Liu L, Li M, Hu G, Zhang L, Xing L
Department of Microbiology, Nankai University, Tianjin 300071, China.
Wei Sheng Wu Xue Bao. 2001 Aug;41(4):397-401.
Using plasmid pTMICL6 containing delta 6-fatty-acid desaturase gene from Mortieralla isabellina M6-22 as a template, 1.38 kb DNA fragment was amplified by PCR. The fragment was subcloned into the yeast-E. coli shuttle vector pYES2.0, then an expression recombinant plamid pYMID6 containing target gene was constructed. The pYMID6 was transformed into Saccharomyces cerevisiae for expression by LiAc method. It was found to exhibit delta 6-fatty acid desaturase activity in the recombinant S. cerevisiae YMID6 in the presence of exogenous fatty acid substrate linoleic acid under introduction of GAL. Expression of the delta 6-fatty acid desaturase gene under appropriate media and temperature conditions led to the production of gamma-linolenic acid reached 8.69% of the total yeast fatty acid by GC-MS detection. It is the first report about expression of M. isabellina D6D gene in S. cerevisiae.
以含有深黄被孢霉M6 - 22的Δ6 - 脂肪酸去饱和酶基因的质粒pTMICL6为模板，通过PCR扩增出1.38 kb的DNA片段。将该片段亚克隆到酵母 - 大肠杆菌穿梭载体pYES2.0中，构建出含有目标基因的表达重组质粒pYMID6。采用LiAc法将pYMID6转化到酿酒酵母中进行表达。发现在导入GAL的情况下，重组酿酒酵母YMID6在存在外源脂肪酸底物亚油酸时表现出Δ6 - 脂肪酸去饱和酶活性。在合适的培养基和温度条件下，Δ6 - 脂肪酸去饱和酶基因的表达通过GC - MS检测导致γ-亚麻酸的产量达到酵母总脂肪酸的8.69%。这是关于深黄被孢霉D6D基因在酿酒酵母中表达的首次报道。
