Enzymatic process optimization for the in vitro production of isoprene from mevalonate

Microbial Cell Factories

Research

Guang Zhao¹  Mo Xian¹  Hui Liu¹  Huibin Zou²  Tao Cheng³  Ningning Chen⁴  Mengxun Shi⁴  Congxia Xie⁵ 

[1] CAS Key Laboratory of Bio-based Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Laoshan District, 266101, Qingdao, China;CAS Key Laboratory of Bio-based Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Laoshan District, 266101, Qingdao, China;College of Chemical Engineering, Qingdao University of Science and Technology, 266042, Qingdao, China;CAS Key Laboratory of Bio-based Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Laoshan District, 266101, Qingdao, China;State Key Laboratory Base of Eco-Chemical Engineering, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, 266042, Qingdao, China;College of Chemical Engineering, Qingdao University of Science and Technology, 266042, Qingdao, China;State Key Laboratory Base of Eco-Chemical Engineering, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, 266042, Qingdao, China;

关键词: Mevalonate;    Isoprene;    Synthetic biochemistry;    Isoprenoids;    Bio-based chemicals;    Enzymatic process;   

DOI  :  10.1186/s12934-016-0622-4

 received in 2016-09-26, accepted in 2016-12-27,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundAs an important bulk chemical for synthetic rubber, isoprene can be biosynthesized by robust microbes. But rational engineering and optimization are often demanded to make the in vivo process feasible due to the complexities of cellular metabolism. Alternative synthetic biochemistry strategies are in fast development to produce isoprene or isoprenoids in vitro.ResultsThis study set up an in vitro enzyme synthetic chemistry process using 5 enzymes in the lower mevalonate pathway to produce isoprene from mevalonate. We found the level and ratio of individual enzymes would significantly affect the efficiency of the whole system. The optimized process using 10 balanced enzyme unites (5.0 µM of MVK, PMK, MVD; 10.0 µM of IDI, 80.0 µM of ISPS) could produce 6323.5 µmol/L/h (430 mg/L/h) isoprene in a 2 ml in vitro system. In a scale up process (50 ml) only using 1 balanced enzyme unit (0.5 µM of MVK, PMK, MVD; 1.0 µM of IDI, 8.0 µM of ISPS), the system could produce 302 mg/L isoprene in 40 h, which showed higher production rate and longer reaction phase with comparison of the in vivo control.ConclusionsBy optimizing the enzyme levels of lower MVA pathway, synthetic biochemistry methods could be set up for the enzymatic production of isoprene or isoprenoids from mevalonate.

【 授权许可】

CC BY   
© The Author(s) 2017