产玉米黄质的人工酵母细胞的构建

doi:10.13523/j.cb.20160809

中国生物工程杂志

2016, Vol. 36

Issue (8): 64-72 DOI: 10.13523/j.cb.20160809

技术与方法

产玉米黄质的人工酵母细胞的构建

梅雪昂¹, 陈艳¹, 王瑞钊¹, 肖文海^1,2, 王颖^1,2, 李霞^1,2, 元英进^1,2

1. 天津大学化工学院制药工程系系统生物工程教育部重点实验室天津 300072;
2. 天津化学化工协同创新中心合成生物学平台天津 300072

Engineered Yeast Cell for Producing Zeaxanthin

MEI Xue-ang¹, CHEN Yan¹, WANG Rui-zhao¹, XIAO Wen-hai^1,2, WANG Ying^1,2, LI Xia^1,2, YUAN Ying-jin^1,2

1. Department of Pharmaceutical Engineering, School of Chemical Engineering and Technology, Tianjin University;Key Laboratory of Systems Bioengineering, Ministry of Education, Tianjin 300072, China;
2. SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering, School of Chemical Engineering and Technology, Tianjin 300072, China

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摘要：

为了实现微生物异源合成天然类胡萝卜素玉米黄质，以一株产β-胡萝卜素的酿酒酵母为底盘细胞，利用合成生物学技术构建人工酵母细胞。通过在染色体整合玉米黄质生物合成关键酶-β-胡萝卜素羟化酶（CrtZ），并对其9种来源进行筛选，发现整合欧文氏菌来源的β-胡萝卜素羟化酶的菌株获得玉米黄质的最高产量。法尼基焦磷酸（FPP）作为合成萜烯类天然产物的重要前体，通过敲除 Lpp1和Dpp1 基因，削减法尼基焦磷酸向法呢醇的转化，为玉米黄质的合成提供更多的前体，使玉米黄质的产量提高了1.27倍（从29 mg/L提高到36.8 mg/L）。在此基础上，通过增加欧文氏菌来源CrtZ的基因拷贝数及调节其启动子的强弱来增强β-胡萝卜素羟化酶的表达强度，使得玉米黄质的摇瓶产量达到96.2 mg/L，是目前公开报道中产量最高的。

关键词： 玉米黄质; 酿酒酵母; 合成生物学; 基因工程

Abstract:

For heterologous synthesize natural carotenoid zeaxanthin in microorganism,a β-carotene producing Saccharomyces cerevisiae was chozen as the host cell to construct engineered yeast with synthetic biology method. The key enzymes β-carotene hydroxylase encoding gene CrtZ from nine sources were integrated in the chromosomal, separately. As the result, the strain carrying CrtZ from Erwinia uredovora achieved the highest titer of zeaxanthin. Moreover, the conversion from farnesyl pyrophosphate (an important precursor for terpenoid natural products) to farnesol was reduced by knocking out gene Lpp1 and Dpp1, providing more precursors for zeaxanthin synthesis. The zeaxanthin yield increased 1.27 fold (from 29 mg/L to 36.8 mg/L) accordingly. Furthermore, a titer of zeaxanthin was achieved as 96.2 mg/L in shake-flask through increasing the CrtZ gene copy number and regulating its promoter's activity,which is the highest reported microbial titer known.

Key words: Synthetic biology Zeaxanthin Gene engineering Saccharomyces cerevisiae

收稿日期: 2016-01-13 出版日期: 2016-03-16

ZTFLH:

Q789

基金资助:

国家高技术研究发展计划资助项目（2012AA02A701）

通讯作者: 李霞 E-mail: lixia01@tju.edu.cn

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引用本文:

梅雪昂, 陈艳, 王瑞钊, 肖文海, 王颖, 李霞, 元英进. 产玉米黄质的人工酵母细胞的构建[J]. 中国生物工程杂志, 2016, 36(8): 64-72.

MEI Xue-ang, CHEN Yan, WANG Rui-zhao, XIAO Wen-hai, WANG Ying, LI Xia, YUAN Ying-jin. Engineered Yeast Cell for Producing Zeaxanthin. China Biotechnology, 2016, 36(8): 64-72.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20160809 或 https://manu60.magtech.com.cn/biotech/CN/Y2016/V36/I8/64

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