CN1279721A - 具有改变的净电荷、在去污剂中使用的多点置换的蛋白酶变体 - Google Patents

具有改变的净电荷、在去污剂中使用的多点置换的蛋白酶变体 Download PDF

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CN1279721A
CN1279721A CN98811435A CN98811435A CN1279721A CN 1279721 A CN1279721 A CN 1279721A CN 98811435 A CN98811435 A CN 98811435A CN 98811435 A CN98811435 A CN 98811435A CN 1279721 A CN1279721 A CN 1279721A
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subtilisin
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A·J·宝露斯
V·夏兰伯格
J·T·小凯利斯
C·培驰
J·纳德尼
D·P·纳奇
R·M·卡尔德威尔
K·D·科里尔
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Abstract

公开了由天然存在的或重组的非人类蛋白酶的DNA序列衍生的新型蛋白酶变体。变异蛋白酶的获得方法通常是,对编码天然存在的或重组的蛋白酶的前体DNA序列进行体外修饰,在前体蛋白酶的氨基酸序列中引起多处氨基酸残基的置换。提供了如下蛋白酶变体,其在一个或多个残基位置处含有氨基酸置换,该置换改变了该位置的电荷。使其电荷与前体蛋白酶相比负电性更强或正电性更弱,因此该蛋白酶变体在低去污剂浓度体系中比前体蛋白酶更有效。也提供了如下蛋白酶变体,其在一个或多个残基位置处含有氨基酸置换,该置换改变了该位置的电荷,使其电荷与前体蛋白酶相比正电性更强或负电性更弱,因此该蛋白酶变体在高去污剂浓度体系中比前体蛋白酶更有效。

Description

具有改变的净电荷、在去污剂中使用的多点置换的蛋白酶变体
相关申请
本申请是1998年10月23日提交的美国专利中请08／956，323、1998年10月23日提交的美国专利申请08／956，564和1998年10月23日提交的美国专利申请08／956，324的部分继续申请，所有这些申请在此都完整引用。
发明背景
丝氨酸蛋白酶是羰基水解酶的一个亚组。其包括多种类别的具有广泛特性和生物功能的酶。Stroud，R．，科学美国人(Sci．Amer．)131：74-88。尽管其功能的多样性，但丝氨酸蛋白酶的催化机制至少由两种遗传分离的酶家族来完成：1)枯草杆菌蛋白酶和2)哺乳动物胰凝乳蛋白酶相关的丝氨酸蛋白酶及同源细菌丝氨酸蛋白酶(例如胰蛋白酶和灰色链霉菌(S．gresius)胰蛋白酶)。这两个丝氨酸蛋白酶家族显示非常相似的催化机理。Kraut，J．(1977)，生物化学年述(Annu．Rev．Biochem．)，46：331-358。此外，尽管一级结构不相关，但这两个酶家族的三级结构存在由丝氨酸、组氨酸和天冬氨酸组成的氨基酸保守催化三联体。
枯草杆茵蛋白酶是由多种芽孢杆菌属的种及其它微生物大量分泌的丝氨酸蛋白酶(分子量约为27，500)。已经测定了芽孢杆茵属至少九个不同种的枯草杆菌蛋白酶的蛋白质序列。Markland，F．S．等人(1983)，Hooppe-Seyler’s Z．Physiol．Chem．，364：1537-1540。曾经报道了来源于解淀粉芽孢杆菌(Bacillus amyloliquefaciens)、地衣芽孢杆茵(Bacilluslicheniformis)和迟缓芽孢杆菌(B．lentus)几种天然变体的枯草杆菌蛋白酶的三维晶体结构。这些研究表明，尽管枯草杆菌蛋白酶与哺乳动物丝氨酸蛋白酶在遗传上是不相关的，但它具有相似的活性部位结构。含有共价结合的肽抑制剂的枯草杆菌蛋白酶(Robertus，J．D．等人(1972)，生物化学(Biochemistry)，11：2439-2449)或产物复合物(Robertus，J．D．等人(1976)，生物化学杂志(J．Biol．Chem．)，251：1097-1103)的x-射线晶体结构也提供了关于枯草杆菌蛋白酶的活性部位和推断的底物结合裂隙的信息。另外，曾经报道了关于枯草杆菌蛋白酶的大量动力学和化学修饰研究(Svendsen，B．(1976)，Carlsberg研究通讯(Carlsberg Res．Commun．)，41：237-291；Markland，F．S．，同前)，并且至少有一条报道，其中枯草杆菌蛋白酶残基222的甲硫氨酸的侧链被过氧化氢转化为甲硫氨酸-亚砜(Stauffer， D．C．等人(1965)，生物化学杂志，244：5333-5338)，并进行了广泛的位点专一诱变(Wells和Estell(1988)TIBS13：291-297)。
在用于去污剂制剂的蛋白酶变体的开发中，一个常见的问题是洗涤条件的多样性，包括可在其中使用蛋白酶变体的不同去污剂制剂。例如，在不同领域中使用的去污剂配方在洗涤用水(wash water)中含有不同浓度的有关成分。例如，欧洲去污剂体系一般在洗涤用水中含有大约4500-5000ppm的去污剂成分，而日本去污剂体系一般在洗涤用水中含有大约667ppm的去污剂成分。在北美洲，尤其在美国，去污剂体系一般在洗涤用水中含有约975ppm的去污剂成分。令人惊奇地，发展了一种可用于低去污剂浓度体系、高去污剂浓度体系和／或中去污剂浓度体系以及可用于所有这三种去污剂浓度体系的蛋白酶变体的合理设计方法。
发明概述
本发明的一个目的在于提供在一个或多个残基位置处含有氨基酸置换的蛋白酶变体，该置换改变了该位置的电荷，使其电荷与前体蛋白酶相比负电性更强或正电性更弱，因此该蛋白酶变体在低去污剂浓度体系中比前体蛋白酶更有效。一种低去污剂浓度体系是在洗涤用水中存在少于约800ppm的去污剂成分的洗涤体系。
本发明的另一个目的在于提供在一个或多个残基位置处含有氨基酸置换的蛋白酶变体，该置换改变了该位置的电荷，使其电荷与前体蛋白酶相比正电性更强或负电性更弱，因此该蛋白酶变体在高去污剂浓度体系中比前体蛋白酶更有效。一种高去污剂浓度体系是在洗涤用水中存在超过约2000ppm的去污剂成分的洗涤体系。
本发明的另一个目的在于提供在一个或多个残基位置处含有氨基酸置换的蛋白酶变体，该置换改变了该位置的电荷，使其电荷与前体蛋白酶相比正电性更强或负电性更弱，因此该蛋白酶变体在中去污剂浓度体系中比前体蛋白酶更有效。一种中去污剂浓度体系是在洗涤用水中存在约800ppm到约2000ppm的去污剂成分的体系。
本发明的另一个目的在于提供在一个或多个残基位置处含有氨基酸置换的蛋白酶变体，该置换改变了该位置的电荷，使其电荷与前体蛋白酶相比负电性更强或正电性更弱，因此该蛋白酶变体在中去污剂浓度体系中比前体蛋白酶更有效。一种中去污剂浓度体系是在洗涤用水中存在约800ppm到约2000ppm的去污剂成分的洗涤体系。
本发明另外一个目的在于提供编码这些蛋白酶变体的DNA序列，以及含有这些变异体DNA序列的表达载体。
更进一步，本发明的另一目的在于提供用这些载体转化的宿主细胞，以及能够表达这种DNA而在细胞内或细胞外产生蛋白酶变体的宿主细胞。
进一步提供一种含有本发明的蛋白酶变体的清洁用组合物。
另外，也提供一种含有本发明的蛋白酶变体的动物饲料。
进一步提供一种产生在低、中及高去污剂浓度体系中都比前体蛋白酶更有效的蛋白酶变体的方法，包括：
a)置换一个或多个残基位置处的氨基酸，其中该置换改变了该位置的电荷，使得其电荷与前体蛋白酶相比正电性更强或负电性更弱；
b)置换一个或多个残基位置处的氨基酸，其中该置换改变了该位置的电荷，使得其电荷与前体蛋白酶相比负电性更强或正电性更弱；
c)检测该变体，以测定它在高、中及低去污剂浓度体系中与前体蛋白酶相比的有效性；及
d)必要时重复步骤a)-c)，以产生在低、中及高去污剂浓度体系中比前体蛋白酶更有效的蛋白酶变体，其中步骤a)和b)能以任意顺序进行。
附图简述
图1A-C描述解淀粉芽孢杆菌枯草杆菌蛋白酶的DNA及氨基酸序列，和该基因的部分限制酶切图谱。
图2描述解淀粉芽孢杆菌(BPN)’和迟缓芽孢杆菌(野生型)枯草杆菌蛋白酶之间的保守氨基酸残基。
图3A和3B描述四种枯草杆菌蛋白酶的氨基酸序列。最上一行代表来源于解淀粉芽孢杆菌枯草杆菌蛋白酶(有时也称为枯草杆菌蛋白酶BPN’)的枯草杆菌蛋白酶的氨基酸序列。第二行描述来源于枯草芽孢杆菌的枯草杆菌蛋白酶的氨基酸序列。第三行描述来源于地衣芽孢杆菌的枯草杆菌蛋白酶的氨基酸序列。第四行描述来源于迟缓芽孢杆菌的枯草杆菌蛋白酶(在PCT WO89／06276中也称为枯草杆菌蛋白酶309)的氨基酸序列。符号*表示与枯草杆菌蛋白酶BPN’相比特定氨基酸残基缺如。
发明详述
如上所述，特定地区具有特定的洗涤条件，因而使用不同类型的去污剂。例如，日本使用低去污剂浓度体系，而欧洲使用高去污剂浓度体系。如前所述，美国使用中去污剂浓度体系。我们发现，在这些不同的去污剂制剂中表现最佳的蛋白酶变体各不相同。然而，由这些观察的结果可以预料，发现一种适用于所有这三种去污剂类型中的蛋白酶是不可能的。令人惊奇地，情况并非如此。已经发展了一种方法，用于合理设计可在低去污剂浓度体系或高去污剂浓度体系乃至中去污剂浓度体系中使用的蛋白酶变体，以及在所有这三种去污剂浓度体系都适用的蛋白酶变体。我们发现，为了产生在低去污剂浓度体系中更有效的蛋白酶变体，用带负电荷残基或中性残基替换带正电残基和／或用带负电残基替换中性残基是必要的。相反，我们注意到，为了产生在高去污剂浓度体系中更有效的蛋白酶变体，用带正电残基或中性残基替换带负电残基和／或用带正电残基替换中性残基是必要的。此外，我们发现，在低去污剂浓度体系和／或高去污剂浓度体系中有用的许多蛋白酶变体在中去污剂浓度体系中也有效。通过平衡这些变化，可能产生在低去污剂浓度体系、低及中去污剂浓度体系、中及高去污剂浓度体系、高去污剂浓度体系或所有这三种去污剂浓度体系中有出色表现的蛋白酶变体。
假定在水溶液中具有酸性或碱性功能的任何离子化氨基酸侧链的静电电荷是pH的函数。酸性残基Glu和Asp在平衡过程中由于在pH3-6之间解离而失去一个质子，由此得到一个负电荷。His、Lys和Arg分别在pH5-8、pH8．5-11．5和pH11-14之间以相似的方式逐步去质子化，由此失去一个正电荷。Tyr OH的质子在pH8．5-11．5之间逐渐解离，Tyr由此得到一个负电荷。羧基端的解离范围是pH1-4，产生一个负电荷，对于氨基端是pH8-11，伴随着一个正电荷的丢失。对于此处列出的氨基酸侧链，解离范围是多种蛋白质的平均值，但公知在某些蛋白质中它受异常结构构型的影响。
所有电荷的累积效应决定了一种蛋白质在特定的pH时是具有净正电荷还是净负电荷。正电荷和负电荷等效并使蛋白质处于静电中性状态时的pH称为等电点(pI)。当pH改变或者添加或去除含有离子化侧链残基的氨基酸时，蛋白质将失去或得到电荷。通过将在特定pH时带负电的残基替换为不带电的或带正电的残基，分别导致+1及+2的形式电荷改变，能实现净正电荷的增加。通过将不带电侧链残基替换为在特定pH时质子化的残基，形式电荷改变将为+1。同样，将在观测pH时带正电及不带电的侧链替换为带负电的侧链，能增加净负电荷，并分别得到-1和-2的负电荷形式增加。
低去污剂浓度体系包括在洗涤用水中存在低于大约800ppm的去污剂成分的去污剂。日本去污剂一般被认为是低去污剂浓度体系，因为它们在洗涤用水中存在大约667ppm的去污剂成分。
中去污剂浓度包括在洗涤用水中存在大约800ppm到约2000ppm的去污剂成分的去污剂。北美去污剂一般被认为是中去污剂浓度体系，因为它们在洗涤用水中存在大约975ppm的去污剂成分。巴西的体系一般在洗涤用水中存在大约1500ppm的去污剂成分。
高去污剂浓度体系包括在洗涤用水中存在高于大约2000ppm的去污剂成分的去污剂。欧洲去污剂一般被认为是高去污剂浓度体系，因为它们在洗涤用水中存在大约4500-5000ppm的去污剂成分。
拉丁美洲去污剂一般是高泡沫磷酸盐助洗去污剂，在拉丁美洲使用的去污剂范围可能处于中及高去污剂浓度，因为在洗涤用水中它们有1500ppm到6000ppm的去污剂成分。如上所述，巴西的体系一般在洗涤用水中含有大约1500ppm的去污剂成分。然而，其它高泡沫磷酸盐助洗去污剂地区，不限于其它拉丁美洲国家，可能有在洗涤用水中存在可达大约6000ppm的去污剂成分的高去污剂浓度体系。
根据前述，显然典型洗涤溶液中去污剂组合物的浓度在全世界范围内各不相同，从低于大约800ppm的去污剂组合物(“低去污剂浓度地区”)，例如在日本为大约667ppm，到约800ppm-约2000ppm(“中去污剂浓度地区”)，例如在美国约为975ppm、在巴西约为1500ppm，到高于大约2000ppm(“高去污剂浓度地区”)，例如在欧洲为大约4500ppm-约5000ppm，在高泡沫助洗剂地区约为6000ppm。
典型洗涤溶液的浓度由经验确定。例如，在美国，一种典型的洗涤机可容纳大约64．4升体积的洗涤溶液。因此，为了获得在洗涤溶液中大约975ppm去污剂的浓度，必须向64．4升洗涤溶液中加入大约62．79克去污剂组合物。这种量是由消费者用去污剂所带量杯测量的加入洗涤用水中的典型量。
蛋白酶是通常用来切割蛋白质或肽的肽键的羰基水解酶。在此使用时，“蛋白酶”意思是天然存在的蛋白酶或重组蛋白酶。天然存在的蛋白酶包括α-氨酰肽水解酶、肽酰氨基酸水解酶、酰基氨基水解酶、丝氨酸羧基肽酶、金属羧基肽酶、巯基蛋白酶、羧基蛋白酶和金属蛋白酶。包括丝氨酸、金属、巯基和酸性蛋白酶，以及内切蛋白酶和外切蛋白酶。
本发明包括非天然存在的羰基水解酶变体的蛋白酶(蛋白酶变体)，与衍生出该变体氨基酸序列的前体羰基水解酶相比，其具有不同的蛋白水解活性、稳定性、底物特异性、pH分布曲线和／或性能特征。特别是，这些蛋白酶变体具有自然界中未知的氨基酸序列，它是由于前体蛋白酶的大多数氨基酸残基被替换为不同氨基酸而产生的。前体蛋白酶可以是天然存在的蛋白酶或重组蛋白酶。
此处有用的蛋白酶变体包含19种天然存在的L-氨基酸中任一种在指定氨基酸残基位置处的置换。这些置换能在任何前体枯草杆菌蛋白酶(原核、真核、哺乳动物等)中进行。在整个申请中，以常用的单字母及三字母密码的形式提及不同的氨基酸。这些密码由Dale，M．W．(1989)，《细茵分子遗传学》(Molecular Genetics of Bacteria)，John Wiley & Sons，Ltd．，附录B确定。
此处有用的蛋白酶变体优选地来源于芽孢杆菌属的枯草杆菌蛋白酶。更优选地，蛋白酶变体来源于迟缓芽孢杆菌枯草杆菌蛋白酶和／或枯草杆菌蛋白酶309。
枯草杆菌蛋白酶是一般用来切割蛋白质或肽的肽键的细菌或真菌蛋白酶。在此使用时，“枯草杆菌蛋白酶”意思是天然存在的枯草杆菌蛋白酶或重组枯草杆菌蛋白酶。已知一系列天然存在的枯草杆菌蛋白酶是由不同微生物种产生且常常为分泌的。该系列成员的氨基酸序列不是完全同源的。然而，该系列中的枯草杆菌蛋白酶显示相同或相似类型的蛋白水解活性。该类丝氨酸蛋白酶具有一种共同的氨基酸序列，该序列定义了有别于胰凝乳蛋白酶相关种类丝氨酸蛋白酶的催化三联体。枯草杆菌蛋白酶和胰凝乳蛋白酶相关的丝氨酸蛋白酶都含有包括天冬氨酸、组氨酸和丝氨酸的催化三联体。在枯草杆菌蛋白酶相关蛋白酶中，这些氨基酸从氨基端向羧基端阅读的相对顺序是天冬氨酸-组氨酸-丝氨酸。但是，在胰凝乳蛋白酶相关蛋白酶中相对顺序为组氨酸-天冬氨酸-丝氨酸。因此，枯草杆菌蛋白酶在此是指含有枯草杆菌蛋白酶相关蛋白酶的催化三联体的丝氨酸蛋白酶。实例包括但不限于此处图3中所确定的枯草杆菌蛋白酶。一般而言，并且对于本发明，蛋白酶中氨基酸的编号对应于分配给图1所示成熟解淀粉芽孢杆菌枯草杆菌蛋白酶序列的编号。
“重组枯草杆菌蛋白酶”或“重组蛋白酶”是指一种枯草杆菌蛋白酶或蛋白酶，其中编码枯草杆菌蛋白酶或蛋白酶的DNA序列受到修饰，产生一种变异(或突变)DNA序列，该序列编码天然存在的氨基酸序列中一种或多种氨基酸的置换、缺失或插入。产生这种修饰、并且可与此处公开的方法结合的适当方法包括美国专利RE34，606、美国专利5，204，015和美国专利5，185，258、美国专利5，700，676、美国专利5，801，038和美国专利5，763，257中公开的方法。
“非人类的枯草杆菌蛋白酶”及其编码DNA可以从多种原核及真核生物中获得。原核生物的合适实例包括革兰氏阴性生物如大肠杆菌或假单胞菌(Pseudomonas)、革兰氏阳性菌如微球菌(Micrococcus)或芽孢杆菌。可从中获得枯草杆菌蛋白酶及其基因的真核生物的实例包括酵母如酿酒酵母(Saccharomyces cerevisiae)、真菌如曲霉(Aspergillus)属的种。
“蛋白酶变体”具有由“前体蛋白酶”的氨基酸序列衍生的氨基酸序列。前体蛋白酶包括天然存在的蛋白酶和重组蛋白酶。蛋白酶变体的氨基酸序列通过前体氨基酸序列中一种或多种氨基酸的置换、缺失或插入而由前体蛋白酶氨基酸序列“衍生”。这种修饰是编码前体蛋白酶氨基酸序列的“前体DNA序列”的修饰，而不是前体蛋白酶自身的操作。对前体DNA序列进行这种操作的适当方法包括此处公开的方法，以及本领域技术人员公知的方法(参见，例如，EP 0 328299、WO89／06279和已经在此引用的美国专利及申请)。
这些氨基酸位置编号是指那些分配给图1所示成熟解淀粉芽孢杆菌枯草杆菌蛋白酶序列的编号。然而，本发明不限于该特定枯草杆菌蛋白酶的突变，而是扩展到在一定位置处含有这样的氨基酸残基的前体蛋白酶，该残基“等同”于解淀粉芽孢杆菌枯草杆菌蛋白酶中特别确定的残基。在本发明的一个优选实施方案中，前体蛋白酶是迟缓芽孢杆菌枯草杆菌蛋白酶，并在迟缓芽孢杆菌中对应于以上所列位点的等同氨基酸残基位置处进行置换。
如果前体蛋白酶的残基(氨基酸)位置与解淀粉芽孢杆菌枯草杆菌蛋白酶中的特定残基或该残基的一部分同源(即，在一级或三级结构中位置相对应)或类似(即，具有相同或相似的化学结合、反应或相互作用的功能能力)，则它相当于解淀粉芽孢杆菌枯草杆菌蛋白酶的残基。
为了建立与一级结构的同源性，将前体蛋白酶的氨基酸序列直接与解淀粉芽孢杆菌枯草杆菌蛋白酶一级序列相比较，特别与公知在序列已知的枯草杆菌蛋白酶中不变的一组残基相比较。例如，在此图2显示解淀粉芽孢杆菌枯草杆菌蛋白酶与迟缓芽孢杆菌枯草杆菌蛋白酶之间的保守残基。对比保守残基，为维持序列对比进行必要的插入和缺失(即，避免保守残基由于任意缺失和插入而消除)之后，确定等同于解淀粉芽孢杆菌枯草杆菌蛋白酶一级序列中特定氨基酸的残基。保守残基的对比优选地应当保持100％的这些残基。然而，超过75％或低至50％的保守残基的对比也足以确定等同的残基。催化三联体Asp32／His64／Ser221的保守性应当保持。Siezen等人(1991)蛋白质工程(Protein Eng．)4(7)：719-737显示了大量丝氨酸蛋白酶的序列对比。Siezen等人将此分组称为枯草杆菌酶(Subtilase)或枯草杆菌蛋白酶样丝氨酸蛋白酶。例如，图3中，排列对比了解淀粉芽孢杆菌、枯草芽孢杆菌、地衣芽孢杆菌(carlsbergensis)和迟缓芽孢杆菌的枯草杆菌蛋白酶之氨基酸序列，以提供氨基酸序列之间最大量的同源性。这些序列的比较显示，在每种序列中含有许多保守残基。图2中确定了这些保守残基(BPN’与迟缓芽孢杆菌之间)。
因此，可用这些保守残基确定其它枯草杆菌蛋白酶，诸如迟缓芽孢杆菌枯草杆菌蛋白酶(1989年7月13日公布的PCT公开文本WO89／06279)、此处优选的蛋白酶前体酶或被称为PB92的枯草杆菌蛋白酶(EP 0 328 299，它与优选的迟缓芽孢杆菌枯草杆菌蛋白酶高度同源)中与解淀粉芽孢杆菌枯草杆菌蛋白酶相应的等同氨基酸残基。在图3A和3B中将这些枯草杆菌蛋白酶中某些氨基酸序列与解淀粉芽孢杆菌枯草杆菌蛋白酶的序列相对比，产生保守残基的最大同源性。如所能看到的，与解淀粉芽孢杆菌枯草杆菌蛋白酶相比，迟缓芽孢杆菌序列中存在大量缺失。因此，例如，在其它枯草杆菌蛋白酶中与解淀粉芽孢杆菌枯草杆菌蛋白酶中的Val165等同的氨基酸对于迟缓芽孢杆菌和地衣芽孢杆菌而言是异亮氨酸。
也可以通过测定已经通过x-射线晶体分析法确定三级结构的前体蛋白酶在三级结构水平上的同源性，来确定“等同的残基”。等同的残基被定义为前体蛋白酶和解淀粉芽孢杆菌枯草杆菌蛋白酶特定氨基酸残基的两个或多个主链原子的原子坐标(N对N、CA对CA、C对C和O对O)经对比后在0．13nm优选地0．1nm之内的残基。在定向并定位最佳模型之后完成对比，得到所述蛋白酶与解淀粉芽孢杆菌枯草杆菌蛋白酶的非氢蛋白质原子的原子坐标的最大重叠。最佳模型是在可获得的最高分辨力时对于实验衍射数据可得到最低R因子的晶体模型。
R因子=(贴11页第4行公式)
与解淀粉芽孢杆菌枯草杆菌蛋白酶特定残基功能类似的等同残基被定义为前体蛋白酶的如下氨基酸，它们可采取一种构象，使得它们以一种确定的方式改变、修饰或贡献于蛋白质结构、底物结合或催化，且其归因于解淀粉芽孢杆菌枯草杆菌蛋白酶的特定残基。此外，它们是前体蛋白酶的那类残基(通过x-射线晶体分析法已获得其三级结构)，其占据了类似位点，使得尽管基于占据同源位点，特定残基的主链原子也许不能满足等同的标准，但该残基的至少两个侧链原子之原子坐标位于解淀粉芽孢杆菌枯草杆菌蛋白酶相应侧链原子的0．13nm内。在EPO公开文本0 251 446(相当于美国专利5，182，204，其公开内容在此引用作为参考)中阐述了解淀粉芽孢杆菌枯草杆菌蛋白酶的三维结构的坐标，并能如上所述用来在三级结构水平上确定等同的残基。
为了置换所确定的某些残基是保守残基，而其它的则不是。对于不保守的残基而言，一种或多种氨基酸的置换限于产生这样一种变体的置换，该变体具有一种与自然中所发现的氨基酸序列不对应的序列。对于保守残基而言，这些置换不能产生天然存在的序列。本发明的蛋白酶变体包括蛋白酶变体的成熟形式，以及这些蛋白酶变体的原-形式和前原-形式。前原-形式是优选的结构，因为其有利于蛋白酶变体的表达、分泌和成熟。
“原序列”是指与蛋白酶成熟形式的N端部分结合的氨基酸序列，当它被去除后，导致该蛋白酶“成熟”形式的出现。实质上发现许多蛋白水解酶为翻译酶原产物，在不存在翻译后加工的情况下以该方式表达。用于产生蛋白酶变体的一种优选原序列是解淀粉芽孢杆菌枯草杆菌蛋白酶的推定的原序列，尽管也可使用其它蛋白酶原序列。
“信号序列”或“前序列”是指与蛋白酶N端部分结合或与原蛋白酶N端部分结合的任何氨基酸序列，它们可能参与蛋白酶成熟形式或原形式的分泌。信号序列的这种定义是一种功能性定义，意为包括蛋白酶基因N端部分所编码的所有氨基酸序列，其在天然条件下参与蛋白酶分泌的实现。本发明利用这些序列实现此处所述蛋白酶变体的分泌。一种可能的信号序列包含枯草芽孢杆菌枯草杆菌蛋白酶信号序列的前7个氨基酸残基，这些残基与迟缓芽孢杆菌枯草杆菌蛋白酶信号序列(ATCC21536)的剩余部分相融合。
蛋白酶变体的“前原”形式由含有与蛋白酶氨基端有效连接的原序列的蛋白酶成熟形式和与原序列氨基端有效连接的“前”序列或“信号”序列组成。
“表达载体”是指一种DNA构建体，其含有与能实现该DNA在适当宿主中表达的适当控制序列有效连接的DNA序列。这类控制序列包括实现转录的启动子、控制这种转录的任意操纵子序列、编码适当mRNA核糖体结合位点的序列和控制转录与翻译终止的序列。载体可以是质粒、噬菌体颗粒，或者仅仅是有效的基因组插入片段。一旦转化到适当宿主中，载体可不依赖于宿主基因组复制并起作用，或者在某些情况中可能整合到基因组自身中。在本说明书中，“质粒”和“载体”有时交换使用，因为质粒是目前最常用的载体形式。然而，本发明意在包括表达载体的其它形式，它们具有等同的功能，并且是或者可成为本领域所公知的。
本发明中所用的“宿主细胞”通常是原核或真核宿主，它们优选地用美国专利RE 34，606中公开的方法操作，以使其不能分泌酶促活性的内切蛋白酶。表达蛋白酶的一种优选宿主细胞是芽孢杆菌株BG2036，它缺乏具有有酶活性的中性蛋白酶和碱性蛋白酶(枯草杆菌蛋白酶)。美国专利5，264，366中详述了菌株BG2036的构建。表达蛋白酶的其它宿主细胞包括枯草芽孢杆菌I168(在美国专利RE 34，606和美国专利5，264，366中也有描述，其公开内容在此引用作为参考)，以及任何合适的芽孢杆菌属的菌株，如地衣芽孢杆菌、迟缓芽孢杆菌等。
用以重组DNA技术构建的载体转化或转染宿主细胞。这些转化的宿主细胞能复制编码蛋白酶变体的载体或表达希望的蛋白酶变体。对于编码蛋白酶变体前-形式或前原-形式的载体而言，这些变体当表达时一般由宿主细胞向宿主细胞培养基中分泌。
当描述两个DNA区域之间的关系时，“有效连接”仅意味着它们在功能上彼此相关。例如，如果一种前序列作为信号序列参与蛋白质成熟形式的分泌，最可能的是参与信号序列的切割，则该前序列与肽有效连接。如果一种启动子控制一种编码序列的转录，则该启动子与该序列有效连接；如果一种核糖体结合位点位于一定位置使得翻译成为可能，则它与编码序列有效连接。
编码天然存在的前体蛋白酶的基因可依照本领域技术人员公知的通用方法获得。这些方法一般包括：合成含有编码目标蛋白酶区推定序列的标记探针、由表达该蛋白酶的生物制备基因组文库，以及通过与该探针杂交为了获得目标基因筛选该文库。然后对阳性杂交克隆作图并测定。
然后用克隆的蛋白酶转化宿主细胞，以表达该蛋白酶。然后将蛋白酶基因连接于高拷贝数质粒中。该质粒在宿主中复制，在此意义上它含有质粒复制所必需的众所周知的元件：与所述基因有效连接的启动子(如果它可被宿主识别，即转录，则可作为基因本身的同源启动子)、转录终止区和聚腺苷酸化区(在某些真核宿主细胞中，它对于由宿主从蛋白酶基因转录的mRNA的稳定性是必需的)，它是外源的或由蛋白酶基因的内源终止区所提供的，质粒中还期望含有选择性基因，如抗生素抗性基因，它使得质粒感染的宿主细胞通过在含有抗生素的培养基中生长以维持连续培养成为可能。高拷贝数质粒也含有宿主的复制起点，从而能在细胞质中不受染色体限制地产生大量质粒。然而，宿主基因组中整合多拷贝的蛋白酶基因也处于本发明之内。利用对同源重组特别敏感的原核和真核生物利于此过程。
该基因可能是天然迟缓芽孢杆菌基因。此外，也可以产生编码天然存在的或突变的前体蛋白酶的合成基因。这种方法中，测定前体蛋白酶的DNA和／或氨基酸序列。之后合成多个、重叠的合成单链DNA片段，杂交并连接后产生编码前体蛋白酶的合成DNA。在美国专利5，204，015的实施例3中描述了合成的基因构建体的一个实例，其公开内容在此引用作为参考。
一旦克隆了天然存在的或合成的前体蛋白酶基因，即进行大量修饰，以增强天然存在的前体蛋白酶合成之外的基因用途。这些修饰包括如美国专利RE 34，606和EPO公布号0 251 446所公开的重组蛋白酶的产生和此处所述的蛋白酶变体的产生。
可用下列盒式诱变法促进本发明的蛋白酶变体的构建，尽管也可使用其它方法。首先，获得天然存在的编码蛋白酶的基因，并全部或部分测序。然后为了寻找希望在编码的酶中进行一个或多个氨基酸突变(缺失、插入或置换)的位点，扫描该序列。评价侧翼于该位点的序列是否存在估计限制酶切位点的存在，用于将基因的短片段替换为表达时编码不同突变体的寡核苷酸集合体。这些限制酶切位点优选地是蛋白酶基因内的唯一位点，以便于基因片段的置换。然而，可使用在蛋白酶基因中不过度丰余的任何适宜的限制酶切位点，只要限制酶切消化产生的基因片段能够在适当的序列中重新装配即可。如果在与所选位点适当距离内(10-15个核苷酸)的位置处不存在限制酶切位点，则可通过以在最终构建中不改变读框也不改变所编码氨基酸的方式置换基因中的核苷酸来产生这些位点。为了改变其序列以符合目标序列，基因突变依照公知的方法通过M13引物延伸来实现。根据遗传密码的丰余性、基因的限制酶图谱和大量不同的限制酶，常规进行定位适当侧翼区并估计所需改变以得到两种适宜的限制酶切位点序列的工作。注意到如果可获得适宜的侧翼限制酶切位点，则上述方法需要仅仅与不合该位点的侧翼区结合应用。
一旦克隆了天然存在的DNA或合成的DNA，即用同源的限制酶消化侧翼于待突变位点的限制酶切位点，并将多数末端互补的寡核苷酸盒连接于该基因中。该方法简化了诱变，因为能合成所有寡核苷酸使其具有相同的限制酶切位点，且合成的接头不是产生限制酶切位点所必需的。
在此使用时，蛋白水解活性被定义为每毫克活性酶的肽键水解率。测定蛋白水解活性有许多众所周知的方法(K．M．Kalisz，“微生物蛋白酶”《生物化学工程／生物技术进展》(Advances in BiochemicalEngineering／Biotechnology)，A．Fiechter编，1988)。除了改进的蛋白水解活性之外，或者作为一种备择，本发明的变异蛋白酶可能具有其它改进的性质，如Km、kcat或kcat／Km比值和／或改进的底物特异性和／或改进的pH活性分布曲线。这些酶能量身定做般地用于例如预计在肽制剂中存在的特定底物，或适于水解过程如洗衣用途。
本发明的一个方面，目的在于获得一种与前体蛋白酶相比具有改变的蛋白水解活性的变异蛋白酶，因为提高这种活性(数字上的变大)使得酶更有效地作用于靶底物的用途成为可能。同样需要的是与前体相比具有改变的热稳定性和／或改变的底物特异性的变异酶。在有些情况中，可能希望较低的蛋白水解活性，例如，在需要蛋白酶合成活性时(对于合成肽)蛋白水解活性的降低可能是有用的。人们可希望降低该蛋白水解活性，该活性能破坏这种合成的产物。相反，在某些情况中，可能希望提高变异酶与其前体相比的蛋白水解活性。此外，变体稳定性-碱稳定性或热稳定性-的提高或降低(改变)可能是所希望的。kcat、Km或kcat／Km的增加或降低对于用来测定这些动力学参数的底物是特异性的。
本发明的另一方面，发现如下的蛋白酶变体在低去污剂浓度中比前体蛋白酶更有效，该蛋白酶变体在一个或多个残基位置处含有氨基酸置换，该置换改变了该位置的电荷，使其电荷与前体蛋白酶相比负电性更强或正电性更弱。
本发明的再另一方面，发现如下的蛋白酶变体在高去污剂浓度中比前体蛋白酶更有效，该蛋白酶变体在一个或多个残基位置处含有氨基酸置换，该置换改变了该位置的电荷，使其电荷与前体蛋白酶相比正电性更强或负电性更弱。
此外，我们发现，在低去污剂浓度体系和／或高去污剂浓度体系中有用的许多蛋白酶变体在中去污剂浓度体系中也是有效的。
这些置换优选地在迟缓芽孢杆菌(重组的或天然型)枯草杆菌蛋白酶中进行，尽管该置换也可在任何芽孢杆菌蛋白酶，优选地在芽孢杆菌枯草杆菌蛋白酶中进行。
根据用变异蛋白酶获得的筛选结果，解淀粉芽孢杆菌枯草杆菌蛋白酶中的显著突变对于这些酶的蛋白水解活性、性能和／或稳定性以及这些变异酶的清洁或洗涤能力至关重要。
本发明的许多蛋白酶变体可用于配制不同的去污剂组合物或个人护理制剂如洗发剂或洗涤剂。许多公知的化合物是在含有本发明之蛋白酶突变体的组合物中有用的适当表面活性剂。其包括非离子、阴离子、阳离子或两性离子去污剂，如Barry J．Anderson的US 4，404，128和Jiri Flora等人的US 4，261，868中所公开的。一种合适的去污剂制剂是美国专利5，204，015(前文引用作为参考)的实施例7中所述的制剂。本领域技术人员熟悉能用作清洁用组合物的不同制剂。除了典型的清洁用组合物之外，易于理解本发明的蛋白酶变体也可用于使用天然或野生型蛋白酶的任何目的。因此，例如，这些变体能用于固体肥皂或液体肥皂的用途、餐具清洁(dishcare)剂、隐形眼镜清洁液或产品、肽水解、废物处理、纺织品用途、用作蛋白质生产中的融合-切割酶，等等。本发明的变体可在去污剂组合物中具有增强的性能(与前体相比)。在此使用时，在去污剂中的增强性能定义为某些酶敏感性污迹如草或血液的清除加强，如在标准洗涤循环后通过通用评估所测定的。
本发明的蛋白酶能以大约0．01％到大约5％(优选地0．1％-0．5％)的重量百分比水平配制于已知的pH6．5-12．0的粉末及液体去污剂中。这些去污剂清洁用组合物也能包含其它的酶如已知的蛋白酶、淀粉酶、纤维素酶、脂肪酶或糖苷内切酶，以及助洗剂和稳定剂。
本发明的蛋白酶向常规清洁用组合物中的添加不产生任何特殊的应用限制。换句话说，适合于去污剂的任何温度和pH也适合于该组合物，只要pH位于上述范围之内，而温度低于所述蛋白酶的变性温度。另外，本发明的蛋白酶也能在不含去污剂的清洁用组合物中使用，单独或与助洗剂和稳定剂结合使用。
本发明也涉及含有本发明的蛋白酶变体的清洁用组合物。清洁用组合物另外也可含有常用于清洁用组合物的添加剂。其能选自但不限于：漂白剂、表面活性剂、助洗剂、酶和漂白催化剂。本领域的一名普通技术人员可容易地理解何种添加剂适合包含于组合物中。此处提供的列表绝不是完全的，而只是作为适当添加剂的实例。本领域的一名普通技术人员可容易地理解，应当只使用那些与该组合物中的酶及其它成分如表面活性剂相适合的添加剂。
制成后，清洁用组合物中存在的添加剂的量为大约0．01％-约99．9％，优选地约1％-约95％，更优选地约1％-约80％。
本发明的变异蛋白酶能包含于动物饲料中，如作为动物饲料添加剂部分，例如，如US 5，612，055、US 5，314，692和US 5，147，642所述。
本发明的一个方面是含有本发明的变异蛋白酶的用于纺织品处理的组合物。如出版物如RD 216，034、EP 134，267、US 4，533，359和EP 344，259中所述，能用该组合物处理如丝绸或毛。
以下以实施例的方式描述，而不应理解为对权利要求书范围的限制。
此处引用的所有出版物和专利在此都完全引入作为参考。实施例1
用本领域众所周知的方法生产并纯化大量蛋白酶变体。所有突变都在迟缓芽孢杆菌GG36枯草杆菌蛋白酶中进行。
用美国系列号60／068，796“一种测定优选的酶和／或优选的去污剂组合物的改进方法”中所述的微观(microswatch)测定法检测产生的蛋白酶变体在两种去污剂及洗涤条件中的表现。
表1-13列出了所测定的变异蛋白酶和在两种不同去污剂中检测的结果。所有数值都是与表中所示第一种蛋白酶比较而得出的(即，1．32的值表示，与表中第一种变体的100％相比释放132％的污物的能力)。
A栏显示变体的电荷差异。对于B栏，去污剂是0．67 g／l过滤的ArielUltra(Procter & Gamble，Cincinnati，OH，美国)，溶于每加仑含有3格令混合Ca2+／Mg2+硬度的溶液中，25℃下每孔使用0．3ppm的酶(低浓度去污剂体系)。对于C栏，去污剂是3．38 g／l过滤的Ariel Futur(Procter &Gamble，cinciNNati，OH，美国)，溶于每加仑含有15格令混合Ca2+／Mg2+硬度的溶液中，40℃下每孔使用0．3ppm的酶(高浓度去污剂体系)。
表1
    A     B     C
 N76D  S103A  V104I  Q109R     1.00     1.00
 N76D  S103A  V104I  Q109R  Q245R     +1     0.48     1.41
表2
 A  B  C
 V68A  N76D  S103A  V104I  G159D  Q236H  Q245R  1.00  1.00
 V68A  N76D  S103A  V104I  G159D  N204D  Q236H  Q245R -1  1.11  0.03
表3
 A     B     C
 V68A  N76D     S103A  V104I     1.00     1.00
 T22K  V68A     N76D  S103A  V104I  +1     0.74     1.85
表4
 A     B     C
 N76D  S103A  V104I  M222S     1.00     1.00
 N76D  S103A  V104I  N173R  M222S  0     0.66     1.84
 Q12R  N76D  S103A  V104I  M222S  Q245R  +1     0.41     5.84
表5
 A  B  C
 Q12R  N76D  S103A  I104T  S130T  M222S  Q245R  1.00  1.00
 Q12R  N76D  S103A  I104T  S130T  M222S  Q245R  N261D  -1  1.79  0.81
 Q12R  N76D  S103A  I104T  S130T  R170S  N185D  M222S  N243D  Q245R  -3  2.87  0.02
表6
 A     B     C
 V68A  N76D  S103A  V104I  G159D  Q236H     1.00     1.00
 V68A  N76D  S103A  V104I  G159D  Q236H  Q245R  +1     0.94     6.80
 V68A  S103A  V104I  G159D  A232V  Q236H  Q245R N252K  +3     0.44     20.60
表7
 A  B  C
 V68A  N76D  S103A  V104I  G159D  A232V  Q236H  Q245R  1.00  1.00
V68A  N76D  S103A  V104I  G159D  P210R  A232V  Q236H  Q245R  +1  0.44  2.66
表8
 A  B     C
V684 S103A  V104I G159D A232V Q236H  Q245R  N252K  1.00     1.00
V68A S103A  V104I G159D A232V Q236H  Q245R  N248D  N252K  -1  1.96     0.65
表9
 A     B     C
 V68A  S103A  V104I  G159D  A232V  Q236H  Q245R  1.00   1.00
 V68A  S103A  V104I  G159D  A232V  Q236H  K237E  Q245R -2  1.27   0.12
表10
 A  B C
 V68A S103A  V104I G159D  A232V  Q236H  Q245R L257V  1.00 1.00
 V68A N76D  S103A V104I  G159D  A232V  Q236H Q245R L257V -1  1.56 0.48
表11
 A  B  C
 S103A  V104I  G159D  A232V Q236H Q245R N248D N252K  1.00  1.00
 S103A  V104I  G159D  L217E A232V Q236H Q245R N248D N252K  -1  1.90  0.15
表12
 A  B  C
 S103A V104I S101G G159D A232V Q236H Q245R  N248D  N252K  1.00 1.00
 N76D S103A V104I S101G G159D A232V Q236H  Q245R  N248D  N252K -1  1.28 0.39
表13
 A  B  C
 N63D S103A  V104I G159D T213R A232V Q236H Q245R  N248D  N252K  1.00  1.00
 N62D S103A  V104I Q109R G159D T213R A232V Q236H  Q245R  N248D  N252K +1  0.40  1.74
实施例2
如实施例1所述制备并检测下列蛋白酶变体。
表14中的变体是具有以下两种类型置换的蛋白酶变体：改变一个位置处的电荷使其电荷与迟缓芽孢杆茵GG36相比负电性更强或正电性更弱的置换，和改变一个位置处的电荷使其电荷正电性更强或负电性更弱的置换，以及不影响特定残基位置电荷的中性置换。这产生在低去污剂浓度体系(A栏；0.67g/l过滤的Ariel Ultra(Procter & Gamble，Cincinnati，OH,美国)，溶于每加仑含有3格令混合Ca2+/Mg2+硬度的溶液中，25℃下每孔使用0.3ppm的酶)和高去污剂浓度体系(B栏；3.38g/l过滤的ArielFutur(Procter & Gamble，Cincinnati,OH，美国)，溶于每加仑含有15格令混合Ca2+/Mg2+硬度的溶液中，40℃下每孔使用0。3ppm的酶)中都比标准表现更好的蛋白酶变体。
表14
A  B
 N76D S103A  V104I 1.00 1.00
 V68A S103A  V104I G159D A232V Q236H  Q245R  N252K 1.41 1.85
 V68A N76D  S103A V104I G159D T213R  A232V  Q236H  Q245R  T260A 1.30 1.73
 V68A S103A  V104I G159D A232V Q236H  Q245R  N248D  N252K 2.77 1.20
 V68A S103A  V104I N140D G159D A232V  Q236H  Q245R  N252K 2％ 1.42
 N43K V68A  S103A V104I G159D A232V  Q236H  Q245R 2.05 1.78
 N43D V68A  S103A V104I G159D A232V  Q236H  Q245R  N252K 2.00 1.34
 V68A N76D  S103A V104I G159D A215R  A232V  Q236H  Q245R 1.67 1.45
 Q12R V68A  N76D S103A V104I G159D  A232V  Q236H  Q245R 2.16 1.72
 N76D S103A  V104I V147I G159D A232V  Q236H  Q245R  N248S  K251R 1.35 1.29
 V68A N76D  S103A V104I G159D A232V  Q236H  Q245R  S256R 2.01 1.72
 V68A N76D  S103A V104I G159D Q206R  A232V  Q236H  Q245R 2.09 1.62
 S103A V104I  G159D A232V Q236H Q245R  N248D  N252K 1.44 1.41
 G20R V68A  S103A V104I G159D A232V  Q236H  Q245R  N248D  N252K 1.81 1.72
 V68A S103A  V104I G159D A232V Q236H  Q245R  N248D  N252K  L257R 1.51 1.41
 V68A S103A  V101I A232V Q236H Q245R  N248D  N252K 1.0 1.50
 N76D S103A  V104I G159D A231V Q236H  Q245R  L257V 1.92 1.09

Claims (24)

1．一种蛋白酶变体，其在一个或多个残基位置处含有氨基酸置换，其中该置换改变了该位置的电荷，使其电荷与前体蛋白酶相比负电性更强或正电性更弱，并且该蛋白酶变体在低去污剂浓度体系中比前体蛋白酶更有效。
2．权利要求1的蛋白酶变体，其中该蛋白酶是枯草杆菌蛋白酶。
3．根据权利要求1的蛋白酶变体，其来源于芽孢杆菌属的枯草杆菌蛋白酶。
4．根据权利要求3的蛋白酶变体，其来源于迟缓芽孢杆茵枯草杆菌蛋白酶。
5．一种编码权利要求1的蛋白酶变体的DNA。
6．一种编码权利要求5的DNA的表达载体。
7．一种用权利要求6的表达载体转化的宿主细胞。
8．一种含有权利要求1的蛋白酶变体的清洁用组合物。
9．一种含有权利要求1的蛋白酶变体的动物饲料。
10．一种含有权利要求1的蛋白酶变体的用于处理纺织品的组合物。
11．一种蛋白酶变体，其在一个或多个残基位置处含有氨基酸置换，其中该置换改变了该位置的电荷，使其电荷与前体蛋白酶相比正电性更强或负电性更弱，并且该蛋白酶变体在高去污剂浓度体系中比前体蛋白酶更有效。
12．权利要求11的蛋白酶变体，其中该蛋白酶是枯草杆菌蛋白酶。
13．根据权利要求11的蛋白酶变体，其来源于芽孢杆菌属的枯草杆菌蛋白酶。
14．根据权利要求13的蛋白酶变体，其来源于迟缓芽孢杆菌枯草杆菌蛋白酶。
15．一种编码权利要求11的蛋白酶变体的DNA。
16．一种编码权利要求15的DNA的表达载体。
17．一种用权利要求16的表达载体转化的宿主细胞。
18．一种含有权利要求11的蛋白酶变体的清洁用组合物。
19．一种含有权利要求11的蛋白酶变体的动物饲料。
20．一种含有权利要求11的蛋白酶变体的用于处理纺织品的组合物。
21．权利要求1的蛋白酶变体，其中高去污剂浓度高于洗涤用水中2000ppm的浓度。
22．一种产生在低、中及高去污剂浓度体系中比前体蛋白酶更有效的蛋白酶变体的方法，包括：
a)置换一个或多个残基位置的氨基酸，其中该置换改变了该位置的电荷，使其电荷与前体蛋白酶相比正电性更强或负电性更弱；
b)置换一个或多个残基位置的氨基酸，其中该置换改变了该位置的电荷，使其电荷与前体蛋白酶相比负电性更强或正电性更弱；
c)检测该变体，测定它在高、中及低去污剂浓度体系中与前体蛋白酶相比的有效性；及
d)必要时重复步骤a)-c)，产生在低、中及高去污剂浓度体系中比前体蛋白酶更有效的蛋白酶变体。
23．一种蛋白酶变体，其在一个或多个残基位置处含有氨基酸置换，其中该置换改变了该位置的电荷，使其电荷与前体蛋白酶相比正电性更强或负电性更弱，并且该蛋白酶变体在中去污剂浓度体系中比前体蛋白酶更有效。
24．一种蛋白酶变体，其在一个或多个残基位置处含有氨基酸置换，其中该置换改变了该位置的电荷，使其电荷与前体蛋白酶相比负电性更强或正电性更弱，并且该蛋白酶变体在中去污剂浓度体系中比前体蛋白酶更有效。
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Families Citing this family (535)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
MA23346A1 (fr) * 1993-10-14 1995-04-01 Genencor Int Variantes de la subtilisine
CA2301851C (en) * 1997-08-29 2012-08-07 Novo Nordisk A/S Protease variants and compositions
ES2368718T3 (es) * 1997-10-23 2011-11-21 Danisco Us Inc. Variantes de subtilisina con múltiples sustituciones.
AR015977A1 (es) * 1997-10-23 2001-05-30 Genencor Int Variantes de proteasa multiplemente substituida con carga neta alterada para su empleo en detergentes
EP1093459B1 (en) 1998-07-02 2008-09-10 The Governing Council Of The University Of Toronto Chemically modified proteins with a carbohydrate moiety
EP1124987A2 (en) * 1998-10-23 2001-08-22 The Procter & Gamble Company Methods for screening protease variants for use in detergent compositions
AU772427B2 (en) 1998-11-10 2004-04-29 Genencor International, Inc. Chemically modified mutant serine hydrolases show improved catalytic activity and chiral selectivity
EP1185302B1 (en) * 1999-04-28 2013-07-24 Genencor International, Inc. Specifically targeted catalytic antagonists and uses thereof
US6627744B2 (en) 1999-07-02 2003-09-30 Genencor International, Inc. Synthesis of glycodendrimer reagents
CN1399677A (zh) * 1999-07-22 2003-02-26 宝洁公司 在确定表位区有氨基酸取代的枯草杆菌蛋白酶变体
BR0012694A (pt) * 1999-07-22 2002-04-09 Procter & Gamble Conjugado de protease, composição de limpeza e composição de tratamento pessoal
MXPA02002682A (es) * 1999-09-09 2002-07-30 Procter & Gamble Una composicion de detergente que contiene una proteasa.
US6899961B2 (en) * 1999-12-15 2005-05-31 Samsung Sdi Co., Ltd. Organic electroluminescence device
US7067049B1 (en) * 2000-02-04 2006-06-27 Exxonmobil Oil Corporation Formulated lubricant oils containing high-performance base oils derived from highly paraffinic hydrocarbons
DE10007608A1 (de) * 2000-02-18 2001-08-30 Henkel Kgaa Protease und Percarbonat enthaltende Wasch- und Reinigungsmittel
WO2001064852A1 (en) * 2000-03-03 2001-09-07 Novozymes A/S Polypeptides having alkaline alpha-amylase activity and nucleic acids encoding same
US6777218B1 (en) * 2000-03-14 2004-08-17 Novozymes A/S Subtilase enzymes having an improved wash performance on egg stains
WO2001075087A2 (en) * 2000-04-03 2001-10-11 Maxygen, Inc. Subtilisin variants
EP2258852A1 (en) * 2000-04-28 2010-12-08 Novozymes A/S Lipolytic enzyme variant
WO2002002725A1 (en) * 2000-06-30 2002-01-10 The Procter & Gamble Company Detergent compositions comprising a cyclodextrin glucanotransferase enzyme
AU2000260631A1 (en) * 2000-06-30 2002-01-14 The Procter & Gamble Company Detergent compositions comprising a maltogenic alpha-amylase enzyme
US20020155574A1 (en) 2000-08-01 2002-10-24 Novozymes A/S Alpha-amylase mutants with altered properties
US6893855B2 (en) * 2000-10-13 2005-05-17 Novozymes A/S Subtilase variants
US6803222B2 (en) 2000-11-22 2004-10-12 Kao Corporation Alkaline proteases
US7303907B2 (en) * 2001-01-08 2007-12-04 Health Protection Agency Degradation and detection of TSE infectivity
EP1241112A3 (en) 2001-03-15 2003-02-26 The Procter & Gamble Company Flexible multiple compartment pouch
RU2311458C2 (ru) * 2001-03-23 2007-11-27 Джененкор Интернэшнл, Инк. Белки, вызывающие измененную иммуногенную реакцию, и способы их получения и использования
DE60234523D1 (de) 2001-05-15 2010-01-07 Novozymes As Alpha-amylasevariante mit veränderten eigenschaften
DK200101090A (da) * 2001-07-12 2001-08-16 Novozymes As Subtilase variants
WO2003057713A2 (en) * 2001-12-31 2003-07-17 Genencor International, Inc. Proteases producing an altered immunogenic response and methods of making and using the same
EP1530631B1 (en) * 2002-01-16 2013-08-07 Genencor International, Inc. Multiply-substituted protease variants
EP1523553B1 (en) * 2002-01-16 2009-12-02 Genencor International, Inc. Multiply-substituted protease variants
US20030180281A1 (en) * 2002-03-11 2003-09-25 Bott Richard R. Preparations for topical skin use and treatment
US7223386B2 (en) * 2002-03-11 2007-05-29 Dow Corning Corporation Preparations for topical skin use and treatment
AU2003228393A1 (en) 2002-03-29 2003-10-13 Genencor International, Inc. Ehanced protein expression in bacillus
JP4693412B2 (ja) 2002-07-29 2011-06-01 アミノテック アーエス １以上の蛋白質を含有する生材料からのペプチド／アミノ酸及び油／脂肪の回収方法、その方法により製造される産物、並びにその産物の使用
US7082689B2 (en) * 2002-10-11 2006-08-01 Black & Decker Inc. Keyless shoe lock for reciprocating saw
TWI319007B (en) * 2002-11-06 2010-01-01 Novozymes As Subtilase variants
US7888093B2 (en) 2002-11-06 2011-02-15 Novozymes A/S Subtilase variants
DE10260903A1 (de) * 2002-12-20 2004-07-08 Henkel Kgaa Neue Perhydrolasen
US20050059567A1 (en) * 2003-09-11 2005-03-17 The Procter & Gamble Company Methods of formulating enzyme cocktails, enzyme cocktails for the removal of egg-based and grass-based stains and/or soils, compositions and products comprising same
CN1906303B (zh) 2003-11-19 2013-06-05 金克克国际有限公司 丝氨酸蛋白酶、编码丝氨酸酶的核酸以及包含它们的载体和宿主细胞
US7985569B2 (en) * 2003-11-19 2011-07-26 Danisco Us Inc. Cellulomonas 69B4 serine protease variants
EP2325318A1 (en) 2004-02-13 2011-05-25 Novozymes A/S Protease variants
DE602005018767D1 (de) 2004-05-17 2010-02-25 Procter & Gamble Bleichmittel mit carbohydratoxidase
US20090060933A1 (en) * 2004-06-14 2009-03-05 Estell David A Proteases producing an altered immunogenic response and methods of making and using the same
JP2006143855A (ja) * 2004-11-18 2006-06-08 Lion Corp 飲料シミ汚れ除去効果を向上させたアミラーゼ含有漂白性組成物
JP2006143853A (ja) * 2004-11-18 2006-06-08 Lion Corp アミラーゼ含有漂白性組成物
US8814861B2 (en) 2005-05-12 2014-08-26 Innovatech, Llc Electrosurgical electrode and method of manufacturing same
US7147634B2 (en) 2005-05-12 2006-12-12 Orion Industries, Ltd. Electrosurgical electrode and method of manufacturing same
EP2385111B1 (en) * 2005-07-08 2016-09-07 Novozymes A/S Subtilase variants
US20070161531A1 (en) * 2005-07-08 2007-07-12 Novozymes A/S Subtilase variants
GB0603775D0 (en) * 2006-02-24 2006-04-05 Health Prot Agency Infectivity assay
CN101415482B (zh) * 2006-03-31 2015-04-15 金克克国际有限公司 用于化合物分离的切向流动过滤设备、系统和方法
US7629158B2 (en) 2006-06-16 2009-12-08 The Procter & Gamble Company Cleaning and/or treatment compositions
WO2008002472A2 (en) 2006-06-23 2008-01-03 Danisco Us Inc., Genencor Division Systematic evaluation of sequence and activity relationships using site evaluation libraries for engineering multiple properties
US20080090747A1 (en) * 2006-07-18 2008-04-17 Pieter Augustinus Protease variants active over a broad temperature range
CN105441353B (zh) 2006-08-11 2019-01-01 诺维信生物股份有限公司 细菌培养物和包含细菌培养物的组合物
JP2010507001A (ja) * 2006-10-16 2010-03-04 ダニスコ・ユーエス・インク、ジェネンコー・ディビジョン 非リン酸系食器洗浄剤
RU2496875C2 (ru) 2007-03-12 2013-10-27 ДАНИСКО ЮЭс ИНК. Мoдифицированные протеазы
US7922970B2 (en) * 2007-04-03 2011-04-12 Kimberly-Clark Worldwide, Inc. Use of sonication to eliminate prions
EP2152732B1 (en) 2007-05-10 2012-03-21 Danisco US Inc. A modified secretion system to increase expression of polypeptides in bacteria
JP5548613B2 (ja) * 2007-06-06 2014-07-16 ダニスコ・ユーエス・インク、ジェネンコー・ディビジョン 複数のタンパク質の性質を改良する方法
JP5238815B2 (ja) 2007-09-12 2013-07-17 ダニスコ・ユーエス・インク 内部汚れ付着が制御された濾過
US8021436B2 (en) 2007-09-27 2011-09-20 The Procter & Gamble Company Cleaning and/or treatment compositions comprising a xyloglucan conjugate
US7618801B2 (en) 2007-10-30 2009-11-17 Danison US Inc. Streptomyces protease
MX2010004676A (es) 2007-11-05 2010-05-20 Danisco Us Inc Variantes de alfa-amilasa con propiedades alteradas.
EP2215202B2 (en) 2007-11-05 2024-01-10 Danisco US Inc. VARIANTS OF BACILLUS sp. TS-23 ALPHA-AMYLASE WITH ALTERED PROPERTIES
WO2009094084A1 (en) 2007-12-21 2009-07-30 Danisco Us Inc., Genencor Division Enhanced protein production in bacillus
JP2011510681A (ja) 2008-02-04 2011-04-07 ダニスコ・ユーエス・インク 改変された特性をもつｔｓ２３アルファ‐アミラーゼ変異体
US8066818B2 (en) * 2008-02-08 2011-11-29 The Procter & Gamble Company Water-soluble pouch
EP2380966B2 (en) * 2008-02-08 2021-11-10 The Procter & Gamble Company Process for making a water-soluble pouch
US20090209447A1 (en) * 2008-02-15 2009-08-20 Michelle Meek Cleaning compositions
EP2100948A1 (en) * 2008-03-14 2009-09-16 The Procter and Gamble Company Automatic dishwashing detergent composition
US20090233830A1 (en) 2008-03-14 2009-09-17 Penny Sue Dirr Automatic detergent dishwashing composition
EP2100947A1 (en) 2008-03-14 2009-09-16 The Procter and Gamble Company Automatic dishwashing detergent composition
JP5973166B2 (ja) 2008-03-26 2016-08-23 ノボザイムス アクティーゼルスカブ 安定化された液体酵素組成物
CN101981193B (zh) 2008-03-28 2016-05-11 丹尼斯科美国公司 在细菌细胞中扩增基因座的方法
US8084240B2 (en) 2008-06-06 2011-12-27 Danisco Us Inc. Geobacillus stearothermophilus α-amylase (AmyS) variants with improved properties
DK2297314T3 (en) * 2008-06-06 2015-07-13 Danisco Us Inc COMPOSITIONS AND METHODS OF COMPREHENSIVE MICROBIAL PROTEASE VARIETIES
EP2166092A1 (en) 2008-09-18 2010-03-24 The Procter and Gamble Company Detergent composition
EP2166075A1 (en) * 2008-09-23 2010-03-24 The Procter and Gamble Company Cleaning composition
EP2166076A1 (en) * 2008-09-23 2010-03-24 The Procter & Gamble Company Cleaning composition
EP2166073A1 (en) 2008-09-23 2010-03-24 The Procter & Gamble Company Cleaning composition
JP5419303B2 (ja) 2008-09-25 2014-02-19 ダニスコ・ユーエス・インク アルファアミラーゼ混合物及びその混合物を使用する方法
BRPI0921041A2 (pt) * 2008-11-11 2019-09-24 Danisco Us Inc composições e métodos que compreendem uma variante de subtilisina
CN103122345A (zh) 2008-11-11 2013-05-29 丹尼斯科美国公司 包含一种或多种可组合的突变的芽孢杆菌属枯草杆菌蛋白酶
WO2010056671A1 (en) * 2008-11-11 2010-05-20 Danisco Us Inc. Compositions and methods comprising a subtilisin variant
EP2589651A3 (en) 2008-11-11 2013-08-28 Danisco US Inc. Compositions and methods comprising serine protease variants
US20100267304A1 (en) * 2008-11-14 2010-10-21 Gregory Fowler Polyurethane foam pad and methods of making and using same
US20100125046A1 (en) 2008-11-20 2010-05-20 Denome Frank William Cleaning products
EP2216390B1 (en) 2009-02-02 2013-11-27 The Procter and Gamble Company Hand dishwashing method
EP2216392B1 (en) * 2009-02-02 2013-11-13 The Procter and Gamble Company Liquid hand dishwashing detergent composition
PL2213713T3 (pl) 2009-02-02 2014-07-31 Procter & Gamble Płynna kompozycja środka czyszczącego do ręcznego mycia naczyń
EP2213715A1 (en) 2009-02-02 2010-08-04 The Procter & Gamble Company Liquid hand dishwashing detergent composition
EP2216391A1 (en) 2009-02-02 2010-08-11 The Procter & Gamble Company Liquid hand dishwashing detergent composition
EP2213714B1 (en) 2009-02-02 2014-06-11 The Procter and Gamble Company Liquid hand dishwashing detergent composition
EP3998328A1 (en) 2009-02-09 2022-05-18 The Procter & Gamble Company Detergent composition
US8153574B2 (en) 2009-03-18 2012-04-10 The Procter & Gamble Company Structured fluid detergent compositions comprising dibenzylidene polyol acetal derivatives and detersive enzymes
US8293697B2 (en) 2009-03-18 2012-10-23 The Procter & Gamble Company Structured fluid detergent compositions comprising dibenzylidene sorbitol acetal derivatives
BRPI1013388A2 (pt) 2009-04-01 2019-04-09 Danisco Us Inc composição de limpeza que compreende uma alfa-amilase e uma protease e método de limpeza de um tecido ou superficie dura
EP2435547A1 (en) 2009-05-26 2012-04-04 The Procter & Gamble Company Aqueous liquid composition for pre-treating soiled dishware
AR076941A1 (es) 2009-06-11 2011-07-20 Danisco Us Inc Cepa de bacillus para una mayor produccion de proteina
US20110009307A1 (en) 2009-07-09 2011-01-13 Alan Thomas Brooker Laundry Detergent Composition Comprising Low Level of Sulphate
BR112012000520A2 (pt) 2009-07-09 2016-02-16 Procter & Gamble composição catalítica detergente para lavagem de roupa que compreende teores relativamente baixos de eletrólito solúvel em água
EP2451922A1 (en) 2009-07-09 2012-05-16 The Procter & Gamble Company Method of laundering fabric using a compacted liquid laundry detergent composition
WO2011005623A1 (en) 2009-07-09 2011-01-13 The Procter & Gamble Company Laundry detergent composition comprising low level of bleach
BR112012000492A2 (pt) 2009-07-09 2019-09-24 Procter & Gamble composição detergente moderadamente alcalina, com baixos teores de coadjuvantes que compreende ácido fitalimido peroxicaproico para tratamento de tecido sólido
EP2451923A1 (en) 2009-07-09 2012-05-16 The Procter & Gamble Company Method of laundering fabric using a liquid laundry detergent composition
JP2012532246A (ja) 2009-07-09 2012-12-13 ザ プロクター アンド ギャンブル カンパニー 比較的低濃度の水溶性電解質を含む触媒性洗濯洗剤組成物
WO2011005804A1 (en) 2009-07-09 2011-01-13 The Procter & Gamble Company Method of laundering fabric using a liquid laundry detergent composition
CN102549034B (zh) 2009-07-31 2014-12-10 阿克佐诺贝尔股份有限公司 用于个人护理方面的杂化共聚物组合物
ES2581916T5 (es) 2009-08-13 2022-11-07 Procter & Gamble Método para lavado de tejidos a baja temperatura
WO2011036263A1 (en) * 2009-09-25 2011-03-31 Novozymes A/S Subtilase variants
AU2010299953B2 (en) * 2009-09-25 2015-02-12 Novozymes A/S Detergent composition
EP2510092A1 (en) 2009-12-09 2012-10-17 The Procter & Gamble Company Fabric and home care products
AR079338A1 (es) 2009-12-09 2012-01-18 Danisco Us Inc Variantes de proteasa de bacillus y acidos nucleicos que codifican dichas variantes
EP2333042B1 (en) 2009-12-10 2015-07-01 The Procter and Gamble Company Automatic dishwashing product and use thereof
EP2333041B1 (en) 2009-12-10 2013-05-15 The Procter & Gamble Company Method and use of a dishwasher composition
ES2423580T5 (es) 2009-12-10 2021-06-17 Procter & Gamble Método y uso de una composición para lavado de vajillas
EP2333040B2 (en) 2009-12-10 2019-11-13 The Procter & Gamble Company Detergent composition
JP2013515139A (ja) 2009-12-21 2013-05-02 ダニスコ・ユーエス・インク サーモビフィダ・フスカのリパーゼを含む洗剤組成物、及びその使用方法
US8741609B2 (en) 2009-12-21 2014-06-03 Danisco Us Inc. Detergent compositions containing Geobacillus stearothermophilus lipase and methods of use thereof
US20120258900A1 (en) 2009-12-21 2012-10-11 Danisco Us Inc. Detergent compositions containing bacillus subtilis lipase and methods of use thereof
RU2012136133A (ru) * 2010-01-22 2014-02-27 ДюПон НЬЮТРИШН БАЙОСАЙЕНСИЗ АпС Способы получения аминозамещенных гликолипидных соединений
CN113186178A (zh) 2010-02-10 2021-07-30 诺维信公司 在螯合剂存在下具有高稳定性的变体和包含变体的组合物
EP2357220A1 (en) 2010-02-10 2011-08-17 The Procter & Gamble Company Cleaning composition comprising amylase variants with high stability in the presence of a chelating agent
ES2401126T3 (es) 2010-02-25 2013-04-17 The Procter & Gamble Company Composición detergente
EP2365058A1 (en) 2010-03-01 2011-09-14 The Procter & Gamble Company Solid laundry detergent composition having an excellent anti-encrustation profile
EP2365059A1 (en) 2010-03-01 2011-09-14 The Procter & Gamble Company Solid laundry detergent composition comprising C.I. fluorescent brightener 260 in alpha-crystalline form
EP2365054A1 (en) 2010-03-01 2011-09-14 The Procter & Gamble Company Solid laundry detergent composition comprising secondary alcohol-based detersive surfactant
EP2380957A1 (en) 2010-04-19 2011-10-26 The Procter & Gamble Company Solid laundry detergent composition having a dynamic in-wash ph profile
EP2377914B1 (en) 2010-04-19 2016-11-09 The Procter & Gamble Company Mildly alkaline, low-built, solid fabric treatment detergent composition comprising perhydrolase
EP2363456A1 (en) 2010-03-01 2011-09-07 The Procter & Gamble Company Solid laundry detergent composition comprising brightener in micronized particulate form
US20120067373A1 (en) 2010-04-15 2012-03-22 Philip Frank Souter Automatic Dishwashing Detergent Composition
AR080886A1 (es) 2010-04-15 2012-05-16 Danisco Us Inc Composiciones y metodos que comprenden proteasas variantes
US8889612B2 (en) 2010-04-19 2014-11-18 The Procter & Gamble Company Method of laundering fabric using a compacted liquid laundry detergent composition
US20110257069A1 (en) 2010-04-19 2011-10-20 Stephen Joseph Hodson Detergent composition
US20110257060A1 (en) 2010-04-19 2011-10-20 Robert Richard Dykstra Laundry detergent composition comprising bleach particles that are suspended within a continuous liquid phase
US20110257062A1 (en) 2010-04-19 2011-10-20 Robert Richard Dykstra Liquid laundry detergent composition comprising a source of peracid and having a ph profile that is controlled with respect to the pka of the source of peracid
EP2380478A1 (en) 2010-04-23 2011-10-26 The Procter & Gamble Company Automatic dishwashing product
ES2565192T3 (es) 2010-04-23 2016-04-01 The Procter & Gamble Company Método para perfumar
EP2383329A1 (en) 2010-04-23 2011-11-02 The Procter & Gamble Company Particle
PL2380961T3 (pl) 2010-04-23 2018-10-31 The Procter & Gamble Company Kompozycja detergentu
ES2533368T3 (es) 2010-04-23 2015-04-09 The Procter & Gamble Company Producto para lavavajillas
ES2579217T3 (es) 2010-04-23 2016-08-08 The Procter & Gamble Company Partícula
DK2566961T3 (en) 2010-05-06 2018-12-03 Danisco Us Inc COMPOSITIONS AND METHODS OF SUBTILISIN VARIETIES
DE102010028951A1 (de) 2010-05-12 2011-11-17 Henkel Ag & Co. Kgaa Lagerstabiles flüssiges Wasch- oder Reinigungsmittel enthaltend Protease und Lipase
AR081423A1 (es) 2010-05-28 2012-08-29 Danisco Us Inc Composiciones detergentes con contenido de lipasa de streptomyces griseus y metodos para utilizarlas
EP2395070A1 (en) 2010-06-10 2011-12-14 The Procter & Gamble Company Liquid laundry detergent composition comprising lipase of bacterial origin
EP2395071A1 (en) 2010-06-10 2011-12-14 The Procter & Gamble Company Solid detergent composition comprising lipase of bacterial origin
WO2011163457A1 (en) 2010-06-23 2011-12-29 The Procter & Gamble Company Product for pre-treatment and laundering of stained fabric
US8685171B2 (en) 2010-07-29 2014-04-01 The Procter & Gamble Company Liquid detergent composition
EP2412792A1 (en) 2010-07-29 2012-02-01 The Procter & Gamble Company Liquid detergent composition
CN101922108B (zh) * 2010-09-14 2012-09-05 东华大学 使用1，4，7-三氮杂环壬烷配合物的活化漂白的方法
WO2012057781A1 (en) 2010-10-29 2012-05-03 The Procter & Gamble Company Cleaning and/or treatment compositions comprising a fungal serine protease
FI123942B (fi) 2010-10-29 2013-12-31 Ab Enzymes Oy Sieniperäisen seriiniproteaasin variantteja
WO2011026154A2 (en) 2010-10-29 2011-03-03 The Procter & Gamble Company Cleaning and/or treatment compositions
DE102010063458A1 (de) * 2010-12-17 2012-06-21 Henkel Ag & Co. Kgaa Lagerstabiles flüssiges Wasch- oder Reinigungsmittel enthaltend Protease und Amylase
AU2012217673B2 (en) 2011-02-15 2017-06-15 Henkel Ag & Co. Kgaa Mitigation of odor in cleaning machines and cleaning processes
DE102011005354A1 (de) 2011-03-10 2012-09-13 Henkel Ag & Co. Kgaa Leistungsverbesserte Proteasevarianten
WO2012149325A1 (en) 2011-04-29 2012-11-01 Danisco Us Inc. Detergent compositions containing geobacillus tepidamans mannanase and methods of use thereof
EP2702152A1 (en) 2011-04-29 2014-03-05 Danisco US Inc. Detergent compositions containing bacillus sp. mannanase and methods of use thereof
BR112013027305A2 (pt) 2011-04-29 2016-11-29 Danisco Us Inc "polipeptídeo recombinante, composição detergente que o compreende, método para limpeza de produto têxtil, vetor de expressão e célula hospedeira".
WO2012151480A2 (en) 2011-05-05 2012-11-08 The Procter & Gamble Company Compositions and methods comprising serine protease variants
AR086281A1 (es) 2011-05-05 2013-12-04 Danisco Us Inc Composiciones y metodos que comprenden variantes de serina proteasas
US20140371435A9 (en) 2011-06-03 2014-12-18 Eduardo Torres Laundry Care Compositions Containing Thiophene Azo Dyes
EP2537918A1 (en) 2011-06-20 2012-12-26 The Procter & Gamble Company Consumer products with lipase comprising coated particles
US20120324655A1 (en) 2011-06-23 2012-12-27 Nalini Chawla Product for pre-treatment and laundering of stained fabric
CN103649307B (zh) 2011-06-30 2020-03-27 诺维信公司 α-淀粉酶变体
EP2540824A1 (en) 2011-06-30 2013-01-02 The Procter & Gamble Company Cleaning compositions comprising amylase variants reference to a sequence listing
EP2551335A1 (en) 2011-07-25 2013-01-30 The Procter & Gamble Company Enzyme stabilized liquid detergent composition
JP2014521770A (ja) 2011-07-27 2014-08-28 ザ プロクター アンド ギャンブル カンパニー 多相液体洗剤組成物
US8679366B2 (en) 2011-08-05 2014-03-25 Ecolab Usa Inc. Cleaning composition containing a polysaccharide graft polymer composition and methods of controlling hard water scale
US8853144B2 (en) 2011-08-05 2014-10-07 Ecolab Usa Inc. Cleaning composition containing a polysaccharide graft polymer composition and methods of improving drainage
US8841246B2 (en) 2011-08-05 2014-09-23 Ecolab Usa Inc. Cleaning composition containing a polysaccharide hybrid polymer composition and methods of improving drainage
US8636918B2 (en) 2011-08-05 2014-01-28 Ecolab Usa Inc. Cleaning composition containing a polysaccharide hybrid polymer composition and methods of controlling hard water scale
US20140187468A1 (en) 2011-08-31 2014-07-03 Danisco Us Inc. Compositions and Methods Comprising a Lipolytic Enzyme Variant
PL2584028T3 (pl) 2011-10-19 2017-10-31 Procter & Gamble Cząstka
CN103889395A (zh) 2011-11-04 2014-06-25 阿克佐诺贝尔化学国际公司 接枝树枝状共聚物及其制备方法
WO2013064647A1 (en) 2011-11-04 2013-05-10 Akzo Nobel Chemicals International B.V. Hybrid dendrite copolymers, compositions thereof and methods for producing the same
DK2788491T3 (en) 2011-12-09 2019-04-01 Danisco Us Inc RIBOSOMAL PROMOTERS FROM B. SUBTILIS FOR PROTEIN PRODUCTION IN MICRO-ORGANISMS
AU2012352763A1 (en) 2011-12-13 2014-04-24 Danisco Us Inc. Enzyme cocktails prepared from mixed cultures
CN104011204A (zh) * 2011-12-20 2014-08-27 诺维信公司 枯草杆菌酶变体和编码它们的多核苷酸
WO2013096653A1 (en) 2011-12-22 2013-06-27 Danisco Us Inc. Compositions and methods comprising a lipolytic enzyme variant
WO2013110766A1 (en) 2012-01-26 2013-08-01 Novozymes A/S Use of polypeptides having protease activity in animal feed and detergents
RU2612215C2 (ru) 2012-02-03 2017-03-03 Дзе Проктер Энд Гэмбл Компани Композиции, содержащие липазы, и способы обработки поверхности
WO2013142486A1 (en) 2012-03-19 2013-09-26 The Procter & Gamble Company Laundry care compositions containing dyes
CN104204198B (zh) 2012-04-02 2018-09-25 诺维信公司 脂肪酶变体以及编码其的多核苷酸
DK2847308T3 (en) 2012-05-07 2017-10-23 Novozymes As Polypeptides with xanthan-degrading activity and polynucleotides encoding them
JP6120953B2 (ja) * 2012-05-30 2017-04-26 クラリアント・ファイナンス・（ビーブイアイ）・リミテッド 界面活性剤溶液中における低温安定剤としてのｎ−メチル−ｎ−アシルグルカミンの使用
EP2674475A1 (en) 2012-06-11 2013-12-18 The Procter & Gamble Company Detergent composition
CN104394708A (zh) 2012-06-20 2015-03-04 诺维信公司 具有蛋白酶活性的多肽在动物饲料和洗涤剂中的用途
US10246692B2 (en) 2012-07-12 2019-04-02 Novozymes A/S Polypeptides having lipase activity and polynucleotides encoding same
US8945314B2 (en) 2012-07-30 2015-02-03 Ecolab Usa Inc. Biodegradable stability binding agent for a solid detergent
US8871699B2 (en) 2012-09-13 2014-10-28 Ecolab Usa Inc. Detergent composition comprising phosphinosuccinic acid adducts and methods of use
US9752105B2 (en) 2012-09-13 2017-09-05 Ecolab Usa Inc. Two step method of cleaning, sanitizing, and rinsing a surface
US9994799B2 (en) 2012-09-13 2018-06-12 Ecolab Usa Inc. Hard surface cleaning compositions comprising phosphinosuccinic acid adducts and methods of use
US20140308162A1 (en) 2013-04-15 2014-10-16 Ecolab Usa Inc. Peroxycarboxylic acid based sanitizing rinse additives for use in ware washing
EP2712915A1 (en) 2012-10-01 2014-04-02 The Procter and Gamble Company Methods of treating a surface and compositions for use therein
CN113105950A (zh) 2012-10-04 2021-07-13 艺康美国股份有限公司 用于洗衣和其他硬表面清洁的预浸泡工艺
US10023851B2 (en) 2012-10-12 2018-07-17 Danisco Us Inc. Compositions and methods comprising a lipolytic enzyme variant
EP2914720B1 (en) 2012-11-05 2022-08-31 Danisco US Inc. Compositions and methods comprising thermolysin protease variants
US20150344858A1 (en) 2012-12-19 2015-12-03 Danisco Us Inc. Novel mannanase, compositions and methods of use thereof
US20140179585A1 (en) 2012-12-20 2014-06-26 The Procter & Gamble Company Detergent composition with silicate coated bleach
US20160053243A1 (en) 2012-12-21 2016-02-25 Danisco Us Inc. Alpha-amylase variants
WO2014099525A1 (en) 2012-12-21 2014-06-26 Danisco Us Inc. Paenibacillus curdlanolyticus amylase, and methods of use, thereof
ES2834373T3 (es) 2013-02-19 2021-06-17 Procter & Gamble Método para lavado de un tejido
EP2767582A1 (en) 2013-02-19 2014-08-20 The Procter and Gamble Company Method of laundering a fabric
EP2767579B1 (en) 2013-02-19 2018-07-18 The Procter and Gamble Company Method of laundering a fabric
MX2015011690A (es) 2013-03-05 2015-12-07 Procter & Gamble Composiciones de azucares mezclados.
ES2676895T5 (es) 2013-03-11 2022-04-27 Danisco Us Inc Variantes combinatorias de alfa-amilasa
BR112015023790B1 (pt) 2013-03-15 2021-09-14 Lubrizol Advanced Materials, Inc Polímeros de ácido itacônico
US9631164B2 (en) 2013-03-21 2017-04-25 Novozymes A/S Polypeptides with lipase activity and polynucleotides encoding same
US9193939B2 (en) 2013-03-28 2015-11-24 The Procter & Gamble Company Cleaning compositions containing a polyetheramine, a soil release polymer, and a carboxymethylcellulose
CN103215151B (zh) * 2013-04-25 2015-01-07 上海巴方精细化工有限公司 矢车菊低刺激护肤香皂
CN105209613A (zh) 2013-05-17 2015-12-30 诺维信公司 具有α淀粉酶活性的多肽
JP6188199B2 (ja) * 2013-05-20 2017-08-30 ライオン株式会社 食器洗い機用洗浄剤
CN103361198B (zh) * 2013-05-22 2015-01-07 上海巴方精细化工有限公司 川芎消炎复合香皂
CN105229116A (zh) 2013-05-28 2016-01-06 宝洁公司 包含光致变色染料的表面处理组合物
EP4159854A1 (en) 2013-05-29 2023-04-05 Danisco US Inc Novel metalloproteases
JP6367930B2 (ja) 2013-05-29 2018-08-01 ダニスコ・ユーエス・インク 新規メタロプロテアーゼ
US20160160202A1 (en) 2013-05-29 2016-06-09 Danisco Us Inc. Novel metalloproteases
EP3004341B1 (en) 2013-05-29 2017-08-30 Danisco US Inc. Novel metalloproteases
FI3013956T3 (fi) 2013-06-27 2023-05-23 Novozymes As Subtilaasivariantteja ja niitä koodittavia polynukleotideja
AU2014286135A1 (en) 2013-07-04 2015-12-03 Novozymes A/S Polypeptides with xanthan lyase activity having anti-redeposition effect and polynucleotides encoding same
US20160160197A1 (en) 2013-07-19 2016-06-09 Danisco Us Inc. Compositions and Methods Comprising a Lipolytic Enzyme Variant
EP3027748B1 (en) * 2013-07-29 2019-09-18 Novozymes A/S Protease variants and polynucleotides encoding same
JP2016533741A (ja) * 2013-07-29 2016-11-04 ダニスコ・ユーエス・インク 酵素の変異体
WO2015038792A1 (en) 2013-09-12 2015-03-19 Danisco Us Inc. Compositions and methods comprising lg12-clade protease variants
CN105555934A (zh) 2013-09-18 2016-05-04 宝洁公司 包含噻吩偶氮羧化物染料的衣物洗涤护理组合物
US9834682B2 (en) 2013-09-18 2017-12-05 Milliken & Company Laundry care composition comprising carboxylate dye
WO2015042086A1 (en) 2013-09-18 2015-03-26 The Procter & Gamble Company Laundry care composition comprising carboxylate dye
EP3047009B1 (en) 2013-09-18 2018-05-16 The Procter and Gamble Company Laundry care composition comprising carboxylate dye
WO2015042013A1 (en) 2013-09-18 2015-03-26 Lubrizol Advanced Materials, Inc. Stable linear polymers
DE102013219467A1 (de) * 2013-09-26 2015-03-26 Henkel Ag & Co. Kgaa Stabilisierte Inhibitor-Proteasevarianten
EP2857485A1 (en) 2013-10-07 2015-04-08 WeylChem Switzerland AG Multi-compartment pouch comprising alkanolamine-free cleaning compositions, washing process and use for washing and cleaning of textiles and dishes
EP2857487A1 (en) 2013-10-07 2015-04-08 WeylChem Switzerland AG Multi-compartment pouch comprising cleaning compositions, washing process and use for washing and cleaning of textiles and dishes
EP2857486A1 (en) 2013-10-07 2015-04-08 WeylChem Switzerland AG Multi-compartment pouch comprising cleaning compositions, washing process and use for washing and cleaning of textiles and dishes
EP3058055A1 (en) 2013-10-15 2016-08-24 Danisco US Inc. Clay granule
DK3068879T3 (da) 2013-11-14 2020-03-16 Danisco Us Inc Stabile enzymer opnået ved glykeringsreduktion
US10533165B2 (en) 2013-12-13 2020-01-14 Danisco Us Inc Serine proteases of the bacillus gibsonii-clade
ES2723948T3 (es) 2013-12-13 2019-09-04 Danisco Us Inc Serina proteasas procedentes de especies de Bacillus
EP4163305A1 (en) 2013-12-16 2023-04-12 Nutrition & Biosciences USA 4, Inc. Use of poly alpha-1,3-glucan ethers as viscosity modifiers
WO2015095358A1 (en) 2013-12-18 2015-06-25 E. I. Du Pont De Nemours And Company Cationic poly alpha-1,3-glucan ethers
DK3089991T3 (da) 2013-12-31 2019-12-02 Danisco Us Inc Forbedret proteinekspression
WO2015112338A1 (en) 2014-01-22 2015-07-30 The Procter & Gamble Company Method of treating textile fabrics
EP3097175B1 (en) 2014-01-22 2018-10-17 The Procter and Gamble Company Fabric treatment composition
WO2015112339A1 (en) 2014-01-22 2015-07-30 The Procter & Gamble Company Fabric treatment composition
EP3097174A1 (en) 2014-01-22 2016-11-30 The Procter & Gamble Company Method of treating textile fabrics
CA2841024C (en) 2014-01-30 2017-03-07 The Procter & Gamble Company Unit dose article
WO2015123323A1 (en) 2014-02-14 2015-08-20 E. I. Du Pont De Nemours And Company Poly-alpha-1,3-1,6-glucans for viscosity modification
WO2015130653A1 (en) 2014-02-25 2015-09-03 The Procter & Gamble Company A process for making renewable surfactant intermediates and surfactants from fats and oils and products thereof
EP3110779A1 (en) 2014-02-25 2017-01-04 The Procter & Gamble Company A process for making renewable surfactant intermediates and surfactants from fats and oils and products thereof
CN106232791B (zh) 2014-02-26 2019-12-10 宝洁公司 包含烷氧基化的聚亚烷基亚胺、有机改性的硅氧烷和基于硅氧烷的稀释剂的清洁组合物
EP2915873A1 (en) * 2014-03-06 2015-09-09 The Procter and Gamble Company Dishwashing composition
WO2015138283A1 (en) 2014-03-11 2015-09-17 E. I. Du Pont De Nemours And Company Oxidized poly alpha-1,3-glucan as detergent builder
EP3116924B1 (en) 2014-03-14 2020-08-19 Lubrizol Advanced Materials, Inc. Itaconic acid polymers and copolymers
MX2016012044A (es) 2014-03-21 2017-06-29 Danisco Us Inc Serina proteasas de especies de bacillus.
US20150275143A1 (en) 2014-03-27 2015-10-01 The Procter & Gamble Company Cleaning compositions containing a polyetheramine
US9719052B2 (en) 2014-03-27 2017-08-01 The Procter & Gamble Company Cleaning compositions containing a polyetheramine
EP2924106A1 (en) 2014-03-28 2015-09-30 The Procter and Gamble Company Water soluble unit dose article
EP2924105A1 (en) 2014-03-28 2015-09-30 The Procter and Gamble Company Water soluble unit dose article
WO2015150457A1 (en) 2014-04-01 2015-10-08 Novozymes A/S Polypeptides having alpha amylase activity
JP2017518407A (ja) 2014-05-06 2017-07-06 ミリケン・アンド・カンパニーＭｉｌｌｉｋｅｎ ＆ Ｃｏｍｐａｎｙ ランドリーケア組成物
US9365805B2 (en) 2014-05-15 2016-06-14 Ecolab Usa Inc. Bio-based pot and pan pre-soak
WO2015187757A1 (en) 2014-06-06 2015-12-10 The Procter & Gamble Company Detergent composition comprising polyalkyleneimine polymers
EP3155097A1 (en) 2014-06-12 2017-04-19 Novozymes A/S Alpha-amylase variants and polynucleotides encoding same
US9714403B2 (en) 2014-06-19 2017-07-25 E I Du Pont De Nemours And Company Compositions containing one or more poly alpha-1,3-glucan ether compounds
US9771548B2 (en) 2014-06-19 2017-09-26 E I Du Pont De Nemours And Company Compositions containing one or more poly alpha-1,3-glucan ether compounds
US10550381B2 (en) 2014-07-04 2020-02-04 Novozymes A/S Variant proteases and amylases having enhanced storage stability
EP3164486B1 (en) * 2014-07-04 2020-05-13 Novozymes A/S Subtilase variants and polynucleotides encoding same
CN106793912A (zh) 2014-08-01 2017-05-31 艺康美国股份有限公司 使用清洁织物的人工表面清洁和洗涤所述清洁织物的方法
EP2987848A1 (en) 2014-08-19 2016-02-24 The Procter & Gamble Company Method of laundering a fabric
EP2987849A1 (en) 2014-08-19 2016-02-24 The Procter and Gamble Company Method of Laundering a Fabric
MX2017003235A (es) 2014-09-10 2017-06-19 Basf Se Composicion de limpieza encapsulada.
US9617502B2 (en) 2014-09-15 2017-04-11 The Procter & Gamble Company Detergent compositions containing salts of polyetheramines and polymeric acid
US20160090552A1 (en) 2014-09-25 2016-03-31 The Procter & Gamble Company Detergent compositions containing a polyetheramine and an anionic soil release polymer
BR112017005767A2 (pt) 2014-09-25 2017-12-12 Procter & Gamble composições de limpeza contendo uma polieteramina
US9388368B2 (en) 2014-09-26 2016-07-12 The Procter & Gamble Company Cleaning compositions containing a polyetheramine
EP3207129B1 (en) 2014-10-17 2019-11-20 Danisco US Inc. Serine proteases of bacillus species
BR112017007949A2 (pt) 2014-10-24 2018-01-23 Danisco Us Inc método para produção de álcool pelo uso de uma tripeptidil-peptidase
WO2016069552A1 (en) 2014-10-27 2016-05-06 Danisco Us Inc. Serine proteases
WO2016069557A1 (en) 2014-10-27 2016-05-06 Danisco Us Inc. Serine proteases of bacillus species
DK3212781T3 (da) 2014-10-27 2019-12-16 Danisco Us Inc Serinproteaser
EP3212782B1 (en) 2014-10-27 2019-04-17 Danisco US Inc. Serine proteases
EP3212783A1 (en) 2014-10-27 2017-09-06 Danisco US Inc. Serine proteases
CN107001994A (zh) 2014-11-17 2017-08-01 宝洁公司 有益剂递送组合物
EP3221447A1 (en) * 2014-11-20 2017-09-27 Novozymes A/S Alicyclobacillus variants and polynucleotides encoding same
US10683528B2 (en) 2014-12-16 2020-06-16 Danisco Us Inc Enhanced protein expression
WO2016100128A1 (en) 2014-12-19 2016-06-23 Danisco Us Inc Enhanced protein expression
CA2969241A1 (en) 2014-12-23 2016-06-30 E.I. Du Pont De Nemours And Company Enzymatically produced cellulose
US20180030456A1 (en) 2015-02-19 2018-02-01 Danisco Us Inc. Enhanced protein expression
CN107454914B (zh) 2015-03-12 2021-09-21 丹尼斯科美国公司 包含lg12进化枝蛋白酶变体的组合物和方法
JP2018521149A (ja) 2015-04-29 2018-08-02 ザ プロクター アンド ギャンブル カンパニー 洗剤組成物
CN107624127A (zh) 2015-04-29 2018-01-23 宝洁公司 处理织物的方法
WO2016176280A1 (en) 2015-04-29 2016-11-03 The Procter & Gamble Company Method of treating a fabric
JP2018517803A (ja) 2015-04-29 2018-07-05 ザ プロクター アンド ギャンブル カンパニー 布地の処理方法
WO2016176296A1 (en) 2015-04-29 2016-11-03 The Procter & Gamble Company Method of laundering a fabric
CN111718806B (zh) 2015-05-04 2022-01-04 美利肯公司 在洗衣护理组合物中作为上蓝剂的隐色三苯甲烷着色剂
EP4212609A1 (en) 2015-05-08 2023-07-19 Novozymes A/S Alpha-amylase variants and polynucleotides encoding same
EP3294882B1 (en) 2015-05-08 2021-07-07 Novozymes A/S Alpha-amylase variants and polynucleotides encoding same
US20180142224A1 (en) 2015-05-08 2018-05-24 Novozymes A/S Alpha-amylase variants having improved performance and stability
JP7274819B2 (ja) 2015-05-13 2023-05-17 ダニスコ・ユーエス・インク ＡｐｒＬ－ＣＬＡＤＥプロテアーゼ変異体及びその使用
WO2016192905A1 (en) 2015-06-02 2016-12-08 Unilever Plc Laundry detergent composition
DK3307427T3 (da) 2015-06-09 2023-11-06 Danisco Us Inc Osmotisk sprængnings-kapsler
EP3307861B1 (en) 2015-06-11 2019-04-03 Unilever PLC, a company registered in England and Wales under company no. 41424 of Laundry detergent composition
EP4234693A3 (en) 2015-06-17 2023-11-01 Danisco US Inc Bacillus gibsonii-clade serine proteases
WO2016206837A1 (en) 2015-06-26 2016-12-29 Unilever Plc Laundry detergent composition
EP3350303B1 (en) 2015-09-17 2020-04-08 Henkel AG & Co. KGaA Detergent compositions comprising polypeptides having xanthan degrading activity
EP3350323B1 (en) 2015-09-17 2021-04-14 Novozymes A/S Polypeptides having xanthan degrading activity and polynucleotides encoding same
BR112018007096A2 (pt) * 2015-10-09 2018-10-23 Novozymes As método de lavagem de roupa, uso de composição detergente e polipeptídeo
WO2017079756A1 (en) 2015-11-05 2017-05-11 Danisco Us Inc Paenibacillus and bacillus spp. mannanases
CN109072208A (zh) 2015-11-05 2018-12-21 丹尼斯科美国公司 类芽孢杆菌属物种甘露聚糖酶
US10822574B2 (en) 2015-11-13 2020-11-03 Dupont Industrial Biosciences Usa, Llc Glucan fiber compositions for use in laundry care and fabric care
EP3374400B1 (en) 2015-11-13 2022-04-13 Nutrition & Biosciences USA 4, Inc. Glucan fiber compositions for use in laundry care and fabric care
WO2017083226A1 (en) 2015-11-13 2017-05-18 E. I. Du Pont De Nemours And Company Glucan fiber compositions for use in laundry care and fabric care
BR112018010475B1 (pt) * 2015-11-25 2023-02-14 Unilever Ip Holdings B.V. Composição de detergente líquido e método de lavagem de roupas utilizando uma composição de detergente líquido para lavar roupas
DK3387124T3 (da) 2015-12-09 2021-08-23 William Cuevas Kombinatoriske alfa-amylasevarianter
JP6591278B2 (ja) * 2015-12-15 2019-10-16 花王株式会社 食器用固体洗浄剤組成物
WO2017106676A1 (en) 2015-12-18 2017-06-22 Danisco Us Inc Polypeptides with endoglucanase activity and uses thereof
EP4219702A3 (en) * 2015-12-30 2023-11-01 Novozymes A/S Enzyme variants and polynucleotides encoding the same
EP3205392A1 (en) 2016-02-12 2017-08-16 Basf Se Microcapsules and process for preparation of microcapsules
EP3205393A1 (en) 2016-02-12 2017-08-16 Basf Se Process for preparation of microcapsules
BR112018016129B1 (pt) 2016-02-17 2022-06-07 Unilever Ip Holdings B.V. Composição de detergente para lavagem de roupas e método doméstico de tratamento de um tecido
WO2017140392A1 (en) 2016-02-17 2017-08-24 Unilever Plc Whitening composition
FI3419991T3 (fi) 2016-03-04 2023-01-31 Modifioidut ribosomaaliset promoottorit proteiinien tuottamiseksi mikro-organismeissa
RU2018135268A (ru) 2016-03-09 2020-04-09 Басф Се Инкапсулированная чистящая композиция для стирки
BR112018068068B1 (pt) 2016-03-21 2023-04-18 Unilever Ip Holdings B.V. Composição aquosa líquida de detergente para lavagem de roupas e método doméstico de tratamento de um tecido
EP3440172B1 (en) 2016-04-08 2019-08-21 Unilever PLC Laundry detergent composition
WO2017182295A1 (en) 2016-04-18 2017-10-26 Basf Se Liquid cleaning compositions
CA3022875A1 (en) * 2016-05-03 2017-11-09 Danisco Us Inc Protease variants and uses thereof
EP3452584B1 (en) 2016-05-05 2021-01-06 Danisco US Inc. Protease variants and uses thereof
JP6067168B1 (ja) * 2016-05-30 2017-01-25 株式会社ニイタカ 自動洗浄機用洗浄剤組成物
JP2019523645A (ja) 2016-05-31 2019-08-29 ダニスコ・ユーエス・インク プロテアーゼ変異体およびその使用
CN106085984B (zh) * 2016-06-02 2019-07-19 天津科技大学 一种新型磷脂酶d及其制备磷脂酸、磷脂酰丝氨酸的方法
CN109312323B (zh) * 2016-06-09 2022-03-08 花王株式会社 突变碱性蛋白酶
BR112018075933A2 (pt) 2016-06-17 2019-10-01 Danisco Us Inc variantes de protease e usos das mesmas
EP3275988B1 (en) 2016-07-26 2020-07-08 The Procter and Gamble Company Automatic dishwashing detergent composition
EP3275989A1 (en) 2016-07-26 2018-01-31 The Procter and Gamble Company Automatic dishwashing detergent composition
EP3275987A1 (en) 2016-07-26 2018-01-31 The Procter and Gamble Company Automatic dishwashing detergent composition
EP3275986B1 (en) 2016-07-26 2020-07-08 The Procter and Gamble Company Automatic dishwashing detergent composition
EP3275985A1 (en) 2016-07-26 2018-01-31 The Procter and Gamble Company Automatic dishwashing detergent composition
CN109844110B (zh) 2016-08-24 2023-06-06 诺维信公司 黄原胶裂解酶变体以及编码它们的多核苷酸
EP3504330A1 (en) 2016-08-24 2019-07-03 Novozymes A/S Gh9 endoglucanase variants and polynucleotides encoding same
US11001827B2 (en) 2016-08-24 2021-05-11 Henkel Ag & Co. Kgaa Detergent compositions comprising xanthan lyase variants I
CN109563451A (zh) 2016-08-24 2019-04-02 汉高股份有限及两合公司 包含gh9内切葡聚糖酶变体i的洗涤剂组合物
BR112019006576A2 (pt) 2016-11-01 2019-07-02 Milliken & Co corantes leuco como agentes de azulamento em composições de cuidados de lavanderia
WO2018085315A1 (en) 2016-11-01 2018-05-11 The Procter & Gamble Company Leuco colorants as bluing agents in laundry care compositions, packaging, kits and methods thereof
MX2019005120A (es) 2016-11-01 2019-06-20 Procter & Gamble Colorantes leuco como agentes de azulado en composiciones para el cuidado de la ropa.
US20180119056A1 (en) 2016-11-03 2018-05-03 Milliken & Company Leuco Triphenylmethane Colorants As Bluing Agents in Laundry Care Compositions
EP3535365A2 (en) 2016-11-07 2019-09-11 Danisco US Inc. Laundry detergent composition
US10577571B2 (en) 2016-11-08 2020-03-03 Ecolab Usa Inc. Non-aqueous cleaner for vegetable oil soils
WO2018102479A1 (en) 2016-12-02 2018-06-07 The Procter & Gamble Company Cleaning compositions including enzymes
US10550443B2 (en) 2016-12-02 2020-02-04 The Procter & Gamble Company Cleaning compositions including enzymes
JP6907318B2 (ja) 2016-12-02 2021-07-21 ザ プロクター アンド ギャンブル カンパニーＴｈｅ Ｐｒｏｃｔｅｒ ＆ Ｇａｍｂｌｅ Ｃｏｍｐａｎｙ 酵素を含む洗浄組成物
EP3555255B1 (en) 2016-12-15 2020-06-24 Unilever PLC Laundry detergent composition
EP3559226B1 (en) 2016-12-21 2023-01-04 Danisco US Inc. Bacillus gibsonii-clade serine proteases
CN110312795A (zh) * 2016-12-21 2019-10-08 丹尼斯科美国公司 蛋白酶变体及其用途
EP3339423A1 (en) 2016-12-22 2018-06-27 The Procter & Gamble Company Automatic dishwashing detergent composition
MX2019009093A (es) 2017-02-01 2019-12-05 Procter & Gamble Composiciones de limpieza que comprenden variantes de amilasa.
KR102375732B1 (ko) 2017-02-24 2022-03-16 다니스코 유에스 인크. 바실러스 리체니포르미스에서 단백질 생산을 증가시키기 위한 조성물 및 방법
WO2018169750A1 (en) 2017-03-15 2018-09-20 Danisco Us Inc Trypsin-like serine proteases and uses thereof
EP3601553A1 (en) 2017-03-31 2020-02-05 Danisco US Inc. Alpha-amylase combinatorial variants
US20200040283A1 (en) 2017-03-31 2020-02-06 Danisco Us Inc Delayed release enzyme formulations for bleach-containing detergents
EP3401385A1 (en) 2017-05-08 2018-11-14 Henkel AG & Co. KGaA Detergent composition comprising polypeptide comprising carbohydrate-binding domain
WO2018206535A1 (en) 2017-05-08 2018-11-15 Novozymes A/S Carbohydrate-binding domain and polynucleotides encoding the same
CA3067837A1 (en) 2017-06-30 2019-01-03 Danisco Us Inc Low-agglomeration, enzyme-containing particles
BR112020000205B1 (pt) 2017-07-07 2023-10-31 Unilever Ip Holdings B.V. Composição de limpeza para a lavagem de tecidos e método doméstico de tratamento de um tecido
EP3649222B1 (en) 2017-07-07 2024-03-13 Unilever IP Holdings B.V. Whitening composition
WO2019036721A2 (en) 2017-08-18 2019-02-21 Danisco Us Inc VARIANTS OF ALPHA-AMYLASES
US11879127B2 (en) 2017-08-23 2024-01-23 Danisco Us Inc. Methods and compositions for efficient genetic modifications of Bacillus licheniformis strains
US11525128B2 (en) 2017-08-24 2022-12-13 Novozymes A/S GH9 endoglucanase variants and polynucleotides encoding same
US11359188B2 (en) 2017-08-24 2022-06-14 Novozymes A/S Xanthan lyase variants and polynucleotides encoding same
US11624059B2 (en) 2017-08-24 2023-04-11 Henkel Ag & Co. Kgaa Detergent compositions comprising GH9 endoglucanase variants II
WO2019038060A1 (en) 2017-08-24 2019-02-28 Henkel Ag & Co. Kgaa DETERGENT COMPOSITION COMPRISING XANTHANE LYASE II VARIANTS
EP3682010A1 (en) 2017-09-13 2020-07-22 Danisco US Inc. Modified 5'-untranslated region (utr) sequences for increased protein production in bacillus
CA3074938A1 (en) 2017-10-12 2019-04-18 The Procter & Gamble Company Leuco colorants as bluing agents in laundry care composition
EP3694973A1 (en) 2017-10-12 2020-08-19 The Procter & Gamble Company Leuco colorants as bluing agents in laundry care compositions
CA3074613A1 (en) 2017-10-12 2019-04-18 The Procter & Gamble Company Leuco colorants in combination with a second whitening agent as bluing agents in laundry care compositions
TWI715878B (zh) 2017-10-12 2021-01-11 美商美力肯及公司 隱色著色劑及組成物
JP7230043B2 (ja) 2017-11-01 2023-02-28 ミリケン・アンド・カンパニー ロイコ化合物、着色剤化合物、およびそれらを含有する組成物
EP3703661A1 (en) 2017-11-02 2020-09-09 Danisco US Inc. Freezing point depressed solid matrix compositions for melt granulation of enzymes
CN111373039A (zh) 2017-11-29 2020-07-03 丹尼斯科美国公司 具有改善的稳定性的枯草杆菌蛋白酶变体
CN111479912B (zh) 2017-11-30 2021-08-10 联合利华知识产权控股有限公司 包含蛋白酶的洗涤剂组合物
DE102017223281A1 (de) * 2017-12-19 2019-06-19 Henkel Ag & Co. Kgaa Reinigugnsmittel enthaltend durch Betain stabilisierte Amylase
EP3502245A1 (en) 2017-12-19 2019-06-26 The Procter & Gamble Company Automatic dishwashing detergent composition
EP3502244A1 (en) 2017-12-19 2019-06-26 The Procter & Gamble Company Automatic dishwashing detergent composition
EP3502246A1 (en) 2017-12-19 2019-06-26 The Procter & Gamble Company Automatic dishwashing detergent composition
EP3502227A1 (en) 2017-12-19 2019-06-26 The Procter & Gamble Company Automatic dishwashing detergent composition
US20210222148A1 (en) 2017-12-21 2021-07-22 Danisco Us Inc. Enzyme-containing, hot-melt granules comprising a thermotolerant desiccant
JP6967447B2 (ja) * 2017-12-27 2021-11-17 小林製薬株式会社 錠剤状義歯洗浄剤
FI3735478T3 (fi) 2018-01-03 2023-10-26 Danisco Us Inc Mutantit ja geneettisesti modifioidut bacillus-solut ja niihin liittyvät menetelmät proteiinituotannon lisäämiseksi
JP7110355B2 (ja) 2018-01-26 2022-08-01 ザ プロクター アンド ギャンブル カンパニー 酵素を含む水溶性単位用量物品
MX2020008302A (es) 2018-02-08 2020-10-14 Danisco Us Inc Partículas de matriz de cera térmicamente resistentes para encapsulación de enzimas.
WO2019162000A1 (en) 2018-02-23 2019-08-29 Henkel Ag & Co. Kgaa Detergent composition comprising xanthan lyase and endoglucanase variants
US20190264139A1 (en) 2018-02-28 2019-08-29 The Procter & Gamble Company Cleaning compositions
EP3533858A1 (en) 2018-02-28 2019-09-04 The Procter & Gamble Company Cleaning composition comprising a glycogen-debranching enzyme and methods of cleaning
EP3768835A1 (en) * 2018-03-23 2021-01-27 Novozymes A/S Subtilase variants and compositions comprising same
EP3775127B1 (en) 2018-05-17 2022-07-20 Unilever IP Holdings B.V. Cleaning composition
WO2019245705A1 (en) 2018-06-19 2019-12-26 Danisco Us Inc Subtilisin variants
WO2019245704A1 (en) 2018-06-19 2019-12-26 Danisco Us Inc Subtilisin variants
CN112189052A (zh) 2018-06-19 2021-01-05 宝洁公司 自动盘碟洗涤剂组合物
EP3810769A1 (en) 2018-06-19 2021-04-28 The Procter & Gamble Company Automatic dishwashing detergent composition
US20200032178A1 (en) 2018-07-27 2020-01-30 The Procter & Gamble Company Water-soluble unit dose articles comprising water-soluble fibrous structures and particles
US20230174962A1 (en) 2018-07-31 2023-06-08 Danisco Us Inc Variant alpha-amylases having amino acid substitutions that lower the pka of the general acid
WO2020047215A1 (en) 2018-08-30 2020-03-05 Danisco Us Inc Enzyme-containing granules
WO2020046613A1 (en) 2018-08-30 2020-03-05 Danisco Us Inc Compositions comprising a lipolytic enzyme variant and methods of use thereof
CN112703246A (zh) 2018-09-17 2021-04-23 联合利华知识产权控股有限公司 洗涤剂组合物
CN113166682A (zh) 2018-09-27 2021-07-23 丹尼斯科美国公司 用于医疗器械清洁的组合物
CN113166745A (zh) 2018-10-12 2021-07-23 丹尼斯科美国公司 在螯合剂存在下具有可增强稳定性的突变的α-淀粉酶
EP3884025B1 (en) 2018-11-20 2022-06-08 Unilever Global Ip Limited Detergent composition
WO2020104155A1 (en) 2018-11-20 2020-05-28 Unilever Plc Detergent composition
CN113056550B (zh) 2018-11-20 2022-10-28 联合利华知识产权控股有限公司 洗涤剂组合物
WO2020104156A1 (en) 2018-11-20 2020-05-28 Unilever Plc Detergent composition
EP3884026A1 (en) 2018-11-20 2021-09-29 Unilever Global Ip Limited Detergent composition
WO2020112599A1 (en) 2018-11-28 2020-06-04 Danisco Us Inc Subtilisin variants having improved stability
EP3898919A1 (en) 2018-12-21 2021-10-27 Novozymes A/S Detergent pouch comprising metalloproteases
EP3702452A1 (en) * 2019-03-01 2020-09-02 Novozymes A/S Detergent compositions comprising two proteases
JP7275298B2 (ja) 2019-03-14 2023-05-17 ザ プロクター アンド ギャンブル カンパニー 酵素を含むクリーニング組成物
WO2020186052A1 (en) 2019-03-14 2020-09-17 The Procter & Gamble Company Method for treating cotton
JP7275297B2 (ja) 2019-03-14 2023-05-17 ザ プロクター アンド ギャンブル カンパニー 酵素を含むクリーニング組成物
JP2022524490A (ja) 2019-03-21 2022-05-06 ノボザイムス アクティーゼルスカブ α－アミラーゼ変異体及びこれをコードするポリヌクレオチド
WO2020229535A1 (en) 2019-05-16 2020-11-19 Unilever Plc Laundry composition
AU2020277383B2 (en) 2019-05-20 2023-03-09 Ecolab Usa Inc. Surfactant package for high foaming detergents with low level of medium to long chain linear alcohols
EP3741283A1 (en) 2019-05-22 2020-11-25 The Procter & Gamble Company Automatic dishwashing method
WO2020242858A1 (en) 2019-05-24 2020-12-03 Danisco Us Inc Subtilisin variants and methods of use
EP3976775A1 (en) 2019-05-24 2022-04-06 The Procter & Gamble Company Automatic dishwashing detergent composition
CN114174486A (zh) 2019-06-06 2022-03-11 丹尼斯科美国公司 用于清洁的方法和组合物
JP7326497B2 (ja) 2019-06-24 2023-08-15 ザ プロクター アンド ギャンブル カンパニー アミラーゼバリアントを含む洗浄組成物
CN113993981A (zh) 2019-06-28 2022-01-28 联合利华知识产权控股有限公司 洗涤剂组合物
CN114008184A (zh) 2019-06-28 2022-02-01 联合利华知识产权控股有限公司 洗涤剂组合物
BR112021022423A2 (pt) 2019-06-28 2022-01-04 Unilever Ip Holdings B V Composição detergente e método doméstico de tratamento de tecidos
US20220372408A1 (en) 2019-06-28 2022-11-24 Conopco, Inc., D/B/A Unilever Detergent composition
US20220364020A1 (en) 2019-06-28 2022-11-17 Conopco, Inc., D/B/A Unilever Detergent composition
WO2020260006A1 (en) 2019-06-28 2020-12-30 Unilever Plc Detergent compositions
EP3770241A1 (de) * 2019-07-22 2021-01-27 Henkel AG & Co. KGaA Reinigungsmittel mit protease zur automatischen dosierung
US11873465B2 (en) 2019-08-14 2024-01-16 Ecolab Usa Inc. Methods of cleaning and soil release of highly oil absorbing substrates employing optimized extended chain nonionic surfactants
BR112022003050A2 (pt) 2019-09-02 2022-05-17 Unilever Ip Holdings B V Composição detergente de lavagem de roupas aquosa e método doméstico para tratar um tecido
BR112022004470A2 (pt) 2019-09-19 2022-05-31 Unilever Ip Holdings B V Composição detergente e método para lavagem não oxidativa de manchas de tecido
AR120142A1 (es) 2019-10-07 2022-02-02 Unilever Nv Composición detergente
US20210122998A1 (en) 2019-10-24 2021-04-29 The Procter & Gamble Company Automatic dishwashing detergent composition comprising an amylase
US11492571B2 (en) 2019-10-24 2022-11-08 The Procter & Gamble Company Automatic dishwashing detergent composition comprising a protease
CN114846023A (zh) 2019-10-24 2022-08-02 丹尼斯科美国公司 成麦芽五糖/麦芽六糖变体α-淀粉酶
KR102316445B1 (ko) * 2019-11-29 2021-10-26 씨제이제일제당 주식회사 신규 세린 프로테아제 변이체
EP3835396A1 (en) 2019-12-09 2021-06-16 The Procter & Gamble Company A detergent composition comprising a polymer
US20230049452A1 (en) 2020-01-13 2023-02-16 Danisco Us Inc Compositions comprising a lipolytic enzyme variant and methods of use thereof
KR20220127844A (ko) 2020-01-15 2022-09-20 다니스코 유에스 인크. 바실러스 리체니포르미스에서 단백질 생산을 향상시키기 위한 조성물 및 방법
EP4097206B1 (en) 2020-01-29 2023-08-09 Unilever IP Holdings B.V. Laundry detergent product
EP3862412A1 (en) 2020-02-04 2021-08-11 The Procter & Gamble Company Detergent composition
WO2021247801A1 (en) 2020-06-05 2021-12-09 The Procter & Gamble Company Detergent compositions containing a branched surfactant
EP4162018B1 (en) 2020-06-08 2024-01-31 Unilever IP Holdings B.V. Method of improving protease activity
US20220000757A1 (en) 2020-07-06 2022-01-06 Ecolab Usa Inc. Foaming mixed alcohol/water compositions comprising a combination of alkyl siloxane and a hydrotrope/solubilizer
EP4176031A1 (en) 2020-07-06 2023-05-10 Ecolab USA Inc. Peg-modified castor oil based compositions for microemulsifying and removing multiple oily soils
EP4175471A1 (en) 2020-07-06 2023-05-10 Ecolab USA Inc. Foaming mixed alcohol/water compositions comprising a structured alkoxylated siloxane
JP2023537336A (ja) 2020-08-04 2023-08-31 ザ プロクター アンド ギャンブル カンパニー 自動食器洗浄方法及びパック
WO2022031310A1 (en) 2020-08-04 2022-02-10 The Procter & Gamble Company Automatic dishwashing method
JP2023535061A (ja) 2020-08-04 2023-08-15 ザ プロクター アンド ギャンブル カンパニー 自動食器洗浄方法
WO2022031309A1 (en) 2020-08-04 2022-02-10 The Procter & Gamble Company Automatic dishwashing method
US20240034960A1 (en) 2020-08-27 2024-02-01 Danisco Us Inc Enzymes and enzyme compositions for cleaning
WO2022043042A1 (en) 2020-08-28 2022-03-03 Unilever Ip Holdings B.V. Detergent composition
CN116018396A (zh) 2020-08-28 2023-04-25 联合利华知识产权控股有限公司 洗涤剂组合物
CN116096703A (zh) 2020-08-28 2023-05-09 联合利华知识产权控股有限公司 表面活性剂和洗涤剂组合物
EP4204530A1 (en) 2020-08-28 2023-07-05 Unilever IP Holdings B.V. Detergent composition
US20230287300A1 (en) 2020-08-28 2023-09-14 Conopco, Inc., D/B/A Unilever Surfactant and detergent composition
EP4225905A2 (en) 2020-10-07 2023-08-16 Novozymes A/S Alpha-amylase variants
WO2022093189A1 (en) 2020-10-27 2022-05-05 Milliken & Company Compositions comprising leuco compounds and colorants
CN116348581A (zh) 2020-10-29 2023-06-27 宝洁公司 含有藻酸盐裂解酶的清洁组合物
US20220169952A1 (en) 2020-11-17 2022-06-02 The Procter & Gamble Company Automatic dishwashing composition comprising amphiphilic graft polymer
EP4001388A1 (en) 2020-11-17 2022-05-25 The Procter & Gamble Company Automatic dishwashing method with amphiphilic graft polymer in the rinse
JP2023547853A (ja) 2020-11-17 2023-11-14 ザ プロクター アンド ギャンブル カンパニー アルカリ性すすぎによる自動食器洗浄方法
EP4006131A1 (en) 2020-11-30 2022-06-01 The Procter & Gamble Company Method of laundering fabric
EP4256020A1 (en) 2020-12-07 2023-10-11 Unilever IP Holdings B.V. Detergent compositions
EP4256019A1 (en) 2020-12-07 2023-10-11 Unilever IP Holdings B.V. Detergent compositions
CA3201033A1 (en) 2020-12-23 2022-06-30 Basf Se Amphiphilic alkoxylated polyamines and their uses
CN112662652A (zh) * 2021-01-20 2021-04-16 天津科技大学 一种胶原降解活力降低的碱性蛋白酶突变体
WO2022165107A1 (en) 2021-01-29 2022-08-04 Danisco Us Inc Compositions for cleaning and methods related thereto
EP4039806A1 (en) 2021-02-04 2022-08-10 Henkel AG & Co. KGaA Detergent composition comprising xanthan lyase and endoglucanase variants with im-proved stability
WO2022171780A2 (en) 2021-02-12 2022-08-18 Novozymes A/S Alpha-amylase variants
WO2022197512A1 (en) 2021-03-15 2022-09-22 The Procter & Gamble Company Cleaning compositions containing polypeptide variants
CA3211422A1 (en) 2021-05-05 2022-11-10 Neil Joseph Lant Methods for making cleaning compositions and detecting soils
EP4086330A1 (en) 2021-05-06 2022-11-09 The Procter & Gamble Company Surface treatment
WO2022243367A1 (en) 2021-05-18 2022-11-24 Nouryon Chemicals International B.V. Polyester polyquats in cleaning applications
EP4341317A1 (en) 2021-05-20 2024-03-27 Nouryon Chemicals International B.V. Manufactured polymers having altered oligosaccharide or polysaccharide functionality or narrowed oligosaccharide distribution, processes for preparing them, compositions containing them, and methods of using them
EP4108767A1 (en) 2021-06-22 2022-12-28 The Procter & Gamble Company Cleaning or treatment compositions containing nuclease enzymes
WO2022268885A1 (en) 2021-06-23 2022-12-29 Novozymes A/S Alpha-amylase polypeptides
WO2023275269A1 (en) 2021-06-30 2023-01-05 Nouryon Chemicals International B.V. Chelate-amphoteric surfactant liquid concentrates and use thereof in cleaning applications
WO2023278297A1 (en) 2021-06-30 2023-01-05 Danisco Us Inc Variant lipases and uses thereof
EP4123007A1 (en) 2021-07-19 2023-01-25 The Procter & Gamble Company Fabric treatment using bacterial spores
EP4123006A1 (en) 2021-07-19 2023-01-25 The Procter & Gamble Company Composition comprising spores and pro-perfume materials
WO2023023644A1 (en) 2021-08-20 2023-02-23 Danisco Us Inc. Polynucleotides encoding novel nucleases, compositions thereof and methods thereof for eliminating dna from protein preparations
WO2023034486A2 (en) 2021-09-03 2023-03-09 Danisco Us Inc. Laundry compositions for cleaning
WO2023039270A2 (en) 2021-09-13 2023-03-16 Danisco Us Inc. Bioactive-containing granules
WO2023041694A1 (en) 2021-09-20 2023-03-23 Unilever Ip Holdings B.V. Detergent composition
WO2023057367A1 (en) 2021-10-08 2023-04-13 Unilever Ip Holdings B.V. Laundry composition
WO2023064749A1 (en) 2021-10-14 2023-04-20 The Procter & Gamble Company A fabric and home care product comprising cationic soil release polymer and lipase enzyme
EP4194536A1 (en) 2021-12-08 2023-06-14 The Procter & Gamble Company Laundry treatment cartridge
EP4194537A1 (en) 2021-12-08 2023-06-14 The Procter & Gamble Company Laundry treatment cartridge
WO2023114939A2 (en) 2021-12-16 2023-06-22 Danisco Us Inc. Subtilisin variants and methods of use
US20230265358A1 (en) 2021-12-16 2023-08-24 The Procter & Gamble Company Home care composition comprising an amylase
WO2023114988A2 (en) 2021-12-16 2023-06-22 Danisco Us Inc. Variant maltopentaose/maltohexaose-forming alpha-amylases
WO2023114795A1 (en) 2021-12-16 2023-06-22 The Procter & Gamble Company Automatic dishwashing composition comprising a protease
US20230365897A1 (en) 2021-12-16 2023-11-16 The Procter & Gamble Company Fabric and home care composition including a protease
WO2023114936A2 (en) 2021-12-16 2023-06-22 Danisco Us Inc. Subtilisin variants and methods of use
WO2023114932A2 (en) 2021-12-16 2023-06-22 Danisco Us Inc. Subtilisin variants and methods of use
US20230272310A1 (en) 2021-12-16 2023-08-31 The Procter & Gamble Company Home care composition
WO2023168234A1 (en) 2022-03-01 2023-09-07 Danisco Us Inc. Enzymes and enzyme compositions for cleaning
EP4273209A1 (en) 2022-05-04 2023-11-08 The Procter & Gamble Company Machine-cleaning compositions containing enzymes
EP4273210A1 (en) 2022-05-04 2023-11-08 The Procter & Gamble Company Detergent compositions containing enzymes
WO2023225459A2 (en) 2022-05-14 2023-11-23 Novozymes A/S Compositions and methods for preventing, treating, supressing and/or eliminating phytopathogenic infestations and infections
EP4279571A1 (en) 2022-05-19 2023-11-22 The Procter & Gamble Company Laundry composition comprising spores
WO2023227421A1 (en) 2022-05-27 2023-11-30 Unilever Ip Holdings B.V. Laundry liquid composition comprising a surfactant, an alkoxylated zwitterionic polyamine polymer, and a fragrance
WO2023227375A1 (en) 2022-05-27 2023-11-30 Unilever Ip Holdings B.V. Laundry liquid composition comprising a surfactant, an aminocarboxylate, an organic acid and a fragrance
WO2023227335A1 (en) 2022-05-27 2023-11-30 Unilever Ip Holdings B.V. Liquid composition comprising linear alkyl benzene sulphonate, methyl ester ethoxylate and alkoxylated zwitterionic polyamine polymer
WO2023227356A1 (en) 2022-05-27 2023-11-30 Unilever Ip Holdings B.V. Composition containing enzyme
WO2023227332A1 (en) 2022-05-27 2023-11-30 Unilever Ip Holdings B.V. Laundry liquid composition comprising a surfactant, an alkoxylated zwitterionic polyamine polymer and a protease
WO2023227331A1 (en) 2022-05-27 2023-11-30 Unilever Ip Holdings B.V. Composition comprising a specific methyl ester ethoxylate surfactant and a lipase
WO2023233026A1 (en) 2022-06-03 2023-12-07 Unilever Ip Holdings B.V. Laundry detergent product
WO2023250301A1 (en) 2022-06-21 2023-12-28 Danisco Us Inc. Methods and compositions for cleaning comprising a polypeptide having thermolysin activity
US20240026248A1 (en) 2022-07-20 2024-01-25 Ecolab Usa Inc. Novel nonionic extended surfactants, compositions and methods of use thereof
EP4321604A1 (en) 2022-08-08 2024-02-14 The Procter & Gamble Company A fabric and home care composition comprising surfactant and a polyester
WO2024050346A1 (en) 2022-09-02 2024-03-07 Danisco Us Inc. Detergent compositions and methods related thereto
WO2024050339A1 (en) 2022-09-02 2024-03-07 Danisco Us Inc. Mannanase variants and methods of use
WO2024050343A1 (en) 2022-09-02 2024-03-07 Danisco Us Inc. Subtilisin variants and methods related thereto
WO2024056334A1 (en) 2022-09-13 2024-03-21 Unilever Ip Holdings B.V. Washing machine and washing method
WO2024056333A1 (en) 2022-09-13 2024-03-21 Unilever Ip Holdings B.V. Washing machine and washing method
WO2024056331A1 (en) 2022-09-13 2024-03-21 Unilever Ip Holdings B.V. Washing machine and washing method
WO2024056278A1 (en) 2022-09-13 2024-03-21 Unilever Ip Holdings B.V. Washing machine and washing method
EP4349946A1 (en) 2022-10-05 2024-04-10 Unilever IP Holdings B.V. Unit dose fabric treatment product
EP4349945A1 (en) 2022-10-05 2024-04-10 Unilever IP Holdings B.V. Laundry liquid composition
EP4349944A1 (en) 2022-10-05 2024-04-10 Unilever IP Holdings B.V. Laundry liquid composition
EP4349947A1 (en) 2022-10-05 2024-04-10 Unilever IP Holdings B.V. Laundry liquid composition
EP4349948A1 (en) 2022-10-05 2024-04-10 Unilever IP Holdings B.V. Laundry liquid composition
EP4349943A1 (en) 2022-10-05 2024-04-10 Unilever IP Holdings B.V. Laundry liquid composition
EP4349942A1 (en) 2022-10-05 2024-04-10 Unilever IP Holdings B.V. Laundry liquid composition

Family Cites Families (94)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE1611703B1 (de) * 1967-10-11 1972-03-09 Windmoeller & Hoelscher Vorrichtung zum Abteilen einer bestimmten Stueckzahl innerhalb einer am Ende einer Sack- oder Beutelmaschine gebildeten kontinuierlichen Folge von Saecken oder Beuteln
US4028263A (en) * 1973-08-24 1977-06-07 Colgate-Palmolive Company Bleaching and brightening detergent composition
NL7904044A (nl) 1978-05-31 1979-12-04 Unilever Nv Werkwijze ter bereiding van homogene, vloeibare compo- sities.
US4261868A (en) 1979-08-08 1981-04-14 Lever Brothers Company Stabilized enzymatic liquid detergent composition containing a polyalkanolamine and a boron compound
US4404128A (en) 1981-05-29 1983-09-13 The Procter & Gamble Company Enzyme detergent composition
GR76237B (zh) 1981-08-08 1984-08-04 Procter & Gamble
JPS58144105A (ja) 1982-02-12 1983-08-27 Kurabo Ind Ltd スケ−ル除去獣毛繊維の製法
GB8310080D0 (en) 1983-04-14 1983-05-18 Interox Chemicals Ltd Bleach composition
US4760025A (en) 1984-05-29 1988-07-26 Genencor, Inc. Modified enzymes and methods for making same
US5264366A (en) 1984-05-29 1993-11-23 Genencor, Inc. Protease deficient bacillus
US5182204A (en) 1984-05-29 1993-01-26 Genencor International, Inc. Non-human carbonyl hydrolase mutants, vectors encoding same and hosts transformed with said vectors
US5204015A (en) 1984-05-29 1993-04-20 Genencor International, Inc. Subtilisin mutants
JPS6141481U (ja) * 1984-08-10 1986-03-17 三菱電機株式会社 シ−ム溶接装置
US4797362A (en) * 1985-06-06 1989-01-10 Lion Corporation Alkaline proteases and microorganisms producing same
US4908773A (en) * 1987-04-06 1990-03-13 Genex Corporation Computer designed stabilized proteins and method for producing same
US4728455A (en) 1986-03-07 1988-03-01 Lever Brothers Company Detergent bleach compositions, bleaching agents and bleach activators
SG30639G (en) 1986-04-30 1995-09-01 Genencor Int Non-human carbonyl hydrolase mutants DNA sequences and vectors encoding same and hosts transformed with said vectors
GB8629837D0 (en) 1986-12-13 1987-01-21 Interox Chemicals Ltd Bleach activation
US4853871A (en) 1987-04-06 1989-08-01 Genex Corporation Computer-based method for designing stablized proteins
US5314692A (en) 1987-08-24 1994-05-24 Cultor Ltd. Enzyme premix for feed and method
WO1989004552A1 (en) 1987-10-30 1989-05-18 Lsi Logic Corporation Method and means of fabricating a semiconductor device package
DK6488D0 (da) * 1988-01-07 1988-01-07 Novo Industri As Enzymer
CN1056187C (zh) * 1988-02-11 2000-09-06 金克克国际有限公司 新的蛋白水解酶及其在洗涤剂中的应用
US5543302A (en) * 1988-05-27 1996-08-06 Solvay Enzymes, Inc. Proteases of altered stability to autolytic degradation
DE3841152A1 (de) 1988-12-07 1990-06-13 Hoechst Ag Verwendung von bakterienlysierendem enzymprodukt als zusatzstoff zur verbesserung der futterverwertung in der tierproduktion
GB8908416D0 (en) 1989-04-13 1989-06-01 Unilever Plc Bleach activation
JPH03505976A (ja) * 1989-05-17 1991-12-26 アムジエン・インコーポレーテツド 多重突然変異スブチリシン
ES2227508T3 (es) * 1989-06-26 2005-04-01 Unilever N.V. Composiciones detergentes enzimaticas.
DK316989D0 (da) * 1989-06-26 1989-06-26 Novo Nordisk As Enzymer
US5665587A (en) * 1989-06-26 1997-09-09 Novo Nordisk A/S Modified subtilisins and detergent compositions containing same
GB9003741D0 (en) 1990-02-19 1990-04-18 Unilever Plc Bleach activation
ES2100925T3 (es) 1990-05-21 1997-07-01 Unilever Nv Activacion de blanqueador.
ES2110488T3 (es) * 1990-12-21 1998-02-16 Novo Nordisk As Mutantes enzimaticos con un bajo grado de variacion de la carga molecular en un intervalo de ph.
US5769630A (en) * 1991-02-25 1998-06-23 Louisiana State University, Subperiosteal bone anchor
GB9108136D0 (en) 1991-04-17 1991-06-05 Unilever Plc Concentrated detergent powder compositions
US5340735A (en) * 1991-05-29 1994-08-23 Cognis, Inc. Bacillus lentus alkaline protease variants with increased stability
US5274147A (en) 1991-07-11 1993-12-28 Lever Brothers Company, Division Of Conopco, Inc. Process for preparing manganese complexes
DE4224125A1 (de) * 1991-07-27 1993-01-28 Solvay Enzymes Gmbh & Co Kg Verfahren zur verbesserung der stabilitaet von enzymen und stabilisierte enzyme
GB9118242D0 (en) 1991-08-23 1991-10-09 Unilever Plc Machine dishwashing composition
GB9124581D0 (en) 1991-11-20 1992-01-08 Unilever Plc Bleach catalyst composition,manufacture and use thereof in detergent and/or bleach compositions
CA2083661A1 (en) 1991-11-26 1993-05-27 Rudolf J. Martens Detergent bleach compositions
US5194416A (en) 1991-11-26 1993-03-16 Lever Brothers Company, Division Of Conopco, Inc. Manganese catalyst for activating hydrogen peroxide bleaching
US5153161A (en) 1991-11-26 1992-10-06 Lever Brothers Company, Division Of Conopco, Inc. Synthesis of manganese oxidation catalyst
GB9127060D0 (en) 1991-12-20 1992-02-19 Unilever Plc Bleach activation
CA2085642A1 (en) 1991-12-20 1993-06-21 Ronald Hage Bleach activation
US5316935A (en) * 1992-04-06 1994-05-31 California Institute Of Technology Subtilisin variants suitable for hydrolysis and synthesis in organic media
EP0571982A1 (en) * 1992-05-27 1993-12-01 Showa Denko Kabushiki Kaisha Alkaline lipase, method for producing the same, microorganism producing the same and detergent composition containing alkaline lipase
DE4218448A1 (de) 1992-06-04 1993-12-09 Solvay Enzymes Gmbh & Co Kg Alkalische Proteasen aus Bacillus pumilus
US5256779A (en) 1992-06-18 1993-10-26 Lever Brothers Company, Division Of Conopco, Inc. Synthesis of manganese oxidation catalyst
US5284944A (en) 1992-06-30 1994-02-08 Lever Brothers Company, Division Of Conopco, Inc. Improved synthesis of 1,4,7-triazacyclononane
PT717778E (pt) * 1992-07-17 2008-01-16 Genencor Int Serina proteases resistentes a alcalinidade elevada
US5280117A (en) 1992-09-09 1994-01-18 Lever Brothers Company, A Division Of Conopco, Inc. Process for the preparation of manganese bleach catalyst
US5567601A (en) * 1993-06-01 1996-10-22 University Of Maryland Subtilisin mutants lacking a primary calcium binding site
WO1995005477A1 (en) * 1993-08-12 1995-02-23 University Of Maryland Thermostable alkaline metalloprotease produced by a hyphomonas, and preparation thereof
US6436690B1 (en) * 1993-09-15 2002-08-20 The Procter & Gamble Company BPN′ variants having decreased adsorption and increased hydrolysis wherein one or more loop regions are substituted
MA23346A1 (fr) 1993-10-14 1995-04-01 Genencor Int Variantes de la subtilisine
ATE245185T1 (de) * 1993-10-14 2003-08-15 Procter & Gamble Proteasen enthaltende bleichmittelzusammensetzungen
JP2888985B2 (ja) * 1993-10-14 1999-05-10 ザ、プロクター、エンド、ギャンブル、カンパニー プロテアーゼ含有クリーニング組成物
US5738120A (en) * 1993-10-23 1998-04-14 Imperial Tobacco Limited Smoking articles
ES2364774T3 (es) * 1994-02-24 2011-09-14 HENKEL AG & CO. KGAA Enzimas mejoradas y detergentes que las contienen.
US5691295A (en) * 1995-01-17 1997-11-25 Cognis Gesellschaft Fuer Biotechnologie Mbh Detergent compositions
ATE512226T1 (de) * 1994-02-24 2011-06-15 Henkel Ag & Co Kgaa Verbesserte enzyme und detergentien damit
AU2067795A (en) * 1994-03-29 1995-10-17 Novo Nordisk A/S Alkaline bacillus amylase
JP3009471B2 (ja) * 1994-04-07 2000-02-14 ザ、プロクター、エンド、ギャンブル、カンパニー 金属含有漂白触媒と酸化防止剤とを含む漂白組成物
US6599730B1 (en) * 1994-05-02 2003-07-29 Procter & Gamble Company Subtilisin 309 variants having decreased adsorption and increased hydrolysis
GB9409336D0 (en) 1994-05-10 1994-06-29 Finnfeeds Int Ltd Use of an enzyme for manufacturing an agent for the treatment and/or prophylaxis of coccidiosis
GB9416841D0 (en) * 1994-08-19 1994-10-12 Finnfeeds Int Ltd An enzyme feed additive and animal feed including it
DE69515331T2 (de) 1994-12-09 2000-10-19 Procter & Gamble Diacylperoxydteilchen enthaltende zusammensetzungen für automatische geschirreinigung
US5534302A (en) 1995-01-05 1996-07-09 National Science Council Method of preparing a fiber reinforced modified phenolic resin composite
US5534179A (en) 1995-02-03 1996-07-09 Procter & Gamble Detergent compositions comprising multiperacid-forming bleach activators
AR000862A1 (es) * 1995-02-03 1997-08-06 Novozymes As Variantes de una ó-amilasa madre, un metodo para producir la misma, una estructura de adn y un vector de expresion, una celula transformada por dichaestructura de adn y vector, un aditivo para detergente, composicion detergente, una composicion para lavado de ropa y una composicion para la eliminacion del
US5837516A (en) * 1995-03-03 1998-11-17 Genentech, Inc. Subtilisin variants capable of cleaving substrates containing basic residues
US5780285A (en) * 1995-03-03 1998-07-14 Genentech, Inc. Subtilisin variants capable of cleaving substrates containing dibasic residues
US6455295B1 (en) * 1995-03-08 2002-09-24 The Procter & Gamble Company Subtilisin Carlsberg variants having decreased adsorption and increased hydrolysis
IL117350A0 (en) * 1995-03-09 1996-07-23 Procter & Gamble Proteinase k variants having decreased adsorption and increased hydrolysis
US6475765B1 (en) * 1995-03-09 2002-11-05 Procter & Gamble Company Subtilisin DY variants having decreased adsorption and increased hydrolysis
IL117352A0 (en) * 1995-03-09 1996-07-23 Procter & Gamble Thermitase variants having decreased adsorption and increased hydrolysis
US5523434A (en) 1995-03-15 1996-06-04 The Procter & Gamble Company Synthesis of bleach activators
BR9608292A (pt) * 1995-04-17 1999-05-11 Procter & Gamble Preparação e uso de partículas compósitas que contém peróxido de diacila
US5837517A (en) * 1995-05-05 1998-11-17 Novo Nordisk A/S Protease variants and compositions
US5597936A (en) 1995-06-16 1997-01-28 The Procter & Gamble Company Method for manufacturing cobalt catalysts
AU5796296A (en) 1995-06-16 1997-01-15 Procter & Gamble Company, The Bleach compositions comprising cobalt catalysts
WO1997000312A1 (en) 1995-06-16 1997-01-03 The Procter & Gamble Company Automatic dishwashing compositions comprising cobalt catalysts
US5576282A (en) 1995-09-11 1996-11-19 The Procter & Gamble Company Color-safe bleach boosters, compositions and laundry methods employing same
US5703034A (en) * 1995-10-30 1997-12-30 The Procter & Gamble Company Bleach catalyst particles
US5762647A (en) * 1995-11-21 1998-06-09 The Procter & Gamble Company Method of laundering with a low sudsing granular detergent composition containing optimally selected levels of a foam control agent bleach activator/peroxygen bleaching agent system and enzyme
WO1997022680A1 (en) * 1995-12-20 1997-06-26 The Procter & Gamble Company Bleach catalyst plus enzyme particles
US5883065A (en) * 1996-01-22 1999-03-16 The Procter & Gamble Company Phase separated detergent composition
JP2000505502A (ja) * 1996-03-07 2000-05-09 ザ、プロクター、エンド、ギャンブル、カンパニー 改良アミラーゼを含んでなる洗剤組成物
US6069122A (en) * 1997-06-16 2000-05-30 The Procter & Gamble Company Dishwashing detergent compositions containing organic diamines for improved grease cleaning, sudsing, low temperature stability and dissolution
US6177392B1 (en) * 1997-01-13 2001-01-23 Ecolab Inc. Stable solid block detergent composition
WO1998055634A1 (en) * 1997-06-04 1998-12-10 The Procter & Gamble Company Protease enzymes for tough cleaning and/or spot and film reduction and compositions incorporating same
US6057171A (en) * 1997-09-25 2000-05-02 Frequency Technology, Inc. Methods for determining on-chip interconnect process parameters
AR015977A1 (es) * 1997-10-23 2001-05-30 Genencor Int Variantes de proteasa multiplemente substituida con carga neta alterada para su empleo en detergentes

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