Abstract
Objectives
To enhance succinic acid production in Corynebacterium glutamicum by increasing the supply of NADH and the rate of glucose consumption by decreasing H+-ATPase activity.
Results
A mutant of C. glutamicum NC-3-1 with decreased H+-ATPase activity was constructed. This increased the rate of glycolysis and the supply of NADH. Fermentation of C. glutamicum NC-3-1 gave 39 % higher succinic acid production (113 and 81 g/l), a 29 % higher succinic acid yield (0.94 and 0.73 g succinic acid/g glucose) and decreased by-products formation compared to that of C. glutamicum NC-3 in 5 l bioreactor.
Conclusion
The point mutation in C. glutamicum NC-3-1 increased the rate of glycolysis and resulted in higher succinic acid production, higher succinic acid yield and significantly decreased formation of by-products.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bernofsky C, Swan M (1973) An improved cycling assay for nicotinamide adenine dinucleotide. Anal Biochem 53:452–458
Chatterjee R, Millard CS, Champion K, Clark DP, Donnelly MI (2001) Mutation of the ptsG gene results in increased production of succinate in fermentation of glucose by Escherichia coli. Appl Environ Microbiol 67:148–154
Chen XJ, Jiang ST, Li XJ, Pan LJ, Zheng Z, Luo SZ (2008) Production of d-lactic acid by Corynebacterium glutamicum under oxygen deprivation. Appl Microbiol Biotechnol 78:449–454
Inui M, Murakami S, Okino S, Kawaguchi H, Vertès AA, Yukawa H (2004) Metabolic analysis of Corynebacterium glutamicum during lactate and succinate productions under oxygen deprivation conditions. J Mol Microbiol Biotechnol 7:182–196
Lee PC, Lee SY, Hong SH, Chang HN (2002) Isolation and characterization of a new succinic acid-producing bacterium, Mannheimia succiniciproducens MBEL55E, from bovine rumen. Appl Microbiol Biotechnol 58:663–668
Lin H, Bennett GN, San KY (2005) Metabolic engineering of aerobic succinate production systems in Escherichia coli to improve process productivity and achieve the maximum theoretical succinate yield. Metab Eng 7:116–127
Litsanov B, Brocker M, Bott M (2012) Toward homosuccinate fermentation: metabolic engineering of Corynebacterium glutamicum for anaerobic production of succinate from glucose and formate. Appl Environ Microbiol 77:90-11
McKinlay JB, Vieille C, Zeikus JG (2007) Prospects for a bio-based succinate industry. Appl Microbiol Biotechnol 76:727–740
Okino S, Inui M, Yukawa H (2005) Production of organic acids by Corynebacterium glutamicum under oxygen deprivation. Appl Microbiol Biotechnol 68:475–480
Sato K, Yoshida Y, Hirahata T (2000) On-line measurement of intracellular ATP of Saccharomyces cerevisiae and pyruvate during sake mashing. J Biosci Bioeng 90:294–301
Schneider J, Niermann K, Wendisch VF (2011) Production of the amino acids l-glutamate, l-lysine, l-ornithine and l-arginine from arabinose by recombinant Corynebacterium glutamicum. J Biotechnol 154:191–198
Sekine H, Shimada T, Hayashi C, Ishiguro A, Tomita F, Yokota A (2001) H+-ATPase defect in Corynebacterium glutamicum abolishes glutamic acid production with enhancement of glucose consumption rate. Appl Microbiol Biotechnol 57:534–540
Wang C, Zhang HL, Cai H, Zhou ZH, Chen YL, Chen YL, Ouyang PK (2013) Succinic acid production from corn cobs hydrolysates by genetically engineered Corynebacterium glutamicum. Appl Biochem Biotechnol 172:340–350
Wieschalka S, Blombach B, Bott M, Eikmanns BJ (2013) Bio-based production of organic acids with Corynebacterium glutamicum. Microb Biotechnol 6:87–102
Yokota A, Terasawa Y, Takaoka N, Shimizu H, Tomita F (1994) Pyruvic acid production by an F1-ATPase-defective mutant of Escherichia coli W1485 lip2. Biosci Biotechnol Biochem 58:2164–2167
Yokota A, Henmi M, Takaoka N, Hayashi C, Takezawa Y, Fukumori Y, Tomita F (1997) Enhancement of glucose metabolism in a pyruvic acid-hyperproducing Escherichia coli mutant defective in F1-ATPase activity. J Ferment Bioeng 83:132–138
Zhou ZH, Wang C, Chen ZJ, Chen YL, Zhang K, Xu HT, Cai H (2015) Increasing available NADH supply during succinic acid production by Corynebacterium glutamicum. Biotechnol Prog 31:12–19
Acknowledgments
This work was supported by the 973 Program of China (grant no. 2011CB707405).
Supporting information
Anaerobic succinic acid production in a seated bottle.
Anaerobic succinic acid production in a 5 l bioreactor.
Supplementary Table 1—Sequences of oligonucleotide primers.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Xu, H., Zhou, Z., Wang, C. et al. Enhanced succinic acid production in Corynebacterium glutamicum with increasing the available NADH supply and glucose consumption rate by decreasing H+-ATPase activity. Biotechnol Lett 38, 1181–1186 (2016). https://doi.org/10.1007/s10529-016-2093-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-016-2093-4