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    • 7. 发明授权
    • Mutant fatty acid desaturase
    • 突变型脂肪酸去饱和酶
    • US06686186B2
    • 2004-02-03
    • US09988929
    • 2001-12-03
    • John ShanklinEdgar B. Cahoon
    • John ShanklinEdgar B. Cahoon
    • C12N902
    • C12N9/0083C12N15/8247
    • The present invention relates to a method for producing mutants of a fatty acid desaturase having a substantially increased activity towards fatty acid substrates with chains containing fewer than 18 carbons relative to an unmutagenized precursor desaturase having an 18 carbon atom chain length substrate specificity. The method involves inducing one or more mutations in the nucleic acid sequence encoding the precursor desaturase, transforming the mutated sequence into an unsaturated fatty acid auxotroph cell such as MH13 E. coli, culturing the cells in the absence of supplemental unsaturated fatty acids, thereby selecting for recipient cells which have received and which express a mutant fatty acid desaturase with an elevated specificity for fatty acid substrates having chain lengths of less than 18 carbon atoms. A variety of mutants having 16 or fewer carbon atom chain length substrate specificities are produced by this method. Mutant desaturases produced by this method can be introduced via expression vectors into prokaryotic and eukaryotic cells and can also be used in the production of transgenic plants which may be used to produce specific fatty acid products.
    • 本发明涉及一种生产脂肪酸去饱和酶突变体的方法,该突变体相对于具有18个碳原子链长基底特异性的未成熟前体去饱和酶,具有对具有少于18个碳的链的脂肪酸底物具有显着增加的活性。 该方法包括在编码前体去饱和酶的核酸序列中诱导一个或多个突变,将突变的序列转化为不饱和脂肪酸营养缺陷型细胞如MH13大肠杆菌,在不存在补充不饱和脂肪酸的情况下培养细胞,从而选择 对于接受并且表达具有链长度小于18个碳原子的脂肪酸底物具有特异性提高的突变脂肪酸去饱和酶的受体细胞。 通过该方法制备具有16个或更少碳原子链长基底特异性的各种突变体。 通过该方法产生的突变型去饱和酶可以通过表达载体导入原核和真核细胞中,并且还可用于生产可用于产生特定脂肪酸产物的转基因植物。