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Succinate Production from Escherichia coli Mutant QQS101 Fermentation |
LI Yi-kui1,2, KANG Jun-hua1, KANG Zhen1, GENG Yan-ping1, WANG Yi-hua2, QI Qing-sheng1 |
1. State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, China;
2. Science Department, Jiangxi Agricultural University, Nanchang 330045, China |
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Abstract Succinate is an important bio-based platform molecule. In the present work, the growth and glucose metabolism of Escherichia coli mutant QQS101 with the deficiency of formate transporter A (focA), formate-pyruvate lyase (pflB) and lactate dehydrogenase (ldhA) under strict anaerobic condition were investigated. The degree of reduction per carbon of glucose and the products of E. coli mix-acids fermentation were compared, and then recognized that non-strict anaerobic condition favored QQS101 producing succinate from glucose. Furthermore, effects of the carbon sources for aerobic growth on fermentation were performed. Results showed that QQS101 could accumulate succinate with a concentration of 31.01 g/L with a yield of 1.258 mol Succinate/mol Glucose, when growing on xylose as the aerobic substrate. During the fermentation, addition of alanine could enhance the molar yield of succinate to glucose utilized.
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Received: 23 March 2010
Published: 25 October 2010
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Corresponding Authors:
QI Qing-sheng
E-mail: qiqingsheng@sdu.edu.cn
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[1] Hong S H, Lee S Y. Importance of redox balance on the production of succinic acid by metabolically engineered Escherichia coli. Appl Microbiol Biotechnol, 2002, 58(3): 286-290.
[2] Akesson M, Karlsson E N, Hagander P, et al. On-line detection of acetate formation in Escherichia coli culture using dissolved oxygen responses to feed transients. Biotechnol Bioeng, 1999, 64(5): 590-598.
[3] 康振, 耿艳萍, 张园园, 等. 好氧发酵生产琥珀酸工程菌株的构建. 生物工程学报, 2008, 24(12): 2081-2085. Kang Z, Geng Y P, Zhang Y Y, et al. Construction of engineered Escherichia coli for aerobic succinate production. Chin J Biotech, 2008, 24(12): 2081-2085.
[4] McKinlay J B, Vieille C, Zeikus J G. Prospects for a bio-based succinate industry. Appl Microbiol Biotechnol, 2007, 76(4): 727-740.
[5] Lin H, Bennett G N, San K Y. Metabolic engineering of aerobic succinate production systems in Escherichia coli to improve process productivity and achieve the maximum theoretical succinate yield. Metab Eng, 2005, 7(2): 116-127.
[6] Sanchez A M, Bennett G N, San K Y. Novel pathway engineering design of the anaerobic central metabolic pathway in Escherichia coli to increase succinate yield and productivity. Metab Eng, 2005, 7(3): 229-239.
[7] Chatterjee R, Millard C S, Champion K, et al. Mutation of the ptsG gene results in increased production of succinate in fermentation of glucose by Escherichia coli. Appl Environ Microbiol, 2001, 67(1): 148-154.
[8] Hong S H, Lee S Y. Metabolic flux analysis for succinic acid production by recombinant Escherichia coli with amplified malic enzyme activity. Biotechnol Bioeng, 2001, 74(2): 89-95.
[9] Sanchez A M, Bennett G N, San K Y. Efficient succinic acid production from glucose through overexpression of pyruvate carboxylase in an Escherichia coli alcohol dehydrogenase and lactate dehydrogenase mutant. Biotechnol Prog, 2005, 21(2): 358-365.
[10] Stols L, Donnelly M I. Production of succinic acid through overexpression of NAD(+)-dependent malic enzyme in an Escherichia coli mutant. Appl Environ Microbiol, 1997, 63(7): 2695-2701.
[11] Mat-Jan F, Alam K Y, Clark D P. Mutants of Escherichia coli deficient in the fermentative lactate dehydrogenase. J Bacteriol, 1989, 171(1): 342-348.
[12] Singh A, Lynch M D, Gill R T. Genes restoring redox balance in fermentation-deficient E. coli NZN111. Metab Eng, 2009, 11(6): 347-354.
[13] Wu H, Li Z M, Zhou L, et al. Improved succinic acid production in the anaerobic culture of an Escherichia coli pflB ldhA double mutant as a result of enhanced anaplerotic activities in the preceding aerobic culture. Appl Environ Microbiol, 2007, 73(24): 7837-7843.
[14] Lara A R, Caspeta L, Gosset G, et al. Utility of an Escherichia coli strain engineered in the substrate uptake system for improved culture performance at high glucose and cell concentrations: an alternative to fed-batch cultures. Biotechnol Bioeng, 2008, 99(4): 893-901.
[15] Becker S, Vlad D, Schuster S, et al. Regulatory O2 tensions for the synthesis of fermentation products in Escherichia coli and relation to aerobic respiration. Arch Microbiology, 1997, 168(4): 290-296.
[16] Nielsen J, Villadsen J, Liden G. Bioreaction Engineering Principles. 2nd ed. New York: Kluwer Academic/Plenum Publishers, 2003.60-73.
[17] Latour D J, Weiner J H. Regulation of in vitro expression of the Escherichia coli frd operon: alanine and Fnr represent positive and negative control elements. Nucleic Acids Research, 1988, 16(14): 6339-6352.
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