|
|
|
| Overproduction of Glutamic Acid in Corynebacterium glutamicum with H+-atp Inactivation |
| ZHANG Bo, LI Tie-min, YANG Zhi-yong, HU Yong-fei, LI Yu |
| School of Life Science, Liaoning University, Shenyang 110036, China |
|
|
|
Abstract In order to confirm the application feasibility of overproduction of glutamic acid in C. glutamicum with H+-ATP inactivation in the construction of genetically modified bacteria. The sequence encoding H+-ATPase γ subunit in C.glutamicum ATCC13032 was partially deleted by crossover PCR, and the mutant, in which H+-atp gene was inactivated, was obtained by insertion inactivation. The glutamic acid production and growth rate of the mutant were tested. The results showed that the maximum production of glutamic acid of the mutant was 51.6 g/L in 100 g/L glucose culture medium, with 42.9 % increase in contrast to the wild parent strain, and that growth rate of the mutant in measurement of the growth curves was lower than wild parent strain. The results suggest that H+-atp gene inactivated in C.glutamicum increases glutamic acid production, and slightly reduces growth rate of the bacteria.
|
|
Received: 02 August 2010
Published: 25 January 2011
|
|
|
|
|
|
[1] Engelbrechft S, Junge W. ATP synthase: a tentative structural model. FEBS Lett, 1997, 414 (3): 485-491.
[2] Senior A E. The proton-translocating ATPase of Escherichia coli. Annu Rev Biophys Chem, 1990, 19: 7-41.
[3] Kobayashi H. A proton-translocating ATPase regulates pH of the bacterial cytoplasm. J Biol Chem, 1985, 260(1): 72-76.
[4] Causey T B, Zhou S, Shanmugam K T. et al. Engineering the metabolism of Escherichia coli W3110 for the conversion of sugar to redox-neutral and oxidized products: Homoacetate production. Proc Natl Acad Sci USA, 2003, 100(3): 825-832.
[5] Causey T B, Shanmugam K T, Yomano L P, et al. Engineering Escherichia coli for efficient conversion of glucose to pyruvate. Proc Natl Acad Sci USA, 2004, 101(8): 2235-2240.
[6] Wada M, Narita K, Yokota A. Alanine production in an H+-ATPase and lactate dehydrogenase defective mutant of Escherichia coli expressing alanine dehydrogenase. Appl Microbiol Biotechnol, 2007, 76(4): 819-825.
[7] Santana M, Ionescu M S, Vertes A, et al. Bacillus subtilis FOF1 ATPase: DNA Sequence of the atp Operon and Characterization of atp Mutants. J Bacteriol, 1994, 176(22): 6802-6811.
[8] Sekine H, Shimada T, Hayashi C, et al. H+-ATPase defect in Corynebacterium glutamicum abolishes glutamic acid production with enhancement of glucose consumption rate. Appl Microbiol Biotechnol, 2001, 57(4):534-540.
[9] Aoki R, Wada M, Takesue N, et al. Enhanced Glutamic Acid Production by a H+-ATPase-Defective Mutant of Corynebacterium glutamicu. Biosci Biotechnol Biochem, 2005, 69 (8): 1466-1472.
[10] Dorella F A, Estevam E M, Cardoso P G, et al. An improved protocol for electrotransformation of Corynebacterium pseudotuberculosis. Vet Microbiol, 2006, 114(3-4): 298-303.
[11] 天津轻工业学院,大连轻工业学院,无锡轻工大学,等.工业发酵分析.北京:中国轻工业出版社,2004.97-99. Tianjin University of Light Industry. Analysis of Industrial Fermentation.Beijing:Light Industry Press of China, 2004.97-99.
[12] 沈萍,范秀容,李广武.微生物学实验.第三版,北京:高等教育出版社,1999.258-261. Shen P,Fan X R, Li G W. Experiment of Microorganism.3rd ed,Beijing: Higher Education Press,1999.258-261.
[13] Mller D, Schauder R, Fuchs G, et al. Acetate oxidation to CO2 via a citric acid cycle involving an ATP-citrate lyase: A mechanism for the synthesis of ATP via substrate level phosphorylation in Desulfobacter postgatei growing on acetate and sulfate. Arch Microbiol, 1987, 148(3): 202-207.
[14] Yokota A, Henmi M, Takaoka N, et al. Enhancement of glucose metabolism in a pyruvic acid-hyperproducing Escherichia coli mutant defective in F1-ATPase activity. J Ferment Bioeng, 1997, 83(2): 132-138.
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
