The impact of PHB accumulation on L-glutamate production by recombinant Corynebacterium glutamicum.
Corynebacterium glutamicum, a gram-positive soil bacterium, has been used extensively for the industrial production of l-glutamate and other amino acids. In this study, an Escherichia coli-C. glutamicum shuttle expression plasmid harboring polyhydroxybutyrate (PHB) synthesis genes, phbCAB from Ralstonia eutropha, was constructed under the Ptrc promoter. C. glutamicum harboring this plasmid accumulated 3-13% PHB with a weight average molecular mass of 125,400 and a polydispersity of 11.3 when grown on glucose. PHB synthesis related enzyme activities including beta-ketothiolase (PhbA), acetoacetyl-CoA reductase (PhbB) and PHB synthase (PhbC) were found to be constitutively produced independent of IPTG. l-Glutamate production increased 39-68% in two C. glutamicum strains harboring PHB synthesis genes compared with their parent strains in shake flask experiments. In fermentor studies, the recombinant produced approximately 23% more l-glutamate compared with that of the wild type, and yielded less intermediate metabolites or by-products including alpha-ketoglutarate, l-glutamine and lactate. These results suggested that the expression of phbCAB genes in C. glutamicum could help regulate glutamate production metabolism. This demonstrated that the expression of PHB synthesis genes has a positive effect on l-glutamate production in C. glutamicum.
Liu
Qian
Q
Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China.
Ouyang
Shao-Ping
SP
Kim
Jonghyok
J
Chen
Guo-Qiang
GQ
D048230:Corynebacterium glutamicum / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D005285:Fermentation; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005822:Genetic Vectors; D018698:Glutamic Acid / Q000096:biosynthesis*; D006885:Hydroxybutyrates / Q000378:metabolism*; D010957:Plasmids; D011091:Polyesters / Q000378:metabolism*; D000094122:Polyhydroxybutyrates
D018698:Glutamic Acid; D006885:Hydroxybutyrates; D011091:Polyesters; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates