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    Cellulase variants
    2.
    发明授权
    Cellulase variants 有权
    纤维素酶变体

    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

    发明人： Kim Vilbour Andersen ,  Martin Schulein ,  Lars Christiansen ,  Bo Damgaard ,  Claus Von der Osten

    IPC分类号： C12N9/42 ,  C12N15/00 ,  C12P21/04 ,  C07H21/04

    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。

    CELLULASE VARIANTS
    3.
    发明申请
    CELLULASE VARIANTS 有权
    纤维素酶变体

    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

    IPC分类号： C11D3/386 ,  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。

    Cellulase Variants
    4.
    发明申请
    Cellulase Variants 有权
    纤维素酶变体

    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。

    CELLULASE VARIANTS
    5.
    发明申请
    CELLULASE VARIANTS 审中-公开
    纤维素酶变体

    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。

    Cellulase Variants
    6.
    发明申请
    Cellulase Variants 审中-公开
    纤维素酶变体

    公开(公告)号：US20120289450A1

    公开(公告)日：2012-11-15

    申请号：US13471757

    申请日：2012-05-15

    申请人： Kim Vilbour Andersen ,  Martin Schülein ,  Lars Christiansen ,  Bo Damgaard ,  Claus Von der Osten

    发明人： Kim Vilbour Andersen ,  Martin Schülein ,  Lars Christiansen ,  Bo Damgaard ,  Claus Von der Osten

    IPC分类号： C12N9/42 ,  C11D3/386

    CPC分类号： C11D3/38645 ,  C12N9/2437 ,  C12Y302/01004

    摘要： The present invention relates to cellulase variants, i.e., endo-beta-1,4-glucanase variants, derived from a parental cellulase, i.e., endo-beta-1,4-glucanase, by substitution, insertion and/or deletion, which variant has a catalytic core domain, in which the variant at position 5 holds an alanine residue (A), a serine residue (S), or a threonine residue (T); at position 8 holds a phenylalanine residue (F), or a tyrosine residue (Y); at position 9 holds a phenylalanine residue (F), a tryptophan residue (W), or a tyrosine residue (Y); at position 10 holds an aspartic acid residue (D); and at position 121 holds an aspartic acid residue (D).

    摘要翻译： 本发明涉及通过取代，插入和/或缺失衍生自亲代纤维素酶（即内切-β-1,4-葡聚糖酶）的纤维素酶变体，即内切-β-1,4-葡聚糖酶变体，该变体 具有催化核心结构域，其中第5位的变体含有丙氨酸残基（A），丝氨酸残基（S）或苏氨酸残基（T）; 在8位保持苯丙氨酸残基（F）或酪氨酸残基（Y）; 位置9含有苯丙氨酸残基（F），色氨酸残基（W）或酪氨酸残基（Y）; 在10位保持天冬氨酸残基（D）; 并且在位置121处保持天冬氨酸残基（D）。

    METHOD FOR PREPARING MODIFIED POLYPEPTIDES
    8.
    发明申请
    METHOD FOR PREPARING MODIFIED POLYPEPTIDES 审中-公开
    制备改性多糖的方法

    公开(公告)号：US20120107852A1

    公开(公告)日：2012-05-03

    申请号：US13272681

    申请日：2011-10-13

    申请人： Torben Halkier ,  Anders Hjelholt Pedersen ,  Jens Sigurd Okkels ,  Kim Vilbour Andersen

    发明人： Torben Halkier ,  Anders Hjelholt Pedersen ,  Jens Sigurd Okkels ,  Kim Vilbour Andersen

    IPC分类号： C12Q1/48

    CPC分类号： C12N15/1058 ,  C07K1/107 ,  C07K14/31 ,  C07K19/00 ,  C12N15/1034

    摘要： Methods for producing polypeptide with altered immunogenicity or improved stability properties are disclosed. The methods involve a) expressing a diversified population of nucleotide sequences encoding a polypeptide of interest, b) screening the polypeptides expressed in step a) for function, immunogenicity and/or stability, c) selecting functional polypeptides having altered immunogenicity and/or increased stability, e.g. functional in vivo half-life as compared to the polypeptide of interest, and d) optionally subjecting the nucleotide sequence encoding the polypeptide selected in step c) to one or more repeated cycles of steps a)-c). In a further step the expressed polypeptides of step a) or c) can be conjugated to at least one non-polypeptide moiety.

    摘要翻译： 公开了产生具有改变的免疫原性或改善的稳定性的多肽的方法。 所述方法包括：a）表达编码感兴趣多肽的核苷酸序列的多样化群体，b）筛选功能，免疫原性和/或稳定性在步骤a）中表达的多肽，c）选择具有改变的免疫原性和/或增加稳定性的功能性多肽 ，例如 与目的多肽相比具有功能的体内半衰期，和d）任选地将编码步骤c）中选择的多肽的核苷酸序列经历步骤a）-c）的一个或多个重复循环。 在另一步骤中，步骤a）或c）的表达多肽可以与至少一个非多肽部分缀合。