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以4-香豆酸为底物转化白藜芦醇大肠杆菌工程菌的构建
翟逸1,李明1,赵秀敏1,张晓利1,南化征1,蒋龙声2,江旭3,阮志勇3
1.山东博奥克生物科技有限公司 重组蛋白基因检测技术国家地方联合工程实验室,山东 聊城 252000;2.宜宾山勾勾农业科技有限公司,四川 宜宾 644000;3.中国农业科学院 农业资源与农业区划研究所,北京 100081
摘要:
白藜芦醇具有多种生物活性,在食品、化妆品及生物制药中具有广阔的应用前景。利用合成生物学生产白藜芦醇是未来的发展趋势。将白藜芦醇合酶(resveratrol synthase,RS)和4-香豆酰-CoA连接酶(4-coumaric acid-CoA ligase,4CL)的编码基因整合到共表达载体pETDuet-1上成功构建了重组质粒pETDuet-rs-4cl,转化大肠杆菌BL21(DE3),在M9培养基中添加4-香豆酸,在诱导条件下培养,发酵液中白藜芦醇含量为8.6 mg/L。本研究在大肠杆菌中实现了白藜芦醇的转化,为微生物法生产白藜芦醇奠定了基础。
关键词:  白藜芦醇  大肠杆菌  重组工程菌  微生物发酵
DOI:10.14188/j.ajsh.2021.02.013
分类号:Q789
基金项目:山东博奥克生物科技有限公司博士后项目(bsh151210);宜宾市科技计划项目(2019YG01)
Construction of engineered Escherichia coli for resveratrol synthesis using 4-coumaric acid as substrate
ZHAI Yi1, LI Ming1, ZHAO Xiumin1, ZHANG Xiaoli1, NAN Huazheng1, JIANG Longsheng2, JIANG Xu3, RUAN Zhiyong3
1.State and Local Joint Engineering Laboratory of Recombinant Protein and Gene Detection Technology, Shandong Boaoke Biotechnology Co., LTD, Liaocheng 252000, Shandong, China;2.Yibin Shan Gougou Agricultural Science and Technology Co. Ltd, Yibin 644000, Sichuan, China;3.Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Abstract:
Resveratrol has multiple biological activities and broad application prospects in food, cosmetics and biopharmaceutical industries. The production of resveratrol using synthetic biology is the trend of development. Resveratrol synthase encoding gene rs and 4-coumaric acid-CoA ligase encoding gene 4cl were inserted into co-expression vector pETDuet-1 and recombinant vector pETDuet-rs-4cl was successfully constructed. The recombinant vector was transformed into Escherichia coli BL21(DE3). The 8.6 mg/L of resveratrol was determined in M9 fermentation broth with additional 4-coumaric acid and inducer. In this study, resveratrol is synthesized in E.coli, which lays a foundation for the production of resveratrol by microbial method.
Key words:  resveratrol  Escherichia coli  recombinant strain  microbial fermentation