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    • 3. 发明申请
    • CELLULASE VARIANTS
    • 纤维素酶变体
    • US20090170747A1
    • 2009-07-02
    • US12394202
    • 2009-02-27
    • Kim Vilbour AndersenMartin SchuleinTorben HenriksenLars ChristensenBo DamgaardClaus Von der Osten
    • Kim Vilbour AndersenMartin SchuleinTorben HenriksenLars ChristensenBo DamgaardClaus Von der Osten
    • C11D3/386C12N9/42
    • C11D3/38645C12N9/2437C12Y302/01004
    • 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.
    • 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。
    • 4. 发明申请
    • Cellulase Variants
    • 纤维素酶变体
    • US20080206836A1
    • 2008-08-28
    • US11830063
    • 2007-07-30
    • Kim Vilbour AndersenMartin SchuleinTorben HenriksenLars ChristiansenBo DamgaardClaus Von der Osten
    • Kim Vilbour AndersenMartin SchuleinTorben HenriksenLars ChristiansenBo DamgaardClaus Von der Osten
    • C12N9/42
    • C11D3/38645C12N9/2437C12Y302/01004
    • 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.
    • 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。
    • 9. 发明申请
    • CELLULASE VARIANTS
    • 纤维素酶变体
    • US20110250674A1
    • 2011-10-13
    • US13162636
    • 2011-06-17
    • Kim Vilbour AndersenMartin SchüleinTorben HenriksenLars ChristiansenBo DamgaardClaus Von der Osten
    • Kim Vilbour AndersenMartin SchüleinTorben HenriksenLars ChristiansenBo DamgaardClaus Von der Osten
    • C12N9/42
    • C11D3/38645C12N9/2437C12Y302/01004
    • 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.
    • 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。