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    • 44. 发明授权
    • Variant lipid acyltransferases and methods of making
    • 变体脂质酰基转移酶及其制备方法
    • US07906307B2
    • 2011-03-15
    • US11852274
    • 2007-09-07
    • Jørn Borch SøeJørn Dalgaard MikkelsonArno de Kreij
    • Jørn Borch SøeJørn Dalgaard MikkelsonArno de Kreij
    • G01N33/53C12Q1/34C12N9/00C12N9/16C12N9/10C07K14/00
    • C12P7/6463C12P7/6454
    • The present invention relates to a method for the production of a variant lipid acyltransferase comprising the steps of: (i) selecting a parent enzyme which is a lipid acyltranserase enzyme; (ii) modifying one or more amino acids to produce a variant lipid acyltransferase; (iii) testing the activity of the variant lipid acyltransferase on a galactolipid and/or phospholipid and/or triglyceride substrate; (iv) selecting a variant enzyme with enhanced activity towards galactolipids compared with the parent enzyme; (v) providing a Bacillus licheniformis cell; (vi) transforming the Bacillus licheniformis cell with a heterologous nucleotide sequence encoding said variant lipid acyltransferase; and (iii) expressing said variant lipid acyltransferase in the cell under the control of a promoter sequence. The variant lipid acyltransferase can undergo post-translations modification, truncation and/or clipping, i.e., to remove a signal peptide. In addition, the present invention further relates to the use of Bacillus licheniformis to express a lipid acyltransferase, a Bacillus licheniformis host cell comprising a heterologous lipid acyltransferase and a vector comprising a nucleotide sequence encoding a lipid acyltransferase operably linked to a promoter sequence homologous to B. licheniformis.
    • 本发明涉及生产变体脂质酰基转移酶的方法,包括以下步骤:(i)选择脂质酰基转移酶的亲本酶; (ii)修饰一个或多个氨基酸以产生变体脂质酰基转移酶; (iii)测试变体脂质酰基转移酶对半乳糖脂和/或磷脂和/或甘油三酯底物的活性; (iv)与亲本酶相比,选择具有增强的对半乳糖脂的活性的变体酶; (v)提供地衣芽孢杆菌细胞; (vi)用编码所述变体脂质酰基转移酶的异源核苷酸序列转化地衣芽孢杆菌细胞; 和(iii)在启动子序列控制下的细胞中表达所述变体脂质酰基转移酶。 变体脂质酰基转移酶可经历翻译后修饰,截短和/或剪切,即去除信号肽。 此外,本发明还涉及地衣芽孢杆菌表达脂质酰基转移酶,包含异源脂质酰基转移酶的地衣芽孢杆菌宿主细胞的使用和包含编码脂质酰基转移酶的核苷酸序列的载体,所述核苷酸序列可操作地连接到与B同源的启动子序列 地衣芽孢杆菌