大肠杆菌中从头合成白藜芦醇途径的设计及优化

doi:10.13523/j.cb.20140212

中国生物工程杂志

2014, Vol. 34

Issue (2): 71-77 DOI: 10.13523/j.cb.20140212

技术与方法

大肠杆菌中从头合成白藜芦醇途径的设计及优化

汪建峰^1,2, 张嗣良¹, 王勇²

1. 华东理工大学生物反应器工程国家重点实验室上海 200237;
2. 中国科学院上海生命科学研究院植物生理生态研究所合成生物学重点实验室上海 200032

Pathway Assembly and Optimization in E. coli for de Novo Biosynthesis of Resveratrol

WANG Jian-feng^1,2, ZHANG Si-liang¹, WANG Yong²

1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China;
2. Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China

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摘要：

白藜芦醇是一种极具药用价值的植物源芪类化合物。为了在E. coli实现白藜芦醇的从头合成，构建了由酪氨酸解氨酶（TAL），香豆酸-CoA合成酶（4CL）和白藜芦醇合成酶（STS）组成的非天然合成途径。经3天发酵后，白藜芦醇产量仅为2.67 mg/L，而其中间体香豆酸的积累达到了95.64 mg/L。为了进一步改善异源途径的效率，对4CL和STS模块采取融合表达、高拷贝表达及启动子工程改造的策略，最终使白藜芦醇产量提高到了9.6倍，达到了25.76 mg/L，同时香豆酸的积累减少到了20.38 mg/L。这些研究结果为更高效白藜芦醇从头合成工程菌的构建及最终实现白藜芦醇的微生物大规模生产奠定了基础。

关键词： 大肠杆菌; 白藜芦醇; 融合表达; 高拷贝表达; 启动子工程

Abstract:

Resveratrol is an important plant stilbene with considerable pharmaceutical values. To achieve the de novo biosynthesis of resveratrol in E. coli, a heterologous resveratrol biosynthetic pathway consisting of tyrosine ammonia lyase (TAL), coumaroyl-CoA synthase (4CL) and stilbene synthase (STS) was constructed. The engineered strain only produced 2.67 mg/L resveratrol after 3 days cultivation. To improve the efficiency of heterologous pathway, the 4CL and STS modules were further engineered using strategies of fusion expression, high-copy expression and promoter engineering. Eventually, the yield of resveratrol reached 25.76 mg/L with a 9.6-fold improvement compared with the initial strain. The useful information for the construction of more efficient recombinant resveratrol producer was provided, and it laid a foundation for the large-scale production of resveratrol through microbial fermentation.

Key words: Escherichia coli Resveratrol Fusion expression High-copy expression Promoter engineering

收稿日期: 2013-10-15 出版日期: 2014-02-25

ZTFLH:

Q819

基金资助:

国家“973”计划（2012C13721104）、国家自然科学基金（31170101，31100073）、中国科学院知识创新项目（ICSCXZ-EW-J-12）资助项目

通讯作者: 王勇 E-mail: yongwang@sibs.ac.cn

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引用本文:

汪建峰, 张嗣良, 王勇. 大肠杆菌中从头合成白藜芦醇途径的设计及优化[J]. 中国生物工程杂志, 2014, 34(2): 71-77.

WANG Jian-feng, ZHANG Si-liang, WANG Yong. Pathway Assembly and Optimization in E. coli for de Novo Biosynthesis of Resveratrol. China Biotechnology, 2014, 34(2): 71-77.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20140212 或 https://manu60.magtech.com.cn/biotech/CN/Y2014/V34/I2/71

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