Development of a plasmid stabilization system in Vibrio natriegens for the high production of 1,3-propanediol and 3-hydroxypropionate

Bioresources and Bioprocessing

Qing Sun¹  Ye Zhang¹  Yu Liu¹  Xuecong Cen¹  Dehua Liu²  Zhen Chen² 

[1] Key Laboratory of Industrial Biocatalysis (Ministry of Education), Department of Chemical Engineering, Tsinghua University, 100084, Beijing, China;Key Laboratory of Industrial Biocatalysis (Ministry of Education), Department of Chemical Engineering, Tsinghua University, 100084, Beijing, China;Tsinghua Innovation Center in Dongguan, 523808, Dongguan, China;Center for Synthetic and Systems Biology, Tsinghua University, 100084, Beijing, China;

关键词: Vibrio natriegens;    1,3-propanediol;    3-hydroxypropionate;    Plasmid maintenance;    Antibiotic free;    Glycerol;   

DOI  :  10.1186/s40643-021-00485-0

来源: Springer

【 摘 要 】

Vibrio natriegens is a promising industrial chassis with a super-fast growth rate and high substrate uptake rates. V. natriegens was previously engineered to produce 1,3-propanediol (1,3-PDO) from glycerol by overexpressing the corresponding genes in a plasmid. However, antibiotic selection pressure for plasmid stability was not satisfactory and plasmid loss resulted in reduced productivity of the bioprocess. In this study, we developed an antibiotic-free plasmid stabilization system for V. natriegens. The system was achieved by shifting the glpD gene, one of the essential genes for glycerol degradation, from the chromosome to plasmid. With this system, engineered V. natriegens can stably maintain a large expression plasmid during the whole fed-batch fermentation and accumulated 69.5 g/L 1,3-PDO in 24 h, which was 23% higher than that based on antibiotic selection system. This system was also applied to engineering V. natriegens for the production of 3-hydroxypropionate (3-HP), enabling the engineered strain to accumulate 64.5 g/L 3-HP in 24 h, which was 30% higher than that based on antibiotic system. Overall, the developed strategy could be useful for engineering V. natriegens as a platform for the production of value-added chemicals from glycerol.Graphic Abstract

【 授权许可】

CC BY