Construction of a novel anaerobic pathway in Escherichia coli for propionate production.

Authors:
Jing Li
Jing Li
National Clinical Research Center of Cardiovascular Diseases
China
Xinna Zhu
Xinna Zhu
Shanghai Jiao Tong University
China
Jing Chen
Jing Chen
University of Kentucky
Lexington | United States
Dongdong Zhao
Dongdong Zhao
Renmin Hospital of Wuhan University
China
Xueli Zhang
Xueli Zhang
Tianjin Institute of Industrial Biotechnology
China
Changhao Bi
Changhao Bi
Joint BioEnergy Institute
United States

BMC Biotechnol 2017 04 14;17(1):38. Epub 2017 Apr 14.

Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, People's Republic of China.

Background: Propionate is widely used as an important preservative and important chemical intermediate for synthesis of cellulose fibers, herbicides, perfumes and pharmaceuticals. Biosynthetic propionate has mainly been produced by Propionibacterium, which has various limitations for industrial application.

Results: In this study, we engineered E. coli by combining reduced TCA cycle with the native sleeping beauty mutase (Sbm) cycle to construct a redox balanced and energy viable fermentation pathway for anaerobic propionate production. As the cryptic Sbm operon was over-expressed in E. coli MG1655, propionate titer reached 0.24 g/L. To increase precursor supply for the Sbm cycle, genetic modification was made to convert mixed fermentation products to succinate, which slightly increased propionate production. For optimal expression of Sbm operon, different types of promoters were examined. A strong constitutive promoter Pbba led to the highest titer of 2.34 g/L. Methylmalonyl CoA mutase from Methylobacterium extorquens AM1 was added to strain T110(pbba-Sbm) to enhance this rate limiting step. With optimized expression of this additional Methylmalonyl CoA mutase, the highest production strain was obtained with a titer of 4.95 g/L and a yield of 0.49 mol/mol glucose.

Conclusions: With various metabolic engineering strategies, the propionate titer from fermentation achieved 4.95 g/L. This is the reported highest anaerobic production of propionate by heterologous host. Due to host advantages, such as non-strict anaerobic condition, mature engineering and fermentation techniques, and low cost minimal media, our work has built the basis for industrial propionate production with E. coli chassis.
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April 2017
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