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    PROTEINS
    6.
    发明申请
    PROTEINS 审中-公开
    蛋白质

    公开(公告)号：WO2005066347A1

    公开(公告)日：2005-07-21

    申请号：PCT/IB2004/004378

    申请日：2004-12-23

    申请人： DANISCO A/S ,  KELLET-SMITH, Anja Hemmingsen ,  LORENTSEN, Rikke, Høegh ,  SØE, jørn, Borch ,  MIKKELSON, Jørn, Dalgaard ,  KREIJ, Arno de

    发明人： KELLET-SMITH, Anja Hemmingsen ,  LORENTSEN, Rikke, Høegh ,  SØE, jørn, Borch ,  MIKKELSON, Jørn, Dalgaard ,  KREIJ, Arno de

    IPC分类号： C12N15/54

    CPC分类号： C12N9/1029 ,  C12Y203/01085

    摘要： The present invention relates to a method of producing a variant glycolipid acyltransferase enzyme comprising: (a) selecting a parent enzyme which is a glycolipid acyltransferase enzyme characterised in that the enzyme comprises the amino acid sequence motif GDSX, wherein X is one or more of the following amino acid residues L, A, V, I, F, Y, H, Q, T, N, M or S; (b) modifying one or more amino acids to produce a variant glycolipid acyltransferase; (c) testing the variant glycolipid acyltransferase for transferase activity, an optionally hydrolytic activity, on a galactolipid substrate, and optionally a phospholipid substrate and/or optionally a triglyceride substrate; (d) selecting a variant enzyme with an enhanced activity towards galactolipids compared with the parent enzyme; and optionally (e) preparing a quantity of the variant enzyme. The present invention further relates to variant lipid acyltransferase enzyme characterised in that the enzyme comprises the amino acid sequence motif GDSX, wherein X is one or more of the following amino acid residues L, A, V, I, F, Y, H, Q, T, N, M or S, and wherein the variant enzyme comprises one or more amino acid modifications compared with a parent sequence at any one or more of the amino acid residues defined in set 2, set 4, set 6 or set 7.

    摘要翻译： 本发明涉及一种生产变体糖脂酰基转移酶的方法，包括：（a）选择亲糖基酶，其为糖脂酰基转移酶，其特征在于该酶包含氨基酸序列基序GDSX，其中X是一个或多个 以下氨基酸残基L，A，V，I，F，Y，H，Q，T，N，M或S; （b）修饰一个或多个氨基酸以产生变体糖脂酰基转移酶; （c）测试变体糖脂酰基转移酶对半乳糖脂底物和任选的磷脂底物和/或任选的甘油三酯底物的转移酶活性，任选水解活性; （d）与母体酶相比，选择对半乳糖脂增强活性的变体酶; 和（e）制备一定量的变体酶。 本发明还涉及变体脂质酰基转移酶，其特征在于该酶包含氨基酸序列基序GDSX，其中X是以下氨基酸残基中的一个或多个L，A，V，I，F，Y，H，Q ，T，N，M或S，并且其中所述变体酶与第2组，第4组，第6组或第7组中定义的任何一个或多个氨基酸残基与母体序列相比包含一个或多个氨基酸修饰。