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    PROTEINS
    1.
    发明申请
    PROTEINS 有权
    蛋白质

    公开(公告)号：US20090181124A1

    公开(公告)日：2009-07-16

    申请号：US12040721

    申请日：2008-02-29

    申请人： Jorn Borch SOE ,  Jorn Dalgaard Mikkelson ,  Arno de Kreij

    发明人： Jorn Borch SOE ,  Jorn Dalgaard Mikkelson ,  Arno de Kreij

    IPC分类号： A21D2/26 ,  A23L1/32 ,  A23K1/00 ,  C12P7/64 ,  C12N9/10

    CPC分类号： C12N9/1029 ,  A21D8/042 ,  A23L15/25 ,  A23L33/10 ,  A23L33/195 ,  C11B3/003 ,  C12P7/62 ,  C12P7/6436 ,  C12P7/6454 ,  C12P7/6481

    摘要： The present invention relates to a method of producing a variant lipid acyltransferase enzyme comprising: (a) selecting a parent enzyme which is a 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; (b) modifying one or more amino acids to produce a variant lipid acyltransferase; (c) testing the variant lipid acyltransferase for 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, X, 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 following amino acid residues when aligned to SEQ ID No. 2: Ser3, Leu17, Ala114, Trp111, Tyr117, Leu118, Pro156, Gly159, Gln160, Asn161, Pro162, Ser163, Ala164, Arg165, Ser166, Gln167, Lys168, Val169, Val170, Glu171, Ala172, Tyr179, Gln182, Lys187, His180, Asn181, Met209, Leu210, Arg211, Asn215, Lys284, Met285, Gln289, Val290, Ala309, Ser310, Lys22, Met23, Gly40, Asn80, Pro81, Lys82, Val112, Asn87, Asn88, −318.

    摘要翻译： 本发明涉及一种生产变体脂质酰基转移酶的方法，包括：（a）选择脂质酰基转移酶的母体酶，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，TN，M或S; （b）修饰一个或多个氨基酸以产生变体脂质酰基转移酶; （c）测试变体脂质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L，A，V，I，F，Y，X，Q中的一个或多个 ，TN，M或S，并且其中当与SEQ ID No.2对应时，所述变体酶与任何一个或多个以下氨基酸残基的亲本序列相比，包含一个或多个氨基酸修饰：Ser3，Leu17，Ala114， Trp111，Tyr117，Leu118，Pro156，Gly159，Gln160，Asn161，Pro162，Ser163，Ala164，Arg165，Ser166，Gln167，Lys168，Val169，Val170，Glu171，Ala172，Tyr179，Gln182，Lys187，His180，Asn181，Met209，Leu210， Arg211，Asn215，Lys284，Met285，Gln289，Val290，Ala309，Ser310，Lys22，Met23，Gly40，Asn80，Pro81，Lys82，Val112，Asn87，Asn88,318。