCIMB 40482和迟缓芽孢杆菌NCIMB40250的芽孢杆菌菌株，或者与这种纤维素酶具有至少60％同源性的纤维素酶。
5.权利要求1或2的方法，其中第一成分是来自真菌菌株、优选腐质霉、最优选H.insolens的第7族纤维素酶。
6.权利要求5的方法，其中第7族纤维素酶是来自H.insolens菌株DSM 1800的内切葡聚糖酶EG I，或与所述EG I具有至少60％同源性的纤维素酶。
7.权利要求1-6任一项的方法，其中第一成分的存在量为0.1-0.5mg/l，或浓度为100-1000ECU/l。
8.权利要求1-7任一项的方法，其中第二成分包括机械磨损剂和磨损性纤维素酶。
9.权利要求1-8任一项的方法，其中第二成分是来自真菌菌株、优选腐质霉菌株、最优选H.insolens的带有纤维素结合域的第45族纤维素酶。
10.权利要求9的方法，其中第45族纤维素酶是来自H.insolens菌株DSM 1800的内切葡聚糖酶EG V，或为与所述EG V有至少60％同源性的纤维素酶。
11.权利要求1-13任一项的方法，其中除指明的第一和第二成分外基本没其他纤维素酶存在。
12.一种纤维素酶组合物，其包括
第一成分，它是
(a)能够水解纤维三糖和/或对硝基苯基-β-1，4-纤维二糖苷的第5族纤维素酶，或
(b)第7族纤维素酶，
和第二成分，它是
(a)机械磨损剂或
(b)具有磨损活性的纤维素酶；
其中每种纤维素酶在pH7下显示其最大活性的至少30％。
13.权利要求15的纤维素酶组合物，其进一步表征如权利要求2-11任一项中所述。
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DK0993/1995 | 1995-09-08 | ||
| DK0993/95 | 1995-09-08 | ||
| DK99395 | 1995-09-08 |
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| Publication Number | Publication Date |
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| CN1242040A true CN1242040A (zh) | 2000-01-19 |
| CN1163577C CN1163577C (zh) | 2004-08-25 |
Family
ID=8099817
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB961975997A Expired - Fee Related CN1163577C (zh) | 1995-09-08 | 1996-09-03 | 防止石洗中回染的方法 |
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| Country | Link |
|---|---|
| EP (1) | EP0850295B2 (zh) |
| JP (1) | JP3943132B2 (zh) |
| CN (1) | CN1163577C (zh) |
| AU (1) | AU6785396A (zh) |
| DE (1) | DE69628311T3 (zh) |
| ES (1) | ES2200070T5 (zh) |
| PL (1) | PL183149B1 (zh) |
| TR (1) | TR199800402T1 (zh) |
| WO (1) | WO1997009410A1 (zh) |
Cited By (1)
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|---|---|---|---|---|
| CN107313273A (zh) * | 2016-04-27 | 2017-11-03 | 尚科纺织企业工业及贸易公司 | 生产含细菌生物聚合物且具有独特外观的染色织物的方法 |
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| JP2006152469A (ja) * | 2004-11-26 | 2006-06-15 | Ochanomizu Univ | 染色繊維製品処理剤及び染色仕上げ処理方法 |
| US7361487B2 (en) | 2006-04-13 | 2008-04-22 | Ab Enzymes Oy | Enzyme fusion proteins and their use |
| BR112013016491A2 (pt) * | 2010-12-30 | 2018-05-22 | Novozymes As | método para tratamento de têxtil |
Family Cites Families (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DK122686D0 (da) | 1986-03-17 | 1986-03-17 | Novo Industri As | Fremstilling af proteiner |
| US4832864A (en) * | 1987-09-15 | 1989-05-23 | Ecolab Inc. | Compositions and methods that introduce variations in color density into cellulosic fabrics, particularly indigo dyed denim |
| US5006126A (en) * | 1988-09-15 | 1991-04-09 | Ecolab Inc. | Cellulase compositions and methods that introduce variations in color density into cellulosic fabrics, particularly indigo dyed denim |
| DK115890D0 (da) * | 1990-05-09 | 1990-05-09 | Novo Nordisk As | Enzym |
| AU639570B2 (en) * | 1990-05-09 | 1993-07-29 | Novozymes A/S | A cellulase preparation comprising an endoglucanase enzyme |
| US5475101A (en) | 1990-10-05 | 1995-12-12 | Genencor International, Inc. | DNA sequence encoding endoglucanase III cellulase |
| US5650322A (en) * | 1990-10-05 | 1997-07-22 | Genencor International, Inc. | Methods for stonewashing fabrics using endoglucanases |
| CA2107206A1 (en) * | 1991-03-29 | 1992-09-30 | Kathleen A. Clarkson | Methods for treating cotton-containing fabrics with cellulase |
| US5565006A (en) | 1993-01-20 | 1996-10-15 | Novo Nordisk A/S | Method for the treatment of dyed fabric |
| EP0702713B1 (en) * | 1993-06-11 | 2002-01-30 | Genencor International, Inc. | Enzymatic methods and use of enzymes for producing stonewashed look on indigo-dyed denim fabric |
| CN1129011A (zh) * | 1993-07-12 | 1996-08-14 | 诺沃挪第克公司 | 含有两种纤维素酶成分的洗涤剂组合物 |
| US5861271A (en) | 1993-12-17 | 1999-01-19 | Fowler; Timothy | Cellulase enzymes and systems for their expressions |
| DE69534513T2 (de) * | 1994-03-08 | 2006-07-27 | Novozymes A/S | Neuartige alkalische zellulasen |
-
1996
- 1996-09-03 EP EP96928351A patent/EP0850295B2/en not_active Expired - Lifetime
- 1996-09-03 JP JP51078297A patent/JP3943132B2/ja not_active Expired - Fee Related
- 1996-09-03 WO PCT/DK1996/000364 patent/WO1997009410A1/en active IP Right Grant
- 1996-09-03 ES ES96928351T patent/ES2200070T5/es not_active Expired - Lifetime
- 1996-09-03 TR TR1998/00402T patent/TR199800402T1/xx unknown
- 1996-09-03 AU AU67853/96A patent/AU6785396A/en not_active Abandoned
- 1996-09-03 DE DE69628311T patent/DE69628311T3/de not_active Expired - Lifetime
- 1996-09-03 CN CNB961975997A patent/CN1163577C/zh not_active Expired - Fee Related
- 1996-09-03 PL PL96325478A patent/PL183149B1/pl unknown
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107313273A (zh) * | 2016-04-27 | 2017-11-03 | 尚科纺织企业工业及贸易公司 | 生产含细菌生物聚合物且具有独特外观的染色织物的方法 |
| CN107313273B (zh) * | 2016-04-27 | 2022-03-11 | 尚科纺织企业工业及贸易公司 | 生产含细菌生物聚合物且具有独特外观的染色织物的方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| ES2200070T5 (es) | 2012-03-27 |
| EP0850295B2 (en) | 2011-11-30 |
| CN1163577C (zh) | 2004-08-25 |
| EP0850295B1 (en) | 2003-05-21 |
| MX9801735A (es) | 1998-05-31 |
| WO1997009410A1 (en) | 1997-03-13 |
| JP3943132B2 (ja) | 2007-07-11 |
| TR199800402T1 (xx) | 1998-06-22 |
| AU6785396A (en) | 1997-03-27 |
| DE69628311T2 (de) | 2004-04-01 |
| EP0850295A1 (en) | 1998-07-01 |
| DE69628311T3 (de) | 2012-05-16 |
| JPH11513081A (ja) | 1999-11-09 |
| PL183149B1 (pl) | 2002-05-31 |
| PL325478A1 (en) | 1998-07-20 |
| ES2200070T3 (es) | 2004-03-01 |
| DE69628311D1 (de) | 2003-06-26 |
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