CN110195354A - 处理聚酯纺织品的方法 - Google Patents
处理聚酯纺织品的方法 Download PDFInfo
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- CN110195354A CN110195354A CN201910213241.7A CN201910213241A CN110195354A CN 110195354 A CN110195354 A CN 110195354A CN 201910213241 A CN201910213241 A CN 201910213241A CN 110195354 A CN110195354 A CN 110195354A
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Abstract
本发明涉及处理聚酯纺织品的方法,具体地涉及在角质酶的存在下糖基水解酶家族61多肽用于聚酯纺织品制造的用途,以及包括糖基水解酶家族61多肽和角质酶的纺织品组合物。
Description
本发明申请是基于申请日为2013年7月18日,申请号为201380038377.X(国际申请号为PCT/CN2013/079609)、名称为“处理聚酯纺织品的方法”的发明专利申请的分案申请。
序列表的引用
本申请包含一个计算机可读形式的序列表。该计算机可读形式通过引用结合在此。
技术领域
本发明涉及糖基水解酶家族61多肽和角质酶在处理聚酯纺织品中的用途,以及包括糖基水解酶家族61多肽和角质酶的纺织品组合物。
背景技术
聚对苯二甲酸乙二酯(缩写为PET)纤维占纺织工业所应用的聚酯的主要部分。这些纤维是由对苯二甲酸和乙二醇的缩聚并从熔化物拉伸纤维产生。
聚酯具有一定的核心优势,包括高强度、柔软的手感、拉伸阻力、耐沾污性、可机洗性能、抗皱性以及抗磨损性。然而,聚酯就其疏水性、起球、静电、可染性、作为粘附介质的钝性表面即软化或湿润度增强化合物、缺乏透气性以及不理想的发光或光泽外观不是那么最佳。
因为其强度,使聚酯织物和/或衣服易经受球粒形成,并且施用至聚酯短纤维材料的精加工布的工艺可能最重要的是为起球控制所设计的那些。所有短纤维材料趋于当在洗涤和穿着的过程中经受轻微磨损时在布表面形成具有缠绕纤维的小球或“球粒”。如果该织物包含相当大的比例的对弯曲磨损具有高耐受性的纤维时,这些球粒将以不足以产生不愉快的手感和外观的数目保留在布的表面。
聚酯的另一个问题是,在合成PET过程中,形成聚(对苯二甲酸乙二酯)的环状或线性低聚物,例如对苯二甲酸-双-2-苯甲酰氧基-乙酯(缩写为BETEB)和/或环状三(对苯二甲酸乙二酯)。这些低聚物部分地沉积在机械上并且部分地留在这些纤维之上和/或之内。低聚物趋于为织物给出浅灰色外观。这是归因于低聚物在该织物表面的沉积,具体地坦率地说该沉积是在高温湿加工像高温染色之后。这些低聚物可通过严格的碱处理去除,其导致纤维材料的显著损失。这些低聚物的有机提取是一种技术可能性,但不是工业上可行的。
工业上已对改进聚酯的特征尤其对减少球粒形成作出了很大努力。
WO 99/001604披露了用一种对苯二甲酸二乙酯水解酶(ETE水解酶)和/或一种乙二醇二苄基酯水解酶(BEB水解酶)降低聚酯织物和/或衣服的起球倾向的方法。
WO 2001/34899披露了用于修饰聚酯的方法,包括用一种酯酶酶处理所述聚酯。
WO 97/27237披露了聚(对苯二甲酸乙二酯)的环状低聚物的酶水解,包括使该环状低聚物经受一种或多种羧酸酯水解酶的作用。
WO 2001/092502披露了用特异腐质霉角质酶变体处理聚酯纺织品。
然而,仍然对酶促聚酯织物和/或衣服处理的改进的益处存在需要,包括增强这些酶对它们的底物的效率。具体地,对更有效的酶组合物存在连续的需要以改进该工艺的经济状况。本发明目标是满足这些需要。
发明内容
本发明涉及一种用于在水性溶液中在角质酶的存在下将聚酯纺织品用糖基水解酶家族61(GH61)多肽进行处理的方法。
本发明还涉及一种包括糖基水解酶家族61多肽和角质酶的纺织品组合物。
在一些实施例中,该聚酯纺织品处理工艺可进一步包括选自下组的一种或多种酶,该组由以下各项组成:脂肪酶、酯酶、漆酶、过氧化物酶和环化酶以及转移酶。
在优选的实施例中,该聚酯纺织品是一种PET纺织品。
在本发明,GH61多肽可增强该角质酶对其底物的效率,具有以下益处的至少一个:减少该聚酯纺织品中的低聚物,减少球粒形成,并且在生物抛光过程中没有织物的大量的重量损失,对聚酯织物的可湿性/亲水性以及抗静电性的改进。
在一个实施例中,可将多种酶与角质酶和GH61一起用于聚酯处理工艺,该多种酶包括选自下组的一种或多种酶,该组由以下各项组成:脂肪酶、酯酶、漆酶、过氧化物酶、环化酶以及转移酶。
在一个实施例中,本发明的方法和组合物可进一步包括一种共物质,例如半胱氨酸和抗坏血酸盐。
在一些实施例中,提供了用于制造聚酯纺织品的方法。在一些实施例中,该纺织品被从织物制备为衣服。
在一些实施例中,用于本发明的角质酶是具有BETEB水解活性的一种角质酶。
具体地,本发明涉及如下各项:
1.一种用于在水性溶液中在角质酶的存在下将聚酯纺织品用糖基水解酶家族61多肽进行处理的方法。
2.如项1所述的方法,其中该纺织品是纱线、织物或衣服。
3.如项1或2所述的方法,其中该聚酯是PET。
4.如项1所述的方法,其中该水性溶液进一步包括选自下组的一种或多种酶,该组由以下各项组成:脂肪酶、酯酶、漆酶、过氧化物酶、环化酶以及转移酶。
5.如以上项中任一项所述的方法,其中与一种糖基水解酶家族61一起使用了一种共物质;优选地该共物质是半胱氨酸。
6.如以上项中任一项所述的方法,其中该糖基水解酶家族61多肽是按以下范围施用的:从0.01至约50毫克蛋白质/克聚酯纺织品,优选0.05-20毫克蛋白质/克聚酯纺织品,优选0.1-15毫克蛋白质/克聚酯纺织品,更优选0.2-8毫克蛋白质/克聚酯纺织品,并且甚至更优选0.2-5毫克蛋白质/克聚酯纺织品。
7.如以上项中任一项所述的方法,其中该角质酶是按以下范围施用的:从约0.01至约50毫克酶蛋白质/克聚酯纺织品,优选0.05-20毫克酶蛋白质/克聚酯纺织品,更优选0.1-15毫克酶蛋白质/克聚酯纺织品,并且甚至更优选0.2-5毫克酶蛋白质/克聚酯纺织品。
8.如以上项中任一项所述的方法,其中该方法是按以下的pH范围进行的:从约pH3至约pH 11,优选从约pH 4至约pH 10的范围,或从约pH 6至约pH 9的范围。
9.如以上项中任一项所述的方法,其中该方法是按以下的温度范围进行的:40℃-100℃,优选50℃-90℃,优选60℃-85℃,更优选65℃-80℃,并且甚至更优选70℃-80℃。
10.如以上项中任一项所述的方法,其中该方法进行了约10分钟至约8小时,优选约20分钟至约180分钟,更优选约30分钟至约150分钟,更优选约45分钟至约120分钟。
11.如以上项中任一项所述的方法,其中处理聚酯纺织品是制造该聚酯纺织品,尤其制造聚酯织物。
12.如项11所述的方法,其中该方法是与现存的聚酯织物制造步骤中的任一个相组合。
13.一种包括糖基水解酶家族61多肽和角质酶的组合物。
14.如项13所述的组合物,其中该组合物进一步包括选自下组的一种或多种酶,该组由以下各项组成:脂肪酶、酯酶、漆酶、过氧化物酶、环化酶以及转移酶。
15.如项13或14所述的组合物,其中该组合物进一步包括一种共物质;优选地该共物质是半胱氨酸。
发明详述
使用以下定义和例子,本发明现在将以参考方式详细说明。在此提到的全部专利和出版物,包括这些专利和出版物内公开的所有序列在内,均明确地通过引用结合在此。
如在此所使用的,单数的术语“一个”、“一种”、和“该”包括复数引用,除非上下文清楚地另外指明。
聚酯纺织品
如在此使用的“聚酯”意指一种包含链内酯基的线性聚合分子,这些酯基是从一种二酸与一种二醇的缩合或从羟基酸的聚合衍生。本发明适用于脂肪族的和芳香族的聚酯两者。特别优选的聚酯是芳香族的聚酯物品,其被用于产生纤维和树脂,并且包括合成地产生的长链聚合物,包括按重量计至少85%、优选至少90%并且最优选至少95%的一种取代的芳香族羧酸的一种酯,例如取代的对苯二甲酸或对位取代的羟苯酸酯或其混合物。其他有用的聚酯物品包括由本体聚合物、纱线、织物、薄膜、树脂以及粉末制成的那些。工业使用中的这些基本的聚酯包括聚对苯二甲酸乙二醇酯(PET)、四亚甲基对苯二甲酸酯(PTMT)、聚对苯二甲酸丁二酯(PBT)、聚对苯二甲酸丙二醇酯(PTT)和聚萘二甲酸乙二醇酯(polyethylenenaphthalate)(PEN)、聚环己烷二亚甲基对苯二甲酸酯(CHDMT)、聚乙烯-4-氧基苯甲酸酯、A-Tell、聚乙交酯、PHBA以及2GN。然而,PET是产生的最常见的线性聚合物并且占现今工业中采用的聚酯的多数。
此处使用的聚酯纺织品意指包括含有聚酯的纤维、纱线、织物以及衣服。该聚酯纱线或织物或衣服可以是以下的任何纱线或织物或衣服:其由纯的聚(对苯二甲酸乙二酯)制成,或其由聚(对苯二甲酸乙二酯)纤维和常规用于制造纺织品例如毛料、棉布、粘胶和丝绸的任何其他材料的共混物制成。
在一个优选实施例中,该聚酯织物是一种包括多于35%(w/w)的聚酯、特别是多于50%、多于65%、多于90%、或多于95%的聚酯的织物共混物。在一个最优选的实施例中,本发明的工艺应用于基本上由聚(对苯二甲酸乙二酯)聚酯材料,即纯的聚(对苯二甲酸乙二酯)聚酯材料组成的织物或衣服。
角质酶
角质酶是根据酶命名法被分类为EC 3.1.1.74的脂肪分解酶。参考国际生物化学与分子生物学联合会的命名委员会的建议(Recommendations of the NomenclatureCommittee of the International Union of Biochemistry and Molecular Biology),学术出版社公司(Academic Press Inc.),1992
为了本发明的目的,根据本发明的实例1,角质酶活性是使用低聚物对苯二甲酸-双-2-苯甲酰氧基-乙酯(BETEB)作为底物确定的。BETEB是PET合成期间的一种副产物,并且通常在纺织品制造期间保留在该织物或衣服中。BETEB是通过例如对苯二甲酸、苯甲酸和乙二醇的缩合产生的,其与PET具有相同的苯甲酰氧基-乙酯单元。
如果在实例1中的测试之后显示透明区的话,所讨论的酶被认为是用于根据本发明使用的一种角质酶。
已知角质酶来自不同的真菌,例如一种丝状真菌角质酶,例如产自于腐质霉属或镰刀菌属或大毁壳属或假单胞菌属的一个菌株,特别是特异腐质霉或豌豆腐皮镰孢或稻梨孢或门多萨假单胞菌,更特别是特异腐质霉菌株DSM 1800(US 5,827,719)、或豌豆腐皮镰孢(WO 90/09446图1;WO94/14964图1D,WO 94/03578图1D,通过引用全部特此结合)或稻梨孢(WO 10/107560SEQ ID NO:1,通过引用合并在此)或门多萨假单胞菌ATCC 53552(US 5,389,536,权利要求1,通过引用特此结合)。
SEQ ID NO:1是特异腐质霉角质酶(对应于US 5,827,719的SEQ ID NO:2的成熟部分)的氨基酸序列。
在一个实施例中,本发明的角质酶与SEQ ID NO:1具有至少70%、或75%、或85%、或90%、或95%、或96%、或97%、或98%、或99%、或100%的一致性。
在一些实施例中,该角质酶可以是SEQ ID NO:1的具有一个或多个(或若干个)氨基酸取代、缺失、和/或插入的变体。优选地,SEQ ID NO:1的氨基酸取代、缺失和/或插入的总数不超过10,例如1、2、3、4、5、6、7、8、或9。将描述于WO 2001/092502中的特异腐质霉角质酶变体通过引用特此结合。角质酶也可以是亲本角质酶的变体,如描述于WO 00/34450(通过引用特此结合)中的那些。
真菌角质酶也可以源自其他真菌菌株,例如丝核菌属例如立枯丝核菌的菌株,或链格孢属例如甘蓝链格孢的菌株(WO 94/03578)。
优选地,该角质酶具有该工艺的pH的1个pH单位内的pH最佳值,例如如果该工艺是在pH 8下运行,该角质酶优选具有7和9之间的pH最佳值。
序列一致性
两个氨基酸序列之间或两个核苷酸序列之间的相关性由参数“序列一致性”描述。
出于本发明的目的,使用如在EMBOSS包(EMBOSS:欧洲分子生物学开放软件套件(The European Molecular Biology Open Software Suite),赖斯(Rice)等人,2000,遗传学趋势(Trends Genet.)16:276-277)(优选3.0.0版或更新版本)的尼德尔(Needle)程序中所实施的尼德尔曼-翁施(Needleman-Wunsch)算法(尼德尔曼和翁施,1970,分子生物学杂志(J.Mol.Biol.)48:443-453)来测定两个氨基酸序列之间的序列一致性的程度。所使用的这些任选参数是空位开放罚分10、空位延伸罚分0.5,及EBLOSUM62(BLOSUM62的EMBOSS版本)取代矩阵。尼德尔标注的“最长的一致性”的输出(使用-非简化选项获得)被用作百分比一致性,并且如下计算:
(一致的残基X 100)/(比对长度-比对中的空位总数)
出于本发明的目的,使用尼德曼-翁施算法(尼德曼(Needleman)和翁施(Wunsch),1970,见上文)来确定两个脱氧核糖核苷酸序列之间的序列一致性的程度,如在EMBOSS软件包(EMBOSS:欧洲分子生物学开放软件套件(The European Molecular Biology OpenSoftware Suite),赖斯(Rice)等人,2000,见上文)(优选3.0.0版或更新版本)的尼德尔程序中所实施的。使用的任选参数是空位开放罚分10,空位延伸罚分0.5及EDNAFULL(NCBINUC4.4的EMBOSS版本)取代矩阵。标记为“最长一致性”的尼德尔的输出(使用–nobrief选项获得)被用作百分比一致性并且被计算如下:
(一致的脱氧核糖核苷酸X 100)/(比对长度-比对中的空位总数)
糖苷水解酶家族61(GH61)多肽
术语“糖苷水解酶家族61”或“GH61”此处被定义为属于糖苷水解酶家族61的一种多肽,根据的是亨利萨塔(Henrissat)B.,1991,生物化学期刊(Biochem.J.)280:309-316,以及亨利萨塔B.和巴洛赫(Bairoch)A.,1996,生物化学期刊,316:695-696。
本发明总体上涉及分离的GH61多肽的用途。可用于本发明的GH61多肽可获得自任何属的微生物。对本发明来说,这里所使用的与指定来源相关的术语“从...中获得”意为由核苷酸序列编码的多肽是由一种天然地具有所述核苷酸序列的来源产生的或者由一种来自于该来源的核苷酸序列已经插入其中的菌株产生的。在一个优选的方面,从指定来源获得的多肽分泌到细胞外。
本发明的GH61多肽可以是细菌多肽。例如,该多肽可以是一种革兰氏阳性细菌多肽,例如芽孢杆菌多肽,例如嗜碱芽孢杆菌、解淀粉芽孢杆菌、短芽孢种菌、坏状芽孢杆菌、凝结芽孢杆菌、灿烂芽孢杆菌、迟缓芽孢杆菌、地衣芽孢杆菌、巨大芽孢杆菌、嗜热脂肪芽孢杆菌、枯草芽孢杆菌或苏云金芽孢杆菌多肽;或链霉菌多肽,例如浅青紫链霉菌或鼠灰链霉菌多肽;或是革兰氏阴性细菌多肽,例如大肠杆菌或假单胞菌多肽。
本发明的GH61多肽也可以是一种真菌多肽,并且更优选酵母多肽,例如假丝酵母属、克鲁维酵母属、毕赤酵母属、酵母属、裂殖酵母属或亚罗酵母属多肽;或更优选丝状真菌多肽,例如支顶孢属、曲霉属、短柄霉属、毛壳菌属、隐球菌属、线黑粉菌科(Filibasidium)、镰孢霉属、腐质霉属、大毁壳属、毛霉属、蚀丝霉属、新美鞭菌、脉孢菌属、拟青霉属、青霉属、梨囊鞭菌属、孔座壳属(Poronia)、裂褶菌属、踝节菌属、嗜热子嚢菌属、梭孢壳菌属、弯颈霉属、木霉属或轮枝孢属多肽。
在本发明中,可以使用具有角质酶增强活性的任何GH61多肽。
在一个实施例中,为了本发明的目的,角质酶增强活性是由PET中低聚物的减少确定的,即通过测量如在实例4中指定的条件下用角质酶和GH61以0.05mg蛋白质/ml的剂量在70℃、pH 8.0水解BETEB 40分钟造成的OD254吸光度的增加来确定。在本发明的优选实施例中,在与当使用角质酶而不使用GH61时的OD结果相比时,OD增加至少0.25,优选至少0.28,更优选至少0.3,更优选至少0.33,更优选至少0.35,更优选至少0.38,更优选至少0.40,甚至更优选地至少0.43,并且最优选地至少0.45。
在一些实施例中,角质酶增强活性是通过测量如在实例6中指定的条件下用角质酶和GH61以2.8mg蛋白质/克织物的剂量在70℃、pH 8.0在洗衣指数计(Launder-O-Meter)中处理PET 2小时造成的球粒形成的减少来确定的。在本发明的优选实施例中,显示在与当使用角质酶而不使用GH61时的起球记录相比时,起球记录增加至少0.125,更优选至少0.250,更优选至少0.375,更优选至少0.500,更优选至少0.625,甚至更优选地至少0.750。
在一个第一方面,具有角质酶增强活性的GH61多肽包括以下基序:
[ILMV]-P-X(4,5)-G-X-Y-[ILMV]-X-R-X-[EQ]-X(4)-[HNQ]和[FW]-[TF]-K-[AIV],
其中X是任何氨基酸,X(4,5)是处于4个或5个连续位置的任何氨基酸,并且X(4)是处于4个连续位置的任何氨基酸。
包括以上提到的基序的分离的多肽可以进一步包括:
H-X(1,2)-G-P-X(3)-[YW]-[AILMV],
[EQ]-X-Y-X(2)-C-X-[EHQN]-[FILV]-X-[ILV],或
H-X(1,2)-G-P-X(3)-[YW]-[AILMV]以及[EQ]-X-Y-X(2)-C-X-[EHQN]-[FILV]-X-[ILV],
其中X是任何氨基酸,X(1,2)是处于1个位置或2个连续位置的任何氨基酸,X(3)是处于3个连续位置的任何氨基酸,并且X(2)是处于2个连续位置的任何氨基酸。在以上基序中,采用可接受的IUPAC单一字母氨基酸缩写。
在一个优选实施例中,具有角质酶增强活性的分离的GH61多肽进一步包括H-X(1,2)-G-P-X(3)-[YW]-[AILMV]。在另一个优选实施例中,具有角质酶增强活性的分离的GH61多肽进一步包括[EQ]-X-Y-X(2)-C-X-[EHQN]-[FILV]-X-[ILV]。在另一个优选实施例中,具有角质酶增强活性的分离的GH61多肽进一步包括H-X(1,2)-G-P-X(3)-[YW]-[AILMV]和[EQ]-X-Y-X(2)-C-X-[EHQN]-[FILV]-X-[ILV]。
在一个第二方面,具有角质酶增强活性的分离的多肽包括以下基序:
[ILMV]-P-X(4,5)-G-X-Y-[ILMV]-X-R-X-[EQ]-X(3)-A-[HNQ],
其中X是任何氨基酸,X(4,5)是处于4个或5个连续位置的任何氨基酸,并且X(3)是处于3个连续位置的任何氨基酸。在以上基序中,采用可接受的IUPAC单一字母氨基酸缩写。
在一个第三方面,该具有角质酶增强活性的GH61多肽包括以下的一个氨基酸序列或由其组成,该氨基酸序列与SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10、SEQ ID NO:11、SEQ IDNO:12、SEQ ID NO:13、SEQ ID NO:14、SEQ ID NO:15、SEQ ID NO:16、SEQ ID NO:17、SEQ IDNO:18、SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:21、SEQ ID NO:22、SEQ ID NO:23、SEQ IDNO:24、SEQ ID NO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ ID NO:28、SEQ ID NO:29、SEQ IDNO:30、SEQ ID NO:31、SEQ ID NO:32、SEQ ID NO:33、SEQ ID NO:34、SEQ ID NO:35、SEQ IDNO:36、SEQ ID NO:37、SEQ ID NO:38、SEQ ID NO:39、SEQ ID NO:40、SEQ ID NO:41、SEQ IDNO:42、SEQ ID NO:43、SEQ ID NO:44、SEQ ID NO:45、SEQ ID NO:46或SEQ ID NO:47的成熟多肽具有至少60%,例如至少65%、至少70%、至少75%、至少80%、至少81%、至少82%、至少83%、至少84%、至少85%、至少86%、至少87%、至少88%、至少89%、至少90%、至少91%、至少92%、至少93%、至少94%、或至少95%、至少96%、至少97%、至少98%、至少99%、或至少100%序列一致性。
在一个实施例中,该成熟多肽包括以下各项或由其组成:SEQ ID NO:2的氨基酸20至326、SEQ ID NO:3的氨基酸18至239、SEQ ID NO:4的氨基酸20至258、SEQ ID NO:5的氨基酸19至226、SEQ ID NO:6的氨基酸20至304、SEQ ID NO:7的氨基酸16至317、SEQ ID NO:8的氨基酸22至249、SEQ ID NO:9的氨基酸20至249、SEQ ID NO:10的氨基酸18至232、SEQ IDNO:11的氨基酸16至235、SEQ ID NO:12的氨基酸19至323、SEQ ID NO:13的氨基酸16至310、SEQ ID NO:14的氨基酸20至246、SEQ ID NO:15的氨基酸22至354、SEQ ID NO:16的氨基酸22至250、SEQ ID NO:17的氨基酸22至322、SEQ ID NO:18的氨基酸24至444、SEQ ID NO:19的氨基酸26至253、SEQ ID NO:20的氨基酸18至246、SEQ ID NO:21的氨基酸20至334、SEQID NO:22的氨基酸18至227、SEQ ID NO:23的氨基酸20至223、SEQ ID NO:24的氨基酸22至368、SEQ ID NO:25的氨基酸25至330、SEQ ID NO:26的氨基酸17至236、SEQ ID NO:27的氨基酸19至250、SEQ ID NO:28的氨基酸23至478、SEQ ID NO:29的氨基酸17至230、SEQ IDNO:30的氨基酸20至257、SEQ ID NO:31的氨基酸23至251、SEQ ID NO:32的氨基酸19至349、SEQ ID NO:33的氨基酸24至436、SEQ ID NO:34的氨基酸21至344、SEQ ID NO:35的氨基酸26至400、SEQ ID NO:36的氨基酸21至389、SEQ ID NO:37的氨基酸22至406、SEQ ID NO:38的氨基酸20至427、SEQ ID NO:39的氨基酸18至267、SEQ ID NO:40的氨基酸21至273、SEQID NO:41的氨基酸21至322、SEQ ID NO:42的氨基酸18至234、SEQ ID NO:43的氨基酸24至233、SEQ ID NO:44的氨基酸17至237、SEQ ID NO:45的氨基酸20至484、SEQ ID NO:46的氨基酸22至320、或SEQ ID NO:47的氨基酸21至330。
优选地,该具有角质酶增强活性的GH61多肽包括以下的一个氨基酸序列或由其组成,该氨基酸序列与SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10、SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13、SEQ ID NO:14、SEQ ID NO:15、SEQ ID NO:16、SEQ ID NO:17、SEQ ID NO:18、SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:21、SEQ ID NO:22、SEQ ID NO:23、SEQ IDNO:24、SEQ ID NO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ ID NO:28、SEQ ID NO:29、SEQ IDNO:30、SEQ ID NO:31、SEQ ID NO:32、SEQ ID NO:33、SEQ ID NO:34、SEQ ID NO:35、SEQ IDNO:36、SEQ ID NO:37、SEQ ID NO:38、SEQ ID NO:39、SEQ ID NO:40、SEQ ID NO:41、SEQ IDNO:42、SEQ ID NO:43、SEQ ID NO:44、SEQ ID NO:45、SEQ ID NO:46或SEQ ID NO:47的成熟多肽具有至少90%一致性。更优选地,与SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4、SEQ IDNO:5、SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10、SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13、SEQ ID NO:14、SEQ ID NO:15、SEQ ID NO:16、SEQ IDNO:17、SEQ ID NO:18、SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:21、SEQ ID NO:22、SEQ IDNO:23、SEQ ID NO:24、SEQ ID NO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ ID NO:28、SEQ IDNO:29、SEQ ID NO:30、SEQ ID NO:31、SEQ ID NO:32、SEQ ID NO:33、SEQ ID NO:34、SEQ IDNO:35、SEQ ID NO:36、SEQ ID NO:37、SEQ ID NO:38、SEQ ID NO:39、SEQ ID NO:40、SEQ IDNO:41、SEQ ID NO:42、SEQ ID NO:43、SEQ ID NO:44、SEQ ID NO:45、SEQ ID NO:46或SEQID NO:47的成熟多肽具有至少95%一致性。最优选地,与SEQ ID NO:2、SEQ ID NO:3、SEQID NO:4、SEQ ID NO:5、SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9、SEQ IDNO:10、SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13、SEQ ID NO:14、SEQ ID NO:15、SEQ IDNO:16、SEQ ID NO:17、SEQ ID NO:18、SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:21、SEQ IDNO:22、SEQ ID NO:23、SEQ ID NO:24、SEQ ID NO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ IDNO:28、SEQ ID NO:29、SEQ ID NO:30、SEQ ID NO:31、SEQ ID NO:32、SEQ ID NO:33、SEQ IDNO:34、SEQ ID NO:35、SEQ ID NO:36、SEQ ID NO:37、SEQ ID NO:38、SEQ ID NO:39、SEQ IDNO:40、SEQ ID NO:41、SEQ ID NO:42、SEQ ID NO:43、SEQ ID NO:44、SEQ ID NO:45、SEQ IDNO:46或SEQ ID NO:47的成熟多肽具有至少100%一致性。
在一个第六方面,该具有角质酶增强活性的GH61多肽是SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9、SEQID NO:10、SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13、SEQ ID NO:14、SEQ ID NO:15、SEQID NO:16、SEQ ID NO:17、SEQ ID NO:18、SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:21、SEQID NO:22、SEQ ID NO:23、SEQ ID NO:24、SEQ ID NO:25、SEQ ID NO:26、SEQ ID NO:27、SEQID NO:28、SEQ ID NO:29、SEQ ID NO:30、SEQ ID NO:31、SEQ ID NO:32、SEQ ID NO:33、SEQID NO:34、SEQ ID NO:35、SEQ ID NO:36、SEQ ID NO:37、SEQ ID NO:38、SEQ ID NO:39、SEQID NO:40、SEQ ID NO:41、SEQ ID NO:42、SEQ ID NO:43、SEQ ID NO:44、SEQ ID NO:45、SEQID NO:46或SEQ ID NO:47的成熟多肽的一个变体,该变体在一个或多个(例如若干个)位置包括一个取代、缺失和/或插入。
优选地,氨基酸改变的性质较小,也就是说不会显著影响蛋白质折叠和/或活性的保守氨基酸取代或插入;典型地是1个至大约30个氨基酸的小段缺失;小段氨基或羧基末端延伸,例如氨基末端的甲硫氨酸残基;多至大约20-25个残基的小段接头肽;或通过改变净电荷或另一功能而协助纯化的小段延伸,例如多组氨酸区段、抗原表位、或结合结构域。
保守取代的实例处于以下组内:碱性氨基酸(精氨酸、赖氨酸以及组氨酸)、酸性氨基酸(谷氨酸和天冬氨酸)、极性氨基酸(谷氨酰胺和天冬酰胺)、疏水性氨基酸(亮氨酸、异亮氨酸以及缬氨酸)、芳香族氨基酸(苯丙氨酸、色氨酸以及酪氨酸)、以及小氨基酸(甘氨酸、丙氨酸、丝氨酸、苏氨酸以及甲硫氨酸)。一般不会改变比活性的氨基酸取代是本领域已知的并且例如在H.诺伊拉特(Neurath)和R.L.希尔(Hill),1979在蛋白质(The Proteins),学术出版社(Academic Press),纽约中描述的。最常发生的交换是Ala/Ser、Val/Ile、Asp/Glu、Thr/Ser、Ala/Gly、Ala/Thr、Ser/Asn、Ala/Val、Ser/Gly、Tyr/Phe、Ala/Pro、Lys/Arg、Asp/Asn、Leu/Ile、Leu/Val、Ala/Glu、以及Asp/Gly。
可替代地,氨基酸改变具有这样一种性质:改变多肽的物理化学特性。例如,氨基酸改变可以提高多肽的热稳定性、改变底物特异性、改变最适pH,等等。
一种母体多肽中的必需氨基酸可以根据本领域中已知的程序来识别,如定点诱变或丙氨酸扫描诱变(坎宁安(Cunningham)和韦尔斯(Wells),1989,科学(Science)244:1081-1085)。在后一项技术中,在该分子中的每个残基处引入单个丙氨酸突变,并且对所得突变体分子的角质酶增强活性进行测试以鉴别对于该分子的活性至关重要的氨基酸残基。还参见,希尔顿(Hilton)等人,1996,生物化学杂志271:4699-4708。也可结合假定接触位点氨基酸的突变,如通过以下技术例如核磁共振、结晶学、电子衍射、或光亲和标记进行确定的对结构进行物理学分析,从而确定酶的活性位点或其他生物学相互作用。参见,例如,德弗斯(de Vos)等人,1992,科学255:306-312;史密斯(Smith)等人,1992,分子生物学杂志224:899-904;乌乐达维尔(Wlodaver)等人,1992,FEBS快报309:59-64。还可以从与亲本多肽相关的多肽的一致性分析推断必需氨基酸的一致性。
可以做出单个或多个氨基酸取代、缺失和/或插入并且使用诱变、重组和/或改组的已知方法进行测试,随后进行相关筛选程序,如由里德哈尔-奥尔森(Reidhaar-Olson)和萨奥尔(Sauer),1988,科学(Science)241:53-57;博维(Bowie)和萨奥尔,1989,美国国家科学院院刊(Proc.Natl.Acad.Sci.USA)86:2152-2156;WO 95/17413;或WO 95/22625披露的那些。其他可以使用的方法包括易错PCR、噬菌体展示(例如洛曼(Lowman)等人,1991,生物化学(Biochemistry)30:10832-10837;美国专利号5,223,409;WO 92/06204)以及区域定向诱变(德比什尔(Derbyshire)等人,1986,基因(Gene)46:145;内尔(Ner)等人,1988,DNA 7:127)。
可以结合诱变/改组方法与高通量自动化筛选方法来检测由宿主细胞表达的克隆的、诱变的多肽的活性(内斯(Ness)等人,1999,自然生物技术(Nature Biotechnology)17:893-896)。编码活性多肽的诱变的DNA分子可以回收自宿主细胞,并且使用本领域的标准方法对其进行迅速测序。这些方法允许迅速确定多肽中单个氨基酸残基的重要性。
SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ ID NO:6、SEQ IDNO:7、SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10、SEQ ID NO:11、SEQ ID NO:12、SEQ IDNO:13、SEQ ID NO:14、SEQ ID NO:15、SEQ ID NO:16、SEQ ID NO:17、SEQ ID NO:18、SEQ IDNO:19、SEQ ID NO:20、SEQ ID NO:21、SEQ ID NO:22、SEQ ID NO:23、SEQ ID NO:24、SEQ IDNO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ ID NO:28、SEQ ID NO:29、SEQ ID NO:30、SEQ IDNO:31、SEQ ID NO:32、SEQ ID NO:33、SEQ ID NO:34、SEQ ID NO:35、SEQ ID NO:36、SEQ IDNO:37、SEQ ID NO:38、SEQ ID NO:39、SEQ ID NO:40、SEQ ID NO:41、SEQ ID NO:42、SEQ IDNO:43、SEQ ID NO:44、SEQ ID NO:45、SEQ ID NO:46或SEQ ID NO:47的成熟多肽的氨基酸取代、缺失和/或插入的总数一直到10个,例如1、2、3、4、5、6、7、8、9或10个。
在一个实施例中,该成熟多肽包括以下各项或由其组成:SEQ ID NO:22的氨基酸20至326、SEQ ID NO:3的氨基酸18至239、SEQ ID NO:4的氨基酸20至258、SEQ ID NO:5的氨基酸19至226、SEQ ID NO:6的氨基酸20至304、SEQ ID NO:7的氨基酸16至317、SEQ ID NO:8的氨基酸22至249、SEQ ID NO:9的氨基酸20至249、SEQ ID NO:10的氨基酸18至232、SEQ IDNO:11的氨基酸16至235、SEQ ID NO:12的氨基酸19至323、SEQ ID NO:13的氨基酸16至310、SEQ ID NO:14的氨基酸20至246、SEQ ID NO:15的氨基酸22至354、SEQ ID NO:16的氨基酸22至250、SEQ ID NO:17的氨基酸22至322、SEQ ID NO:18的氨基酸24至444、SEQ ID NO:19的氨基酸26至253、SEQ ID NO:20的氨基酸18至246、SEQ ID NO:21的氨基酸20至334、SEQID NO:22的氨基酸18至227、SEQ ID NO:23的氨基酸20至223、SEQ ID NO:24的氨基酸22至368、SEQ ID NO:25的氨基酸25至330、SEQ ID NO:26的氨基酸17至236、SEQ ID NO:27的氨基酸19至250、SEQ ID NO:28的氨基酸23至478、SEQ ID NO:29的氨基酸17至230、SEQ IDNO:30的氨基酸20至257、SEQ ID NO:31的氨基酸23至251、SEQ ID NO:32的氨基酸19至349、SEQ ID NO:33的氨基酸24至436、SEQ ID NO:34的氨基酸21至344、SEQ ID NO:35的氨基酸26至400、SEQ ID NO:36的氨基酸21至389、SEQ ID NO:37的氨基酸22至406、SEQ ID NO:38的氨基酸20至427、SEQ ID NO:39的氨基酸18至267、SEQ ID NO:40的氨基酸21至273、SEQID NO:41的氨基酸21至322、SEQ ID NO:42的氨基酸18至234、SEQ ID NO:43的氨基酸24至233、SEQ ID NO:44的氨基酸17至237、SEQ ID NO:45的氨基酸20至484、SEQ ID NO:46的氨基酸22至320、或SEQ ID NO:47的氨基酸1至20。
共物质
与GH61多肽一起添加一种共物质甚至可以进一步增强该酶促效率。
在一个方面,根据WO 2008/151043,具有角质酶增强活性的GH61多肽是在一种可溶性活化二价金属阳离子的存在下使用。在一个优选方面,该可溶性活化二价金属阳离子选自周期表中的碱金属或过渡金属。在一个更优选方面,该可溶性活化二价金属阳离子选自下组,该组由以下各项组成:Mn++、Co++、Mg++、Ca++及其组合。在一个更优选的方面,该可溶性活化二价金属阳离子是Mn++。在另一个更优选的方面,该可溶性活化二价金属阳离子是Co++。在另一个更优选的方面,该可溶性活化二价金属阳离子是Mg++。在另一个更优选的方面,该可溶性活化二价金属阳离子是Ca++。在另一个更优选方面,该可溶性活化二价金属阳离子是选自下组的两种或更多种(若干种)阳离子,该组由以下各项组成:Mn++、Co++、Mg++、以及Ca++。在一个最优选的方面,该可溶性活化二价金属阳离子处于硫酸锰的形式。
在一个方面,该具有角质酶增强活性的GH61多肽在一种二氧基化合物、一种双环化合物、一种杂环化合物、一种含氮化合物或一种含硫化合物存在下使用。
二氧基化合物可以包括包含两个或更多个氧原子的任何适合化合物。在一些方面,二氧基化合物包含一个如在此所述的被取代的芳基部分。二氧基化合物可以包含一个或多个(例如若干个)羟基和/或羟基衍生物,而且包括缺乏羟基和羟基衍生物的取代的芳基部分。二氧基化合物的非限制性实例包括邻苯二酚或儿茶酚;咖啡酸;3,4-二羟基苯甲酸;4-叔丁基-5-甲氧基-1,2-苯二醇;连苯三酚;没食子酸;3,4,5-三羟基苯甲酸甲酯;2,3,4-三羟基二苯甲酮;2,6-二甲氧基苯酚;芥子酸;3,5-二羟基苯甲酸;4-氯-1,2-苯二醇;4-硝基-1,2-苯二醇;丹宁酸;没食子酸乙酯;羟乙酸甲酯;二羟基富马酸;2-丁炔-1,4-二醇;克酮酸;1,3-丙二醇;酒石酸;2,4-戊二醇;3-乙氧基-1,2-丙二醇;2,4,4'-三羟基二苯甲酮;顺-2-丁烯-1,4-二醇;3,4-二羟基-3-环丁烯-1,2-二酮;二羟基丙酮;丙烯醛缩醛;4-羟基苯甲酸甲酯;4-羟基苯甲酸;以及3,5-二甲氧基-4-羟基苯甲酸甲酯;或其盐或溶剂化物。
双环化合物可以包括如在此所描述的任何适合的取代的稠合环系统。这些化合物可以包含一个或多个(例如,数个)另外的环,并且除非另外说明,否则不限于具体数目的环。在一方面,双环化合物是一种类黄酮。在另一方面,双环化合物是一种任选取代的异类黄酮。在另一方面,双环化合物是一种任选取代的花色离子(flavylium ion),如一种任选取代的花色素或任选取代的花色素苷、或其衍生物。双环化合物的非限制性实例包括:表儿茶素、槲皮素、杨梅黄酮、紫衫叶素、堪非醇、桑色素、刺槐素、柚皮素、异鼠李素、芹黄素、矢车菊素、矢车菊素苷、黑豆多酚、花青素鼠李葡糖苷、或其盐或溶剂合物。
杂环化合物可以是如在此所描述的任何适合的化合物,如包含一个杂原子的一种任选取代的芳香族或非芳香族环。在一方面,杂环是包含一个任选取代的杂环烷基部分或一个任选取代的杂芳基部分的一种化合物。在另一方面,任选取代的杂环烷基部分或任选取代的杂芳基部分是一个任选取代的5元杂环烷基或一个任选取代的5元杂芳基部分。在另一方面,任选取代的杂环烷基或任选取代的杂芳基部分是选自以下各项的一个任选取代的部分:吡唑基、呋喃基、咪唑基、异噁唑基、噁二唑基、噁唑基、吡咯基、吡啶基、嘧啶基、哒嗪基、噻唑基、三唑基、噻吩基、二氢噻吩并-吡唑基、硫茚基、咔唑基、苯并咪唑基、苯并噻吩基、苯并呋喃基、吲哚基、喹啉基、苯并三唑基、苯并噻唑基、苯并噁唑基、苯并咪唑基、异喹啉基、异吲哚基、吖啶基、苯并异噁唑基、二甲基乙内酰脲、吡嗪基、四氢呋喃基、吡咯啉基、吡咯烷基、吗啉基、吲哚基、二氮杂卓基、氮杂卓基、硫杂卓基、哌啶基、以及氧杂卓基。在另一方面,任选取代的杂环烷基部分或任选取代的杂芳基部分是一个任选取代的呋喃基。杂环化合物的非限制性实例包括:(1,2-二羟乙基)-3,4-二羟呋喃-2(5H)-酮、4-羟基-5-甲基-3-呋喃酮、5-羟基-2(5H)-呋喃酮、[1,2-二羟乙基]呋喃-2,3,4(5H)-三酮、α-羟基-γ-丁内酯、核糖酸γ-内酯、己醒糖酸γ-内酯(aldohexuronicaldohexuronicacidγ-lactone)、葡糖酸δ-内酯、4-羟基香豆素、二氢苯并呋喃、5-(羟甲基)糠醛、联糠醛、2(5H)-呋喃酮、5,6-二氢-2H-吡喃-2-酮、以及5,6-二氢-4-羟基-6-甲基-2H-吡喃-2-酮、或其盐或溶剂化物。
含氮化合物可以是具有一个或多个(例如若干个)氮原子的任何合适的化合物。在一个方面,含氮化合物包含一个胺、亚胺、羟胺或氮氧化物部分。含氮化合物的非限制性实例包括:丙酮肟、紫尿酸、吡啶-2-醛肟、2-氨基苯酚、1,2-苯二胺、2,2,6,6-四甲基-1-哌啶基氧基、5,6,7,8-四氢生物蝶呤、6,7-二甲基-5,6,7,8-四氢蝶呤、以及马来酰胺酸、或其盐或溶剂化物。
醌化合物可以是如在此所描述的包含一个醌部分的任何适合的化合物。醌化合物的非限制性实例包括:1,4-苯醌、1,4-萘醌、2-羟基-1,4-萘醌、2,3-二甲氧基-5-甲基-1,4-苯醌或辅酶Q0、2,3,5,6-四甲基-1,4-苯醌或杜醌、1,4-二羟基蒽醌、3-羟基-1-甲基-5,6-吲哚啉二酮或肾上腺素红、4-叔丁基-5-甲氧基-1,2-苯醌、吡咯并喹啉醌、或其盐或溶剂化物。
含硫化合物可以是包括一个或多个(例如若干个)硫原子的任何适合化合物。在一个方面,含硫化合物包含一个选自以下各项的部分:亚硫酰基、硫醚、亚磺酰基、磺酰基、硫酰胺、磺酰胺、磺酸以及磺酸酯。含硫化合物的非限制性实例包括乙硫醇;2-丙硫醇;2-丙烯-1-硫醇;2-巯基乙磺酸;苯硫酚;苯-1,2-二硫醇;半胱氨酸;蛋氨酸;谷胱甘肽;胱氨酸;或其盐或溶剂化物。
在一方面,以上所描述的这种化合物对聚酯纺织品材料的量,作为与纤维素的葡糖基单元的摩尔比是约10-6至约10,例如,约10-6至约7.5、约10-6至约5、约10-6至约2.5、约10-6至约1、约10-5至约1、约10-5至约10-1、约10-4至约10-1、约10-3至约10-1、或约10-3至约10-2。在另一方面,以上所描述的这种化合物的量是约0.1μM至约1M,例如,约0.5μM至约0.75M、约0.75μM至约0.5M、约1μM至约0.25M、约1μM至约0.1M、约5μM至约50mM、约10μM至约25mM、约50μM至约25mM、约10μM至约10mM、约5μM至约5mM、或约0.1mM至约1mM。
术语“液体”意指该溶液相,无论是水性的、有机的、还是其组合。
聚酯织物制造工艺
聚酯例如聚(对苯二甲酸乙二酯)是通过以下合成:缩合,从熔化物拉伸为纤维,可能地切割为短纤维,可能地与其他纤维类型混合,并且纺成纱线。
在编织纱线或织成织物之后,一般对该织物进行处理以去除纺丝油(spin finishoil),例如在一个工艺中其中首先将该织物在180℃进行热定型并且然后在80℃-100℃用表面活性剂(有时也添加碱)进行预处理,并且然后任选地接着通过使用剧烈的碱在高达130℃水解聚酯织物以引起重量减少过程,以使其具有更柔软和更有光泽的外观。然后将该聚酯织物进行热定型并且用分散染料在pH 4.5-6、高达130℃下进行染色,接着用硫代硫酸钠在60℃-80℃、pH 10下进行还原洗净。如果必要,这些过程后面可接着精加工(后处理)步骤以进一步改进纺织品特性,例如抗起球、可湿性改进或抗静电处理。
在合成和拉伸过程中,在这些纤维之上或之中形成聚对苯二甲酸乙二酯的环状或线性低聚物。环状和/或线性低聚物的去除可通过用一种或多种角质酶水解来完成。该角质酶打破该环状低聚物的环结构并打破BETEB链以通过水解酯键产生苯甲酸、对苯二酸(terephathalate acid)以及乙二醇。可将所得的产物在温和条件下去除。
本发明的用GH61多肽和角质酶处理聚酯纺织品的方法发生在预处理、重量减少、分散染色或后精加工的一个或多个后续步骤期间,以赋予该聚酯织物以下效果中的至少一个:减少该聚酯纺织品中的低聚物,减少球粒形成,改进亲水性和抗静电特性等。本发明的方法可以作为分离的步骤或与现存的聚酯加工步骤中的任一个组合进行。
本发明的工艺易于使用于纺织品工业中,因为它可以使用现存的湿加工设备进行,例如在经轴染色机、浸轧(Pad-Roll)、轱辘/绞车(Jigger/Winch)、J-盒(J-Box)、或轧蒸(Pad-Steam)型设备中进行。该工艺优选发生在精加工(后处理)步骤过程中。
如此处使用的,术语“生物抛光”、“去球”、“减少球粒形成”以及“抗起球”是可互换的。
不应用精加工组件的情况下,聚酯织物具有相当硬和坚硬的手感外观。一些织物表面不光滑,因为小的有绒毛的微纤维从其中突出。另外,在相对短期的穿着之后,起球发生于该织物表面上,从而给予它无吸引力的磨损的样子。
生物抛光是在聚酯织物制造过程中对其进行处理的方法,该方法改进了针对“减少球粒形成”的织物品质。生物抛光的最重要效果的特征可在于较少起毛和起球,增加的光彩/光泽(gloss/luster),改进的纺织品手感,增加的持久柔软性,抗静电特性和/或改进的吸水性。在本背景下,术语“减少球粒形成”旨在意为根据本发明的方法处理的织物表面对表面上球粒形成的耐受性。
出于本发明的目的,可根据材料与方法部分中的“起球记录测试”的描述来测试球粒形成。该测试的结果是依照“起球记录”来表示,该记录在标度上是从起球记录1(严重球粒形成)至起球记录5(无球粒形成)的评级,允许1/4起球记录。
由于生物抛光的方法催化聚酯纤维表面的水解,最终酶作用将导致纤维或织物的重量损失。在一个优选实施例中,生物抛光是以这样一种方式进行,以至于获得受控的、部分的纤维表面水解,即获得适当的抛光效果而不用过度损失织物强度。
出于本发明的目的,该生物抛光效果是如在实例6中指定的条件下通过用2.8mg蛋白质/克织物的角质酶和2.8mg蛋白质/克织物的GH61在70℃、pH 8.0在洗衣指数计(Launder-O-Meter)中处理PET 2小时测量的。在本发明的一个优选实施例中,这种用角质酶和GH61的处理导致如下的起球记录:至少2.00、优选至少2.25、并且甚至更优选至少2.5,而同时优选显示少于5%、优选少于4%、更优选少于3%、更优选少于2%并且最优选少于1%的重量损失。在优选的实施例中,与用2.8mg蛋白质/克织物的角质酶而不用GH61的处理相比,如在实例6中指定的条件下用2.8mg蛋白质/克的角质酶和2.8mg蛋白质/克的GH61在洗衣指数计(Launder-O-Meter)中的PET处理导致0.25的起球记录增加。
工艺条件
可在聚酯纺织品制造工艺中使用GH61多肽与角质酶的组合,无论作为现存聚酯制造步骤中任一个之后的单独步骤还是与现存聚酯制造步骤像预处理、重量减少、分散染色或后精加工中任一个的组合。
建议有待用于本方法中的适合的液体/纺织品比率可处于从约20:1至约1:1的范围中,优选处于从约15:1至约3:1的范围中,更优选处于从15:1至5:1的范围中(体积/重量,ml/mg)。
对于本发明的反应时间通常处于从约10分钟至约8小时的范围中。优选地,该反应时间处于从约20分钟至约180分钟的范围中,更优选该反应时间处于从约30分钟至约150分钟的范围中,最优选地该反应时间处于从约45分钟至约120分钟的范围中。
该反应介质的pH很大程度上取决于所讨论的一种或多种酶。优选地本发明的工艺是在从该角质酶的pH最佳值+/-1个pH单位下进行的。优选地,本发明的工艺是在处于从约pH 3至约pH 11的范围中的、优选处于从约pH 4至约pH 10范围中的、或处于从约pH 6至约pH 9的范围中的pH下进行。
本发明的处理温度优选地根据该角质酶的最佳温度+/-10℃选择。优选地该工艺能够在100℃以下、优选90℃以下、更优选80℃以下、并且甚至更优选75℃以下的温度运行。
在一些实施例中,本发明的工艺在40℃-100℃、优选50℃-90℃、优选60℃-85℃、更优选65℃-80℃、并且甚至更优选70℃-80℃的温度范围进行。
酶剂量很大程度上取决于酶反应时间,即相对短的酶反应时间使相对增加的酶剂量成为必需,并且反之亦然。通常,酶剂量可根据可用的反应时间来确定。
根据本发明的方法有待使用的GH61多肽的量取决于许多因素并且优选地应当由技术人员优化。根据本发明,该GH61多肽在该水性介质中的优选浓度是从约0.01至约50毫克蛋白质/克聚酯纺织品,优选0.05-20毫克(mg)蛋白质/克(g)聚酯纺织品,优选0.1-15毫克蛋白质/克聚酯纺织品,更优选0.2-8毫克蛋白质/克聚酯纺织品,并且甚至更优选0.2-5毫克蛋白质/克聚酯纺织品。
根据本发明的方法有待使用的角质酶的量取决于许多因素并且优选地应当由技术人员优化。根据本发明,该角质酶在该水性介质中的优选浓度是从约0.01至约50毫克酶蛋白质/克聚酯纺织品,优选0.05-20毫克酶蛋白质/克聚酯纺织品,更优选0.1-15毫克酶蛋白质/克聚酯纺织品,并且甚至更优选0.2-5毫克酶蛋白质/克聚酯纺织品。优选地,角质酶和GH61之间的剂量比率是1:1至1:0.5。
本发明的工艺可进一步包括添加一种或多种能够改进酶-底物相互作用的化学品(为了改进该底物的可达到性和/或溶解反应产物),这些化学品可在酶处理之前或同时添加。这类化学品可以具体是如上所述的共物质、表面活性剂、润湿剂、抗起球剂以及分散剂,或其混合物。
本发明的工艺可任选地包括一个漂洗步骤,在此期间使这些水解的低聚物经受漂洗,具体地用碱溶液漂洗。碱溶液溶解这些低聚物的线性片段,并且在一定程度上可进一步水解这些线性片段。
在本发明的方法中使用的水性组合物可进一步包括选自下组的一种或多种酶,该组由以下各项组成:脂肪酶、酯酶、漆酶、过氧化物酶等。
用于处理纺织品的组合物
本发明还囊括适于处理纺织品的一种组合物,其中该组合物包括一种GH61多肽和一种角质酶。
本发明的组合物的使用可为该聚酯织物提供以下效果中的至少一个:减少该聚酯纺织品中的低聚物,减少球粒形成,改进亲水性和抗静电特性等。
本发明的纺织品组合物适于该聚酯制造过程例如预处理、重量减少、分散染色和后精加工中的一个或多个,无论是以分离的步骤还是与这些步骤中的任一个组合。
在本发明中,GH61多肽通过减少达到相同程度的去球所需要的角质酶的量来增强该角质酶活性。
在本发明的一些实施例中,这种包含一种GH61多肽和一种角质酶的组合物进一步包含其他组分,包括而不限于其他酶,以及一种或多种表面活性剂、漂白剂、消泡剂、构建剂系统以及类似的。
适合于在本发明中使用的酶包括而不限于脂肪酶、酯酶、漆酶、过氧化物酶、环化酶以及转移酶。
在一个实施例中,该纺织品组合物包括一种或多种选自下组的GH 61多肽,该组由一个氨基酸序列组成,该氨基酸序列与SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4、SEQ IDNO:5、SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10、SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13、SEQ ID NO:14、SEQ ID NO:15、SEQ ID NO:16、SEQ IDNO:17、SEQ ID NO:18、SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:21、SEQ ID NO:22、SEQ IDNO:23、SEQ ID NO:24、SEQ ID NO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ ID NO:28、SEQ IDNO:29、SEQ ID NO:30、SEQ ID NO:31、SEQ ID NO:32、SEQ ID NO:33、SEQ ID NO:34、SEQ IDNO:35、SEQ ID NO:36、SEQ ID NO:37、SEQ ID NO:38、SEQ ID NO:39、SEQ ID NO:40、SEQ IDNO:41、SEQ ID NO:42、SEQ ID NO:43、SEQ ID NO:44、SEQ ID NO:45、SEQ ID NO:46或SEQID NO:47的成熟多肽具有至少60%,例如至少65%、至少70%、至少75%、至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、或至少95%、至少96%、至少97%、至少98%、至少99%、或至少100%的程度的一致性。
在一个甚至更优选的方面,该纺织品组合物进一步包括如在以上的“共物质”部分中所述的一种共物质。在一个优选实施例中,该共物质是半胱氨酸。
该纺织品组合物可处于任何形式,例如固体、液体、膏体、凝胶或其任何组合。
表面活性剂
在处理聚酯纺织品中,可使用一种常规的表面活性剂来改进与酶的接触。
本发明的纺织品组合物可以包括一种或多种表面活性剂,它们可以是阴离子的和/或阳离子的和/或非离子的和/或半极性的和/或兼性离子的,或其混合物。这种或这些表面活性剂典型地以组合物的重量计是从约0.001%至20%的水平存在,例如约0.005%至约10%、或约0.01%至约5%、或约0.02%至约1%。
更特别地,在本发明的工艺或组合物中使用的表面活性剂包括一种非离子表面活性剂。非离子型表面活性剂的非限制性实例包括醇乙氧基化物(AE或AEO)、醇丙氧基化物、丙氧基化的脂肪醇(PFA)、烷氧基化的脂肪酸烷基酯(例如乙氧基化的和/或丙氧基化的脂肪酸烷基酯)、烷基酚乙氧基化物(APE)、Triton、壬基酚乙氧基化物(NPE)、烷基多糖苷(APG)、烷氧基化胺、脂肪酸单乙醇酰胺(FAM)、脂肪酸二乙醇酰胺(FADA)、乙氧基化的脂肪酸单乙醇酰胺(EFAM)、丙氧基化的脂肪酸单乙醇酰胺(PFAM)、多羟基烷基脂肪酸酰胺、或葡萄糖胺的N-酰基N-烷基衍生物(葡糖酰胺(GA)、或脂肪酸葡糖酰胺(FAGA))、以及在SPAN和TWEEN商品名下可获得的产品、以及其组合。
其他酶类
该酶促聚酯制造工艺以及该纺织品组合物可包括一种或多种另外的酶,例如脂肪酶、酯酶、漆酶、过氧化物酶、环化酶以及转移酶。
脂肪酶:适合的脂肪酶包括细菌或真菌来源的脂肪酶。包括化学修饰的或蛋白工程化的突变体。该脂肪酶例如可以是三酰甘油脂肪酶(EC3.1.1.3)、磷脂酶A2(EC3.1.1.4)、溶血磷脂酶(EC 3.1.1.5)、单酰甘油脂肪酶(EC 3.1.1.23)、半乳糖脂酶(EC3.1.1.26)、磷脂酶A1(EC 3.1.1.32)、脂蛋白脂肪酶(EC 3.1.1.34)。实例包括来自嗜热真菌属的脂肪酶,例如来自如在EP 258 068和EP 305 216中描述的疏棉状嗜热丝孢菌(T.lanuginosus)(之前命名为柔毛腐质霉(Humicola lanuginosa));一种假单胞菌属脂肪酶,例如来自产碱假单胞菌(P.alcaligenes)或类产碱假单胞菌(P.pseudoalcaligenes)(EP 218 272)、洋葱假单胞菌(P.cepacia)(EP 331 376)、斯氏假单胞菌(P.stutzeri)(GB1,372,034)、萤光假单胞菌(P.fluorescens)、假单胞菌属菌株SD 705(WO 95/06720和WO96/27002)、威斯康星假单胞菌(P.wisconsinensis)(WO 96/12012);一种芽孢杆菌属脂肪酶,例如来自枯草芽孢杆菌(达托斯(Dartois)等人,1993,生物化学与生物物理学学报(Biochemica etBiophysica Acta),1131:253-360)、嗜热脂肪芽杆菌(JP 64/744992)或短小芽孢杆菌(WO 91/16422)。
其他实例是脂肪酶变体,例如WO 92/05249、WO 94/01541、EP 407225、EP 260105、WO 95/35381、WO 96/00292、WO 95/30744、WO 94/25578、WO 95/14783、WO 95/22615、WO 97/04079、WO 97/07202、WO 00/060063、WO 2007/087508以及WO 2009/109500中描述的那些。
优选的可商购获得的脂肪酶包括LipolaseTM、Lipolase UltraTM、以及LipexTM;LecitaseTM、LipolexTM;LipocleanTM、LipoprimeTM(诺维信公司)。其他可商购的脂肪酶包括Lumafast(Genencor Int Inc(杰能科国际公司));Lipomax(Gist-Brocades公司/GenencorInt Inc(杰能科国际公司))以及来自Solvay(苏威公司)的芽孢杆菌脂肪酶。
过氧化物酶/氧化酶:适合的过氧化物酶/氧化酶包括那些植物、细菌或真菌来源的过氧化物酶/氧化酶。包括化学修饰的或蛋白工程化的突变体。有用的过氧化物酶的实例包括来自鬼伞属,例如来自灰盖鬼伞的过氧化物酶,以及其变体,如在WO 93/24618、WO 95/10602、以及WO 98/15257中描述的那些。
可商购获得的过氧化物酶包括GuardzymeTM(诺维信公司)。
环化酶:术语“环化酶”意指根据EC 1.11.2.1的一种“非特异性环化酶”活性,其催化来自H2O2的一个氧原子插入多种底物(例如硝基苯并二茂)中。有用的环化酶的实例包括WO 2008/119780中所述的环化酶。
在以下编号的段落中进一步描述本方法和组合物。
1.一种用于在水性溶液中在角质酶的存在下将聚酯纺织品用糖基水解酶家族61多肽进行处理的方法。
2.在段落1所述的方法的一些实施例中,其中该纺织品是纱线、织物或衣服。
3.在段落1或2所述的方法的一些实施例中,其中该聚酯是PET。
4.在段落1所述的方法的一些实施例中,其中该水性溶液进一步包括选自下组的一种或多种酶,该组由以下各项组成:脂肪酶、酯酶、漆酶、过氧化物酶、环化酶以及转移酶。
5.在以上段落中任一项所述的方法的一些实施例中,其中与一种糖基水解酶家族61一起使用了一种共物质,优选地该共物质是半胱氨酸。
6.在以上段落中任一项所述的方法的一些实施例中,其中该糖基水解酶家族61多肽是按以下范围施用的:从0.01至约50毫克蛋白质/克聚酯纺织品,优选0.05-20毫克蛋白质/克聚酯纺织品,优选0.1-15毫克蛋白质/克聚酯纺织品,更优选0.2-8毫克蛋白质/克聚酯纺织品,并且甚至更优选0.25-5毫克蛋白质/克聚酯纺织品。
7.在以上段落中任一项所述的方法的一些实施例中,其中该角质酶是按以下范围施用的:从约0.01至约50毫克酶蛋白质/克聚酯纺织品,优选0.05-20毫克酶蛋白质/克聚酯纺织品,更优选0.1-15毫克酶蛋白质/克聚酯纺织品,并且甚至更优选0.2-5毫克酶蛋白质/克聚酯纺织品。
8.在以上段落中任一项所述的方法的一些实施例中,其中该方法是在以下的pH范围进行的:从约pH 3至约pH 11,优选以从约pH 4至约pH 10的范围,或以从约pH 6至约pH 9的范围。
9.在以上段落中任一项所述的方法的一些实施例中,其中该方法是在以下的温度范围进行的:40℃-100℃,优选50℃-90℃,优选60℃-85℃,更优选65℃-80℃,并且甚至更优选70℃-80℃。
10.在以上段落中任一项所述的方法的一些实施例中,其中该方法进行了约10分钟至约8小时,优选约20分钟至约180分钟,更优选约30分钟至约150分钟,更优选约45分钟至约120分钟。
11.在以上段落中任一项所述的方法的一些实施例中,其中用于处理聚酯纺织品的方法是制造一种聚酯纺织品,尤其制造一种聚酯织物。
12.在段落11所述的方法的一些实施例中,其中该方法与现存的聚酯织物制造步骤中的任一个相组合。
13.在以上段落中任一项所述的方法的一些实施例中,其中该角质酶具有BETEB水解活性。
14.在以上段落中任一项所述的方法的一些实施例中,其中该角质酶与SEQ IDNO:1具有至少90%序列一致性,或包括SEQ ID NO:1的一个或多个(或若干个)氨基酸取代、缺失、和/或插入。
15.在以上段落中任一项所述的方法的一些实施例中,其中当根据实例4的条件测量时,该糖基水解酶家族61多肽具有角质酶增强活性。
16.在以上段落中任一项所述的方法的一些实施例中,其中该糖基水解酶家族61多肽与SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:9、SEQ ID NO:10、SEQ ID NO:11、SEQ ID NO:12、SEQ ID NO:13、SEQ ID NO:14、SEQ ID NO:15、SEQ ID NO:16、SEQ ID NO:17、SEQ ID NO:18、SEQ IDNO:19、SEQ ID NO:20、SEQ ID NO:21、SEQ ID NO:22、SEQ ID NO:23、SEQ ID NO:24、SEQ IDNO:25、SEQ ID NO:26、SEQ ID NO:27、SEQ ID NO:28、SEQ ID NO:29、SEQ ID NO:30、SEQ IDNO:31、SEQ ID NO:32、SEQ ID NO:33、SEQ ID NO:34、SEQ ID NO:35、SEQ ID NO:36、SEQ IDNO:37、SEQ ID NO:38、SEQ ID NO:39、SEQ ID NO:40、SEQ ID NO:41、SEQ ID NO:42、SEQ IDNO:43、SEQ ID NO:44、SEQ ID NO:45、SEQ ID NO:46或SEQ ID NO:47的成熟多肽具有至少90%序列一致性。
17.一种用于处理纺织品的组合物,包括一种糖基水解酶家族61多肽和一种角质酶。
18.在段落17所述的组合物的一些实施例中,其中该组合物进一步包括选自下组的一种或多种酶,该组由以下各项组成:脂肪酶、酯酶、漆酶、过氧化物酶、环化酶以及转移酶。
19.在段落17或18所述的组合物的一些实施例中,其中该组合物进一步包括一种共物质;优选地该共物质是半胱氨酸。
20.在段落17-19中任一项所述的组合物的一些实施例中,其中该组合物进一步包括一种表面活性剂,优选一种非离子表面活性剂。
21.一种糖基水解酶家族61多肽促进角质酶对一种聚酯纺织品的作用的用途。
22.在段落21所述的用途的一些实施例中,其中该作用是减少聚酯纺织品上的球粒形成。
23.在段落21或22所述的用途的一些实施例中,其中在如实例6所指定的条件下,与当使用角质酶而不使用GH61时的起球记录相比时,起球记录增加至少0.125,更优选至少0.250,更优选至少0.375,更优选至少0.500,更优选至少0.625,甚至更优选地至少0.750。
24.在段落21所述的用途的一些实施例中,其中该作用是当与相同条件下不使用GH61的相同工艺运行相比时,减少聚对苯二甲酸乙二酯的环状或线性低聚物在机械和/或纺织品上的沉积。
25.在段落24所述的用途的一些实施例中,其中该低聚物是酸-双-2-苯甲酰氧基-乙酯和/或对苯二甲酸三乙二酯。
实例
材料与方法
蛋白质
角质酶A:来自特异腐质霉的角质酶的变体,具有对SEQ ID NO:1的亲本特异腐质霉角质酶(WO 2001/092502中所述的角质酶A)的取代E6Q+A14P+E47K+R51P+E179Q+G8D+N15D+S48E+A88H+N91H+A130V+R189V
角质酶B:来自特异腐质霉的角质酶的变体,具有对SEQ ID NO:1的亲本特异腐质霉角质酶(WO 2001/092502中所述的角质酶B)的取代E6Q+A14P+E47K+R51P+E179Q+G8D+N15D+T29M+S48E+A88H+N91H+A130V+T166I+L167P+R189V
Af GH61的成熟多肽:烟曲霉GH61B多肽,示为SEQ ID NO:16的氨基酸22至250(描述于US 2010124769)
Ta GH61的成熟多肽:橙色嗜热子囊菌GH61A多肽,示为SEQ ID NO:8的氨基酸22至249(描述于WO 2005/074656)
Nc GH61的成熟多肽:粗糙脉孢菌GH61多肽,示为SEQ ID NO:47的氨基酸21-330(描述于WO 2011080267)
Ts GH61的成熟多肽:柄篮状菌GH61多肽,示为SEQ ID NO:46的氨基酸22至320(UNIPROT:B8M2G3)
化学品
Triton X-100(中国北京克劳泽生物技术有限公司(Beijing KehaozeBiotechnology Co.,Ltd.China))
BETEB(对苯二甲酸-双-2-苯甲酰氧基-乙酯)
PET(聚对苯二甲酸乙二酯,100%型64风格,短纤维织PET织物,可商购于SDL.)
试剂/底物
布里顿-罗宾逊缓冲液(Britton-Robinson Buffer):将0.04M H3BO3、0.04M H3PO4和0.04M CH3COOH的酸性混合物用0.2M NaOH滴定到所希望的pH。
通过10倍稀释上述布里顿-罗宾逊缓冲液并且然后将该溶液用NaOH滴定到所希望的pH获得4mM布里顿-罗宾逊缓冲液。
2.5%BETEB底物:2.5g BETEB+100ml去离子水+0.5ml 1%Triton-X 100
OD吸光度和pH测量
使用角质酶A和B水解在埃普多夫(eppendorf)管中的PET或BETEB。水解产物是对苯二甲酸盐及其酯,其具有254nm(UV)左右的特征吸收峰。因此在254nm的OD吸光度反映酶的向着聚酯的水解活性。在254nm的OD吸光度越高,向着PET或BETEB的酶活性越强。在254nm处的OD是在SpectraMax M2酶标仪(分子器件有限公司(Molecular Devices,LLC.))中读取。如果该吸光度超过该酶标仪的有效范围1.5的话,将该溶液进行稀释。稀释x15意指该溶液已被稀释15倍。
水解产物对苯二甲酸盐是酸性的并且将因此降低溶液的pH,因此在该反应之前和之后的pH变化是用于测试酶活性的一个参数。
重量损失确定
将小块布样在编号之前置于控制室(65%+/-5%湿度,20℃+/-1℃)中24小时,通过分析天平(对于100g以下的样品)或精密天平(对于100g以上的样品)称重并记录。在处理之后,将所有样品滚动干燥(AEG,LAVATHERM 37700,德国)1小时并在与以上相同的控制室中调节24小时。对于每份样品,重量损失定义如下:
重量损失=(处理之前的重量-处理之后的重量)/处理之前的重量X(100%)
起球记录测试
将已经在标准气候(65%湿度,20℃)下预调节至少24小时的织物(包括处理的和未处理的)针对起球记录用Nu-马丁代尔(Martindale)测试仪(詹姆斯(James)H.希尔有限公司(Heal Co.Ltd),英格兰)进行测试,用相同类型的未处理的织物作为磨损的织物。在2000回转之后进行标准的起球测试(瑞士标准(SN)198525),通过从具有如下的定义的含义的1-5标记,其中1显示不良的抗起球而5显示优异的抗起球特性。因此马丁代尔起球记录得分越高,该生物抛光处理越有效。
记录5:无起球
记录4:轻微起球
记录3:中等起球
记录2:明显起球
记录1:严重起球
允许1/2、1/4记录
为了使结果更可靠,对每个样品由不同的人进行3个分开读数,并且这3个读数的平均值被采用作为起球记录的最终结果。
蛋白质含量
在这些实例中使用的酶产品或多肽产品中的蛋白质浓度可用BCATM蛋白质测定试剂盒(产品编号23225,可商购于赛默飞世尔科技公司(ThermoFisher Scientific Inc.))根据产品手册进行测量。
实例1:BETEB-琼脂板用于评价该角质酶活性
BETEB被角质酶水解为更易溶的试剂。因此,在通过酶水解之后,在倾倒了琼脂和BETEB的混合物的板上存在透明区。
BETEB的水解会产生
通过以下过程测量角质酶活性:
a)BETEB溶液制剂:将5ml 100%乙醇添加到具有塞子的玻璃瓶中,将20mg BETEB添加到该乙醇中并且然后将该瓶置于60℃水浴以溶解该BETEB。
b)通过以下制备1.5%琼脂溶液:将0.75g琼脂添加到45ml Tris-HCl缓冲液(25mM,pH 7.0)中,并且然后将烘干器置于微波炉中加热两次持续30秒以溶解该琼脂。
c)将该琼脂溶液冷却至60℃并与步骤a中制备的BETEB溶液混合。将该混合物倾倒皮氏培养皿中。
d)用6mm直径的尖端或打孔器在该皮氏培养皿中挖小孔。
e)对于每个孔将30微克/ml的酶样品由带有75微升(ul)酶样品的尖端添加到皮氏培养皿中。将该皮氏培养皿置于37℃过夜。
在孔周围的区域中角质酶A和角质酶B均显示透明区,因为BETEB被该角质酶水解。
实例2:角质酶A与GH61用于PET处理。
在此实例中,将Af GH61和Ta GH61两种GH61分别与角质酶A组合使用,以在1.5ml埃普多夫管中水解PET点。
将PET织物切成0.5cm直径、0.005g/块的小块,并且将两块加入每个埃普多夫管中。将布里顿-罗宾逊缓冲液(4mM,pH 8)和1%Triton X 100置于恒温混匀仪中在70℃持续5分钟以预热。在预热之后,将角质酶和GH61添加到该管中以得到1ml的总体积,其中TritonX 100的终浓度是0.2g/l,并且角质酶和GH61在该溶液中的终浓度是如表1中所示。如在材料与方法部分中所述的测试OD 254吸光度和pH,直接示为表1中0小时的数据。将这些管置于恒温混匀仪中以在1000rpm和70℃启动该反应。在反应持续如在表1中指定的某一时间段之后,通过将这些埃普多夫管转移到冰浴中持续10分钟来停止该反应。然后将这些埃普多夫管在13000g/min离心10秒以得到上清液用于OD和pH确定。将这些上清液稀释5倍用于OD测试。
表1.角质酶A与两种GH61用于PET处理的结果(70℃,pH 8.0,1000rpm,0-4小时)
注:表1中每个酶组合的三次重复样品的平均值。
如可从表1看到的,在2小时反应之后,对于单独的角质酶A在254nm处的吸光度是0.629,并且对于角质酶A与Af GH61的组合是0.716。2小时之后的pH变化(“pH变化”意指在0小时处的初始pH与反应之后的最终pH之间的差异),当单独使用角质酶A时是0.03(即6.91减6.88);而当一起使用角质酶A和Af GH61时,导致0.07的pH变化(即6.91减6.84)。添加相同剂量的Ta GH61使254nm处的吸光度从0.629增加到0.774,并略微使pH变化从0.03增加到0.06(即6.85减6.79)。
当该反应时间延长到4小时时,Af GH61和角质酶A的组合使254nm处的吸光度从0.806增加到0.899并使pH变化从0.08增加到0.11。在用角质酶A和Ta GH61 4小时之后,吸光度从0.806增加到0.909并且pH变化从0.08增加到0.10。
还发现,单独用GH61时254nm处的吸光度将轻微变化。然而,来自单独GH61的吸光度变化少于当将GH61与角质酶A组合时的值。例如,在单独用Af GH61 4小时反应后,略微地使254nm处的吸光度增加0.01(即0.384减0.374),并且角质酶A使该吸光度增加0.388(即0.806减0.418),并且AfGH61和角质酶A的组合导致0.505(即0.899减0.394)的显著的吸光度增加。因此,将GH61添加到角质酶中导致对PET处理过程中增加PET水解的协同效应。
实例3:角质酶B与GH61用于PET处理。
在此实例中,将两种GH61分别与角质酶B组合使用以在埃普多夫管中水解PET织物。处理方案与实例2中所述的相同。
表2.角质酶B与GH61用于PET处理的结果(70℃,pH 7.0,1000rpm,0-4小时)
注:表2中每个酶组合的三次重复样品的平均值。
如可从表2看到的,在2小时处,将Af GH61或Ta GH61添加到角质酶B使254nm处的吸光度分别增加0.057和0.028,并且在4小时处,该吸光度分别增加0.082和0.058。经2至4小时,在反应之前和之后在pH变化上也存在轻微增加。总之,GH61显示出对角质酶B的促进效应。
实例4:角质酶A与两种GH61用于低聚物处理。
在PET合成和PET的处理过程中BETEB以一种低聚物产生,其可能保留在该纺织品织物中。
将GH61(AfGH61或TaGH61)与角质酶A在1.5ml埃普多夫管中组合使用。添加40mM布里顿-罗宾逊缓冲液(pH 8)以得到该酶和GH61的在0.05mg酶蛋白质/ml浓度的溶液,如表3中所示。将这些管置于恒温混匀仪中在70℃持续5分钟以预热。在预热后,将100ul 2.5%BETEB底物添加到该酶溶液中以起始该反应。将埃普多夫管置于恒温混匀仪中在70℃,1000rpm持续表3中指定的时间。通过将该埃普多夫管转移至冰浴持续10分钟来停止该反应。将这些埃普多夫管在13000g/min离心10秒以得到上清液用于OD和pH确定。将源自0h、20分钟和40分钟反应的上清液稀释5倍,而将源自1小时反应的上清液稀释75倍用于OD测试。表3中针对0小时处取样时间的数据意指添加BETEB之前测试的数据。
表3.角质酶A与两种GH61用于BETEB处理的结果(70℃,pH 8.0,1000rpm,0-1小时)
注:每个酶组合的三次重复样品的平均值。
如表3所示的,在20min反应之后,对于单独的角质酶A 254nm处的吸光度是1.126并且对于角质酶A与Af GH61的组合是1.200。当单独使用角质酶A时20min之后的pH变化是0.16(即7.51减7.35);而当一起使用角质酶A和Af GH61时导致0.17的pH变化(即7.52减7.35)。添加相同剂量的TaGH61使254nm处的吸光度从1.126增加到1.312并略微使pH变化从0.16增加到0.19(即7.5减7.31)。
在40min反应之后,对于单独的角质酶A 254nm处的吸光度是2.293并且对于角质酶A与Af GH61的组合是2.760。当单独使用角质酶A时40min之后的pH变化是0.34(即7.51减7.17);而当一起使用角质酶A和Af GH61时导致0.37的pH变化(即7.52减7.15)。添加相同剂量的Ta GH61使254nm处的吸光度从2.293增加到2.840并使pH变化从0.34增加到0.67(即7.5减6.83)。当与角质酶A组合时,GH61的显著促进效应可用BETEB作为底物进行检测。
实例5:角质酶B与四种GH61用于低聚物处理。
将四种GH61与角质酶B组合,以在以0.01mg酶蛋白质/ml溶液和0.01mg GH61/ml溶液的剂量水解低聚物BETEB。处理方案与实例4中所述的相同。将源自0h和20分钟的上清液稀释5倍,而将源自40分钟和1小时反应的上清液稀释75倍用于OD测试。
表4.角质酶B与四种GH61用于BETEB处理的结果(70℃,pH 8.0,1000rpm,0-1小时)
注:每个酶组合的三次重复样品的平均值。
如可从表4看到的,在20分钟反应之后,对于单独的角质酶B 254nm处的吸光度是1.713并且对于角质酶B与Af GH61的组合是1.787。
在40分钟反应之后,对于单独的角质酶B 254nm处的吸光度是0.507并且对于角质酶B与Af GH61的组合是0.559。当单独使用角质酶B时40分钟之后的pH变化是0.28(即7.93减7.65);而当一起使用角质酶B和Af GH61时导致0.35的pH变化(即7.93减7.58)。在1小时反应之后获得相似的结果,OD吸光度从0.748增加到0.832,pH变化从0.35增加到0.47(即7.93减7.46)。
总之,当与角质酶B一起使用时,4种不同的GH61显示出对水解BETEB的促进效应。
实例6:在LOM中角质酶A与两种GH61用于PET生物抛光
在洗衣指数计(LOM,SDL-阿特拉斯(Atlas)LP2)中用角质酶和GH61进行PET生物抛光。
将PET织物切成矩形块,5cm宽并且10cm长并且重约1g。将该织物通过缝合锁边。将这些块在编号之前置于控制室(65%相对湿度,20℃)中24小时,通过分析天平称重并记录。在每个烧杯中放置一个调节过的块。对于每个烧杯,使用10个小的钢球(M6M-SR-A4-80,抗酸性)来提供机械辅助。然后根据表5,基于对实际织物重量的计算,以液体与织物10:1(v/w)的比率添加该缓冲液(布里顿-罗宾逊缓冲液,pH=8)和这些酶溶液。测量254nm处的OD吸光度和溶液的初始pH(对于0小时处取样时间指定的数据)。
在选择温度之后启动该LOM机器。将该机器设置为当温度达到70℃时暂停。每个烧杯装配一种内衬有2个neoprin垫片的盖子并且用金属夹紧装置紧密闭合。将这些烧杯加载到预热的LOM中。在垂直位,在这4个鼓位中的每个中,使用金属架来接纳并固定5个烧杯。闭合该LOM盖子并继续洗涤程序,并且启动计时。2小时之后,去除所有烧杯并将这些PET样品转移到钝化溶液(2g/L碳酸钠)在95℃下持续10分钟。然后将这些织物在热水中漂洗2次并在冷水中漂洗2次。将这些PET样品滚动干燥(AEG,LAVATHERM 37700,德国)1小时,并且然后在评价之前,将这些样品在20℃,65%相对湿度调节24小时。
还将来自处理浴的来自每个烧杯的溶液进行收集,并在13000rpm离心1分钟,以进一步收集上清液用于pH测量和254nm处吸光度测定。织物评价包括重量损失和起球记录。
表5:在LOM中角质酶A与两种GH61用于PET处理的结果
从上表看,显然与单独使用角质酶相比,当使用角质酶与TaGH61或AfGH61的组合时,LOM中就起球记录而言的应用性能已得以显著地改进。同时,当与未用角质酶和GH61处理的织物相比时,重量损失仍处于0.8%或0.56%的低水平。所以,在用于PET生物抛光的角质酶A与TaGH61或AfGH61之间存在协同。
在此描述并且要求的本发明不应限于在此披露的具体方面的范围,因为这些方面旨在作为本发明若干方面的说明。预期任何等效方面都处于本发明的范围内。实际上,除在此所示和描述的那些之外,本发明的不同修改对于本领域普通技术人员而言从前述描述将变得清楚。这类修改也旨在落入所附权利要求书的范围内。在有冲突的情况下,以包括定义的本披露为准。
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Gln Gly Pro Ile Leu Val Trp Met Tyr Lys Cys Pro Gly Ser Phe Ser
100 105 110
Ser Cys Asp Gly Ser Gly Ala Gly Trp Phe Lys Ile Asp Glu Ala Gly
115 120 125
Phe His Gly Asp Gly Val Lys Val Phe Leu Asp Thr Glu Asn Pro Ser
130 135 140
Gly Trp Asp Ile Ala Lys Leu Val Gly Gly Asn Lys Gln Trp Ser Ser
145 150 155 160
Lys Val Pro Glu Gly Leu Ala Pro Gly Asn Tyr Leu Val Arg His Glu
165 170 175
Leu Ile Ala Leu His Gln Ala Asn Asn Pro Gln Phe Tyr Pro Glu Cys
180 185 190
Ala Gln Val Val Ile Thr Gly Ser Gly Thr Ala Gln Pro Asp Ala Ser
195 200 205
Tyr Lys Ala Ala Ile Pro Gly Tyr Cys Asn Gln Asn Asp Pro Asn Ile
210 215 220
Lys Val Pro Ile Asn Asp His Ser Ile Pro Gln Thr Tyr Lys Ile Pro
225 230 235 240
Gly Pro Pro Val Phe Lys Gly Thr Ala Ser Lys Lys Ala Arg Asp Phe
245 250 255
Thr Ala
<210> 5
<211> 226
<212> PRT
<213> 土生梭孢壳
<400> 5
Met Leu Ala Asn Gly Ala Ile Val Phe Leu Ala Ala Ala Leu Gly Val
1 5 10 15
Ser Gly His Tyr Thr Trp Pro Arg Val Asn Asp Gly Ala Asp Trp Gln
20 25 30
Gln Val Arg Lys Ala Asp Asn Trp Gln Asp Asn Gly Tyr Val Gly Asp
35 40 45
Val Thr Ser Pro Gln Ile Arg Cys Phe Gln Ala Thr Pro Ser Pro Ala
50 55 60
Pro Ser Val Leu Asn Thr Thr Ala Gly Ser Thr Val Thr Tyr Trp Ala
65 70 75 80
Asn Pro Asp Val Tyr His Pro Gly Pro Val Gln Phe Tyr Met Ala Arg
85 90 95
Val Pro Asp Gly Glu Asp Ile Asn Ser Trp Asn Gly Asp Gly Ala Val
100 105 110
Trp Phe Lys Val Tyr Glu Asp His Pro Thr Phe Gly Ala Gln Leu Thr
115 120 125
Trp Pro Ser Thr Gly Lys Ser Ser Phe Ala Val Pro Ile Pro Pro Cys
130 135 140
Ile Lys Ser Gly Tyr Tyr Leu Leu Arg Ala Glu Gln Ile Gly Leu His
145 150 155 160
Val Ala Gln Ser Val Gly Gly Ala Gln Phe Tyr Ile Ser Cys Ala Gln
165 170 175
Leu Ser Val Thr Gly Gly Gly Ser Thr Glu Pro Pro Asn Lys Val Ala
180 185 190
Phe Pro Gly Ala Tyr Ser Ala Thr Asp Pro Gly Ile Leu Ile Asn Ile
195 200 205
Tyr Tyr Pro Val Pro Thr Ser Tyr Gln Asn Pro Gly Pro Ala Val Phe
210 215 220
Ser Cys
225
<210> 6
<211> 304
<212> PRT
<213> 土生梭孢壳
<400> 6
Met Lys Gly Leu Phe Ser Ala Ala Ala Leu Ser Leu Ala Val Gly Gln
1 5 10 15
Ala Ser Ala His Tyr Ile Phe Gln Gln Leu Ser Ile Asn Gly Asn Gln
20 25 30
Phe Pro Val Tyr Gln Tyr Ile Arg Lys Asn Thr Asn Tyr Asn Ser Pro
35 40 45
Val Thr Asp Leu Thr Ser Asp Asp Leu Arg Cys Asn Val Gly Ala Gln
50 55 60
Gly Ala Gly Thr Asp Thr Val Thr Val Lys Ala Gly Asp Gln Phe Thr
65 70 75 80
Phe Thr Leu Asp Thr Pro Val Tyr His Gln Gly Pro Ile Ser Ile Tyr
85 90 95
Met Ser Lys Ala Pro Gly Ala Ala Ser Asp Tyr Asp Gly Ser Gly Gly
100 105 110
Trp Phe Lys Ile Lys Asp Trp Gly Pro Thr Phe Asn Ala Asp Gly Thr
115 120 125
Ala Thr Trp Asp Met Ala Gly Ser Tyr Thr Tyr Asn Ile Pro Thr Cys
130 135 140
Ile Pro Asp Gly Asp Tyr Leu Leu Arg Ile Gln Ser Leu Ala Ile His
145 150 155 160
Asn Pro Trp Pro Ala Gly Ile Pro Gln Phe Tyr Ile Ser Cys Ala Gln
165 170 175
Ile Thr Val Thr Gly Gly Gly Asn Gly Asn Pro Gly Pro Thr Ala Leu
180 185 190
Ile Pro Gly Ala Phe Lys Asp Thr Asp Pro Gly Tyr Thr Val Asn Ile
195 200 205
Tyr Thr Asn Phe His Asn Tyr Thr Val Pro Gly Pro Glu Val Phe Ser
210 215 220
Cys Asn Gly Gly Gly Ser Asn Pro Pro Pro Pro Val Ser Ser Ser Thr
225 230 235 240
Pro Ala Thr Thr Thr Leu Val Thr Ser Thr Arg Thr Thr Ser Ser Thr
245 250 255
Ser Ser Ala Ser Thr Pro Ala Ser Thr Gly Gly Cys Thr Val Ala Lys
260 265 270
Trp Gly Gln Cys Gly Gly Asn Gly Tyr Thr Gly Cys Thr Thr Cys Ala
275 280 285
Ala Gly Ser Thr Cys Ser Lys Gln Asn Asp Tyr Tyr Ser Gln Cys Leu
290 295 300
<210> 7
<211> 317
<212> PRT
<213> 土生梭孢壳
<400> 7
Met Lys Gly Leu Ser Leu Leu Ala Ala Ala Ser Ala Ala Thr Ala His
1 5 10 15
Thr Ile Phe Val Gln Leu Glu Ser Gly Gly Thr Thr Tyr Pro Val Ser
20 25 30
Tyr Gly Ile Arg Asp Pro Ser Tyr Asp Gly Pro Ile Thr Asp Val Thr
35 40 45
Ser Asp Ser Leu Ala Cys Asn Gly Pro Pro Asn Pro Thr Thr Pro Ser
50 55 60
Pro Tyr Ile Ile Asn Val Thr Ala Gly Thr Thr Val Ala Ala Ile Trp
65 70 75 80
Arg His Thr Leu Thr Ser Gly Pro Asp Asp Val Met Asp Ala Ser His
85 90 95
Lys Gly Pro Thr Leu Ala Tyr Leu Lys Lys Val Asp Asp Ala Leu Thr
100 105 110
Asp Thr Gly Ile Gly Gly Gly Trp Phe Lys Ile Gln Glu Ala Gly Tyr
115 120 125
Asp Asn Gly Asn Trp Ala Thr Ser Thr Val Ile Thr Asn Gly Gly Phe
130 135 140
Gln Tyr Ile Asp Ile Pro Ala Cys Ile Pro Asn Gly Gln Tyr Leu Leu
145 150 155 160
Arg Ala Glu Met Ile Ala Leu His Ala Ala Ser Thr Gln Gly Gly Ala
165 170 175
Gln Leu Tyr Met Glu Cys Ala Gln Ile Asn Val Val Gly Gly Ser Gly
180 185 190
Ser Ala Ser Pro Gln Thr Tyr Ser Ile Pro Gly Ile Tyr Gln Ala Thr
195 200 205
Asp Pro Gly Leu Leu Ile Asn Ile Tyr Ser Met Thr Pro Ser Ser Gln
210 215 220
Tyr Thr Ile Pro Gly Pro Pro Leu Phe Thr Cys Ser Gly Ser Gly Asn
225 230 235 240
Asn Gly Gly Gly Ser Asn Pro Ser Gly Gly Gln Thr Thr Thr Ala Lys
245 250 255
Pro Thr Thr Thr Thr Ala Ala Thr Thr Thr Ser Ser Ala Ala Pro Thr
260 265 270
Ser Ser Gln Gly Gly Ser Ser Gly Cys Thr Val Pro Gln Trp Gln Gln
275 280 285
Cys Gly Gly Ile Ser Phe Thr Gly Cys Thr Thr Cys Ala Ala Gly Tyr
290 295 300
Thr Cys Lys Tyr Leu Asn Asp Tyr Tyr Ser Gln Cys Gln
305 310 315
<210> 8
<211> 249
<212> PRT
<213> 橙色嗜热子囊菌
<400> 8
Met Ser Phe Ser Lys Ile Ile Ala Thr Ala Gly Val Leu Ala Ser Ala
1 5 10 15
Ser Leu Val Ala Gly His Gly Phe Val Gln Asn Ile Val Ile Asp Gly
20 25 30
Lys Tyr Tyr Gly Gly Tyr Leu Val Asn Gln Tyr Pro Tyr Met Ser Asn
35 40 45
Pro Pro Glu Val Ile Ala Trp Ser Thr Thr Ala Thr Asp Leu Gly Phe
50 55 60
Val Asp Gly Thr Gly Tyr Gln Thr Pro Asp Ile Ile Cys His Arg Gly
65 70 75 80
Ala Lys Pro Gly Ala Leu Thr Ala Pro Val Ser Pro Gly Gly Thr Val
85 90 95
Glu Leu Gln Trp Thr Pro Trp Pro Asp Ser His His Gly Pro Val Ile
100 105 110
Asn Tyr Leu Ala Pro Cys Asn Gly Asp Cys Ser Thr Val Asp Lys Thr
115 120 125
Gln Leu Glu Phe Phe Lys Ile Ala Glu Ser Gly Leu Ile Asn Asp Asp
130 135 140
Asn Pro Pro Gly Ile Trp Ala Ser Asp Asn Leu Ile Ala Ala Asn Asn
145 150 155 160
Ser Trp Thr Val Thr Ile Pro Thr Thr Ile Ala Pro Gly Asn Tyr Val
165 170 175
Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gln Asn Gln Asp Gly
180 185 190
Ala Gln Asn Tyr Pro Gln Cys Ile Asn Leu Gln Val Thr Gly Gly Gly
195 200 205
Ser Asp Asn Pro Ala Gly Thr Leu Gly Thr Ala Leu Tyr His Asp Thr
210 215 220
Asp Pro Gly Ile Leu Ile Asn Ile Tyr Gln Lys Leu Ser Ser Tyr Ile
225 230 235 240
Ile Pro Gly Pro Pro Leu Tyr Thr Gly
245
<210> 9
<211> 249
<212> PRT
<213> 里氏木霉
<400> 9
Met Lys Ser Cys Ala Ile Leu Ala Ala Leu Gly Cys Leu Ala Gly Ser
1 5 10 15
Val Leu Gly His Gly Gln Val Gln Asn Phe Thr Ile Asn Gly Gln Tyr
20 25 30
Asn Gln Gly Phe Ile Leu Asp Tyr Tyr Tyr Gln Lys Gln Asn Thr Gly
35 40 45
His Phe Pro Asn Val Ala Gly Trp Tyr Ala Glu Asp Leu Asp Leu Gly
50 55 60
Phe Ile Ser Pro Asp Gln Tyr Thr Thr Pro Asp Ile Val Cys His Lys
65 70 75 80
Asn Ala Ala Pro Gly Ala Ile Ser Ala Thr Ala Ala Ala Gly Ser Asn
85 90 95
Ile Val Phe Gln Trp Gly Pro Gly Val Trp Pro His Pro Tyr Gly Pro
100 105 110
Ile Val Thr Tyr Val Val Glu Cys Ser Gly Ser Cys Thr Thr Val Asn
115 120 125
Lys Asn Asn Leu Arg Trp Val Lys Ile Gln Glu Ala Gly Ile Asn Tyr
130 135 140
Asn Thr Gln Val Trp Ala Gln Gln Asp Leu Ile Asn Gln Gly Asn Lys
145 150 155 160
Trp Thr Val Lys Ile Pro Ser Ser Leu Arg Pro Gly Asn Tyr Val Phe
165 170 175
Arg His Glu Leu Leu Ala Ala His Gly Ala Ser Ser Ala Asn Gly Met
180 185 190
Gln Asn Tyr Pro Gln Cys Val Asn Ile Ala Val Thr Gly Ser Gly Thr
195 200 205
Lys Ala Leu Pro Ala Gly Thr Pro Ala Thr Gln Leu Tyr Lys Pro Thr
210 215 220
Asp Pro Gly Ile Leu Phe Asn Pro Tyr Thr Thr Ile Thr Ser Tyr Thr
225 230 235 240
Ile Pro Gly Pro Ala Leu Trp Gln Gly
245
<210> 10
<211> 232
<212> PRT
<213> 嗜热毁丝霉
<400> 10
Met Lys Phe Thr Ser Ser Leu Ala Val Leu Ala Ala Ala Gly Ala Gln
1 5 10 15
Ala His Tyr Thr Phe Pro Arg Ala Gly Thr Gly Gly Ser Leu Ser Gly
20 25 30
Glu Trp Glu Val Val Arg Met Thr Glu Asn His Tyr Ser His Gly Pro
35 40 45
Val Thr Asp Val Thr Ser Pro Glu Met Thr Cys Tyr Gln Ser Gly Val
50 55 60
Gln Gly Ala Pro Gln Thr Val Gln Val Lys Ala Gly Ser Gln Phe Thr
65 70 75 80
Phe Ser Val Asp Pro Ser Ile Gly His Pro Gly Pro Leu Gln Phe Tyr
85 90 95
Met Ala Lys Val Pro Ser Gly Gln Thr Ala Ala Thr Phe Asp Gly Thr
100 105 110
Gly Ala Val Trp Phe Lys Ile Tyr Gln Asp Gly Pro Asn Gly Leu Gly
115 120 125
Thr Asp Ser Ile Thr Trp Pro Ser Ala Gly Lys Thr Glu Val Ser Val
130 135 140
Thr Ile Pro Ser Cys Ile Asp Asp Gly Glu Tyr Leu Leu Arg Val Glu
145 150 155 160
His Ile Ala Leu His Ser Ala Ser Ser Val Gly Gly Ala Gln Phe Tyr
165 170 175
Ile Ala Cys Ala Gln Leu Ser Val Thr Gly Gly Ser Gly Thr Leu Asn
180 185 190
Thr Gly Ser Leu Val Ser Leu Pro Gly Ala Tyr Lys Ala Thr Asp Pro
195 200 205
Gly Ile Leu Phe Gln Leu Tyr Trp Pro Ile Pro Thr Glu Tyr Ile Asn
210 215 220
Pro Gly Pro Ala Pro Val Ser Cys
225 230
<210> 11
<211> 235
<212> PRT
<213> 嗜热毁丝霉
<400> 11
Met Lys Ala Leu Ser Leu Leu Ala Ala Ala Ser Ala Val Ser Ala His
1 5 10 15
Thr Ile Phe Val Gln Leu Glu Ala Asp Gly Thr Arg Tyr Pro Val Ser
20 25 30
Tyr Gly Ile Arg Asp Pro Ser Tyr Asp Gly Pro Ile Thr Asp Val Thr
35 40 45
Ser Asn Asp Val Ala Cys Asn Gly Gly Pro Asn Pro Thr Thr Pro Ser
50 55 60
Ser Asp Val Ile Thr Val Thr Ala Gly Thr Thr Val Lys Ala Ile Trp
65 70 75 80
Arg His Thr Leu Gln Ser Gly Pro Asp Asp Val Met Asp Ala Ser His
85 90 95
Lys Gly Pro Thr Leu Ala Tyr Leu Lys Lys Val Gly Asp Ala Thr Lys
100 105 110
Asp Ser Gly Val Gly Gly Gly Trp Phe Lys Ile Gln Glu Asp Gly Tyr
115 120 125
Asn Asn Gly Gln Trp Gly Thr Ser Thr Val Ile Ser Asn Gly Gly Glu
130 135 140
His Tyr Ile Asp Ile Pro Ala Cys Ile Pro Glu Gly Gln Tyr Leu Leu
145 150 155 160
Arg Ala Glu Met Ile Ala Leu His Ala Ala Gly Ser Pro Gly Gly Ala
165 170 175
Gln Leu Tyr Met Glu Cys Ala Gln Ile Asn Ile Val Gly Gly Ser Gly
180 185 190
Ser Val Pro Ser Ser Thr Val Ser Phe Pro Gly Ala Tyr Ser Pro Asn
195 200 205
Asp Pro Gly Leu Leu Ile Asn Ile Tyr Ser Met Ser Pro Ser Ser Ser
210 215 220
Tyr Thr Ile Pro Gly Pro Pro Val Phe Lys Cys
225 230 235
<210> 12
<211> 323
<212> PRT
<213> 嗜热毁丝霉
<400> 12
Met Lys Ser Phe Ala Leu Thr Thr Leu Ala Ala Leu Ala Gly Asn Ala
1 5 10 15
Ala Ala His Ala Thr Phe Gln Ala Leu Trp Val Asp Gly Val Asp Tyr
20 25 30
Gly Ala Gln Cys Ala Arg Leu Pro Ala Ser Asn Ser Pro Val Thr Asp
35 40 45
Val Thr Ser Asn Ala Ile Arg Cys Asn Ala Asn Pro Ser Pro Ala Arg
50 55 60
Gly Lys Cys Pro Val Lys Ala Gly Ser Thr Val Thr Val Glu Met His
65 70 75 80
Gln Gln Pro Gly Asp Arg Ser Cys Ser Ser Glu Ala Ile Gly Gly Ala
85 90 95
His Tyr Gly Pro Val Met Val Tyr Met Ser Lys Val Ser Asp Ala Ala
100 105 110
Ser Ala Asp Gly Ser Ser Gly Trp Phe Lys Val Phe Glu Asp Gly Trp
115 120 125
Ala Lys Asn Pro Ser Gly Gly Ser Gly Asp Asp Asp Tyr Trp Gly Thr
130 135 140
Lys Asp Leu Asn Ser Cys Cys Gly Lys Met Asn Val Lys Ile Pro Ala
145 150 155 160
Asp Leu Pro Ser Gly Asp Tyr Leu Leu Arg Ala Glu Ala Leu Ala Leu
165 170 175
His Thr Ala Gly Ser Ala Gly Gly Ala Gln Phe Tyr Met Thr Cys Tyr
180 185 190
Gln Leu Thr Val Thr Gly Ser Gly Ser Ala Ser Pro Pro Thr Val Ser
195 200 205
Phe Pro Gly Ala Tyr Lys Ala Thr Asp Pro Gly Ile Leu Val Asn Ile
210 215 220
His Ala Pro Leu Ser Gly Tyr Thr Val Pro Gly Pro Ala Val Tyr Ser
225 230 235 240
Gly Gly Ser Thr Lys Lys Ala Gly Ser Ala Cys Thr Gly Cys Glu Ser
245 250 255
Thr Cys Ala Val Gly Ser Gly Pro Thr Ala Thr Val Ser Gln Ser Pro
260 265 270
Gly Ser Thr Ala Thr Ser Ala Pro Gly Gly Gly Gly Gly Cys Thr Val
275 280 285
Gln Lys Tyr Gln Gln Cys Gly Gly Glu Gly Tyr Thr Gly Cys Thr Asn
290 295 300
Cys Ala Ser Gly Ser Thr Cys Ser Ala Val Ser Pro Pro Tyr Tyr Ser
305 310 315 320
Gln Cys Val
<210> 13
<211> 310
<212> PRT
<213> 嗜热毁丝霉
<400> 13
Met Lys Pro Phe Ser Leu Val Ala Leu Ala Thr Ala Val Ser Gly His
1 5 10 15
Ala Ile Phe Gln Arg Val Ser Val Asn Gly Gln Asp Gln Gly Gln Leu
20 25 30
Lys Gly Val Arg Ala Pro Ser Ser Asn Ser Pro Ile Gln Asn Val Asn
35 40 45
Asp Ala Asn Met Ala Cys Asn Ala Asn Ile Val Tyr His Asp Ser Thr
50 55 60
Ile Ile Lys Val Pro Ala Gly Ala Arg Val Gly Ala Trp Trp Gln His
65 70 75 80
Val Ile Gly Gly Pro Gln Gly Ala Asn Asp Pro Asp Asn Pro Ile Ala
85 90 95
Ala Ser His Lys Gly Pro Ile Gln Val Tyr Leu Ala Lys Val Asp Asn
100 105 110
Ala Ala Thr Ala Ser Pro Ser Gly Leu Arg Trp Phe Lys Val Ala Glu
115 120 125
Arg Gly Leu Asn Asn Gly Val Trp Ala Val Asp Glu Leu Ile Ala Asn
130 135 140
Asn Gly Trp His Tyr Phe Asp Leu Pro Ser Cys Val Ala Pro Gly Gln
145 150 155 160
Tyr Leu Met Arg Val Glu Leu Leu Ala Leu His Ser Ala Ser Ser Pro
165 170 175
Gly Gly Ala Gln Phe Tyr Met Gly Cys Ala Gln Ile Glu Val Thr Gly
180 185 190
Ser Gly Thr Asn Ser Gly Ser Asp Phe Val Ser Phe Pro Gly Ala Tyr
195 200 205
Ser Ala Asn Asp Pro Gly Ile Leu Leu Ser Ile Tyr Asp Ser Ser Gly
210 215 220
Lys Pro Thr Asn Gly Gly Arg Ser Tyr Pro Ile Pro Gly Pro Arg Pro
225 230 235 240
Ile Ser Cys Ser Gly Ser Gly Asp Gly Gly Asn Asn Gly Gly Gly Gly
245 250 255
Asp Asp Asn Asn Asn Asn Asn Gly Gly Gly Asn Asn Gly Gly Gly Gly
260 265 270
Gly Gly Ser Val Pro Leu Tyr Gly Gln Cys Gly Gly Ile Gly Tyr Thr
275 280 285
Gly Pro Thr Thr Cys Ala Gln Gly Thr Cys Lys Val Ser Asn Glu Tyr
290 295 300
Tyr Ser Gln Cys Leu Pro
305 310
<210> 14
<211> 246
<212> PRT
<213> 嗜热毁丝霉
<400> 14
Met Lys Leu Ser Leu Phe Ser Val Leu Ala Thr Ala Leu Thr Val Glu
1 5 10 15
Gly His Ala Ile Phe Gln Lys Val Ser Val Asn Gly Ala Asp Gln Gly
20 25 30
Ser Leu Thr Gly Leu Arg Ala Pro Asn Asn Asn Asn Pro Val Gln Asp
35 40 45
Val Asn Ser Gln Asp Met Ile Cys Gly Gln Ser Gly Ser Thr Ser Asn
50 55 60
Thr Ile Ile Glu Val Lys Ala Gly Asp Arg Ile Gly Ala Trp Tyr Gln
65 70 75 80
His Val Ile Gly Gly Ala Gln Phe Pro Asn Asp Pro Asp Asn Pro Ile
85 90 95
Ala Lys Ser His Lys Gly Pro Val Met Ala Tyr Leu Ala Lys Val Asp
100 105 110
Asn Ala Ala Thr Ala Ser Lys Thr Gly Leu Lys Trp Phe Lys Ile Trp
115 120 125
Glu Asp Thr Phe Asn Pro Ser Thr Lys Thr Trp Gly Val Asp Asn Leu
130 135 140
Ile Asn Asn Asn Gly Trp Val Tyr Phe Asn Leu Pro Gln Cys Ile Ala
145 150 155 160
Asp Gly Asn Tyr Leu Leu Arg Val Glu Val Leu Ala Leu His Ser Ala
165 170 175
Tyr Ser Gln Gly Gln Ala Gln Phe Tyr Gln Ser Cys Ala Gln Ile Asn
180 185 190
Val Ser Gly Gly Gly Ser Phe Thr Pro Pro Ser Thr Val Ser Phe Pro
195 200 205
Gly Ala Tyr Ser Ala Ser Asp Pro Gly Ile Leu Ile Asn Ile Tyr Gly
210 215 220
Ala Thr Gly Gln Pro Asp Asn Asn Gly Gln Pro Tyr Thr Ala Pro Gly
225 230 235 240
Pro Ala Pro Ile Ser Cys
245
<210> 15
<211> 354
<212> PRT
<213> 橙色嗜热子囊菌
<400> 15
Met Ser Phe Ser Lys Ile Ala Ala Ile Thr Gly Ala Ile Thr Tyr Ala
1 5 10 15
Ser Leu Ala Ala Ala His Gly Tyr Val Thr Gly Ile Val Ala Asp Gly
20 25 30
Thr Tyr Tyr Gly Gly Tyr Ile Val Thr Gln Tyr Pro Tyr Met Ser Thr
35 40 45
Pro Pro Asp Val Ile Ala Trp Ser Thr Lys Ala Thr Asp Leu Gly Phe
50 55 60
Val Asp Pro Ser Ser Tyr Ala Ser Ser Asp Ile Ile Cys His Lys Gly
65 70 75 80
Ala Glu Pro Gly Ala Leu Ser Ala Lys Val Ala Ala Gly Gly Thr Val
85 90 95
Glu Leu Gln Trp Thr Asp Trp Pro Glu Ser His Lys Gly Pro Val Ile
100 105 110
Asp Tyr Leu Ala Ala Cys Asn Gly Asp Cys Ser Thr Val Asp Lys Thr
115 120 125
Lys Leu Glu Phe Phe Lys Ile Asp Glu Ser Gly Leu Ile Asp Gly Ser
130 135 140
Ser Ala Pro Gly Thr Trp Ala Ser Asp Asn Leu Ile Ala Asn Asn Asn
145 150 155 160
Ser Trp Thr Val Thr Ile Pro Ser Thr Ile Ala Pro Gly Asn Tyr Val
165 170 175
Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Thr Asn Gly
180 185 190
Ala Gln Asn Tyr Pro Gln Cys Ile Asn Leu Glu Val Thr Gly Ser Gly
195 200 205
Thr Asp Thr Pro Ala Gly Thr Leu Gly Thr Glu Leu Tyr Lys Ala Thr
210 215 220
Asp Pro Gly Ile Leu Val Asn Ile Tyr Gln Thr Leu Thr Ser Tyr Asp
225 230 235 240
Ile Pro Gly Pro Ala Leu Tyr Thr Gly Gly Ser Ser Gly Ser Ser Gly
245 250 255
Ser Ser Asn Thr Ala Lys Ala Thr Thr Ser Thr Ala Ser Ser Ser Ile
260 265 270
Val Thr Pro Thr Pro Val Asn Asn Pro Thr Val Thr Gln Thr Ala Val
275 280 285
Val Asp Val Thr Gln Thr Val Ser Gln Asn Ala Ala Val Ala Thr Thr
290 295 300
Thr Pro Ala Ser Thr Ala Val Ala Thr Ala Val Pro Thr Gly Thr Thr
305 310 315 320
Phe Ser Phe Asp Ser Met Thr Ser Asp Glu Phe Val Ser Leu Met Arg
325 330 335
Ala Thr Val Asn Trp Leu Leu Ser Asn Lys Lys His Ala Arg Asp Leu
340 345 350
Ser Tyr
<210> 16
<211> 250
<212> PRT
<213> 烟曲霉
<400> 16
Met Thr Leu Ser Lys Ile Thr Ser Ile Ala Gly Leu Leu Ala Ser Ala
1 5 10 15
Ser Leu Val Ala Gly His Gly Phe Val Ser Gly Ile Val Ala Asp Gly
20 25 30
Lys Tyr Tyr Gly Gly Tyr Leu Val Asn Gln Tyr Pro Tyr Met Ser Asn
35 40 45
Pro Pro Asp Thr Ile Ala Trp Ser Thr Thr Ala Thr Asp Leu Gly Phe
50 55 60
Val Asp Gly Thr Gly Tyr Gln Ser Pro Asp Ile Ile Cys His Arg Asp
65 70 75 80
Ala Lys Asn Gly Lys Leu Thr Ala Thr Val Ala Ala Gly Ser Gln Ile
85 90 95
Glu Phe Gln Trp Thr Thr Trp Pro Glu Ser His His Gly Pro Leu Ile
100 105 110
Thr Tyr Leu Ala Pro Cys Asn Gly Asp Cys Ala Thr Val Asp Lys Thr
115 120 125
Thr Leu Lys Phe Val Lys Ile Ala Ala Gln Gly Leu Ile Asp Gly Ser
130 135 140
Asn Pro Pro Gly Val Trp Ala Asp Asp Glu Met Ile Ala Asn Asn Asn
145 150 155 160
Thr Ala Thr Val Thr Ile Pro Ala Ser Tyr Ala Pro Gly Asn Tyr Val
165 170 175
Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Leu Asn Gly
180 185 190
Ala Gln Asn Tyr Pro Gln Cys Phe Asn Ile Gln Ile Thr Gly Gly Gly
195 200 205
Ser Ala Gln Gly Ser Gly Thr Ala Gly Thr Ser Leu Tyr Lys Asn Thr
210 215 220
Asp Pro Gly Ile Lys Phe Asp Ile Tyr Ser Asp Leu Ser Gly Gly Tyr
225 230 235 240
Pro Ile Pro Gly Pro Ala Leu Phe Asn Ala
245 250
<210> 17
<211> 322
<212> PRT
<213> 嗜松青霉
<400> 17
Met Pro Ser Thr Lys Val Ala Ala Leu Ser Ala Val Leu Ala Leu Ala
1 5 10 15
Ser Thr Val Ala Gly His Gly Phe Val Gln Asn Ile Val Ile Asp Gly
20 25 30
Lys Ser Tyr Ser Gly Tyr Leu Val Asn Gln Phe Pro Tyr Glu Ser Asn
35 40 45
Pro Pro Ala Val Ile Gly Trp Ala Thr Thr Ala Thr Asp Leu Gly Phe
50 55 60
Val Ala Pro Ser Glu Tyr Thr Asn Ala Asp Ile Ile Cys His Lys Asn
65 70 75 80
Ala Thr Pro Gly Ala Leu Ser Ala Pro Val Ala Ala Gly Gly Thr Val
85 90 95
Glu Leu Gln Trp Thr Thr Trp Pro Asp Ser His His Gly Pro Val Ile
100 105 110
Ser Tyr Leu Ala Asn Cys Asn Gly Asn Cys Ser Thr Val Asp Lys Thr
115 120 125
Lys Leu Asp Phe Val Lys Ile Asp Gln Gly Gly Leu Ile Asp Asp Thr
130 135 140
Thr Pro Pro Gly Thr Trp Ala Ser Asp Lys Leu Ile Ala Ala Asn Asn
145 150 155 160
Ser Trp Thr Val Thr Ile Pro Ser Thr Ile Ala Pro Gly Asn Tyr Val
165 170 175
Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Ala Asp Gly
180 185 190
Ala Gln Asn Tyr Pro Gln Cys Ile Asn Leu Glu Ile Thr Gly Ser Gly
195 200 205
Thr Ala Ala Pro Ser Gly Thr Ala Gly Glu Lys Leu Tyr Thr Ser Thr
210 215 220
Asp Pro Gly Ile Leu Val Asn Ile Tyr Gln Ser Leu Ser Thr Tyr Val
225 230 235 240
Ile Pro Gly Pro Thr Leu Trp Ser Gly Ala Ala Asn Gly Ala Val Ala
245 250 255
Thr Gly Ser Ala Thr Ala Val Ala Thr Thr Ala Thr Ala Ser Ala Thr
260 265 270
Ala Thr Pro Thr Thr Leu Val Thr Ser Val Ala Pro Ala Ser Ser Thr
275 280 285
Phe Ala Thr Ala Val Val Thr Thr Val Ala Pro Ala Val Thr Asp Val
290 295 300
Val Thr Val Thr Asp Val Val Thr Val Thr Thr Val Ile Thr Thr Thr
305 310 315 320
Val Leu
<210> 18
<211> 444
<212> PRT
<213> 嗜热子嚢菌属
<400> 18
Met Leu Ser Phe Ala Ser Ala Lys Ser Ala Val Leu Thr Thr Leu Leu
1 5 10 15
Leu Leu Gly Ser Ala Gln Ala His Thr Leu Met Thr Thr Leu Phe Val
20 25 30
Asp Gly Val Asn Gln Gly Asp Gly Val Cys Ile Arg Met Asn Asn Asn
35 40 45
Gly Ser Thr Ala Asn Thr Tyr Ile Gln Pro Val Thr Ser Lys Asp Ile
50 55 60
Ala Cys Gly Ile Gln Gly Glu Ile Gly Ala Ala Arg Val Cys Pro Ala
65 70 75 80
Lys Ala Ser Ser Thr Leu Thr Phe Gln Phe Arg Glu Gln Pro Ser Asn
85 90 95
Pro Asn Ser Ala Pro Leu Asp Pro Ser His Lys Gly Pro Ala Ala Val
100 105 110
Tyr Leu Lys Lys Val Asp Ser Ala Ile Ala Ser Asn Asn Ala Ala Gly
115 120 125
Asp Gly Trp Phe Lys Ile Trp Glu Ser Val Tyr Asp Glu Ser Thr Gly
130 135 140
Lys Trp Gly Thr Thr Lys Met Ile Glu Asn Asn Gly His Ile Ser Val
145 150 155 160
Lys Val Pro Asp Asp Ile Glu Gly Gly Tyr Tyr Leu Ala Arg Thr Glu
165 170 175
Leu Leu Ala Leu His Ala Ala Asn Glu Gly Asp Pro Gln Phe Tyr Val
180 185 190
Gly Cys Ala Gln Leu Phe Ile Asp Ser Ala Gly Thr Ala Lys Pro Pro
195 200 205
Thr Val Ser Ile Gly Glu Gly Thr Tyr Asp Leu Ser Met Pro Ala Met
210 215 220
Thr Tyr Asn Ile Tyr Gln Thr Pro Leu Ala Leu Pro Tyr Pro Met Tyr
225 230 235 240
Gly Pro Pro Val Tyr Thr Pro Gly Ser Gly Ser Gly Ser Gly Ser Gly
245 250 255
Ser Gly Ser Ala Ser Ala Thr Arg Ser Ser Ala Ile Pro Thr Ala Thr
260 265 270
Ala Val Thr Asp Cys Ser Ser Glu Glu Asp Arg Glu Asp Ser Val Met
275 280 285
Ala Thr Gly Val Pro Val Ala Arg Ser Thr Leu Arg Thr Trp Val Asp
290 295 300
Arg Leu Ser Trp His Gly Lys Ala Arg Glu Asn Val Lys Pro Ala Ala
305 310 315 320
Arg Arg Ser Ala Leu Val Gln Thr Glu Gly Leu Lys Pro Glu Gly Cys
325 330 335
Ile Phe Val Asn Gly Asn Trp Cys Gly Phe Glu Val Pro Asp Tyr Asn
340 345 350
Asp Ala Glu Ser Cys Trp Ala Ala Ser Asp Asn Cys Trp Lys Gln Ser
355 360 365
Asp Ser Cys Trp Asn Gln Thr Gln Pro Thr Gly Tyr Asn Asn Cys Gln
370 375 380
Ile Trp Gln Asp Gln Lys Cys Lys Pro Ile Gln Asp Ser Cys Ser Gln
385 390 395 400
Ser Asn Pro Thr Gly Pro Pro Asn Lys Gly Lys Asp Ile Thr Pro Thr
405 410 415
Trp Pro Pro Leu Glu Gly Ser Met Lys Thr Phe Thr Lys Arg Thr Val
420 425 430
Ser Tyr Arg Asp Trp Ile Met Lys Arg Lys Gly Ala
435 440
<210> 19
<211> 253
<212> PRT
<213> 青霉菌属
<400> 19
Met Leu Ser Ser Thr Thr Arg Thr Leu Ala Phe Thr Gly Leu Ala Gly
1 5 10 15
Leu Leu Ser Ala Pro Leu Val Lys Ala His Gly Phe Val Gln Gly Ile
20 25 30
Val Ile Gly Asp Gln Phe Tyr Ser Gly Tyr Ile Val Asn Ser Phe Pro
35 40 45
Tyr Glu Ser Asn Pro Pro Pro Val Ile Gly Trp Ala Thr Thr Ala Thr
50 55 60
Asp Leu Gly Phe Val Asp Gly Thr Gly Tyr Gln Gly Pro Asp Ile Ile
65 70 75 80
Cys His Arg Asn Ala Thr Pro Ala Pro Leu Thr Ala Pro Val Ala Ala
85 90 95
Gly Gly Thr Val Glu Leu Gln Trp Thr Pro Trp Pro Asp Ser His His
100 105 110
Gly Pro Val Ile Thr Tyr Leu Ala Pro Cys Asn Gly Asn Cys Ser Thr
115 120 125
Val Asp Lys Thr Thr Leu Glu Phe Phe Lys Ile Asp Gln Gln Gly Leu
130 135 140
Ile Asp Asp Thr Ser Pro Pro Gly Thr Trp Ala Ser Asp Asn Leu Ile
145 150 155 160
Ala Asn Asn Asn Ser Trp Thr Val Thr Ile Pro Asn Ser Val Ala Pro
165 170 175
Gly Asn Tyr Val Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Asn
180 185 190
Asn Lys Asp Gly Ala Gln Asn Tyr Pro Gln Cys Ile Asn Ile Glu Val
195 200 205
Thr Gly Gly Gly Ser Asp Ala Pro Glu Gly Thr Leu Gly Glu Asp Leu
210 215 220
Tyr His Asp Thr Asp Pro Gly Ile Leu Val Asp Ile Tyr Glu Pro Ile
225 230 235 240
Ala Thr Tyr Thr Ile Pro Gly Pro Pro Glu Pro Thr Phe
245 250
<210> 20
<211> 246
<212> PRT
<213> 土生梭孢壳
<400> 20
Met Lys Phe Ser Leu Val Ser Leu Leu Ala Tyr Gly Leu Ser Val Glu
1 5 10 15
Ala His Ser Ile Phe Gln Arg Val Ser Val Asn Gly Gln Asp Gln Gly
20 25 30
Leu Leu Thr Gly Leu Arg Ala Pro Ser Asn Asn Asn Pro Val Gln Asp
35 40 45
Val Asn Ser Gln Asn Met Ile Cys Gly Gln Ser Gly Ser Lys Ser Gln
50 55 60
Thr Val Ile Asn Val Lys Ala Gly Asp Arg Ile Gly Ser Leu Trp Gln
65 70 75 80
His Val Ile Gly Gly Ala Gln Phe Ser Gly Asp Pro Asp Asn Pro Ile
85 90 95
Ala His Ser His Lys Gly Pro Val Met Ala Tyr Leu Ala Lys Val Asp
100 105 110
Asn Ala Ala Ser Ala Ser Gln Thr Gly Leu Lys Trp Phe Lys Ile Trp
115 120 125
Gln Asp Gly Phe Asp Thr Ser Ser Lys Thr Trp Gly Val Asp Asn Leu
130 135 140
Ile Lys Asn Asn Gly Trp Val Tyr Phe His Leu Pro Gln Cys Leu Ala
145 150 155 160
Pro Gly Gln Tyr Leu Leu Arg Val Glu Val Leu Ala Leu His Ser Ala
165 170 175
Tyr Gln Gln Gly Gln Ala Gln Phe Tyr Gln Ser Cys Ala Gln Ile Asn
180 185 190
Val Ser Gly Ser Gly Ser Phe Ser Pro Ser Gln Thr Val Ser Ile Pro
195 200 205
Gly Val Tyr Ser Ala Thr Asp Pro Ser Ile Leu Ile Asn Ile Tyr Gly
210 215 220
Ser Thr Gly Gln Pro Asp Asn Gly Gly Lys Ala Tyr Asn Pro Pro Gly
225 230 235 240
Pro Ala Pro Ile Ser Cys
245
<210> 21
<211> 334
<212> PRT
<213> 土生梭孢壳
<400> 21
Met Arg Thr Thr Phe Ala Ala Ala Leu Ala Ala Phe Ala Ala Gln Glu
1 5 10 15
Val Ala Gly His Ala Ile Phe Gln Gln Leu Trp His Gly Ser Ser Cys
20 25 30
Val Arg Met Pro Leu Ser Asn Ser Pro Val Thr Asn Val Gly Ser Arg
35 40 45
Asp Met Ile Cys Asn Ala Gly Thr Arg Pro Val Ser Gly Lys Cys Pro
50 55 60
Val Lys Ala Gly Gly Thr Val Thr Val Glu Met His Gln Gln Pro Gly
65 70 75 80
Asp Arg Ser Cys Asn Asn Glu Ala Ile Gly Gly Ala His Trp Gly Pro
85 90 95
Val Gln Val Tyr Leu Ser Lys Val Glu Asp Ala Ser Thr Ala Asp Gly
100 105 110
Ser Thr Gly Trp Phe Lys Ile Phe Ala Asp Thr Trp Ser Lys Lys Ala
115 120 125
Gly Ser Ser Val Gly Asp Asp Asp Asn Trp Gly Thr Arg Asp Leu Asn
130 135 140
Ala Cys Cys Gly Lys Met Gln Val Lys Ile Pro Ala Asp Ile Pro Ser
145 150 155 160
Gly Asp Tyr Leu Leu Arg Ala Glu Ala Leu Ala Leu His Thr Ala Gly
165 170 175
Gln Val Gly Gly Ala Gln Phe Tyr Met Ser Cys Tyr Gln Ile Thr Val
180 185 190
Ser Gly Gly Gly Ser Ala Ser Pro Ala Thr Val Lys Phe Pro Gly Ala
195 200 205
Tyr Ser Ala Asn Asp Pro Gly Ile His Ile Asn Ile His Ala Ala Val
210 215 220
Ser Asn Tyr Val Ala Pro Gly Pro Ala Val Tyr Ser Gly Gly Thr Thr
225 230 235 240
Lys Val Ala Gly Ser Gly Cys Gln Gly Cys Glu Asn Thr Cys Lys Val
245 250 255
Gly Ser Ser Pro Thr Ala Thr Ala Pro Ser Gly Lys Ser Gly Ala Gly
260 265 270
Ser Asp Gly Gly Ala Gly Thr Asp Gly Gly Ser Ser Ser Ser Ser Pro
275 280 285
Asp Thr Gly Ser Ala Cys Ser Val Gln Ala Tyr Gly Gln Cys Gly Gly
290 295 300
Asn Gly Tyr Ser Gly Cys Thr Gln Cys Ala Pro Gly Tyr Thr Cys Lys
305 310 315 320
Ala Val Ser Pro Pro Tyr Tyr Ser Gln Cys Ala Pro Ser Ser
325 330
<210> 22
<211> 227
<212> PRT
<213> 土生梭孢壳
<400> 22
Met Lys Leu Ser Val Ala Ile Ala Val Leu Ala Ser Ala Leu Ala Glu
1 5 10 15
Ala His Tyr Thr Phe Pro Ser Ile Gly Asn Thr Ala Asp Trp Gln Tyr
20 25 30
Val Arg Ile Thr Thr Asn Tyr Gln Ser Asn Gly Pro Val Thr Asp Val
35 40 45
Thr Ser Asp Gln Ile Arg Cys Tyr Glu Arg Asn Pro Gly Thr Gly Ala
50 55 60
Gln Gly Ile Tyr Asn Val Thr Ala Gly Gln Thr Ile Asn Tyr Asn Ala
65 70 75 80
Lys Ala Ser Ile Ser His Pro Gly Pro Met Ser Phe Tyr Ile Ala Lys
85 90 95
Val Pro Ala Gly Gln Thr Ala Ala Thr Trp Asp Gly Lys Gly Ala Val
100 105 110
Trp Thr Lys Ile Tyr Gln Asp Met Pro Lys Phe Gly Ser Ser Leu Thr
115 120 125
Trp Pro Thr Met Gly Ala Lys Ser Val Pro Val Thr Ile Pro Arg Cys
130 135 140
Leu Gln Asn Gly Asp Tyr Leu Leu Arg Ala Glu His Ile Ala Leu His
145 150 155 160
Ser Ala Ser Ser Val Gly Gly Ala Gln Phe Tyr Leu Ser Cys Ala Gln
165 170 175
Leu Thr Val Ser Gly Gly Ser Gly Thr Trp Asn Pro Lys Asn Arg Val
180 185 190
Ser Phe Pro Gly Ala Tyr Lys Ala Thr Asp Pro Gly Ile Leu Ile Asn
195 200 205
Ile Tyr Tyr Pro Val Pro Thr Ser Tyr Ser Pro Pro Gly Pro Pro Ala
210 215 220
Glu Thr Cys
225
<210> 23
<211> 223
<212> PRT
<213> 土生梭孢壳
<400> 23
Met Lys Leu Ser Ser Gln Leu Ala Ala Leu Thr Leu Ala Ala Ala Ser
1 5 10 15
Val Ser Gly His Tyr Ile Phe Glu Gln Ile Ala His Gly Gly Thr Lys
20 25 30
Phe Pro Pro Tyr Glu Tyr Ile Arg Arg Asn Thr Asn Tyr Asn Ser Pro
35 40 45
Val Thr Ser Leu Ser Ser Asn Asp Leu Arg Cys Asn Val Gly Gly Glu
50 55 60
Thr Ala Gly Asn Thr Thr Val Leu Asp Val Lys Ala Gly Asp Ser Phe
65 70 75 80
Thr Phe Tyr Ser Asp Val Ala Val Tyr His Gln Gly Pro Ile Ser Leu
85 90 95
Tyr Met Ser Lys Ala Pro Gly Ser Val Val Asp Tyr Asp Gly Ser Gly
100 105 110
Asp Trp Phe Lys Ile His Asp Trp Gly Pro Thr Phe Ser Asn Gly Gln
115 120 125
Ala Ser Trp Pro Leu Arg Asp Asn Tyr Gln Tyr Asn Ile Pro Thr Cys
130 135 140
Ile Pro Asn Gly Glu Tyr Leu Leu Arg Ile Gln Ser Leu Ala Ile His
145 150 155 160
Asn Pro Gly Ala Thr Pro Gln Phe Tyr Ile Ser Cys Ala Gln Val Arg
165 170 175
Val Ser Gly Gly Gly Ser Ala Ser Pro Ser Pro Thr Ala Lys Ile Pro
180 185 190
Gly Ala Phe Lys Ala Thr Asp Pro Gly Tyr Thr Ala Asn Ile Tyr Asn
195 200 205
Asn Phe His Ser Tyr Thr Val Pro Gly Pro Ala Val Phe Gln Cys
210 215 220
<210> 24
<211> 368
<212> PRT
<213> 土生梭孢壳
<400> 24
Met Pro Ser Phe Ala Ser Lys Thr Leu Leu Ser Thr Leu Ala Gly Ala
1 5 10 15
Ala Ser Val Ala Ala His Gly His Val Ser Asn Ile Val Ile Asn Gly
20 25 30
Val Ser Tyr Gln Gly Tyr Asp Pro Thr Ser Phe Pro Tyr Met Gln Asn
35 40 45
Pro Pro Ile Val Val Gly Trp Thr Ala Ala Asp Thr Asp Asn Gly Phe
50 55 60
Val Ala Pro Asp Ala Phe Ala Ser Gly Asp Ile Ile Cys His Lys Asn
65 70 75 80
Ala Thr Asn Ala Lys Gly His Ala Val Val Ala Ala Gly Asp Lys Ile
85 90 95
Phe Ile Gln Trp Asn Thr Trp Pro Glu Ser His His Gly Pro Val Ile
100 105 110
Asp Tyr Leu Ala Ser Cys Gly Ser Ala Ser Cys Glu Thr Val Asp Lys
115 120 125
Thr Lys Leu Glu Phe Phe Lys Ile Asp Glu Val Gly Leu Val Asp Gly
130 135 140
Ser Ser Ala Pro Gly Val Trp Gly Ser Asp Gln Leu Ile Ala Asn Asn
145 150 155 160
Asn Ser Trp Leu Val Glu Ile Pro Pro Thr Ile Ala Pro Gly Asn Tyr
165 170 175
Val Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Glu Asn Ala Asp
180 185 190
Gly Ala Gln Asn Tyr Pro Gln Cys Phe Asn Leu Gln Ile Thr Gly Thr
195 200 205
Gly Thr Ala Thr Pro Ser Gly Val Pro Gly Thr Ser Leu Tyr Thr Pro
210 215 220
Thr Asp Pro Gly Ile Leu Val Asn Ile Tyr Ser Ala Pro Ile Thr Tyr
225 230 235 240
Thr Val Pro Gly Pro Ala Leu Ile Ser Gly Ala Val Ser Ile Ala Gln
245 250 255
Ser Ser Ser Ala Ile Thr Ala Ser Gly Thr Ala Leu Thr Gly Ser Ala
260 265 270
Thr Ala Pro Ala Ala Ala Ala Ala Thr Thr Thr Ser Thr Thr Asn Ala
275 280 285
Ala Ala Ala Ala Thr Ser Ala Ala Ala Ala Ala Gly Thr Ser Thr Thr
290 295 300
Thr Thr Ser Ala Ala Ala Val Val Gln Thr Ser Ser Ser Ser Ser Ser
305 310 315 320
Ala Pro Ser Ser Ala Ala Ala Ala Ala Thr Thr Thr Ala Ala Ala Ser
325 330 335
Ala Arg Pro Thr Gly Cys Ser Ser Gly Arg Ser Arg Lys Gln Pro Arg
340 345 350
Arg His Ala Arg Asp Met Val Val Ala Arg Gly Ala Glu Glu Ala Asn
355 360 365
<210> 25
<211> 330
<212> PRT
<213> 土生梭孢壳
<400> 25
Met Pro Pro Ala Leu Pro Gln Leu Leu Thr Thr Val Leu Thr Ala Leu
1 5 10 15
Thr Leu Gly Ser Thr Ala Leu Ala His Ser His Leu Ala Tyr Ile Ile
20 25 30
Val Asn Gly Lys Leu Tyr Gln Gly Phe Asp Pro Arg Pro His Gln Ala
35 40 45
Asn Tyr Pro Ser Arg Val Gly Trp Ser Thr Gly Ala Val Asp Asp Gly
50 55 60
Phe Val Thr Pro Ala Asn Tyr Ser Thr Pro Asp Ile Ile Cys His Ile
65 70 75 80
Ala Gly Thr Ser Pro Ala Gly His Ala Pro Val Arg Pro Gly Asp Arg
85 90 95
Ile His Val Gln Trp Asn Gly Trp Pro Val Gly His Ile Gly Pro Val
100 105 110
Leu Ser Tyr Leu Ala Arg Cys Glu Ser Asp Thr Gly Cys Thr Gly Gln
115 120 125
Asn Lys Thr Ala Leu Arg Trp Thr Lys Ile Asp Asp Ser Ser Pro Thr
130 135 140
Met Gln Asn Val Ala Gly Ala Gly Thr Gln Gly Glu Gly Thr Pro Gly
145 150 155 160
Lys Arg Trp Ala Thr Asp Val Leu Ile Ala Ala Asn Asn Ser Trp Gln
165 170 175
Val Ala Val Pro Ala Gly Leu Pro Thr Gly Ala Tyr Val Leu Arg Asn
180 185 190
Glu Ile Ile Ala Leu His Tyr Ala Ala Arg Lys Asn Gly Ala Gln Asn
195 200 205
Tyr Pro Leu Cys Met Asn Leu Trp Val Asp Ala Ser Gly Asp Asn Ser
210 215 220
Ser Val Ala Ala Thr Thr Ala Ala Val Thr Ala Gly Gly Leu Gln Met
225 230 235 240
Asp Ala Tyr Asp Ala Arg Gly Phe Tyr Lys Glu Asn Asp Pro Gly Val
245 250 255
Leu Val Asn Val Thr Ala Ala Leu Ser Ser Tyr Val Val Pro Gly Pro
260 265 270
Thr Val Ala Ala Gly Ala Thr Pro Val Pro Tyr Ala Gln Gln Ser Pro
275 280 285
Ser Val Ser Thr Ala Ala Gly Thr Pro Val Val Val Thr Arg Thr Ser
290 295 300
Glu Thr Ala Pro Tyr Thr Gly Ala Met Thr Pro Thr Val Ala Ala Arg
305 310 315 320
Met Lys Gly Arg Gly Tyr Asp Arg Arg Gly
325 330
<210> 26
<211> 236
<212> PRT
<213> 土生梭孢壳
<400> 26
Met Lys Thr Phe Thr Ala Leu Leu Ala Ala Ala Gly Leu Val Ala Gly
1 5 10 15
His Gly Tyr Val Asp Asn Ala Thr Ile Gly Gly Gln Phe Tyr Gln Asn
20 25 30
Pro Ala Val Leu Thr Phe Phe Gln Pro Asp Arg Val Ser Arg Ser Ile
35 40 45
Pro Gly Asn Gly Pro Val Thr Asp Val Thr Leu Ile Asp Leu Gln Cys
50 55 60
Asn Ala Asn Ser Thr Pro Ala Lys Leu His Ala Thr Ala Ala Ala Gly
65 70 75 80
Ser Asp Val Ile Leu Arg Trp Thr Leu Trp Pro Glu Ser His Val Gly
85 90 95
Pro Val Ile Thr Tyr Met Ala Arg Cys Pro Asp Thr Gly Cys Gln Asp
100 105 110
Trp Met Pro Gly Thr Ser Ala Val Trp Phe Lys Ile Lys Glu Gly Gly
115 120 125
Arg Asp Gly Thr Ser Asn Thr Trp Ala Asp Thr Pro Leu Met Thr Ala
130 135 140
Pro Thr Ser Tyr Thr Tyr Thr Ile Pro Ser Cys Leu Lys Lys Gly Tyr
145 150 155 160
Tyr Leu Val Arg His Glu Ile Ile Ala Leu His Ala Ala Tyr Thr Tyr
165 170 175
Pro Gly Ala Gln Phe Tyr Pro Gly Cys His Gln Leu Asn Val Thr Gly
180 185 190
Gly Gly Ser Thr Val Pro Ser Ser Gly Leu Val Ala Phe Pro Gly Ala
195 200 205
Tyr Lys Gly Ser Asp Pro Gly Ile Thr Tyr Asp Ala Tyr Lys Ala Gln
210 215 220
Thr Tyr Gln Ile Pro Gly Pro Ala Val Phe Thr Cys
225 230 235
<210> 27
<211> 250
<212> PRT
<213> 土生梭孢壳
<400> 27
Met Ala Leu Leu Leu Leu Ala Gly Leu Ala Ile Leu Ala Gly Pro Ala
1 5 10 15
His Ala His Gly Gly Leu Ala Asn Tyr Thr Val Gly Asn Thr Trp Tyr
20 25 30
Arg Gly Tyr Asp Pro Phe Thr Pro Ala Ala Asp Gln Ile Gly Gln Pro
35 40 45
Trp Met Ile Gln Arg Ala Trp Asp Ser Ile Asp Pro Ile Phe Ser Val
50 55 60
Asn Asp Lys Ala Leu Ala Cys Asn Thr Pro Ala Thr Ala Pro Thr Ser
65 70 75 80
Tyr Ile Pro Ile Arg Ala Gly Glu Asn Ile Thr Ala Val Tyr Trp Tyr
85 90 95
Trp Leu His Pro Val Gly Pro Met Thr Ala Trp Leu Ala Arg Cys Asp
100 105 110
Gly Asp Cys Arg Asp Ala Asp Val Asn Glu Ala Arg Trp Phe Lys Ile
115 120 125
Trp Glu Ala Gly Leu Leu Ser Gly Pro Asn Leu Ala Glu Gly Met Trp
130 135 140
Tyr Gln Lys Ala Phe Gln Asn Trp Asp Gly Ser Pro Asp Leu Trp Pro
145 150 155 160
Val Thr Ile Pro Ala Gly Leu Lys Ser Gly Leu Tyr Met Ile Arg His
165 170 175
Glu Ile Leu Ser Ile His Val Glu Asp Lys Pro Gln Phe Tyr Pro Glu
180 185 190
Cys Ala His Leu Asn Val Thr Gly Gly Gly Asp Leu Leu Pro Pro Asp
195 200 205
Glu Phe Leu Val Lys Phe Pro Gly Ala Tyr Lys Glu Asp Asn Pro Ser
210 215 220
Ile Lys Ile Asn Ile Tyr Ser Asp Gln Tyr Ala Asn Thr Thr Asn Tyr
225 230 235 240
Thr Ile Pro Gly Gly Pro Ile Trp Asp Gly
245 250
<210> 28
<211> 478
<212> PRT
<213> 土生梭孢壳
<400> 28
Met Met Pro Ser Leu Val Arg Phe Ser Met Gly Leu Ala Thr Ala Phe
1 5 10 15
Ala Ser Leu Ser Thr Ala His Thr Val Phe Thr Thr Leu Phe Ile Asn
20 25 30
Gly Val Asp Gln Gly Asp Gly Thr Cys Ile Arg Met Ala Lys Lys Gly
35 40 45
Ser Val Cys Thr His Pro Ile Ala Gly Gly Leu Asp Ser Pro Asp Met
50 55 60
Ala Cys Gly Arg Asp Gly Gln Gln Ala Val Ala Phe Thr Cys Pro Ala
65 70 75 80
Pro Ala Gly Ser Lys Leu Ser Phe Glu Phe Arg Met Trp Ala Asp Ala
85 90 95
Ser Gln Pro Gly Ser Ile Asp Pro Ser His Leu Gly Ser Thr Ala Ile
100 105 110
Tyr Leu Lys Gln Val Ser Asn Ile Ser Ser Asp Ser Ala Ala Gly Pro
115 120 125
Gly Trp Phe Lys Ile Tyr Ala Glu Gly Tyr Asp Thr Ala Ala Lys Lys
130 135 140
Trp Ala Thr Glu Lys Leu Ile Asp Asn Gly Gly Leu Leu Ser Ile Glu
145 150 155 160
Leu Pro Pro Thr Leu Pro Ala Gly Tyr Tyr Leu Ala Arg Ser Glu Ile
165 170 175
Val Thr Ile Gln Asn Val Thr Asn Asp His Val Asp Pro Gln Phe Tyr
180 185 190
Val Gly Cys Ala Gln Leu Phe Val Gln Gly Pro Pro Thr Thr Pro Thr
195 200 205
Val Pro Pro Asp Arg Leu Val Ser Ile Pro Gly His Val His Ala Ser
210 215 220
Asp Pro Gly Leu Thr Phe Asn Ile Trp Arg Asp Asp Pro Ser Lys Thr
225 230 235 240
Ala Tyr Thr Val Val Gly Pro Ala Pro Phe Ser Pro Thr Ala Ala Pro
245 250 255
Thr Pro Thr Ser Thr Asn Thr Asn Gly Gln Gln Gln Gln Gln Gln Gln
260 265 270
Gln Ala Ile Lys Gln Thr Asp Gly Val Ile Pro Ala Asp Cys Gln Leu
275 280 285
Lys Asn Ala Asn Trp Cys Gly Ala Glu Val Pro Ala Tyr Ala Asp Glu
290 295 300
Ala Gly Cys Trp Ala Ser Ser Ala Asp Cys Phe Ala Gln Leu Asp Ala
305 310 315 320
Cys Tyr Thr Ser Ala Pro Pro Thr Gly Ser Arg Gly Cys Arg Leu Trp
325 330 335
Glu Asp Trp Cys Thr Gly Ile Gln Gln Gly Cys Arg Ala Gly Arg Trp
340 345 350
Arg Gly Pro Pro Pro Phe His Gly Glu Gly Ala Ala Ala Glu Thr Ala
355 360 365
Ser Ala Gly Arg Gly Gly Ala Arg Ile Ala Ala Val Ala Gly Cys Gly
370 375 380
Gly Gly Thr Gly Asp Met Val Glu Glu Val Phe Leu Phe Tyr Trp Asp
385 390 395 400
Ala Cys Ser Gly Trp Arg Arg Ser Arg Gly Gly Gly Ser Ile Leu Ala
405 410 415
Arg Leu Ile Leu His Val Leu Leu Pro Leu Leu Arg Pro Arg Arg Ala
420 425 430
Pro Arg Val His Leu Leu Leu Phe His Leu Tyr Leu Asn Phe Cys Tyr
435 440 445
Pro Gly Thr Ser Gly Phe Tyr Asn Arg Leu Ser Ile Lys Leu Gly Ile
450 455 460
Trp Pro Ser Lys Met Ser Pro Asp Val Ala His Tyr Val Lys
465 470 475
<210> 29
<211> 230
<212> PRT
<213> 土生梭孢壳
<400> 29
Met Gln Leu Leu Val Gly Leu Leu Leu Ala Ala Val Ala Ala Arg Ala
1 5 10 15
His Tyr Thr Phe Pro Arg Leu Val Val Asn Gly Gln Pro Glu Asp Lys
20 25 30
Asp Trp Ser Val Thr Arg Met Thr Lys Asn Ala Gln Ser Lys Gln Gly
35 40 45
Val Gln Asp Pro Thr Ser Pro Asp Ile Arg Cys Tyr Thr Ser Gln Thr
50 55 60
Ala Pro Asn Val Ala Thr Val Pro Ala Gly Ala Thr Val His Tyr Ile
65 70 75 80
Ser Thr Gln Gln Ile Asn His Pro Gly Pro Thr Gln Tyr Tyr Leu Ala
85 90 95
Lys Val Pro Ala Gly Ser Ser Ala Lys Thr Trp Asp Gly Ser Gly Ala
100 105 110
Val Trp Phe Lys Ile Ser Thr Thr Met Pro Tyr Leu Asp Asn Asn Lys
115 120 125
Gln Leu Val Trp Pro Asn Gln Asn Thr Tyr Thr Thr Val Asn Thr Thr
130 135 140
Ile Pro Ala Asp Thr Pro Ser Gly Glu Tyr Leu Leu Arg Val Glu Gln
145 150 155 160
Ile Ala Leu His Leu Ala Ser Gln Pro Asn Gly Ala Gln Phe Tyr Leu
165 170 175
Ala Cys Ser Gln Ile Gln Ile Thr Gly Gly Gly Asn Gly Thr Pro Gly
180 185 190
Pro Leu Val Ala Leu Pro Gly Ala Tyr Lys Ser Asn Asp Pro Gly Ile
195 200 205
Leu Val Asn Ile Tyr Ser Met Gln Pro Gly Asp Tyr Lys Pro Pro Gly
210 215 220
Pro Pro Val Trp Ser Gly
225 230
<210> 30
<211> 257
<212> PRT
<213> 土生梭孢壳
<400> 30
Met Lys Leu Tyr Leu Ala Ala Phe Leu Gly Ala Val Ala Thr Pro Gly
1 5 10 15
Ala Phe Ala His Gln Ile His Gly Ile Leu Leu Val Asn Gly Thr Glu
20 25 30
Thr Pro Glu Trp Lys Tyr Val Arg Asp Val Ala Trp Glu Gly Ala Tyr
35 40 45
Glu Pro Glu Lys Tyr Pro Asn Thr Glu Phe Phe Lys Thr Pro Pro Gln
50 55 60
Thr Asp Ile Asn Asn Pro Asn Ile Thr Cys Gly Arg Asn Ala Phe Asp
65 70 75 80
Ser Ala Ser Lys Thr Glu Thr Ala Asp Ile Leu Ala Gly Ser Glu Val
85 90 95
Gly Phe Arg Val Ser Trp Asp Gly Asn Gly Lys Tyr Gly Val Phe Trp
100 105 110
His Pro Gly Pro Gly Gln Ile Tyr Leu Ser Arg Ala Pro Asn Asp Asp
115 120 125
Leu Glu Asp Tyr Arg Gly Asp Gly Asp Trp Phe Lys Ile Ala Thr Gly
130 135 140
Ala Ala Val Ser Asn Thr Glu Trp Leu Leu Trp Asn Lys His Asp Phe
145 150 155 160
Asn Phe Thr Ile Pro Lys Thr Thr Pro Pro Gly Lys Tyr Leu Met Arg
165 170 175
Ile Glu Gln Phe Met Pro Ser Thr Val Glu Tyr Ser Gln Trp Tyr Val
180 185 190
Asn Cys Ala His Val Asn Ile Ile Gly Pro Gly Gly Gly Thr Pro Thr
195 200 205
Gly Phe Ala Arg Phe Pro Gly Thr Tyr Thr Val Asp Asp Pro Gly Ile
210 215 220
Lys Val Pro Leu Asn Gln Ile Val Asn Ser Gly Glu Leu Pro Gln Asp
225 230 235 240
Gln Leu Arg Leu Leu Glu Tyr Lys Pro Pro Gly Pro Ala Leu Trp Thr
245 250 255
Gly
<210> 31
<211> 251
<212> PRT
<213> 甲壳嗜热子囊菌
<400> 31
Met Ala Phe Ser Gln Ile Met Ala Ile Thr Gly Val Phe Leu Ala Ser
1 5 10 15
Ala Ser Leu Val Ala Gly His Gly Phe Val Gln Asn Ile Val Ile Asp
20 25 30
Gly Lys Ser Tyr Gly Gly Tyr Ile Val Asn Gln Tyr Pro Tyr Met Ser
35 40 45
Asp Pro Pro Glu Val Val Gly Trp Ser Thr Thr Ala Thr Asp Leu Gly
50 55 60
Phe Val Asp Gly Thr Gly Tyr Gln Gly Pro Asp Ile Ile Cys His Arg
65 70 75 80
Gly Ala Lys Pro Ala Ala Leu Thr Ala Gln Val Ala Ala Gly Gly Thr
85 90 95
Val Lys Leu Glu Trp Thr Pro Trp Pro Asp Ser His His Gly Pro Val
100 105 110
Ile Asn Tyr Leu Ala Pro Cys Asn Gly Asp Cys Ser Thr Val Asp Lys
115 120 125
Thr Gln Leu Lys Phe Phe Lys Ile Ala Gln Ala Gly Leu Ile Asp Asp
130 135 140
Asn Ser Pro Pro Gly Ile Trp Ala Ser Asp Asn Leu Ile Ala Ala Asn
145 150 155 160
Asn Ser Trp Thr Val Thr Ile Pro Thr Thr Thr Ala Pro Gly Asn Tyr
165 170 175
Val Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Lys Asp
180 185 190
Gly Ala Gln Asn Tyr Pro Gln Cys Ile Asn Leu Lys Val Thr Gly Asn
195 200 205
Gly Ser Gly Asn Pro Pro Ala Gly Ala Leu Gly Thr Ala Leu Tyr Lys
210 215 220
Asp Thr Asp Pro Gly Ile Leu Ile Asn Ile Tyr Gln Lys Leu Ser Ser
225 230 235 240
Tyr Val Ile Pro Gly Pro Ala Leu Tyr Thr Gly
245 250
<210> 32
<211> 349
<212> PRT
<213> 甲壳嗜热子囊菌
<400> 32
Met Ser Phe Ser Lys Ile Leu Ala Ile Ala Gly Ala Ile Thr Tyr Ala
1 5 10 15
Ser Ser Ala Ala Ala His Gly Tyr Val Gln Gly Ile Val Val Asp Gly
20 25 30
Ser Tyr Tyr Gly Gly Tyr Met Val Thr Gln Tyr Pro Tyr Thr Ala Gln
35 40 45
Pro Pro Glu Leu Ile Ala Trp Ser Thr Lys Ala Thr Asp Leu Gly Phe
50 55 60
Val Asp Gly Ser Gly Tyr Thr Ser Pro Asp Ile Ile Cys His Lys Gly
65 70 75 80
Ala Glu Pro Gly Ala Gln Ser Ala Lys Val Ala Ala Gly Gly Thr Val
85 90 95
Glu Leu Gln Trp Thr Ala Trp Pro Glu Ser His Lys Gly Pro Val Ile
100 105 110
Asp Tyr Leu Ala Ala Cys Asp Gly Asp Cys Ser Ser Val Asp Lys Thr
115 120 125
Ala Leu Lys Phe Phe Lys Ile Asp Glu Ser Gly Leu Ile Asp Gly Asn
130 135 140
Gly Ala Gly Thr Trp Ala Ser Asp Thr Leu Ile Lys Asn Asn Asn Ser
145 150 155 160
Trp Thr Val Thr Ile Pro Ser Thr Ile Ala Ser Gly Asn Tyr Val Leu
165 170 175
Arg His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Lys Asp Gly Ala
180 185 190
Gln Asn Tyr Pro Gln Cys Ile Asn Leu Glu Val Thr Gly Ser Gly Thr
195 200 205
Glu Asn Pro Ala Gly Thr Leu Gly Thr Ala Leu Tyr Thr Asp Thr Asp
210 215 220
Pro Gly Leu Leu Val Asn Ile Tyr Gln Gly Leu Ser Asn Tyr Ser Ile
225 230 235 240
Pro Gly Pro Ala Leu Tyr Ser Gly Asn Ser Asp Asn Ala Gly Ser Leu
245 250 255
Asn Pro Thr Thr Thr Pro Ser Ile Gln Asn Ala Ala Ala Ala Pro Ser
260 265 270
Thr Ser Thr Ala Ser Val Val Thr Asp Ser Ser Ser Ala Thr Gln Thr
275 280 285
Ala Ser Val Ala Ala Thr Thr Pro Ala Ser Thr Ser Ala Val Thr Ala
290 295 300
Ser Pro Ala Pro Asp Thr Gly Ser Asp Val Thr Lys Tyr Leu Asp Ser
305 310 315 320
Met Ser Ser Asp Glu Val Leu Thr Leu Val Arg Gly Thr Leu Ser Trp
325 330 335
Leu Val Ser Asn Lys Lys His Ala Arg Asp Leu Ser His
340 345
<210> 33
<211> 436
<212> PRT
<213> 甲壳嗜热子囊菌
<400> 33
Met Leu Ser Phe Ile Pro Thr Lys Ser Ala Ala Leu Thr Thr Leu Leu
1 5 10 15
Leu Leu Gly Thr Ala His Ala His Thr Leu Met Thr Thr Met Phe Val
20 25 30
Asp Gly Val Asn Gln Gly Asp Gly Val Cys Ile Arg Met Asn Asn Asp
35 40 45
Gly Gly Thr Ala Asn Thr Tyr Ile Gln Pro Ile Thr Ser Lys Asp Ile
50 55 60
Ala Cys Gly Ile Gln Gly Glu Ile Gly Ala Ser Arg Val Cys Pro Val
65 70 75 80
Lys Ala Ser Ser Thr Leu Thr Phe Gln Phe Arg Glu Gln Pro Asn Asn
85 90 95
Pro Asn Ser Ser Pro Leu Asp Pro Ser His Lys Gly Pro Ala Ala Val
100 105 110
Tyr Leu Lys Lys Val Asp Ser Ala Ile Ala Ser Asn Asn Ala Ala Gly
115 120 125
Asp Ser Trp Phe Lys Ile Trp Glu Ser Val Tyr Asp Glu Ser Thr Gly
130 135 140
Lys Trp Gly Thr Thr Lys Met Ile Glu Asn Asn Gly His Ile Ser Val
145 150 155 160
Lys Val Pro Asp Asp Ile Glu Gly Gly Tyr Tyr Leu Ala Arg Thr Glu
165 170 175
Leu Leu Ala Leu His Ser Ala Asp Gln Gly Asp Pro Gln Phe Tyr Val
180 185 190
Gly Cys Ala Gln Leu Phe Ile Asp Ser Asp Gly Thr Ala Lys Pro Pro
195 200 205
Thr Val Ser Ile Gly Glu Gly Thr Tyr Asp Leu Ser Met Pro Ala Met
210 215 220
Thr Tyr Asn Ile Trp Glu Thr Pro Leu Ala Leu Pro Tyr Pro Met Tyr
225 230 235 240
Gly Pro Pro Val Tyr Thr Pro Gly Ser Gly Ser Gly Ser Val Arg Ala
245 250 255
Thr Ser Ser Ser Ala Val Pro Thr Ala Thr Glu Ser Ser Phe Val Glu
260 265 270
Glu Arg Ala Asn Pro Val Thr Ala Asn Ser Val Tyr Ser Ala Arg Gly
275 280 285
Lys Phe Lys Thr Trp Ile Asp Lys Leu Ser Trp Arg Gly Lys Val Arg
290 295 300
Glu Asn Val Arg Gln Ala Ala Gly Arg Arg Ser Thr Leu Val Gln Thr
305 310 315 320
Val Gly Leu Lys Pro Lys Gly Cys Ile Phe Val Asn Gly Asn Trp Cys
325 330 335
Gly Phe Glu Val Pro Asp Tyr Asn Asp Ala Glu Ser Cys Trp Ala Ala
340 345 350
Ser Asp Asn Cys Trp Lys Gln Ser Asp Ala Cys Trp Asn Lys Thr Gln
355 360 365
Pro Thr Gly Tyr Asn Asn Cys Gln Ile Trp Gln Asp Lys Lys Cys Lys
370 375 380
Val Ile Gln Asp Ser Cys Ser Gly Pro Asn Pro His Gly Pro Pro Asn
385 390 395 400
Lys Gly Lys Asp Leu Thr Pro Glu Trp Pro Pro Leu Lys Gly Ser Met
405 410 415
Asp Thr Phe Ser Lys Arg Thr Ile Gly Tyr Arg Asp Trp Ile Val Arg
420 425 430
Arg Arg Gly Ala
435
<210> 34
<211> 344
<212> PRT
<213> 棘孢曲霉
<400> 34
Met Lys Tyr Ile Pro Leu Val Ile Ala Val Ala Ala Gly Leu Ala Arg
1 5 10 15
Pro Ala Thr Ala His Tyr Ile Phe Ser Lys Leu Val Leu Asn Gly Glu
20 25 30
Ala Ser Ala Asp Trp Gln Tyr Ile Arg Glu Thr Thr Arg Ser Ile Val
35 40 45
Tyr Glu Pro Thr Lys Tyr Thr Ser Thr Phe Asp Asn Leu Thr Pro Ser
50 55 60
Asp Ser Asp Phe Arg Cys Asn Leu Gly Ser Phe Ser Asn Ala Ala Lys
65 70 75 80
Thr Glu Val Ala Glu Val Ala Ala Gly Asp Thr Ile Ala Met Lys Leu
85 90 95
Phe Tyr Asp Thr Ser Ile Ala His Pro Gly Pro Gly Gln Val Tyr Met
100 105 110
Ser Lys Ala Pro Thr Gly Asn Val Gln Glu Tyr Gln Gly Asp Gly Asp
115 120 125
Trp Phe Lys Ile Trp Glu Lys Thr Leu Cys Asn Thr Asp Gly Asp Leu
130 135 140
Thr Thr Glu Ala Trp Cys Thr Trp Gly Met Ser Gln Phe Glu Phe Gln
145 150 155 160
Ile Pro Ala Ala Thr Pro Ala Gly Glu Tyr Leu Val Arg Ala Glu His
165 170 175
Ile Gly Leu His Gly Ala Gln Ala Asn Glu Ala Glu Phe Phe Tyr Ser
180 185 190
Cys Ala Gln Ile Lys Val Thr Gly Ser Gly Thr Gly Ser Pro Ser Leu
195 200 205
Thr Tyr Gln Ile Pro Gly Leu Tyr Asn Asp Thr Met Thr Leu Phe Asn
210 215 220
Gly Leu Asn Leu Trp Thr Asp Ser Ala Glu Lys Val Gln Leu Asp Phe
225 230 235 240
Leu Glu Thr Pro Ile Gly Asp Asp Val Trp Ser Gly Ala Gly Ser Gly
245 250 255
Ser Pro Ser Ala Ala Thr Ser Ser Thr Ser Gly Ala Thr Leu Ala Ala
260 265 270
Gln Gly Thr Thr Thr Ser Ala Ala His Ala Gln Ala Gln Thr Thr Ile
275 280 285
Thr Thr Ser Thr Ser Thr Ile Thr Ser Leu Glu Ser Ala Ser Ser Thr
290 295 300
Asp Leu Val Ala Gln Tyr Gly Gln Cys Gly Gly Leu Asn Trp Ser Gly
305 310 315 320
Pro Thr Glu Cys Glu Thr Pro Tyr Thr Cys Val Gln Gln Asn Pro Tyr
325 330 335
Tyr His Gln Cys Val Asn Ser Cys
340
<210> 35
<211> 400
<212> PRT
<213> 棘孢曲霉
<400> 35
Met Ser Val Ala Lys Phe Ala Gly Val Ile Leu Gly Ser Ala Ala Leu
1 5 10 15
Val Ala Gly His Gly Tyr Val Ser Gly Ala Val Val Asp Gly Thr Tyr
20 25 30
Tyr Gly Gly Tyr Ile Val Thr Ser Tyr Pro Tyr Ser Ser Asp Pro Pro
35 40 45
Glu Thr Ile Gly Trp Ser Thr Glu Ala Thr Asp Leu Gly Phe Val Asp
50 55 60
Gly Ser Glu Tyr Ala Asp Ala Asp Ile Ile Cys His Lys Ser Ala Lys
65 70 75 80
Pro Gly Ala Ile Ser Ala Glu Val Lys Ala Gly Gly Thr Val Glu Leu
85 90 95
Gln Trp Thr Thr Trp Pro Asp Ser His His Gly Pro Val Leu Thr Tyr
100 105 110
Leu Ala Asn Cys Asn Gly Asp Cys Ser Ser Val Thr Lys Thr Asp Leu
115 120 125
Glu Phe Phe Lys Ile Asp Glu Ser Gly Leu Ile Asn Asp Asp Asp Val
130 135 140
Pro Gly Thr Trp Ala Ser Asp Asn Leu Ile Ala Asn Asn Asn Ser Trp
145 150 155 160
Thr Val Thr Ile Pro Ser Asp Ile Ala Ala Gly Asn Tyr Val Leu Arg
165 170 175
His Glu Ile Ile Ala Leu His Ser Ala Gly Asn Lys Asp Gly Ala Gln
180 185 190
Asn Tyr Pro Gln Cys Leu Asn Leu Lys Val Thr Gly Gly Gly Asp Leu
195 200 205
Ala Pro Ser Gly Thr Ala Gly Glu Ser Leu Tyr Lys Asp Thr Asp Ala
210 215 220
Gly Ile Leu Val Asn Ile Tyr Gln Ser Leu Ser Ser Tyr Asp Ile Pro
225 230 235 240
Gly Pro Ala Met Tyr Asn Ala Thr Ser Ser Ser Ser Ser Ser Ser Ser
245 250 255
Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Gly Ser Ser Ser Ser
260 265 270
Ala Ala Ala Ser Ser Ser Ser Ser Ser Ser Ser Thr Thr Ala Ala Ala
275 280 285
Ala Ala Ala Thr Ser Ala Ala Ser Ser Val Thr Ser Ala Ala Gly Ser
290 295 300
Val Val Thr Gln Thr Ala Thr Ala Val Glu Thr Asp Thr Ala Thr Ala
305 310 315 320
Tyr Gln Thr Ser Thr Glu Val Ala Gln Val Thr Val Thr Gly Ser Ala
325 330 335
Pro Gln Gln Thr Tyr Val Ala Thr Pro Ser Ser Ser Ser Ser Ala Ser
340 345 350
Ser Ser Ser Ser Ala Ser Val Ser Thr Ser Thr Ser Leu Thr Ser Tyr
355 360 365
Phe Glu Ser Leu Ser Ala Asp Gln Phe Leu Ser Val Leu Lys Gln Thr
370 375 380
Phe Thr Trp Leu Val Ser Glu Lys Lys His Ala Arg Asp Leu Ser Ala
385 390 395 400
<210> 36
<211> 389
<212> PRT
<213> 棘孢曲霉
<400> 36
Met Lys Ser Ser Thr Phe Gly Met Leu Ala Leu Ala Ala Ala Ala Lys
1 5 10 15
Met Val Asp Ala His Thr Thr Val Phe Ala Val Trp Ile Asn Gly Glu
20 25 30
Asp Gln Gly Leu Gly Asn Ser Ala Ser Gly Tyr Ile Arg Ser Pro Pro
35 40 45
Ser Asn Ser Pro Val Lys Asp Val Thr Ser Thr Asp Ile Thr Cys Asn
50 55 60
Val Asn Gly Asp Gln Ala Ala Ala Lys Thr Leu Ser Val Lys Gly Gly
65 70 75 80
Asp Val Val Thr Phe Glu Trp His His Asp Ser Arg Asp Ala Ser Asp
85 90 95
Asp Ile Ile Ala Ser Ser His Lys Gly Pro Val Met Val Tyr Met Ala
100 105 110
Pro Thr Thr Ala Gly Ser Ser Gly Lys Asn Trp Val Lys Ile Ala Glu
115 120 125
Asp Gly Tyr Ser Asp Gly Thr Trp Ala Val Asp Thr Leu Ile Ala Asn
130 135 140
Ser Gly Lys His Asn Ile Thr Val Pro Asp Val Pro Ala Gly Asp Tyr
145 150 155 160
Leu Phe Arg Pro Glu Ile Ile Ala Leu His Glu Ala Glu Asn Glu Gly
165 170 175
Gly Ala Gln Phe Tyr Met Glu Cys Val Gln Phe Lys Val Thr Ser Asp
180 185 190
Gly Ala Asn Thr Leu Pro Asp Gly Val Ser Leu Pro Gly Ala Tyr Ser
195 200 205
Ala Thr Asp Pro Gly Ile Leu Phe Asn Met Tyr Gly Ser Phe Asp Ser
210 215 220
Tyr Pro Ile Pro Gly Pro Ser Val Trp Asp Gly Thr Ser Ser Gly Ser
225 230 235 240
Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ala Ala Ala Ala
245 250 255
Val Val Ala Thr Ser Ser Ser Ser Ser Ser Ala Ser Ile Glu Ala Val
260 265 270
Thr Thr Lys Gly Ala Val Ala Ala Val Ser Thr Ala Ala Ala Val Ala
275 280 285
Pro Thr Thr Thr Thr Ala Ala Pro Thr Thr Phe Ala Thr Ala Val Ala
290 295 300
Ser Thr Lys Lys Ala Thr Ala Cys Arg Asn Lys Thr Lys Ser Ser Ser
305 310 315 320
Ala Ala Thr Thr Ala Ala Ala Val Ala Glu Thr Thr Ser Ser Thr Ala
325 330 335
Ala Ala Thr Ala Ala Ala Ser Ser Ala Ser Ser Ala Ser Gly Thr Ala
340 345 350
Gly Lys Tyr Glu Arg Cys Gly Gly Gln Gly Trp Thr Gly Ala Thr Thr
355 360 365
Cys Val Asp Gly Trp Thr Cys Lys Gln Trp Asn Pro Tyr Tyr Tyr Gln
370 375 380
Cys Val Glu Ser Ala
385
<210> 37
<211> 406
<212> PRT
<213> 棘孢曲霉
<400> 37
Met Arg Gln Ala Gln Ser Leu Ser Leu Leu Thr Ala Leu Leu Ser Ala
1 5 10 15
Thr Arg Val Ala Gly His Gly His Val Thr Asn Val Val Val Asn Gly
20 25 30
Val Tyr Tyr Glu Gly Phe Asp Ile Asn Ser Phe Pro Tyr Glu Ser Asp
35 40 45
Pro Pro Lys Val Ala Ala Trp Thr Thr Pro Asn Thr Gly Asn Gly Phe
50 55 60
Ile Ser Pro Ser Asp Tyr Gly Thr Asp Asp Ile Ile Cys His Gln Asn
65 70 75 80
Ala Thr Asn Ala Gln Ala His Ile Val Val Ala Ala Gly Asp Lys Ile
85 90 95
Asn Ile Gln Trp Thr Ala Trp Pro Asp Ser His His Gly Pro Val Leu
100 105 110
Asp Tyr Leu Ala Arg Cys Asp Gly Glu Cys Glu Thr Val Asp Lys Thr
115 120 125
Thr Leu Glu Phe Phe Lys Ile Asp Gly Val Gly Leu Ile Ser Asp Thr
130 135 140
Glu Val Pro Gly Thr Trp Gly Asp Asp Gln Leu Ile Ala Asn Asn Asn
145 150 155 160
Ser Trp Leu Val Glu Ile Pro Pro Thr Ile Ala Pro Gly Asn Tyr Val
165 170 175
Leu Arg His Glu Leu Ile Ala Leu His Ser Ala Gly Thr Glu Asp Gly
180 185 190
Ala Gln Asn Tyr Pro Gln Cys Phe Asn Leu Gln Val Thr Gly Ser Gly
195 200 205
Thr Asp Glu Pro Ala Gly Thr Leu Gly Thr Lys Leu Tyr Thr Glu Asp
210 215 220
Glu Ala Gly Ile Val Val Asn Ile Tyr Thr Ser Leu Ser Ser Tyr Ala
225 230 235 240
Val Pro Gly Pro Thr Gln Tyr Ser Gly Ala Val Ser Val Ser Gln Ser
245 250 255
Thr Ser Ala Ile Thr Ser Thr Gly Thr Ala Val Val Gly Ser Gly Ser
260 265 270
Ala Val Ala Thr Ser Ala Ala Ala Ala Thr Thr Ser Ala Ala Ala Ser
275 280 285
Ser Ala Ala Ala Ala Thr Thr Ala Ala Ala Val Thr Ser Ala Asn Ala
290 295 300
Asn Thr Gln Ile Ala Gln Pro Ser Ser Ser Ser Ser Tyr Ser Gln Ile
305 310 315 320
Ala Val Gln Val Pro Ser Ser Trp Thr Thr Leu Val Thr Val Thr Pro
325 330 335
Pro Ala Ala Ala Ala Thr Thr Pro Ala Ala Val Pro Glu Pro Gln Thr
340 345 350
Pro Ser Ala Ser Ser Gly Ala Thr Thr Thr Ser Ser Ser Ser Gly Ala
355 360 365
Ala Gln Ser Leu Tyr Gly Gln Cys Gly Gly Ile Asn Trp Thr Gly Ala
370 375 380
Thr Ser Cys Val Glu Gly Ala Thr Cys Tyr Gln Tyr Asn Pro Tyr Tyr
385 390 395 400
Tyr Gln Cys Ile Ser Ala
405
<210> 38
<211> 427
<212> PRT
<213> 棘孢曲霉
<400> 38
Met Ser Leu Ser Lys Ile Ala Thr Leu Leu Leu Gly Ser Val Ser Leu
1 5 10 15
Val Ala Gly His Gly Tyr Val Ser Ser Ile Glu Val Asp Gly Thr Thr
20 25 30
Tyr Gly Gly Tyr Leu Val Asp Thr Tyr Tyr Tyr Glu Ser Asp Pro Pro
35 40 45
Glu Leu Ile Ala Trp Ser Thr Asn Ala Thr Asp Asp Gly Tyr Val Ser
50 55 60
Pro Ser Asp Tyr Glu Ser Val Asn Ile Ile Cys His Lys Gly Ser Ala
65 70 75 80
Pro Gly Ala Leu Ser Ala Pro Val Ala Pro Gly Gly Trp Val Gln Met
85 90 95
Thr Trp Asn Thr Trp Pro Thr Asp His His Gly Pro Val Ile Thr Tyr
100 105 110
Met Ala Asn Cys His Gly Ser Cys Ala Asp Val Asp Lys Thr Thr Leu
115 120 125
Glu Phe Phe Lys Ile Asp Ala Gly Gly Leu Ile Asp Asp Thr Asp Val
130 135 140
Pro Gly Thr Trp Ala Thr Asp Glu Leu Ile Glu Asp Ser Tyr Ser Arg
145 150 155 160
Asn Ile Thr Ile Pro Ser Asp Ile Ala Pro Gly Tyr Tyr Val Leu Arg
165 170 175
His Glu Ile Ile Ala Leu His Ser Ala Glu Asn Leu Asp Gly Ala Gln
180 185 190
Asn Tyr Pro Gln Cys Ile Asn Leu Glu Val Thr Gly Ser Glu Thr Ala
195 200 205
Thr Pro Ser Gly Thr Leu Gly Thr Ala Leu Tyr Lys Glu Thr Asp Pro
210 215 220
Gly Ile Tyr Val Asp Ile Trp Asn Thr Leu Ser Thr Tyr Thr Ile Pro
225 230 235 240
Gly Pro Ala Leu Tyr Thr Ala Gly Ser Thr Ala Thr Ala Ala Ala Ala
245 250 255
Ala Asp Thr Thr Thr Thr Ser Ala Gly Thr Thr Ala Glu Ala Thr Thr
260 265 270
Ala Ala Ala Ala Val Ser Thr Thr Ala Asp Ala Val Pro Thr Glu Ser
275 280 285
Ser Ala Pro Ser Glu Thr Ser Ala Thr Thr Ala Asn Pro Ala Arg Pro
290 295 300
Thr Ala Gly Ser Asp Ile Arg Phe Gln Pro Gly Gln Val Lys Ala Gly
305 310 315 320
Ala Ser Val Asn Asn Ser Ala Thr Glu Thr Ser Ser Gly Glu Ser Ala
325 330 335
Thr Thr Thr Thr Thr Ser Val Ala Thr Ala Ala Ser Ser Ala Asp Ser
340 345 350
Ser Thr Thr Ser Gly Val Leu Ser Gly Ala Cys Ser Gln Glu Gly Tyr
355 360 365
Trp Tyr Cys Asn Gly Gly Thr Ala Phe Gln Arg Cys Val Asn Gly Glu
370 375 380
Trp Asp Ala Ser Gln Ser Val Ala Ala Gly Thr Val Cys Thr Ala Gly
385 390 395 400
Ile Ser Glu Thr Ile Thr Ile Ser Ala Ala Ala Thr Arg Arg Asp Ala
405 410 415
Met Arg Arg His Leu Ala Arg Pro Lys Arg His
420 425
<210> 39
<211> 267
<212> PRT
<213> 棘孢曲霉
<400> 39
Met Leu Val Lys Leu Ile Ser Phe Leu Ser Ala Ala Thr Ser Val Ala
1 5 10 15
Ala His Gly His Val Ser Asn Ile Val Ile Asn Gly Val Ser Tyr Arg
20 25 30
Gly Trp Asp Ile Asn Ser Asp Pro Tyr Asn Ser Asn Pro Pro Val Val
35 40 45
Val Ala Trp Gln Thr Pro Asn Thr Ala Asn Gly Phe Ile Ser Pro Asp
50 55 60
Ala Tyr Asp Thr Asp Asp Val Ile Cys His Leu Ser Ala Thr Asn Ala
65 70 75 80
Arg Gly His Ala Val Val Ala Ala Gly Asp Lys Ile Ser Leu Gln Trp
85 90 95
Thr Thr Trp Pro Asp Ser His His Gly Pro Val Ile Ser Tyr Leu Ala
100 105 110
Asn Cys Gly Ser Ser Cys Glu Thr Val Asp Lys Thr Thr Leu Glu Phe
115 120 125
Phe Lys Ile Asp Gly Val Gly Leu Val Asp Glu Ser Asn Pro Pro Gly
130 135 140
Ile Trp Gly Asp Asp Glu Leu Ile Ala Asn Asn Asn Ser Trp Leu Val
145 150 155 160
Glu Ile Pro Ala Ser Ile Ala Pro Gly Tyr Tyr Val Leu Arg His Glu
165 170 175
Leu Ile Ala Leu His Gly Ala Gly Ser Glu Asn Gly Ala Gln Asn Tyr
180 185 190
Met Gln Cys Phe Asn Leu Gln Val Thr Gly Thr Gly Thr Val Gln Pro
195 200 205
Ser Gly Val Leu Gly Thr Glu Leu Tyr Lys Pro Thr Asp Ala Gly Ile
210 215 220
Leu Val Asn Ile Tyr Gln Ser Leu Ser Thr Tyr Val Val Pro Gly Pro
225 230 235 240
Thr Leu Ile Pro Gln Ala Val Ser Leu Val Gln Ser Ser Ser Thr Ile
245 250 255
Thr Ala Ser Gly Thr Ala Val Thr Thr Thr Ala
260 265
<210> 40
<211> 273
<212> PRT
<213> 棘孢曲霉
<400> 40
Met Lys Tyr Leu Ala Ile Phe Ala Ala Ala Ala Ala Gly Leu Ala Arg
1 5 10 15
Pro Thr Ala Ala His Tyr Ile Phe Ser Lys Leu Ile Leu Asp Gly Glu
20 25 30
Val Ser Glu Asp Trp Gln Tyr Ile Arg Lys Thr Thr Arg Glu Thr Cys
35 40 45
Tyr Leu Pro Thr Lys Phe Thr Asp Thr Phe Asp Asn Leu Thr Pro Asn
50 55 60
Asp Gln Asp Phe Arg Cys Asn Leu Gly Ser Phe Ser Asn Ala Ala Lys
65 70 75 80
Thr Glu Val Ala Glu Val Glu Ala Gly Ser Thr Ile Gly Met Gln Leu
85 90 95
Phe Ala Gly Ser His Met Arg His Pro Gly Pro Ala Gln Val Phe Met
100 105 110
Ser Lys Ala Pro Ser Gly Asn Val Gln Ser Tyr Glu Gly Asp Gly Ser
115 120 125
Trp Phe Lys Ile Trp Glu Arg Thr Leu Cys Asp Lys Ser Gly Asp Leu
130 135 140
Thr Gly Asp Ala Trp Cys Thr Tyr Gly Gln Thr Glu Ile Glu Phe Gln
145 150 155 160
Ile Pro Glu Ala Thr Pro Thr Gly Glu Tyr Leu Val Arg Ala Glu His
165 170 175
Ile Gly Leu His Arg Ala Gln Ser Asn Gln Ala Glu Phe Tyr Tyr Ser
180 185 190
Cys Ala Gln Val Lys Val Thr Gly Asn Gly Thr Gly Val Pro Ser Gln
195 200 205
Thr Tyr Gln Ile Pro Gly Met Tyr Asn Asp Arg Ser Glu Leu Phe Asn
210 215 220
Gly Leu Asn Leu Trp Ser Tyr Ser Val Glu Asn Val Glu Ala Ala Met
225 230 235 240
Lys Asn Ser Ile Val Gly Asp Glu Ile Trp Asn Gly Ser Ser Val Pro
245 250 255
Ser Glu Ser His Val Pro Lys Tyr Lys Lys Ser His Ala Cys Arg Val
260 265 270
Tyr
<210> 41
<211> 322
<212> PRT
<213> 浅红孔菌(Aurantiporus alborubescens)
<400> 41
Met Arg Thr Ile Ala Thr Phe Val Thr Leu Val Ala Ser Val Leu Pro
1 5 10 15
Ala Val Leu Ala His Gly Gly Val Leu Ser Tyr Ser Asn Gly Gly Asn
20 25 30
Trp Tyr Trp Gly Trp Lys Pro Tyr Asn Ser Pro Asp Gly Gln Thr Thr
35 40 45
Ile Gln Arg Pro Trp Ala Thr Tyr Asn Pro Ile Thr Asp Ala Thr Asp
50 55 60
Pro Thr Ile Ala Cys Asn Asn Asp Gly Thr Ser Gly Ala Leu Gln Leu
65 70 75 80
Thr Ala Thr Val Ala Ala Gly Ser Ala Ile Thr Ala Tyr Trp Asn Gln
85 90 95
Val Trp Pro His Asp Lys Gly Pro Met Thr Thr Tyr Leu Ala Gln Cys
100 105 110
Pro Gly Ser Thr Cys Thr Gly Val Asn Ala Lys Thr Leu Lys Trp Phe
115 120 125
Lys Ile Asp His Ala Gly Leu Leu Ser Gly Thr Val Tyr Ser Gly Ser
130 135 140
Trp Ala Ser Gly Lys Met Ile Ala Gln Asn Ser Thr Trp Thr Thr Thr
145 150 155 160
Ile Pro Ala Thr Val Pro Ser Gly Asn Tyr Leu Ile Arg Phe Glu Thr
165 170 175
Ile Ala Leu His Ser Leu Pro Ala Gln Phe Tyr Pro Glu Cys Ala Gln
180 185 190
Ile Gln Ile Thr Gly Gly Gly Ser Arg Ala Pro Thr Ala Ala Glu Leu
195 200 205
Val Ser Phe Pro Gly Ala Tyr Ser Asn Asn Asp Pro Gly Val Asn Ile
210 215 220
Asp Ile Tyr Ser Asn Ala Ala Gln Ser Ala Thr Thr Tyr Val Ile Pro
225 230 235 240
Gly Pro Pro Leu Tyr Gly Gly Ala Ser Gly Ser Gly Pro Ser Ser Ala
245 250 255
Pro Pro Ser Ser Thr Pro Gly Ser Ser Ser Thr Ser His Gly Pro Thr
260 265 270
Ser Val Ser Thr Ser Ser Ser Ala Ala Pro Ser Thr Thr Gly Thr Val
275 280 285
Thr Gln Tyr Gly Gln Cys Gly Gly Ile Gly Trp Ala Gly Ala Thr Gly
290 295 300
Cys Ile Ser Pro Phe Lys Cys Thr Val Ile Asn Asp Tyr Tyr Tyr Gln
305 310 315 320
Cys Leu
<210> 42
<211> 234
<212> PRT
<213> 浅红孔菌
<400> 42
Met Lys Ala Ile Leu Ala Ile Phe Ser Ala Leu Ala Pro Leu Ala Ala
1 5 10 15
Ala His Tyr Thr Phe Pro Asp Phe Ile Val Asn Gly Thr Thr Thr Ala
20 25 30
Asp Trp Val Tyr Ile Arg Glu Thr Ala Asn His Tyr Ser Asn Gly Pro
35 40 45
Val Thr Asn Val Asn Asp Pro Glu Phe Arg Cys Tyr Glu Leu Asp Leu
50 55 60
Gln Asn Thr Ala Ala Ser Thr Leu Thr Ala Thr Val Ser Ala Gly Ser
65 70 75 80
Ser Val Gly Phe Lys Ala Asn Ser Ala Leu Tyr His Pro Gly Tyr Leu
85 90 95
Asp Val Tyr Met Ser Lys Ala Thr Pro Ala Ala Asn Ser Pro Ser Ala
100 105 110
Gly Thr Asp Gln Ser Trp Phe Lys Val Tyr Glu Ser Ala Pro Val Phe
115 120 125
Ala Asn Gly Ala Leu Ser Phe Pro Ser Glu Asn Ile Gln Ser Phe Thr
130 135 140
Phe Thr Ile Pro Lys Ser Leu Pro Ser Gly Gln Tyr Leu Ile Arg Val
145 150 155 160
Glu His Ile Ala Leu His Ser Ala Ser Ser Tyr Gly Gly Ala Gln Phe
165 170 175
Tyr Ile Ser Cys Ala Gln Val Asn Val Val Asn Gly Gly Asn Gly Asn
180 185 190
Pro Gly Pro Leu Val Lys Ile Pro Gly Val Tyr Thr Gly Asn Glu Pro
195 200 205
Gly Ile Leu Ile Asn Ile Tyr Ser Phe Pro Pro Gly Phe Ser Gly Tyr
210 215 220
Gln Ser Pro Gly Pro Ala Val Trp Arg Gly
225 230
<210> 43
<211> 233
<212> PRT
<213> 囊状长毛盘菌
<400> 43
Met Thr Pro Leu Lys Leu Arg Pro Leu Leu Leu Leu Val Leu Ser Thr
1 5 10 15
Thr Leu Ser Leu Val His Ala His Tyr Arg Phe Tyr Glu Leu Ile Ala
20 25 30
Asn Gly Ala Thr His Ala Ser Phe Glu Tyr Ile Arg Gln Trp Val Pro
35 40 45
Ile Tyr Ser Asn Ser Pro Val Thr Asp Val Thr Ser Val Asn Leu Arg
50 55 60
Cys Asn Val Asn Ala Thr Pro Ala Ala Glu Val Ile Thr Val Ala Ala
65 70 75 80
Gly Ser Thr Val Gly Phe Val Ala Asp Thr Thr Val Thr His Pro Gly
85 90 95
Ala Phe Thr Ala Tyr Met Ala Lys Ala Pro Glu Asp Ile Thr Glu Trp
100 105 110
Asp Gly Asn Gly Asp Trp Phe Lys Ile Trp Glu Lys Gly Pro Thr Ser
115 120 125
Ile Thr Ser Ser Gly Ile Thr Trp Asp Val Thr Asp Thr Gln Trp Thr
130 135 140
Phe Thr Ile Pro Ser Ala Thr Pro Asn Gly Gln Tyr Leu Leu Arg Phe
145 150 155 160
Glu His Ile Ala Leu His Ala Ala Ser Thr Val Gly Gly Ala Gln Phe
165 170 175
Tyr Met Ser Cys Ala Gln Ile Gln Val Thr Asn Gly Gly Asn Gly Ser
180 185 190
Pro Gly Pro Thr Ile Lys Phe Pro Gly Gly Tyr Ser Ala Thr Asp Pro
195 200 205
Gly Ile Leu Ile Asn Ile Tyr Tyr Pro Ile Pro Thr Ser Tyr Thr Ile
210 215 220
Pro Gly Pro Pro Val Trp Thr Gly Lys
225 230
<210> 44
<211> 237
<212> PRT
<213> 囊状长毛盘菌
<400> 44
Met Lys Cys Leu Leu Ser Leu Leu Leu Ala Ala Thr Ala Val Ser Ala
1 5 10 15
His Thr Ile Phe Gln Glu Ile Gly Ile Asn Gly Val Met Gln Ala Arg
20 25 30
Tyr Asp Tyr Met Arg Leu Pro Ser Tyr Asp Gly Pro Ile Thr Asp Val
35 40 45
Thr Ser Thr Tyr Met Ala Cys Asn Gly Gly Pro Asn Pro Leu Val Gln
50 55 60
Ile Ser Asn Asp Val Ala Phe Val Lys Ala Gly Asp Ser Ile Thr Leu
65 70 75 80
Gln Trp Ala Gln Thr Leu Thr Thr Asp Phe Asn Thr Gly Leu Ile Ile
85 90 95
Asp Pro Ser His Leu Gly Pro Val Met Val Tyr Met Ala Lys Val Pro
100 105 110
Ser Ala Thr Gly Pro Ile Pro Asn Ser Gly Trp Phe Lys Ile Tyr Glu
115 120 125
Asp Gly Tyr Asp Pro Thr Thr Lys Thr Trp Ala Val Thr Lys Leu Ile
130 135 140
Asn Asn Lys Gly Lys Val Thr Val Thr Ile Pro Ser Cys Leu Pro Ala
145 150 155 160
Gly Asp Tyr Leu Leu Arg Gly Glu Ile Ile Ala Leu His Ala Ala Ser
165 170 175
Thr Tyr Pro Gly Ala Gln Phe Tyr Met Glu Cys Ala Gln Leu Arg Leu
180 185 190
Thr Ser Gly Gly Thr Lys Met Pro Thr Thr Tyr Asn Ile Pro Gly Ile
195 200 205
Tyr Ser Pro Thr Asp Pro Gly Val Thr Phe Asn Leu Tyr Asn Gly Phe
210 215 220
Thr Ser Tyr Thr Ile Pro Gly Pro Arg Pro Phe Thr Cys
225 230 235
<210> 45
<211> 484
<212> PRT
<213> 托姆青霉
<400> 45
Met Ser Leu Ser Lys Ile Ser Gly Leu Ile Leu Gly Ser Ala Ala Leu
1 5 10 15
Val Ala Gly His Gly Tyr Val Ser Gly Ile Val Val Asp Asp Thr Tyr
20 25 30
Tyr Gly Gly Tyr Leu Val Thr Gln Tyr Pro Tyr Glu Ser Asp Ala Pro
35 40 45
Glu Leu Ile Ala Trp Ser Glu Gln Glu Thr Asp Leu Gly Tyr Ile Asp
50 55 60
Gly Ser Glu Tyr Ala Asn Ser Asn Ile Ile Cys His Lys Glu Ala Lys
65 70 75 80
Pro Gly Ala Leu Glu Ala Pro Val Lys Ala Gly Gly Ser Val Glu Leu
85 90 95
Gln Trp Thr Thr Trp Pro Thr Ser His His Gly Pro Val Ile Thr Tyr
100 105 110
Met Ala Asn Cys Asn Gly Asp Cys Asp Asp Val Asp Lys Thr Thr Leu
115 120 125
Gln Phe Phe Lys Ile Asp Gln Gly Gly Leu Ile Ser Asp Thr Thr Glu
130 135 140
Pro Gly Thr Trp Ala Thr Asp Asn Leu Ile Ala Asn Asn Asn Ser Arg
145 150 155 160
Thr Val Thr Val Pro Ser Asp Ile Ala Asp Gly Asn Tyr Val Leu Arg
165 170 175
His Glu Ile Ile Ala Leu His Ser Ala Gly Glu Thr Asn Gly Ala Gln
180 185 190
Asn Tyr Pro Gln Cys Ile Asn Leu Lys Val Thr Gly Gly Gly Ser Ala
195 200 205
Thr Pro Ser Gly Thr Leu Gly Thr Ala Leu Tyr Lys Asn Thr Asp Pro
210 215 220
Gly Ile Leu Ile Asn Ile Tyr Thr Ser Leu Ser Thr Tyr Asp Ile Pro
225 230 235 240
Gly Pro Thr Leu Tyr Thr Ala Gly Ala Ala Ala Ala Thr Ala Ala Ser
245 250 255
Thr Ala Ala Ser Ser Thr Ala Ala Ala Val Thr Thr Ala Asp Ala Val
260 265 270
Thr Thr Ala Ala Ala Val Thr Ser Ser Ser Ala Ser Val Glu Val Val
275 280 285
Pro Thr Thr Thr Pro Ser Ser Ser Ile Val Ser Ala Phe Pro Thr Trp
290 295 300
Ser Pro Ser Ser Thr Pro Pro Phe Ser Asn Ser Ser Asn Gly Trp Arg
305 310 315 320
Pro Ser Phe Ser Arg Gly Pro Gly Gly Pro Arg Phe Thr Ser Ala Pro
325 330 335
Ala Pro Gln Phe Ser Ala Pro Ser Gly Ala Gln Gln Lys Gln Ser Ala
340 345 350
Thr Ala Thr Pro Ile Val Ala Thr Pro Val Val Ile Thr Met Thr Glu
355 360 365
Thr Ser Thr Ser Trp Val Thr Glu Met Val Thr Leu Thr Asp Lys Ser
370 375 380
Val Val Gln Thr Thr Ser Ala Val Pro Val Val Val Ala Ala Thr Thr
385 390 395 400
Thr Leu Thr Glu Gly Ser Glu Pro Ala Gln Thr Ala Ser Pro Ser Val
405 410 415
Val Ser Gly Ser Ser Ser Ser Gly Ser Ser Ser Ser Ser Thr Thr Thr
420 425 430
Thr Ser Lys Thr Ser Thr Gly Ser Asp Tyr Val Ser Ser Asp Trp Met
435 440 445
Ser Tyr Leu Ser Ser Leu Ser Ala Ala Glu Val Leu Gln Met Leu Arg
450 455 460
Gln Thr Phe Arg Trp Met Val Ser Asn Asp Lys Val His Ala Arg Asp
465 470 475 480
Ile Thr Ile Asn
<210> 46
<211> 320
<212> PRT
<213> 柄篮状菌
<400> 46
Met Pro Ser Thr Lys Val Ala Ala Leu Ser Ala Val Leu Ala Leu Ala
1 5 10 15
Ser Thr Val Ala Gly His Gly Phe Val Gln Asn Ile Val Ile Asp Gly
20 25 30
Lys Ser Tyr Thr Gly Tyr Leu Val Asn Gln Tyr Pro Tyr Gln Ser Asn
35 40 45
Pro Pro Ala Val Ile Gly Trp Ser Thr Thr Ala Thr Asp Leu Gly Phe
50 55 60
Val Asp Gly Ser Gly Tyr Thr Asn Pro Asp Ile Ile Cys His Lys Asn
65 70 75 80
Ala Lys Pro Gly Gln Leu Ser Ala Pro Val Ala Ala Gly Gly Lys Val
85 90 95
Glu Leu Glu Trp Thr Thr Trp Pro Glu Ser His His Gly Pro Val Ile
100 105 110
Ser Tyr Leu Ala Asn Cys Asn Gly Asp Cys Thr Thr Val Asp Lys Thr
115 120 125
Lys Leu Glu Phe Val Lys Ile Asp Gln Arg Gly Leu Ile Asp Asp Ser
130 135 140
Asn Pro Pro Gly Thr Trp Ala Ala Asp Gln Leu Ile Ala Ala Asn Asn
145 150 155 160
Ser Trp Thr Val Thr Ile Pro Glu Ser Ile Ala Pro Gly Asn Tyr Val
165 170 175
Leu Arg His Glu Ile Ile Ala Leu His Ser Ala Asn Asn Ala Thr Gly
180 185 190
Ala Gln Asn Tyr Pro Gln Cys Ile Asn Leu Gln Ile Thr Gly Ser Gly
195 200 205
Thr Ala Asn Pro Ser Gly Thr Pro Gly Glu Lys Leu Tyr Thr Pro Thr
210 215 220
Asp Pro Gly Ile Leu Val Asn Ile Tyr Gln Ser Leu Ser Ser Tyr Val
225 230 235 240
Ile Pro Gly Pro Thr Leu Trp Ser Gly Ala Ala Ala His Val Val Ala
245 250 255
Thr Ala Ala Gly Ser Ala Thr Gly Val Ala Ser Ala Thr Ala Thr Pro
260 265 270
Thr Thr Leu Val Thr Ala Val Ser Ser Pro Thr Gly Ala Pro Ser Val
275 280 285
Val Thr Pro Glu Ala Pro Ser Val Thr Ser Phe Ala Pro Val Val Thr
290 295 300
Val Thr Asp Val Val Thr Val Thr Thr Val Ile Thr Thr Thr Ile Ser
305 310 315 320
<210> 47
<211> 330
<212> PRT
<213> 粗糙脉孢菌
<400> 47
Met Arg Ser Thr Leu Val Thr Gly Leu Ile Ala Gly Leu Leu Ser Gln
1 5 10 15
Gln Ala Ala Ala His Ala Thr Phe Gln Ala Leu Trp Val Asp Gly Ala
20 25 30
Asp Tyr Gly Ser Gln Cys Ala Arg Val Pro Pro Ser Asn Ser Pro Val
35 40 45
Thr Asp Val Thr Ser Asn Ala Met Arg Cys Asn Thr Gly Thr Ser Pro
50 55 60
Val Ala Lys Lys Cys Pro Val Lys Ala Gly Ser Thr Val Thr Val Glu
65 70 75 80
Met His Gln Gln Ala Asn Asp Arg Ser Cys Ser Ser Glu Ala Ile Gly
85 90 95
Gly Ala His Tyr Gly Pro Val Leu Val Tyr Met Ser Lys Val Ser Asp
100 105 110
Ala Ala Ser Ala Asp Gly Ser Ser Gly Trp Phe Lys Ile Phe Glu Asp
115 120 125
Thr Trp Ala Lys Lys Pro Ser Ser Ser Ser Gly Asp Asp Asp Phe Trp
130 135 140
Gly Val Lys Asp Leu Asn Ser Cys Cys Gly Lys Met Gln Val Lys Ile
145 150 155 160
Pro Ser Asp Ile Pro Ala Gly Asp Tyr Leu Leu Arg Ala Glu Val Ile
165 170 175
Ala Leu His Thr Ala Ala Ser Ala Gly Gly Ala Gln Leu Tyr Met Thr
180 185 190
Cys Tyr Gln Ile Ser Val Thr Gly Gly Gly Ser Ala Thr Pro Ala Thr
195 200 205
Val Ser Phe Pro Gly Ala Tyr Lys Ser Ser Asp Pro Gly Ile Leu Val
210 215 220
Asp Ile His Ser Ala Met Ser Thr Tyr Val Ala Pro Gly Pro Ala Val
225 230 235 240
Tyr Ser Gly Gly Ser Ser Lys Lys Ala Gly Ser Gly Cys Val Gly Cys
245 250 255
Glu Ser Thr Cys Lys Val Gly Ser Gly Pro Thr Gly Thr Ala Ser Ala
260 265 270
Val Pro Val Ala Ser Thr Ser Ala Ala Ala Gly Gly Gly Gly Gly Gly
275 280 285
Gly Ser Gly Gly Cys Ser Val Ala Lys Tyr Gln Gln Cys Gly Gly Thr
290 295 300
Gly Tyr Thr Gly Cys Thr Ser Cys Ala Ser Gly Ser Thr Cys Ser Ala
305 310 315 320
Val Ser Pro Pro Tyr Tyr Ser Gln Cys Val
325 330
Claims (16)
1.一种减少聚酯纺织品中的低聚物的方法,包括在水性溶液中在角质酶的存在下将聚酯纺织品用糖基水解酶家族61多肽进行处理,其中该方法是按以下的温度范围进行的:40℃-100℃。
2.如权利要求1所述的方法,其中该纺织品是纱线、织物或衣服。
3.如权利要求1或2所述的方法,其中该聚酯是PET。
4.如权利要求1所述的方法,其中该水性溶液进一步包括选自下组的一种或多种酶,该组由以下各项组成:脂肪酶、酯酶、漆酶、过氧化物酶、环化酶以及转移酶。
5.如以上权利要求中任一项所述的方法,其中与一种糖基水解酶家族61一起使用了一种共物质;优选地该共物质是半胱氨酸。
6.如以上权利要求中任一项所述的方法,其中该糖基水解酶家族61多肽是按以下范围施用的:从0.01至约50毫克蛋白质/克聚酯纺织品,优选0.05-20毫克蛋白质/克聚酯纺织品,优选0.1-15毫克蛋白质/克聚酯纺织品,更优选0.2-8毫克蛋白质/克聚酯纺织品,并且甚至更优选0.2-5毫克蛋白质/克聚酯纺织品。
7.如以上权利要求中任一项所述的方法,其中该角质酶是按以下范围施用的:从约0.01至约50毫克酶蛋白质/克聚酯纺织品,优选0.05-20毫克酶蛋白质/克聚酯纺织品,更优选0.1-15毫克酶蛋白质/克聚酯纺织品,并且甚至更优选0.2-5毫克酶蛋白质/克聚酯纺织品。
8.如以上权利要求中任一项所述的方法,其中该方法是按以下的pH范围进行的:从约pH 3至约pH 11,优选从约pH 4至约pH 10的范围,或从约pH 6至约pH 9的范围。
9.如以上权利要求中任一项所述的方法,其中该方法是按以下的温度范围进行的:50℃-90℃,优选60℃-85℃,更优选65℃-80℃,并且甚至更优选70℃-80℃。
10.如以上权利要求中任一项所述的方法,其中该方法进行了约10分钟至约8小时,优选约20分钟至约180分钟,更优选约30分钟至约150分钟,更优选约45分钟至约120分钟。
11.如以上权利要求中任一项所述的方法,其中处理聚酯纺织品是制造该聚酯纺织品,尤其制造聚酯织物。
12.如权利要求11所述的方法,其中该方法是与现存的聚酯织物制造步骤中的任一个相组合。
13.如以上权利要求中任一项所述的方法,其中该角质酶与SEQ ID NO:1具有至少90%序列一致性,或包括SEQ ID NO:1的一个或多个(或若干个)氨基酸取代、缺失、和/或插入。
14.一种糖基水解酶家族61多肽促进角质酶减少聚酯纺织品中的低聚物的用途。
15.权利要求14所述的用途,其中该用途是当与相同条件下不使用糖基水解酶家族61多肽的相同工艺运行相比时,减少聚对苯二甲酸乙二酯的环状或线性低聚物在机械和/或纺织品上的沉积。
16.权利要求14或15所述的用途,其中该低聚物是酸-双-2-苯甲酰氧基-乙酯和/或对苯二甲酸三乙二酯。
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WO2012061517A1 (en) * | 2010-11-02 | 2012-05-10 | Novozymes, Inc. | Methods of pretreating cellulosic material with a gh61 polypeptide |
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WO2001034899A1 (en) * | 1999-11-05 | 2001-05-17 | Genencor International, Inc. | Enzymes useful for changing the properties of polyester |
CN101910391A (zh) * | 2008-01-04 | 2010-12-08 | 宝洁公司 | 包含糖基水解酶和包含有益剂递送颗粒的衣物洗涤剂组合物 |
WO2011080267A2 (en) * | 2009-12-29 | 2011-07-07 | Novozymes A/S | Polypetides having detergency enhancing effect |
WO2012061517A1 (en) * | 2010-11-02 | 2012-05-10 | Novozymes, Inc. | Methods of pretreating cellulosic material with a gh61 polypeptide |
WO2012089023A1 (en) * | 2010-12-30 | 2012-07-05 | Novozymes A/S | Processes for treating textile with polypeptide having cellulolytic enzyme enhancing activity |
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