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公开(公告)号：US08017372B2
公开(公告)日：2011-09-13
申请号：US11830063
申请日：2007-07-30
申请人： Kim Vilbour Andersen , Martin Schulein , Torben Henriksen, legal representative , Lars Christiansen , Bo Damgaard , Claus Von der Osten
CPC分类号： C11D3/38645 , C12N9/2437 , C12Y302/01004
摘要： The present invention relates to a method for improving the properties of a cellulolytic enzyme by amino acid substitution, deletion or insertion, the method comprising the steps of: a. constructing a multiple alignment of at least two amino acid sequences known to have three-dimensional structures similar to endoglucanase V (EGV) from Humicola insolens known from Protein Data Bank entry 4ENG; b. constructing a homology-built three-dimensional structure of the cellulolytic enzyme based on the structure of the EGV; c. identifying amino acid residue positions present in a distance from the substrate binding cleft of not more than 5 Å; d. identifying surface-exposed amino acid residues of the enzyme; e. identifying all charged or potentially charged amino acid residue positions of the enzyme; f. choosing one or more positions wherein the amino acid residue is to be substituted, deleted or where an insertion is to be provided; and g. carrying out the substitution, deletion or insertion by using conventional protein engineering techniques. Also described are cellulase variants obtained by this method.
摘要翻译： 本发明涉及通过氨基酸取代，缺失或插入改进纤维素分解酶的性质的方法，该方法包括以下步骤：a。 构建已知具有类似于蛋白质数据库条目4ENG中已知的Humicola insolens的内切葡聚糖酶V（EGV）的三维结构的至少两个氨基酸序列的多重比对; b。 基于EGV的结构构建纤维素分解酶的同源构建的三维结构; C。 鉴定存在于与所述基质结合裂缝一定距离的氨基酸残基位置不大于5埃; d。 鉴定酶的表面暴露的氨基酸残基; e。 鉴定酶的所有带电荷或潜在带电的氨基酸残基位置; F。 选择其中氨基酸残基被取代，缺失或提供插入的一个或多个位置; 和g。 通过使用常规蛋白质工程技术进行取代，缺失或插入。 还描述了通过该方法获得的纤维素酶变体。
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公开(公告)号：US20090170747A1
公开(公告)日：2009-07-02
申请号：US12394202
申请日：2009-02-27
申请人： Kim Vilbour Andersen , Martin Schulein , Torben Henriksen , Lars Christensen , Bo Damgaard , Claus Von der Osten
发明人： Kim Vilbour Andersen , Martin Schulein , Torben Henriksen , Lars Christensen , Bo Damgaard , Claus Von der Osten
CPC分类号： C11D3/38645 , C12N9/2437 , C12Y302/01004
摘要： The present invention relates to a method for improving the properties of a cellulolytic enzyme by amino acid substitution, deletion or insertion, the method comprising the steps of:a. constructing a multiple alignment of at least two amino acid sequences known to have three-dimensional structures similar to endoglucanase V (EGV) from Humicola insolens known from Protein Data Bank entry 4ENG; b. constructing a homology-built three-dimensional structure of the cellulolytic enzyme based on the structure of the EGV; c. identifying amino acid residue positions present in a distance from the substrate binding cleft of not more than 5 Å; d. identifying surface-exposed amino acid residues of the enzyme; e. identifying all charged or potentially charged amino acid residue positions of the enzyme; f. choosing one or more positions wherein the amino acid residue is to be substituted, deleted or where an insertion is to be provided; and g. carrying out the substitution, deletion or insertion by using conventional protein engineering techniques. Also described are cellulase variants obtained by this method.
摘要翻译： 本发明涉及通过氨基酸取代，缺失或插入改进纤维素分解酶的性质的方法，该方法包括以下步骤：a。 构建已知具有类似于蛋白质数据库条目4ENG中已知的Humicola insolens的内切葡聚糖酶V（EGV）的三维结构的至少两个氨基酸序列的多重比对; b。 基于EGV的结构构建纤维素分解酶的同源构建的三维结构; C。 鉴定存在于与所述基质结合裂缝一定距离的氨基酸残基位置不大于5埃; d。 鉴定酶的表面暴露的氨基酸残基; e。 鉴定酶的所有带电荷或潜在带电的氨基酸残基位置; F。 选择其中氨基酸残基被取代，缺失或提供插入的一个或多个位置; 和g。 通过使用常规蛋白质工程技术进行取代，缺失或插入。 还描述了通过该方法获得的纤维素酶变体。
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公开(公告)号：US20080206836A1
公开(公告)日：2008-08-28
申请号：US11830063
申请日：2007-07-30
申请人： Kim Vilbour Andersen , Martin Schulein , Torben Henriksen , Lars Christiansen , Bo Damgaard , Claus Von der Osten
发明人： Kim Vilbour Andersen , Martin Schulein , Torben Henriksen , Lars Christiansen , Bo Damgaard , Claus Von der Osten
IPC分类号： C12N9/42
CPC分类号： C11D3/38645 , C12N9/2437 , C12Y302/01004
摘要： The present invention relates to a method for improving the properties of a cellulolytic enzyme by amino acid substitution, deletion or insertion, the method comprising the steps of: a. constructing a multiple alignment of at least two amino acid sequences known to have three-dimensional structures similar to endoglucanase V (EGV) from Humicola insolens known from Protein Data Bank entry 4ENG; b. constructing a homology-built three-dimensional structure of the cellulolytic enzyme based on the structure of the EGV; c. identifying amino acid residue positions present in a distance from the substrate binding cleft of not more than 5 Å; d. identifying surface-exposed amino acid residues of the enzyme; e. identifying all charged or potentially charged amino acid residue positions of the enzyme; f. choosing one or more positions wherein the amino acid residue is to be substituted, deleted or where an insertion is to be provided; and g. carrying out the substitution, deletion or insertion by using conventional protein engineering techniques. Also described are cellulase variants obtained by this method.
摘要翻译： 本发明涉及通过氨基酸取代，缺失或插入改进纤维素分解酶的性质的方法，该方法包括以下步骤：a。 构建已知具有类似于蛋白质数据库条目4ENG中已知的Humicola insolens的内切葡聚糖酶V（EGV）的三维结构的至少两个氨基酸序列的多重比对; b。 基于EGV的结构构建纤维素分解酶的同源构建的三维结构; C。 鉴定存在于与所述基质结合裂缝一定距离的氨基酸残基位置不大于5埃; d。 鉴定酶的表面暴露的氨基酸残基; e。 鉴定酶的所有带电荷或潜在带电的氨基酸残基位置; F。 选择其中氨基酸残基被取代，缺失或提供插入的一个或多个位置; 和g。 通过使用常规蛋白质工程技术进行取代，缺失或插入。 还描述了通过该方法获得的纤维素酶变体。
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公开(公告)号：US08642730B2
公开(公告)日：2014-02-04
申请号：US12567302
申请日：2009-09-25
申请人： Lene Nonboe Andersen , Soren Flensted Lassen , Markus Sakari Kauppinen , Lene Lange , Ruby Ilum Nielsen , Shinobu Takagi , Michiko Ihara , Torben Henriksen
发明人： Martin Schulein , Lene Nonboe Andersen , Soren Flensted Lassen , Markus Sakari Kauppinen , Lene Lange , Ruby Ilum Nielsen , Shinobu Takagi , Michiko Ihara
IPC分类号： C07K1/00
CPC分类号： C12N9/2437 , A61K38/00 , C05F11/08 , C05F17/0036 , C07K2319/00 , C11D3/386 , C12P19/14 , C12Y302/01004 , D06M16/003 , D06P5/02 , D06P5/158 , D21C5/005 , D21H17/005 , D21H21/10 , Y02P20/145 , Y02W30/43
摘要： The present invention relates to enzyme preparations consisting essentially of an enzyme which has cellulytic activity and comprises a first amino acid sequence having the following sequence (SEQ ID NO: 79) Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa 1 2 3 4 5 6 7 8 9 10 11 12 Trp Xaa 13 14 and a second amino acid sequence having the following sequence Trp Cys Cys Xaa Cys(SEQ ID NO: 80) 1 2 3 4 5 wherein, at position 3 of the first sequence, the amino acid is Trp, Tyr or Phe; at position 4 of the first sequence, the amino acid is Trp, Tyr or Phe; at position 8 of the first sequence, the amino acid is Arg, Lys or His; at positions 9, 10, 12 and 14, respectively, of the first sequence, and at position 4 of the second sequence, the amino acid is any of the 20 naturally occurring amino acid residues with the provisos that, in the first amino acid sequence, (i) when the amino residue at position 12 is Ser, then the amino acid residue at position 14 is not Ser, and (ii) when the amino residue at position 12 is Gly, then the amino acid residue at position 14 is not Ala, performs very well in industrial applications such as laundry compositions, for biopolishing of newly manufactured textiles, for providing an abraded look of cellulosic fabric or garment, and for treatment of paper pulp. Further, the invention relates to DNA constructs encoding such enzymes, a method for providing a gene encoding for such enzymes, a method of producing the enzymes, enzyme preparations containing such enzymes, and the use of these enzymes for a number of industrial applications.
摘要翻译： 本发明涉及基本上由具有纤维素分解活性的酶组成并包含具有以下序列的第一氨基酸序列的酶制剂（SEQ ID NO：79）Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa 123456789101112 Trp Xaa 1314 和具有以下序列的第二氨基酸序列Trp Cys Cys Xaa Cys（SEQ ID NO：80）12345其中，在第一序列的3位，氨基酸为Trp，Tyr或Phe; 在第一序列的位置4处，氨基酸是Trp，Tyr或Phe; 在第一序列的位置8，氨基酸是Arg，Lys或His; 在第一序列的第9,10,12和14位，第2个序列的第4位，氨基酸是20个天然存在的氨基酸残基中的任一个，条件是在第一个氨基酸序列 ，（i）当第12位的氨基残基为Ser时，第14位的氨基酸残基不为Ser，（ii）当第12位的氨基残基为Gly时，第14位的氨基酸残基为 Ala在工业应用中表现良好，例如洗衣组合物，用于生物抛光新制造的纺织品，用于提供纤维素织物或衣服的磨损外观以及纸浆处理。 此外，本发明涉及编码这些酶的DNA构建体，提供编码这种酶的基因的方法，生产酶的方法，含有这些酶的酶制剂，以及这些酶用于许多工业应用的用途。
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公开(公告)号：US20110250674A1
公开(公告)日：2011-10-13
申请号：US13162636
申请日：2011-06-17
申请人： Kim Vilbour Andersen , Martin Schülein , Torben Henriksen , Lars Christiansen , Bo Damgaard , Claus Von der Osten
发明人： Kim Vilbour Andersen , Martin Schülein , Torben Henriksen , Lars Christiansen , Bo Damgaard , Claus Von der Osten
IPC分类号： C12N9/42
CPC分类号： C11D3/38645 , C12N9/2437 , C12Y302/01004
摘要： The present invention relates to a method for improving the properties of a cellulolytic enzyme by amino acid substitution, deletion or insertion, the method comprising the steps of: a. constructing a multiple alignment of at least two amino acid sequences known to have three-dimensional structures similar to endoglucanase V (EGV) from Humicola insolens known from Protein Data Bank entry 4ENG; b. constructing a homology-built three-dimensional structure of the cellulolytic enzyme based on the structure of the EGV; c. identifying amino acid residue positions present in a distance from the substrate binding cleft of not more than 5 Å; d. identifying surface-exposed amino acid residues of the enzyme; e. identifying all charged or potentially charged amino acid residue positions of the enzyme; f. choosing one or more positions wherein the amino acid residue is to be substituted, deleted or where an insertion is to be provided; and g. carrying out the substitution, deletion or insertion by using conventional protein engineering techniques. Also described are cellulase variants obtained by this method.
摘要翻译： 本发明涉及通过氨基酸取代，缺失或插入改进纤维素分解酶的性质的方法，该方法包括以下步骤：a。 构建已知具有类似于蛋白质数据库条目4ENG中已知的Humicola insolens的内切葡聚糖酶V（EGV）的三维结构的至少两个氨基酸序列的多重比对; b。 基于EGV的结构构建纤维素分解酶的同源构建的三维结构; C。 鉴定存在于与所述基质结合裂缝一定距离的氨基酸残基位置不大于5埃; d。 鉴定酶的表面暴露的氨基酸残基; e。 鉴定酶的所有带电荷或潜在带电的氨基酸残基位置; F。 选择其中氨基酸残基被取代，缺失或提供插入的一个或多个位置; 和g。 通过使用常规蛋白质工程技术进行取代，缺失或插入。 还描述了通过该方法获得的纤维素酶变体。
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公开(公告)号：US20100107342A1
公开(公告)日：2010-05-06
申请号：US12567302
申请日：2009-09-25
申请人： MARTIN SCHULEIN , TORBEN HENRIKSEN , LENE NONBOE ANDERSEN , SOREN FLENSTED LASSEN , MARKUS SAKARI KAUPPINEN , LENE LANGE , RUBY IIUM NIELSEN , SHINOBU TAKAGI , MICHIKO IHARA
发明人： MARTIN SCHULEIN , TORBEN HENRIKSEN , LENE NONBOE ANDERSEN , SOREN FLENSTED LASSEN , MARKUS SAKARI KAUPPINEN , LENE LANGE , RUBY IIUM NIELSEN , SHINOBU TAKAGI , MICHIKO IHARA
CPC分类号： C12N9/2437 , A61K38/00 , C05F11/08 , C05F17/0036 , C07K2319/00 , C11D3/386 , C12P19/14 , C12Y302/01004 , D06M16/003 , D06P5/02 , D06P5/158 , D21C5/005 , D21H17/005 , D21H21/10 , Y02P20/145 , Y02W30/43
摘要： The present invention relates to enzyme preparations consisting essentially of an enzyme which has cellulytic activity and comprises a first amino acid sequence having the following sequence (SEQ ID NO: 79) Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa 1 2 3 4 5 6 7 8 9 10 11 12 Trp Xaa 13 14 and a second amino acid sequence having the following sequence Trp Cys Cys Xaa Cys(SEQ ID NO: 80) 1 2 3 4 5 wherein, at position 3 of the first sequence, the amino acid is Trp, Tyr or Phe; at position 4 of the first sequence, the amino acid is Trp, Tyr or Phe; at position 8 of the first sequence, the amino acid is Arg, Lys or His; at positions 9, 10, 12 and 14, respectively, of the first sequence, and at position 4 of the second sequence, the amino acid is any of the 20 naturally occurring amino acid residues with the provisos that, in the first amino acid sequence, (i) when the amino residue at position 12 is Ser, then the amino acid residue at position 14 is not Ser, and (ii) when the amino residue at position 12 is Gly, then the amino acid residue at position 14 is not Ala, performs very well in industrial applications such as laundry compositions, for biopolishing of newly manufactured textiles, for providing an abraded look of cellulosic fabric or garment, and for treatment of paper pulp. Further, the invention relates to DNA constructs encoding such enzymes, a method for providing a gene encoding for such enzymes, a method of producing the enzymes, enzyme preparations containing such enzymes, and the use of these enzymes for a number of industrial applications.
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公开(公告)号：US07611882B2
公开(公告)日：2009-11-03
申请号：US10478840
申请日：2002-05-14
申请人： Mads Eskelund Bjørnvad , Martin Schülein , Torben Henriksen, legal representative , Vibeke Skovgaard Nielsen , Johan Smets , Carsten Andersen , Thomas Thisted
发明人： Mads Eskelund Bjørnvad , Martin Schülein , Vibeke Skovgaard Nielsen , Johan Smets , Carsten Andersen , Thomas Thisted
CPC分类号： A61K8/66 , A61Q11/00 , C11D3/38636 , C12N9/88
摘要： Detergent composition comprising a surfactant and a pectate lyase (EC 4.2.2.2) enzyme en-coded by a DNA sequence endogeneous to a strain of Bacillus subtilis or a stabilized variant thereof provides superior cleaning and stain removal.
摘要翻译： 包含表面活性剂和由枯草芽孢杆菌菌株或其稳定变体的内切DNA序列编码的果胶酸裂解酶（EC 4.2.2.2）酶的洗涤剂组合物提供了优异的清洁和去污。
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公开(公告)号：US07226773B2
公开(公告)日：2007-06-05
申请号：US10965499
申请日：2004-10-14
申请人： Martin Schulein, deceased , Torben Henriksen, legal representative , Lene Nonboe Andersen , Soren Flensted Lassen , Markus Sakari Kauppinen , Lene Lange , Ruby Ilum Nielsen , Shinobu Takagi , Michiko Ihara
发明人： Martin Schulein, deceased , Torben Henriksen, legal representative , Lene Nonboe Andersen , Soren Flensted Lassen , Markus Sakari Kauppinen , Lene Lange , Ruby Ilum Nielsen , Shinobu Takagi , Michiko Ihara
CPC分类号： C12N9/2437 , A61K38/00 , C05F11/08 , C05F17/0036 , C07K2319/00 , C11D3/386 , C12P19/14 , C12Y302/01004 , D06M16/003 , D06P5/02 , D06P5/158 , D21C5/005 , D21H17/005 , D21H21/10 , Y02P20/145 , Y02W30/43
摘要： The present invention relates to enzyme preparations consisting essentially of an enzyme which has cellulytic activity and comprises a first amino acid sequence having the following sequence (SEQ ID NO: 79) ThrArgXaaXaaAspCysCysXaaXaaXaaCysXaaTrpXaa 1234567891011121314 and a second amino acid sequence having the following sequence TrpCysCysXaaCys(SEQ ID NO: 80) 12345 wherein, at position 3 of the first sequence, the amino acid is Trp, Tyr or Phe; at position 4 of the first sequence, the amino acid is Trp, Tyr or Phe; at position 8 of the first sequence, the amino acid is Arg, Lys or His; at positions 9, 10, 12 and 14, respectively, of the first sequence, and at position 4 of the second sequence, the amino acid is any of the 20 naturally occurring amino acid residues with the provisos that, in the first amino acid sequence, (i) when the amino residue at position 12 is Ser, then the amino acid residue at position 14 is not Ser, and (ii) when the amino residue at position 12 is Gly, then the amino acid residue at position 14 is not Ala, performs very well in industrial applications such as laundry compositions, for biopolishing of newly manufactured textiles, for providing an abraded look of cellulosic fabric or garment, and for treatment of paper pulp. Further, the invention relates to DNA constructs encoding such enzymes, a method for providing a gene encoding for such enzymes, a method of producing the enzymes, enzyme preparations containing such enzymes, and the use of these enzymes for a number of industrial applications.
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公开(公告)号：US07041488B2
公开(公告)日：2006-05-09
申请号：US10479446
申请日：2002-06-06
发明人： Martin Schülein
CPC分类号： C11D3/386
摘要： An enzyme exhibiting endo-beta-1,4-glucanase activity (EC 3.2.1.4), which is selected from one of a) a polypeptide encoded by the DNA sequence of positions 1 to 2322 of SEQ ID NO:1,b) a polypeptide produced by culturing a cell comprising the sequence of SEQ ID NO:1 under conditions wherein the DNA sequence is expressed; c) an endo-beta-1,4-glucanase enzyme having a sequence of at least 97% identity to the amino acid sequence of position 1 to position 773 of SEQ ID NO:2, and fragments thereof exhibitng endo-beta-1,4-glucanase activity, and d) a polypeptide having endo-beta-1,4-glucanase activity that is encoded by a polynucleo-tide that hybridizes with the nucleotide sequence shown in positions 1-2322 of SEQ ID NO:1, is useful for detergent and textile applications.
摘要翻译： 显示内β-1,4-葡聚糖酶活性的酶（EC 3.2.1.4），其选自a）由SEQ ID NO：1的1至2322位的DNA序列编码的多肽之一，b）a 通过在表达DNA序列的条件下培养包含SEQ ID NO：1的序列的细胞产生的多肽; c）具有与SEQ ID NO：2的位置1至位置773的氨基酸序列具有至少97％同一性的序列的内切-β-1,4-葡聚糖酶，并且其片段表现出内切-β-1， 4-葡聚糖酶活性，和d）具有与SEQ ID NO：1所示1-2322位所示的核苷酸序列杂交的多核苷酸编码的具有内切-β-1,4-葡聚糖酶活性的多肽是有用的 用于洗涤剂和纺织品应用。
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公开(公告)号：US20080145912A1
公开(公告)日：2008-06-19
申请号：US11740076
申请日：2007-04-25
申请人： Martin Schulein , Torben Henriksen , Lene Nonboe Andersen , Soren Flensted Lassen , Markus Sakari Kauppinen , Lene Lange , Ruby Ilum Nielsen , Shinobu Takagi , Michiko Ihara
发明人： Martin Schulein , Torben Henriksen , Lene Nonboe Andersen , Soren Flensted Lassen , Markus Sakari Kauppinen , Lene Lange , Ruby Ilum Nielsen , Shinobu Takagi , Michiko Ihara
摘要： The present invention relates to enzyme preparations consisting essentially of an enzyme which has cellulytic activity and comprises a first amino acid sequence having the following sequence (SEQ ID NO: 79) Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa Xaa Cys Xaa 1 2 3 4 5 6 7 8 9 10 11 12 Trp Xaa 13 14 and a second amino acid sequence having the following sequence Trp Cys Cys Xaa Cys(SEQ ID NO: 80) 1 2 3 4 5 wherein, at position 3 of the first sequence, the amino acid is Trp, Tyr or Phe; at position 4 of the first sequence, the amino acid is Trp, Tyr or Phe; at position 8 of the first sequence, the amino acid is Arg, Lys or His; at positions 9, 10, 12 and 14, respectively, of the first sequence, and at position 4 of the second sequence, the amino acid is any of the 20 naturally occurring amino acid residues with the provisos that, in the first amino acid sequence, (i) when the amino residue at position 12 is Ser, then the amino acid residue at position 14 is not Ser, and (ii) when the amino residue at position 12 is Gly, then the amino acid residue at position 14 is not Ala, performs very well in industrial applications such as laundry compositions, for biopolishing of newly manufactured textiles, for providing an abraded look of cellulosic fabric or garment, and for treatment of paper pulp. Further, the invention relates to DNA constructs encoding such enzymes, a method for providing a gene encoding for such enzymes, a method of producing the enzymes, enzyme preparations containing such enzymes, and the use of these enzymes for a number of industrial applications.
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