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    • 5. 发明申请
    • Cell-wall degrading enzyme variants
    • 细胞壁降解酶变体
    • US20030017575A1
    • 2003-01-23
    • US09910505
    • 2001-07-19
    • Novozymes A/S
    • Sanne O. Schroder GladCarsten AndersenMartin SchuleinHanne DelaTorben Peter Frandsen
    • C12N009/88C12P019/04C07H021/04C12P021/02C12N005/06
    • C11D3/38636C12N9/88
    • A variant of a cell-wall degrading enzyme having a beta-helix structure, which variant holds at least one substituent in a position determined by identifying all residues potentially belonging to a stack; characterizing the stack as interior or exterior; characterizing the stack as polar, hydrophobic or aromatic/heteroaromatic based on the dominating characteristics of the parent or wild-type enzyme stack residues and/or its orientation relative to the beta-helix (interior or exterior); optimizing all stack positions of a stack either to hydrophobic aliphatic amino acids, hydrophobic aromatic or polar amino acids by allowing mutations within one or all positions to amino acids belonging to one of these groups; measuring thermostability of the variants by DSC or an application-related assay such as a Pad-Steam application test; and selecting the stabilized variants. Variant of a wild-type parent pectate lyase (EC 4.2.2.2) having the conserved amino acid residues D111, D141 or E141, D145, K165, R194 and R199 when aligned with the pectate lyase comprising the amino acid sequence of SEQ ID NO: 2 are preferred.
    • 具有β-螺旋结构的细胞壁降解酶的变体,该变体保持在通过鉴定潜在属于堆叠的所有残基而确定的位置中的至少一个取代基; 将堆栈表征为内部或外部; 基于母体或野生型酶堆残基的主导特征和/或其相对于β-螺旋(内部或外部)的取向,将所述叠层表征为极性,疏水性或芳族/杂芳族化合物; 通过在属于这些基团之一的氨基酸的一个或所有位置内允许突变将堆叠的所有堆叠位置优化为疏水性脂族氨基酸,疏水性芳族或极性氨基酸; 通过DSC测量变体的热稳定性或应用相关测定,例如Pad-Steam应用测试; 并选择稳定的变体。 当与包含SEQ ID NO:1的氨基酸序列的果胶酸裂解酶比对时,具有保守氨基酸残基D111,D141或E141,D145,K165,R194和R199的野生型亲本果胶酸裂合酶(EC 4.2.2.2) 2是优选的。
    • 6. 发明申请
    • Cell-wall degrading enzyme variants
    • 细胞壁降解酶变体
    • US20030175940A1
    • 2003-09-18
    • US10403192
    • 2003-03-31
    • Novozymes A/S
    • Sanne O. Schroder GladCarsten AndersenMartin SchuleinHanne DelaTorben Peter
    • C12N009/88G01N033/53G06F019/00G01N033/48G01N033/50C07H021/04C12N001/21C11D003/386C12N015/74
    • C11D3/38636C12N9/88
    • The present invention relates to variants of a cell-wall degrading enzyme having a beta-helix structure, which variant has at least one substituent in a position determined by identifying all residues potentially belonging to a stack; characterizing the stack as interior or exterior; characterizing the stack as polar, hydrophobic or aromatic/heteroaromatic based on the dominating characteristics of the parent or wild-type enzyme stack residues and/or its orientation relative to the beta-helix (interior or exterior); optimizing all stack positions of a stack either to hydrophobic aliphatic amino acids, hydrophobic aromatic or polar amino acids by allowing mutations within one or all positions to amino acids belonging to one of these groups; measuring thermostability of the variants by DSC or an application-related assay such as a Pad-Steam application test; and selecting the stabilized variants. Variant of a wild-type parent pectate lyase (EC 4.2.2.2) having the conserved amino acid residues D111, D141 or E141, D145, K165, R194 and R199 when aligned with the pectate lyase comprising the amino acid sequence of SEQ ID NO: 2 are preferred.
    • 本发明涉及具有β-螺旋结构的细胞壁降解酶的变体,该变体具有至少一个取代基,其位置通过鉴定潜在属于堆叠的所有残基而确定; 将堆栈表征为内部或外部; 基于母体或野生型酶堆残基的主导特征和/或其相对于β-螺旋(内部或外部)的取向,将所述叠层表征为极性,疏水性或芳族/杂芳族化合物; 通过在属于这些基团之一的氨基酸的一个或所有位置内允许突变将堆叠的所有堆叠位置优化为疏水性脂族氨基酸,疏水性芳族或极性氨基酸; 通过DSC测量变体的热稳定性或应用相关测定,例如Pad-Steam应用测试; 并选择稳定的变体。 当与包含SEQ ID NO:1的氨基酸序列的果胶酸裂解酶比对时,具有保守氨基酸残基D111,D141或E141,D145,K165,R194和R199的野生型亲本果胶酸裂合酶(EC 4.2.2.2) 2是优选的。
    • 7. 发明申请
    • Endoglucanases
    • 内切葡聚糖酶
    • US20030054539A1
    • 2003-03-20
    • US10007521
    • 2001-12-10
    • Novozymes A/S
    • Martin SchuleinHanne DelaLene Nonboe AndersenSoren Flensted LassenMarkus Sakari KauppinenLene LangeRuby Ilum NielsenShinobu TakagiMichiko Ihara
    • C12N009/44D06M016/00
    • C12N9/2437A61K38/00C05F11/08C05F17/0036C07K2319/00C11D3/386C12P19/14C12Y302/01004D06M16/003D06P5/02D06P5/158D21C5/005D21H17/005D21H21/10Y02P20/145Y02W30/43
    • 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 1 (SEQ ID NO:79) Thr Arg Xaa Xaa Asp Cys Cys Xaa Xaa 1nullnullnull2nullnullnull3nullnullnull4nullnullnull5nullnullnull6nullnullnull7nullnullnull8nullnullnull9 Xaa Cys Xaa Trp Xaa 10nullnull11nullnull12nullnull13nullnull14 and a second amino acid sequence having the following sequence 2 Trp Cys Cys Xaa Cys (SEQ ID NO:80) 1nullnullnull2nullnullnull3nullnullnull4nullnullnull5 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 good 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.
    • 本发明涉及酶制剂,其基本上由具有纤维素分解活性的酶组成,并且包含具有以下序列的第一氨基酸序列和具有以下序列的第二个氨基酸序列,其中在第一序列的3位是氨基酸 是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在工业应用中表现非常好,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服装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。