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    • 4. 发明申请
    • AMYLASE VARIANTS
    • WO1996023873A1
    • 1996-08-08
    • PCT/DK1996000056
    • 1996-02-05
    • NOVO NORDISK A/SBISGÅRD-FRANTZEN, HenrikSVENDSEN, AllanBORCHERT, Torben, Vedel
    • NOVO NORDISK A/S
    • C12N09/28
    • C12C5/004C11D3/386C11D3/38672C11D3/38681C12N9/2417D06L1/14D21C5/005D21C5/022D21C5/027D21H17/005D21H17/28Y02E50/17Y02W30/648
    • A variant of a parent alpha -amylase, which parent alpha -amylase (i) has an amino acid sequence selected from the amino acid sequences shown in SEQ ID No. 1, SEQ ID No. 2, SEQ ID No. 3, and SEQ ID No. 7, respectively; or (ii) displays at least 80 % homology with one or more of these amino acid sequences; and/or displays immunological cross-reactivity with an antibody raised against a alpha -amylase having one of these amino acid sequences; and/or is encoded by a DNA sequence which hybridizes with the same probe as a DNA sequence encoding a alpha -amylase having one of these amino acid sequences; in which variant: (a) at least one amino acid residue of the parent alpha -amylase has been deleted; and/or (b) at least one amino acid residue of the parent alpha -amylase has been replaced by a different amino acid residue; and/or (c) at least one amino acid residue has been inserted relative to the parent alpha -amylase; the variant having alpha -amylase activity and exhibiting at least one of the following properties relative to the parent alpha -amylase: increased thermostability; increased stability towards oxidation; and reduced Ca dependency; with the proviso that the amino acid sequence of the variant is not identical to any of the amino acid sequences shown in SEQ ID No. 1, SEQ ID No. 2, SEQ ID No. 3 and SEQ ID No. 7, respectively.
    • 母体α-淀粉酶的变体,其母体α-淀粉酶(i)具有选自SEQ ID No.1,SEQ ID No.2,SEQ ID No.3和SEQ SEQ ID No.3所示的氨基酸序列的氨基酸序列 编号7; 或(ii)与这些氨基酸序列中的一个或多个显示至少80%的同源性; 和/或显示与针对具有这些氨基酸序列之一的α-淀粉酶产生的抗体的免疫交叉反应性; 和/或由与编码具有这些氨基酸序列之一的α-淀粉酶的DNA序列相同的探针杂交的DNA序列编码; 在该变体中:(a)母体α-淀粉酶的至少一个氨基酸残基已被缺失; 和/或(b)母体α-淀粉酶的至少一个氨基酸残基被不同的氨基酸残基替代; 和/或(c)相对于母体α-淀粉酶插入至少一个氨基酸残基; 所述变体具有α-淀粉酶活性并且相对于母体α-淀粉酶显示出至少一种以下性质:增加的热稳定性; 提高氧化稳定性; 并减少Ca 2+依赖性; 条件是变体的氨基酸序列分别与SEQ ID No.1,SEQ ID No.2,SEQ ID No.3和SEQ ID No.7所示的任何氨基酸序列不同。
    • 9. 发明申请
    • A METHOD OF DESIGNING ALPHA-AMYLASE MUTANTS WITH PREDETERMINED PROPERTIES
    • 设计具有预定特性的α-淀粉酶突变体的方法
    • WO1996023874A1
    • 1996-08-08
    • PCT/DK1996000057
    • 1996-02-05
    • NOVO NORDISK A/SSVENDSEN, AllanBISGÅRD-FRANTZEN, HenrikBORCHERT, Torben, Vedel
    • NOVO NORDISK A/S
    • C12N09/28
    • C12N9/2417C11D3/386
    • A method of constructing a variant of a parent Termamyl-like alpha -amylase, which variant has alpha -amylase activity and at least one altered property as compared to the parent alpha -amylase, comprises i) analysing the structure of the parent Termamyl-like alpha -amylase to identify at least one amino acid residue or at least one structural part of the Termamyl-like alpha -amylase structure, which amino acid residue or structural part is believed to be of relevance for altering the property of the parent Termamyl-like alpha -amylase (as evaluated on the basis of structural or functional considerations), ii) constructing a Termamyl-like alpha -amylase variant, which as compared to the parent Termamyl-like alpha -amylase, has been modified in the amino acid residue or structural part identified in i) so as to alter the property, and iii) testing the resulting Termamyl-like alpha -amylase variant for the property in question.
    • 构建母体类戊酰样α-淀粉酶的变体的方法,该变体与亲本α-淀粉酶相比具有α-淀粉酶活性和至少一种改变的性质,其包括:i)分析亲本类戊酰样的结构 以鉴定至少一个氨基酸残基或至少一个类甲酰样α-淀粉酶结构的结构部分,该氨基酸残基或结构部分被认为与改变母体类戊酰样物质的性质相关 α-淀粉酶(基于结构或功能考虑评估),ii)构建与母体类戊酰样α-淀粉酶相比的类戊酰样α-淀粉酶变体,其在氨基酸残基或 在i)中鉴定的结构部分,以便改变该性质,和iii)测试所得到的类戊酰样α-淀粉酶变体。