代谢工程改造大肠杆菌对中碳脂肪酸生产的加强作用

doi:10.7506/spkx1002-6630-201321027

食品科学 ›› 2013, Vol. 34 ›› Issue (21): 131-135.doi: 10.7506/spkx1002-6630-201321027

代谢工程改造大肠杆菌对中碳脂肪酸生产的加强作用

刘强1,2，郑艳宁2,3，姜兴林2,3，刘东杰1，秦文1,*，咸漠2

1.四川农业大学食品学院，四川雅安 625014；2.中国科学院青岛生物能源与过程研究所，山东青岛 266101；
3.中国科学院大学，北京 100049

收稿日期:2012-09-26 修回日期:2013-10-11 出版日期:2013-11-15 发布日期:2013-10-28
通讯作者: 秦文 E-mail:qinwen1967@yahoo.com.cn
基金资助:
国家自然科学基金项目(21106170)；中国科学院知识创新工程重要方向性项目(KSCX2-EW-G-13)；
广东省中国科学院全面战略合作项目(2009B091300146)

Metabolic Engineering of Escherich coli to Enhance the Production of Medium-chain Fatty Acids

LIU Qiang1,2，ZHENG Yan-ning2,3，JIANG Xing-lin2,3，LIU Dong-jie1，QIN Wen1,*，XIAN Mo2

1. College of Food Science, Sichuan Agricultural University, Ya’an 625014, China；2. Qingdao Institute of Biomass Energy and Bioprocess
Technology, Chinese Academy of Sciences, Qingdao 266101, China；3. University of Chinese Academy of Sciences, Beijing 100049, China

Received:2012-09-26 Revised:2013-10-11 Online:2013-11-15 Published:2013-10-28
Contact: qin wen E-mail:qinwen1967@yahoo.com.cn

摘要/Abstract

摘要：

通过在大肠杆菌(Escherichia coli)中过量表达来自加州月桂(Umbellulana california)的硫酯酶基因BTE，极大地加强了大肠杆菌中碳脂肪酸(MCFAs)的生产。并对MCFAs在大肠杆菌胞内外的分布，以及是否对大肠杆菌细胞膜磷脂组成产生影响等进行了系统的分析。结果发现，产生的MCFAs绝大部分存在于大肠杆菌细胞内，且未对其细胞膜磷脂组成产生明显影响。为了继续提高MCFAs的产量，又进一步对大肠杆菌进行了代谢工程改造，通过结合表达乙酰-CoA羧化酶(ACCase)，优化硫酯酶的表达水平等，最终获得了188.9mg/L的总MCFAs产量，并且产生的MCFAs的比例高达85%以上，实现了MCFAs的高特异性生产。

关键词: 中碳脂肪酸, 大肠杆菌, 硫酯酶, 代谢工程

Abstract:

Medium-chain fatty acids (MCFAs) are the major components of medium-chain triglycerides (MCT), which
play an important role in the food additive industry. In this study, the production of MCFAs was greatly enhanced by
overexpressed thioesterase gene of Umbellulana california (BTE) in E. coli. In addition, the location of the produced
MCFAs and the effect of synthesized MCFAs on the composition of membrane phospholipids were also determined. The
results indicated that MCFAs were mainly located in E. coli, and their production had little effect on the composition of
membrane phospholipids. To further improve the production of MCFAs, strategies including co-overexpression of acetyl-
CoA carboxylase (ACCase) and the optimization of the expression level of thioesterase BTE were also carried out. Finally,
188.9 mg/L MCFAs were achieved, representing a rather high production of MCFAs. Meanwhile, the MCFAs accounted for
over 85% of the total free fatty acids, suggesting the high-specificity production of MCFAs.

Key words: medium-chain fatty acid, Escherichia coli, thioesterase, metabolic engineering

中图分类号:

Q815

刘强1,2，郑艳宁2,3，姜兴林2,3，刘东杰1，秦文1,*，咸漠2. 代谢工程改造大肠杆菌对中碳脂肪酸生产的加强作用[J]. 食品科学, 2013, 34(21): 131-135.

LIU Qiang1,2，ZHENG Yan-ning2,3，JIANG Xing-lin2,3，LIU Dong-jie1，QIN Wen1,*，XIAN Mo2. Metabolic Engineering of Escherich coli to Enhance the Production of Medium-chain Fatty Acids[J]. FOOD SCIENCE, 2013, 34(21): 131-135.

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代谢工程改造大肠杆菌对中碳脂肪酸生产的加强作用

Metabolic Engineering of Escherich coli to Enhance the Production of Medium-chain Fatty Acids

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