生物学杂志

生物学杂志 ›› 2022, Vol. 39 ›› Issue (4): 18-.doi: 10.3969/j.issn.2095-1736.2022.04.018

• 研究报告 • 上一篇    下一篇

RNA支架系统介导大肠杆菌生产甲羟戊酸

  

  1. 1. 江南大学 粮食发酵工艺与技术国家工程实验室， 无锡 214122; 2. 江南大学 
    工业生物技术教育部重点实验室， 无锡 214122; 3. 江南大学 生物工程学院， 无锡 214122
  • 出版日期:2022-08-18 发布日期:2022-08-15
  • 通讯作者: 白仲虎,教授,博士生导师,主要从事发酵工程和生物医药过程工程等研究,E-mail:baizhonghu@jiangnan.edu.cn;刘春立，助理研究员，主要从事代谢工程和合成生物学研究，E-mail: liuchunli201774@126.com
  • 作者简介:董洪钢，硕士研究生，研究方向为细胞代谢工程，E-mail:deserce@163.com
  • 基金资助:
    江苏省青年基金项目（BK20190610）

Increasing mevalonate production mediated by RNA scaffolds system in Escherichia coli

  1. 1. National Engineering Laboratory of Cereal Fermentation Technology, Wuxi 214122, China; 2. Key Laboratory
    of Industrial Biotechnology, Ministry of Education, Wuxi 214122, China; 3. School of Biotechnology,
    Jiangnan University, Wuxi 214122, China
  • Online:2022-08-18 Published:2022-08-15

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摘要： 1. National Engineering Laboratory of Cereal Fermentation Technology, Wuxi 214122, China; 2. Key Laboratory
of Industrial Biotechnology, Ministry of Education, Wuxi 214122, China; 3. School of Biotechnology,
Jiangnan University, Wuxi 214122, China

关键词: 甲羟戊酸, MVA途径, RNA支架系统, 大肠杆菌, GFP

Abstract: In this study, the advantage of 0D RNA scaffold system was used to co-localize the three-step reaction of mevalonate to improve the yield of mevalonate in E. coli. Firstly, RNA-EMSA was used to demonstrate the interaction between RNA aptamers and RBDs (RNA binding domains). Subsequently, RBDs were fused with fluorescent protein to evaluate the efficiency of RNA scaffold system. Compared with that of the control strain BLCGG, the relative fluorescence intensities of BLPCG and BLCPG were increased by 126% and 129%, respectively. The fluorescence intensity of BLPPG was increased by 375% in the strain expressed 0D PP7 RNA scaffold system. Finally, mevalonate was produced by 0D RNA scaffold system. Compared with that of the control strain BLCCES, the mevalonate production of BLPPES expressing 0D PP7 RNA scaffold system was increased by 84%, achieved 3.13 g/L. Therefore, RNA scaffold system is an effective tool to improve the efficiency of multi-enzyme metabolic pathway.

Key words: mevalonate, MVA pathway, RNA scaffold system, E. coli, GFP

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