生物合成7-脱氢胆甾醇酵母底盘的初探

摘要

7-脱氢胆甾醇是生产维生素D₃的关键前体，目前利用化学合成和生物转化生产7-脱氢胆甾醇的方法无法适应大规模工业生产的需要，合成生物学的出现为次级代谢物的异源合成提供了新的选择，酿酒酵母和解脂酵母都是适用于工业生产的底盘生物。在酿酒酵母中，麦角固醇合成途径会抑制7-脱氢胆甾醇的合成，通过敲除麦角固醇合成途径上的非必需基因erg5并引入人源的C24还原酶基因dhcr24，可以实现在酿酒酵母中生产7-脱氢胆甾醇，摇瓶发酵产量为2.62 mg·L^-1。解脂酵母中7-脱氢胆甾醇的合成不受麦角固醇的影响，通过将人源的dhcr24基因整合到基因组rDNA位点上，7-脱氢胆甾醇的产量最高达到27.91 mg·L^-1。为寻找适合于异源表达甾醇类次级代谢物的酵母底盘提供了可供借鉴的研究方法，具有一定的指导意义。

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	苏皖
	刘悦
	周晓
	王颖

关键词： 7-脱氢胆甾醇；合成生物学；底盘细胞；酵母

Abstract：

7-Dehydrocholesterol is an important intermediate in synthesizing vitamin D₃, while the chemical synthesis and the biological transformation techniques could not satisfy the requirement of industrial production so far. The emerge of synthetic biology has provided another choice for the heterologous synthesis of secondary metabolites. Saccharomyces cerevisiae and Yarrowia lipolytica are model organisms in industry. The ergosterol biosynthesis may suppress the 7-dehydrocholesterol production in S.cerevisiae. The combined strategy of knocking out erg5, a non-essential gene for ergosterol synthesis, and introducing human dhcr24 gene encoding C24 reductase resulted in 2.62 mg·L^-1 7-dehydrocholesterol production in shake flask culture. The ergosterol did not show any influence on the synthesis of 7-dehydrosterol in Y.lipolytica. After integrating the human dhcr24 gene into rDNA site of the genome, the highest production of 7-dehydrosterol in Y.lipolytica was 27.91 mg·L^-1. The results in this study provide a reference of the discovery of suitable chassis for the biosynthesis of sterol metabolites.

Keywords： 7-dehydrocholesterol； synthetic biology； chassis； yeast

Received 2015-05-19;

Corresponding Authors: 周晓,E-mail:zhouriyao@tju.edu.cn。 Email: zhouriyao@tju.edu.cn

About author: 苏皖(1989-),女,硕士研究生,现从事生物制药方面的研究。

引用本文:

苏皖, 刘悦, 周晓, 王颖.生物合成7-脱氢胆甾醇酵母底盘的初探[J]. 化学工业与工程, 2017,34(4): 83-90

Su Wan, Liu Yue, Zhou Xiao, Wang Ying.The Exploration of Yeast Chassis for 7-Dehydrocholesterol Biosynthesis[J]. Chemcial Industry and Engineering, 2017,34(4): 83-90

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