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| Cloning, Expression of the Pyruvate Kinase Gene from Corynebacterium crenatum and Its Effect on L-Arginine Synthesis |
| LU Yan1, RAO Zhi-ming1, XU Mei-juan1, MAN Zai-wei1, ZHANG Xian1, GENG Yan2, XU Zheng-hong2 |
1. The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China;
2. Lab of Pharmaceutical Engineering, School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China |
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Abstract In order to increase glocuse consumption rate and shorten fermentation time, pyruvate kinase(EC 2.7.1.40, PYK)gene from L-arginine producing mutant Corynebacterium crenatum SYPA 5-5 was expressed in Corynebacterium crenatum SYPA 5-5. The results of fermentive character showed that compared with wild-type strain, the average glocuse consumption rate was improved 33.5% to 1.71 g/L/h, the L-arginine concentration was improved 20% to 43.32 g/L, and the maximum yield of L-Arg was improved 19.5% to 0.55 g/L/h. Overexpression of pyruvate kinase could increased the glocuse consumption and improved the yield of L-arginine.
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Received: 25 November 2013
Published: 25 March 2014
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[1] 徐美娟, 陆元修, 周晨, 等. 钝齿棒杆菌精氨酸琥珀酸酶编码基因argH的克隆表达及其重组菌发酵产精氨酸研究.中国生物工程杂志, 2010, 30(9):49-55. Xu M J, Lu Y X, Zhou CH, et al. Cloning, expression and analysis of argH gene encoding argininosuccinate lyase from Corynebacterium crenatum. China Biotechnology, 2010, 30(9):49-55.
[2] 许正宏, 熊筱晶, 窦文芳, 等. L-精氨酸高产菌的选育及基于代谢流量分布的育种机制.工业微生物, 2006, 36(4):1-6. Xu ZH H, Xiong X J, Dou W F, et al. Improvement of high L-arginine producing strain and the mechanism analysis based on metabolic flux distribution. Ind Microbiol, 2006, 36(4): 1-6.
[3] 徐美娟, 张显, 饶志明, 等. 钝齿棒杆菌N-乙酰鸟氨酸转氨酶的克隆表达分析及其重组菌的精氨酸发酵.生物工程学报, 2011, 27(7):1013-1023. Xu M J, Zhang X, Rao ZH M, et al. Cloning, expression and characterization of N-Acetylornithine aminotransferase from Corynebacterium crenatum and its effects on L-arginine fermentation. Chin J Biotech, 2011, 27(7):1013-1023.
[4] Dou W F, Xu M J, Cai D M, et al. Improvement of L-argnine production by overexpression of a bifunctional ornithine acetyltransferase in Corynebacterium crenatum. Appl Biochem Biotechnol, 2011, 165(3-4):845-855.
[5] Xu M J, Rao ZH M, Yang J, et al. Heterologous and homologous expression of the arginine biosynthetic argC~H cluster from Corynebacterium crenatum for improvement of L-arginine production. J Ind Microbiol Biotechnol, 2012, 39 (3):495-502.
[6] Xu M J, Rao Zh M, Dou W F, et al. Site-directed mutagenesis and feedback-resistant N-acetyl-L-glutamate kinase (NAGK) increase Corynebacterium crenatum L-arginine production. Amino Acids, 2012, 43(1):255-266.
[7] Kayne F. Pyruvate kinase. The Enzymes, 1973, 8(10):353-382.
[8] David C, Rachel W. Mining for allosteric information:natual mutations and positional sequence conservation in pyruvate kinase. Iubmb Life, 2006, 58(1):31-38.
[9] Marcel G, Mike J, Sang H L, et al. Cloing of the pyruvate kinase gene (pyk) of Corynebacterium glutamicum and site-specific inactivation of pyk in a lysine-producing Corynebacterium lactofermentum strain. Applied and Environmental Microbiology, 1994, 60(7):2492-2500.
[10] Judith B, Corinna K, Christoph W.Metabolic responses to pyruvate kinase deletion in lysine producing Corynebacterium glutamicum. Microbial Cell Factories, 2008, 7(8):1475-2859.
[11] 李鸿艳, 周思甜, 马建辉, 等. 大肠杆菌Ⅱ型丙酮酸激酶的纯化分析及其作为偶联酶的应用. 中国生物工程杂志, 2013, 33(5):68-74. Li H Y, Zhou S T, Ma J H, et al. Purification and characterization of pyruvate kinaseⅡfrom Escherichia coli and its application as a coupling enzyme. China Biotechnology, 2013, 33(5): 68-74.
[12] 徐美娟, 张显, 饶志明, 等. 钝齿棒杆菌N-乙酰鸟氨酸转氨酶的克隆表达分析及其重组菌的精氨酸发酵. 生物工程学报, 2011, 27(7):1013-1023. Xu M J, Zhang X, Rao Zh M, et al. Cloning, expression and characterization of N-acetylornithine aminotransferase from Corynebacterium crenatum and its effects on L-arginine fermentation. Chinese Journal of Biotechnology, 2011, 27(7):1013-1023.
[13] Kirchner O, Tauch A. Tools for genetic engineering in the amino acid-producing bacterium Corynebacterium glutamicum. J Biotechnol, 2003, 104(1/3):287-299.
[14] Joseph S, David W R.Molecular Cloning.3rd ed.New York: Cold Spring Harbor Laboratory Press, 2001, 1523-1574.
[15] Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 1976, 72(12):248-254.
[16] Jetten M S, Gubler M E, Lee S H, et al. Structural and functional analysis of pyruvate kinase from Corynebacterium glutamicum. Applied Environment Microbiology, 1994, 60(7):2501-2507.
[17] Xu H, Dou W F, Xu H Y, et al. A two-stage oxygen supply strategy for enhanced l-arginine production by orynebacterium crenatum based on metabolic fluxes analysis. Biochem Eng J, 2009, 43(1):41-51.
[18] 王镜岩, 沈同. 生物化学.第三版. 北京:高等教育出版社, 2001:78 -278. Wang J Y, Shen T. Biochemistry. 3rd. Beijing: High Education Press, 2001:78 -278.
[19] 宁正祥. 食品成分分析手册. 北京:中国轻工业出版社, 2001:50-55. Ning ZH X. Handbook of Food Composition Analysis. Beijing: China Light Industry Press, 2001:50-55.
[20] 俞海青, 伍时华, 张克旭, 等. α-萘酚法定量测定L-Arg的研究. 天津科技大学学报, 2001, 26(2):30-33. Yu H Q, Wu SH H, Zhang K X, et al. The study on quantitative determination of L-arginine content with α-naphtnol. J Tianjing Univ Sci Technol, 2001, 26(2):30-33.
[21] 徐美娟.钝齿棒杆菌SYPA5-5发酵产L-精氨酸的代谢工程改造.无锡: 江南大学, 生物工程学院, 2012. Xu M J. Metabolic Engineering of Corynebacterium crenatum SYPA 5-5 for the L-arginine Production. Wuxi: Jiangnan University, College of Biotechnology, 2012. |
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