Improved production of propionic acid in Propionibacterium jensenii via combinational overexpression of glycerol dehydrogenase and malate dehydrogenase from Klebsiella pneumoniae

• Published in: Applied and Environmental Microbiology Vol. 81; no. 7; pp. 2256 - 2264
• Main Authors: Liu, Long, Zhuge, Xin, Shin, Hyun-Dong, Chen, Rachel R, Li, Jianghua, Du, Guocheng, Chen, Jian
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 01.04.2015
• Subjects: Anaerobiosis; Biochemistry; Bioengineering; Biosynthesis; Biotechnology; Carbon - metabolism; Fumarate Hydratase - genetics; Fumarate Hydratase - metabolism; Gene Expression; Gene Expression Profiling; Genetic Vectors; Glycerol; Glycerol - metabolism; Gram-negative bacteria; Klebsiella pneumoniae; Klebsiella pneumoniae - enzymology; Klebsiella pneumoniae - genetics; Malate Dehydrogenase - genetics; Malate Dehydrogenase - metabolism; Metabolic Engineering; Metabolites; Propionates - metabolism; Propionibacterium - genetics; Propionibacterium - metabolism; Propionibacterium jensenii; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Sugar Alcohol Dehydrogenases - genetics; Sugar Alcohol Dehydrogenases - metabolism; Transcription, Genetic
• ISSN: 0099-2240; 1098-5336; 1098-6596
• DOI: 10.1128/AEM.03572-14

Microbial production of propionic acid (PA), an important chemical building block used as a preservative and chemical intermediate, has gained increasing attention for its environmental friendliness over traditional petrochemical processes. In previous studies, we constructed a shuttle vector as a useful tool for engineering Propionibacterium jensenii, a potential candidate for efficient PA synthesis. In this study, we identified the key metabolites for PA synthesis in P. jensenii by examining the influence of metabolic intermediate addition on PA synthesis with glycerol as a carbon source under anaerobic conditions. We also further improved PA production via the overexpression of the identified corresponding enzymes, namely, glycerol dehydrogenase (GDH), malate dehydrogenase (MDH), and fumarate hydratase (FUM). Compared to those in wild-type P. jensenii, the activities of these enzymes in the engineered strains were 2.91- ± 0.17- to 8.12- ± 0.37-fold higher. The transcription levels of the corresponding enzymes in the engineered strains were 2.85- ± 0.19- to 8.07- ± 0.63-fold higher than those in the wild type. The coexpression of GDH and MDH increased the PA titer from 26.95 ± 1.21 g/liter in wild-type P. jensenii to 39.43 ± 1.90 g/liter in the engineered strains. This study identified the key metabolic nodes limiting PA overproduction in P. jensenii and further improved PA titers via the coexpression of GDH and MDH, making the engineered P. jensenii strain a potential industrial producer of PA.