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    METHOD FOR DESIGNING MUTATED ENZYME, METHOD FOR PREPARING THE SAME, AND MUTATED ENZYME
    1.
    发明申请
    METHOD FOR DESIGNING MUTATED ENZYME, METHOD FOR PREPARING THE SAME, AND MUTATED ENZYME 审中-公开
    用于设计突变酶的方法，其制备方法和突变酶

    公开(公告)号：US20090280553A1

    公开(公告)日：2009-11-12

    申请号：US12309929

    申请日：2007-06-28

    申请人： Bunzo Mikami ,  Hiroyuki Iwamoto ,  Shotaro Yamaguchi

    发明人： Bunzo Mikami ,  Hiroyuki Iwamoto ,  Shotaro Yamaguchi

    IPC分类号： C12N9/24 ,  C12N9/44 ,  C12N9/26 ,  C07H21/04 ,  C12N1/21

    CPC分类号： C12Y302/01041 ,  C07K2299/00 ,  C12N9/2451 ,  C12N9/2457 ,  C12N9/246 ,  C12Y302/01068

    摘要： It is intended to provide a novel method for improving an enzyme hydrolyzing an a-1,6-glycosidic linkage. A mutated enzyme is designed by specifying one or more amino acids selected from the group shown below in an amino acid sequence of an enzyme (an enzyme to be mutated) that hydrolyzes an a-1,6-glycosidic linkage, that is, the group consisting of an amino acid corresponding to an amino acid at the 292 position, an amino acid corresponding to an amino acid at the 371 position, an amino acid corresponding to an amino acid at the 406 position, an amino acid corresponding to an amino acid at the 407 position, an amino acid corresponding to an amino acid at the 437 position, an amino acid corresponding to an amino acid at the 465 position, an amino acid corresponding to an amino acid at the 475 position, an amino acid corresponding to an amino acid at the 476 position; an amino acid corresponding to an amino acid at the 525 position, an amino acid corresponding to an amino acid at the 526 position, an amino acid corresponding to an amino acid at the 580 position and an amino acid corresponding to an amino acid at the 582 position of the amino acid represented by in SEQ ID NO: 2 (step (1)) and constructing an amino acid sequence in which the amino acid(s) specified in the step (1) is/are substituted with another amino acid or deleted based on the amino acid sequence of the enzyme to be mutated (step (2)).

    摘要翻译： 旨在提供一种改善水解a-1,6-糖苷键的酶的新方法。 突变酶通过在水解a-1,6-糖苷键的酶（待突变的酶）的氨基酸序列中指定一个或多个选自下文所示的氨基酸来设计，即，该基团 由对应于292位氨基酸的氨基酸，对应于371位氨基酸的氨基酸，对应于406位氨基酸的氨基酸，对应于氨基酸的氨基酸 407位，对应于437位氨基酸的氨基酸，对应于465位氨基酸的氨基酸，对应于475位氨基酸的氨基酸，对应于氨基酸的氨基酸 酸在476位; 对应于525位氨基酸的氨基酸，对应于526位氨基酸的氨基酸，对应于580位氨基酸的氨基酸和对应于582位氨基酸的氨基酸 并且构建其中在步骤（1）中规定的氨基酸被其它氨基酸所取代的氨基酸序列的氨基酸序列（步骤（1））所示的氨基酸的位置） 基于要突变的酶的氨基酸序列（步骤（2））。

    METHOD FOR DESIGNING MUTANT ENZYME, METHOD FOR PREPARING MUTANT ENZYME, AND MUTANT ENZYME
    2.
    发明申请
    METHOD FOR DESIGNING MUTANT ENZYME, METHOD FOR PREPARING MUTANT ENZYME, AND MUTANT ENZYME 有权

    公开(公告)号：US20110165605A1

    公开(公告)日：2011-07-07

    申请号：US13062798

    申请日：2009-08-12

    申请人： Ryota Hashizume ,  Bunzo Mikami ,  Hirotaka Matsubara ,  Akiko Matsunaga ,  Shotaro Yamaguchi

    发明人： Ryota Hashizume ,  Bunzo Mikami ,  Hirotaka Matsubara ,  Akiko Matsunaga ,  Shotaro Yamaguchi

    IPC分类号： C12Q1/34 ,  C12N9/00 ,  C12N9/80 ,  C12N9/78 ,  C07H21/04 ,  C12N1/00

    CPC分类号： C12N9/80 ,  C12N9/48

    摘要： An object is to provide a novel method of improving an enzyme capable of deamidating a protein. A mutant enzyme is designed by the following steps: (1) specifying one or more amino acids selected from the following group, namely, consisting of an amino acid corresponding to the amino acid at position 35, an amino acid corresponding to the amino acid at position 38, an amino acid corresponding to the amino acid at position 39, an amino acid corresponding to the amino acid at position 40, an amino acid corresponding to the amino acid at position 41, an amino acid corresponding to the amino acid at position 42, an amino acid corresponding to the amino acid at position 43, an amino acid corresponding to the amino acid at position 45, an amino acid corresponding to the amino acid at position 46, an amino acid corresponding to the amino acid at position 49, an amino acid corresponding to the amino acid at position 79, an amino acid corresponding to the amino acid at position 80, an amino acid corresponding to the amino acid at position 81, an amino acid corresponding to the amino acid at position 82, an amino acid corresponding to the amino acid at position 83, an amino acid corresponding to the amino acid at position 84, an amino acid corresponding to the amino acid at position 103, an amino acid corresponding to the amino acid at position 104, an amino acid corresponding to the amino acid at position 105, an amino acid corresponding to the amino acid at position 106, an amino acid corresponding to the amino acid at position 117, an amino acid corresponding to the amino acid at position 142, an amino acid corresponding to the amino acid at position 143, an amino acid corresponding to the amino acid at position 146, an amino acid corresponding to the amino acid at position 166, and an amino acid corresponding to the amino acid at position 185 in an amino acid sequence set forth in SEQ ID NO: 2, in a protein deamidase (an enzyme to be mutated); and (2) constructing an amino acid sequence having substitution of the amino acid(s) specified in the step (1) by another amino acid(s) or having deletion of the amino acid(s) specified in the step (1) using the amino acid sequence for an enzyme to be mutated as a base sequence.

    Method for designing mutant protein deamidase
    3.
    发明授权
    Method for designing mutant protein deamidase 有权
    设计突变蛋白脱酰胺酶的方法

    公开(公告)号：US08735131B2

    公开(公告)日：2014-05-27

    申请号：US13062798

    申请日：2009-08-12

    申请人： Ryota Hashizume ,  Bunzo Mikami ,  Hirotaka Matsubara ,  Akiko Matsunaga ,  Shotaro Yamaguchi

    发明人： Ryota Hashizume ,  Bunzo Mikami ,  Hirotaka Matsubara ,  Akiko Matsunaga ,  Shotaro Yamaguchi

    IPC分类号： C12P21/00 ,  C12N15/00 ,  C12N9/78

    CPC分类号： C12N9/80 ,  C12N9/48

    摘要： An object is to provide a novel method for improving an enzyme capable of deamidating a protein. A mutant protein deamidase is designed by the following steps: (A) identifying one or more amino acid in an amino acid sequence for a protein deamidase which corresponds to the amino acid at position 35, 38 to 43, 45, 46, 49, 79 to 84, 103 to 106, 117, 142, 143, 146, 166, or 185 in the amino acid sequence set forth in SEQ ID NO: 2; and (B) constructing a mutant amino acid sequence of the protein deamidase by substituting the one or more amino acid identified in step (A) with another amino acid or other amino acids or by deleting the one or more amino acid identified in step (A).

    摘要翻译： 目的是提供一种改善能够使蛋白质脱酰胺化的酶的新方法。 通过以下步骤设计突变型蛋白质脱酰胺酶：（A）鉴定氨基酸序列中一个或多个对应于第35,38,43,45,46,49,79位的氨基酸的蛋白质脱酰胺酶的氨基酸 至SEQ ID NO：2所示氨基酸序列中的84,103至106,117,142,143,146,166或185; 和（B）通过用步骤（A）中鉴定的一个或多个氨基酸用另一个氨基酸或其他氨基酸取代蛋白质脱酰胺酶的突变氨基酸序列，或通过删除步骤（A）中鉴定的一个或多个氨基酸 ）。