Effect of co-expression of nicotinic acid phosphoribosyl transferase and pyruvate carboxylase on succinic acid production in Escherichia coli BA002.
- Author:
Weijia CAO
;
Dongmei GOU
;
Liya LIANG
;
Rongming LIU
;
Kequan CHEN
;
Jiangfeng MA
;
Min JIANG
- Publication Type:Journal Article
- MeSH:
Anaerobiosis;
Escherichia coli;
enzymology;
genetics;
metabolism;
Fermentation;
Genetic Engineering;
Glucose;
metabolism;
Industrial Microbiology;
Lactococcus lactis;
enzymology;
NAD;
metabolism;
Pentosyltransferases;
biosynthesis;
genetics;
Pyruvate Carboxylase;
biosynthesis;
genetics;
Succinic Acid;
metabolism
- From:
Chinese Journal of Biotechnology
2013;29(12):1855-1859
- CountryChina
- Language:Chinese
-
Abstract:
Escherichia coli BA002, in which the ldhA and pflB genes are deleted, cannot utilize glucose anaerobically due to the inability to regenerate NAD+. To restore glucose utilization, overexpression of nicotinic acid phosphoribosyltransferase (NAPRTase) encoded by the pncB gene, a rate-limiting enzyme of NAD(H) synthesis pathway, resulted in a significant increase in cell mass and succinate production under anaerobic conditions. However, a high concentration of pyruvate was accumulated. Thus, co-expression of NAPRTase and the heterologous pyruvate carboxylase (PYC) of Lactococcus lactis subsp. cremoris NZ9000 in recombinant E. coli BA016 was investigated. Results in 3 L fermentor showed that OD600 is 4.64 and BA016 consumed 35.00 g/L glucose and produced 25.09 g/L succinate after 112 h under anaerobic conditions. Overexpression of pncB and pyc in BA016, the accumulation of pyruvic acid was further decreased, and the formation of succinic acid was further increased.
