高山被孢霉二酰甘油酰基转移酶1的鉴定和功能表征
首发时间:2019-02-06
摘要:高山被孢霉是商业化生产花生四烯酸的产油微生物,其脂质主要以甘油三酯形式储存,然而至今还未见关于该菌株中甘油三酯合成途径的关键限速酶二酰甘油酰基转移酶1(DGAT1)功能特性的研究报道。为研究高山被孢霉甘油三酯合成过程中该关键酶的功能,本文通过同源性比对筛选出编码高山被孢霉中DGAT1的基因Madgat1a,通过在酿酒酵母中异源表达鉴定其蛋白表达和重组菌脂质合成情况。结果表明,Madgat1a表现出催化甘油三酯合成的功能,重组菌中甘油三酯含量提升7.2倍,总脂肪酸含量提升2.4倍。该结果为通过基因工程进一步提升高山被孢霉等产油微生物脂质含量提供理论支持。
关键词: 产油微生物 高山被孢霉 二酰甘油酰基转移酶 脂肪酸 甘油三酯 异源表达
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Identification and functional characterization of diacylglycerol acyltransferase 1 from Mortierella alpina
Abstract:Mortierella alpina is an oil-producing microorganism for the commercial production of arachidonic acid. Its lipids are mainly stored in the form of triglycerides. However, there have been no reports on the functional properties of the diacylglycerol acyltransferase 1 (DGAT1) , which is the key rate-limiting enzyme in the triglyceride synthesis pathway in this strain. In order to study the function of DGAT1 in the synthesis of triglyceride of Mortierella alpina, this study screened the homologous gene Madgat1 by homology alignment, and identify its protein expression and its impact on lipid synthesis by heterologous expression in S. cerevisiae. As a result, Madgat1 showed the function of catalyzing the synthesis of TAG. The TAG content of the recombinants was increased by 7.2 times and the total fatty acid content was increased by 2.4 times. The results provide theoretical support for further improving the lipid production of oil-producing microorganisms such as Mortierella alpina by genetic engineering.
Keywords: Oil-producing microorganisms Mortierella alpina diacylglycerol acyltransferase fatty acid triglyceride heterologous expression
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