米曲霉6-磷酸葡萄糖脱氢酶基因gsdA的克隆及生物信息学分析

doi:10.3969/j.issn.1009-7791.2012.03.003

亚热带植物科学 ›› 2012, Vol. 41 ›› Issue (03): 11-15.DOI: 10.3969/j.issn.1009-7791.2012.03.003

米曲霉6-磷酸葡萄糖脱氢酶基因gsdA的克隆及生物信息学分析

刘薇,吴晶晶,陈宏文

华侨大学工业生物技术福建省高等学校重点实验室，福建厦门 361021

收稿日期:2012-04-25 修回日期:2012-04-25 出版日期:2012-09-10 发布日期:2012-09-10
通讯作者: 陈宏文
作者简介:刘薇(1988-)，女，安徽潜山人，硕士研究生，从事基因工程研究。
基金资助:
教育部留学回国人员科研启动基金(2010)|福建省高等学校新世纪优秀人才支持计划(07FJRCO3)|中央高校基本科研业务费专项资金(JB-ZR1112)

Cloning and Bioinformatic Analysis of Glucose-6-phosphate 1-dehydrogenase Gene(gsdA) from Aspergillus oryzae

LIU Wei,WU Jing-jing,CHEN Hong-wen

Key Laboratory of Industrial Biotechnology, Fujian Province, Huaqiao University, Xiamen 361021, Fujian China

Received:2012-04-25 Revised:2012-04-25 Online:2012-09-10 Published:2012-09-10
Contact: CHEN Hong-wen

摘要/Abstract

摘要： 6-磷酸葡萄糖脱氢酶催化6-磷酸葡萄糖生成6-磷酸葡萄糖酸，并生成NADPH，是微生物胞内磷酸戊糖途径（PPP）的关键酶。本研究以食品安全菌米曲霉CICC2012为材料，克隆获得6-磷酸葡萄糖脱氢酶基因(GenBank登录号：JN123468)。序列分析表明，该酶是由222个氨基酸组成的亲水性蛋白；128～134位氨基酸序列DHYLGKE为活性区域；170～176位氨基酸序列GTEGRGG可能为辅因子结合位点。进化树分析表明，米曲霉6-磷酸葡萄糖脱氢酶同其他丝状真菌及酵母的G6PDH较相似。

关键词: 6-磷酸葡萄糖脱氢酶, 克隆, 生物信息学分析

Abstract: Glucose-6-phosphate 1-dehydrogenase(G6PDH) is one of the key enzymes in pentose phosphate pathway(PPP). Here, the gene encoding G6PDH is cloned from Aspergillus oryzae CICC2012. This gene was sequenced and submitted to GenBank(accession number: JN123468). The sequence was analyzed bioinformatically. The results show that G6PDH from A. oryzae is a hydrophilic enzyme consisting of 222 amino acids. The sequence from 128 to 134(DHYLGKE) is the active site, and the sequences from 170 to 176(GTEGRGG) is the possible site of coenzyme binding. The phylogenetic tree shows that G6PDH of A. oryzae is similar to other filamentous fungi and the yeast one, while distinguished from the bacterial type, the plant one, and the human one.

Key words: glucose-6-phosphate 1-dehydrogenase (G6PDH), cloning, bioinformatic analysis

中图分类号:

S852

刘薇,吴晶晶,陈宏文. 米曲霉6-磷酸葡萄糖脱氢酶基因gsdA的克隆及生物信息学分析[J]. 亚热带植物科学, 2012, 41(03): 11-15.

LIU Wei,WU Jing-jing,CHEN Hong-wen. Cloning and Bioinformatic Analysis of Glucose-6-phosphate 1-dehydrogenase Gene(gsdA) from Aspergillus oryzae[J]. Subtropical Plant Science, 2012, 41(03): 11-15.

参考文献

[1] Van A F, Cardinaels C, Clijsters H. Induction of enzyme capacity in plants as a result of heavy metal toxicity: Dose-response relations in Phaseolus vulgaris treated with zinc and cadmium[J]. Environmental Pollution, 1988,52(2): 103-115.
[2] Nemoto Y, Sasakuma T. Specific expression of glucose-6-phosphate dehydrogenase (G6PDH) gene by salt stress in wheat Triticum aestivum[J]. Plant Science, 2000,158(1): 53-60.
[3] Huang J, Zhang H, Wang J, et al. Molecular cloning and characterization of rice 6-phosphogluconate dehydrogenase gene that is up-regulated by salt stress[J]. Molecular Biology Reports, 2003,30(4): 223-227.
[4] Sauer U, Canonaco F, Heri S, et al. The soluble and membrane-bound transhydrogenases UdhA and PntAB have divergent functions in NADPH metabolism of Escherichia coli[J]. Journal of Biological Chemistry, 2004,279(8): 6 613-6 619.
[5] Hou J, Vemuri G N, Bao X, et al. Impact of overexpressing NADH kinase on glucose and xylose metabolism in recombinant xylose-utilizing Saccharomyces cerevisiae[J]. Applied Microbiology and Biotechnology, 2009,82(5): 909-919.
[6] Lee W H, Park J B, Park K, et al. Enhanced production of ?-caprolactone by overexpression of NADPH-regenerating glucose 6-phosphate dehydrogenase in recombinant Escherichia coli harboring cyclohexanone monooxygenase gene[J]. Applied Microbiology and Biotechnology, 2007,76(2): 329-338.
[7] Georgi T, Rittmann D, Wendisch V F. Lysine and glutamate production by Corynebacterium glutamicum on glucose, fructose and sucrose: roles of malic enzyme and fructose-1, 6-bisphosphatase[J]. Metabolic Engineering, 2005,7(4): 291-301.
[8] Kwon D H, Kim M D, Lee T H, et al. Elevation of glucose 6-phosphate dehydrogenase activity increases xylitol production in recombinant Saccharomyces cerevisiae[J]. Journal of Molecular Catalysis B: Enzymatic, 2006,43(1-4): 86-89.
[9] Lee W H, Chin Y W, Han N S, et al. Enhanced production of GDP-L-fucose by overexpression of NADPH regenerator in recombinant Escherichia coli[J]. Applied Microbiology and Biotechnology, 2011,91(4): 967-976.
[10] Broek P, Goosen T, Wennekes B, et al. Isolation and characterization of the glucose-6-phosphate dehydrogenase encoding gene (gsdA) from Aspergillus niger[J]. Molecular and General Genetics MGG, 1995,247(2): 229-239.
[11] 袁洪水,李术娜,张爱莲,等. 石英砂研磨法快速提取顶头孢霉染色体DNA[J]. 河北农业大学学报, 2007(5): 8-11.
[12] 黄骥,王建飞,张红生. 植物戊糖磷酸途径及其两个关键酶的研究进展[J]. 植物学通报, 2004(2): 139-145.
[13] Jeffery J, Persson B, Wood I, et al. Glucose-6-phosphate dehydrogenase. Structure-function relationships and the Pichia jadinii enzyme structure[J]. European Journal of Biochemistry, 1993,212(1): 41-49.
[14] Persson B, Jornvall H, Wood I, et al. Functionally important regions of glucose-6-phosphate dehydrogenase defined by the Saccharomyces cerevisiae enzyme and its differences from the mammalian and insect forms[J]. European Journal of Biochemistry, 1991,198(2): 485-491.

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米曲霉6-磷酸葡萄糖脱氢酶基因gsdA的克隆及生物信息学分析

Cloning and Bioinformatic Analysis of Glucose-6-phosphate 1-dehydrogenase Gene(gsdA) from Aspergillus oryzae

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