Isoprene production by Escherichia coli through the exogenous mevalonate pathway with reduced formation of fermentation byproducts

Microbial Cell Factories

Research

Ju-Eon Park¹  Seon-Yeong Jo¹  Myeong-Seok Cha¹  Chonglong Wang¹  Seon-Won Kim¹  Hui-Jung Jang²  Jung-Hun Kim³  Eui-Sung Choi⁴ 

[1] Division of Applied Life Science (BK21 Plus), PMBBRC, Institute of Agricultural and Life Science, Gyeongsang National University, 52828, Jinju, South Korea;Division of Applied Life Science (BK21 Plus), PMBBRC, Institute of Agricultural and Life Science, Gyeongsang National University, 52828, Jinju, South Korea;Life Science Research Institute, Daewoong Pharmaceutical Corporation, Yongin, South Korea;Division of Applied Life Science (BK21 Plus), PMBBRC, Institute of Agricultural and Life Science, Gyeongsang National University, 52828, Jinju, South Korea;Research Center for Industrial Chemical Biotechnology, KRICT, 44468, Ulsan, South Korea;Industrial Biotechnology Research Center, KRIBB, 305-806, Daejeon, South Korea;

关键词: Bioisoprene;    Mevalonate pathway;    Isoprene synthase;    Escherichia coli;    Carbon utilization;   

DOI  :  10.1186/s12934-016-0612-6

 received in 2016-06-13, accepted in 2016-12-04,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundIsoprene, a volatile C5 hydrocarbon, is an important platform chemical used in the manufacturing of synthetic rubber for tires and various other applications, such as elastomers and adhesives.ResultsIn this study, Escherichia coli MG1655 harboring Populus trichocarpa isoprene synthase (PtispS) and the exogenous mevalonate (MVA) pathway produced 80 mg/L isoprene. Codon optimization and optimal expression of the ispS gene via adjustment of the RBS strength and inducer concentration increased isoprene production to 199 and 337 mg/L, respectively. To augment expression of MVA pathway genes, the MVA pathway was cloned on a high-copy plasmid (pBR322 origin) with a strong promoter (Ptrc), which resulted in an additional increase in isoprene production up to 956 mg/L. To reduce the formation of byproducts derived from acetyl-CoA (an initial substrate of the MVA pathway), nine relevant genes were deleted to generate the E. coli AceCo strain (E. coli MG1655 ΔackA-pta, poxB, ldhA, dld, adhE, pps, and atoDA). The AceCo strain harboring the ispS gene and MVA pathway showed enhanced isoprene production of 1832 mg/L in flask culture with reduced accumulation of byproducts.ConclusionsWe achieved a 23-fold increase in isoprene production by codon optimization of PtispS, augmentation of the MVA pathway, and deletion of genes involved in byproduct formation.

【 授权许可】

CC BY   
© The Author(s) 2016