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Cloning and Optimizing Expression of Enoate Reductase Gene from Clostridium acetobutylicum in Escherichia coli(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
2011 01
Page:
87-90
Research Field:
Articles
Publishing date:

Info

Title:
Cloning and Optimizing Expression of Enoate Reductase Gene from Clostridium acetobutylicum in Escherichia coli
Author(s):
LÜ Tong LIU Yang ZHAO Qing REN Jie WANG Min WU Qiaqing & ZHU Dunming
(1College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China)
(2Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China)
Keywords:
enoate reductase optimizing expression asymmetric reduction C=C bond Clostridium acetobutylicum
CLC:
Q936 : Q78
PACS:
DOI:
10.3724/SP.J.1145.2011.00087
DocumentCode:

Abstract:
Enone reductase is a class of biocatalysts which catalyze the reduction of C=C bond of enones and/or enoates. The gene which encodes enone reductase from Clostridium acetobutylicum ATCC824 was cloned. The cloned sequence consisted of 1 995 bp and encoded a protein of 664 amino acids, with a molecular weight of 73 ×103. The gene was cloned into the expression vector pET-32a(+), and the resulting plasmid DNA was transformed into the Escherichia coli Rosetta (DE3) pLysS strain. The heterologous expression was optimized and the best results were obtained for induction at an D600 nm of 0.6~0.8 with 0.1 mmol/L IPTG 12 h at 18 ℃. The SDS-PAGE analysis indicated that the ERs was expressed in the supernatant. Fig 5, Ref 16

References

1 Hall M, Stueckler C, Hauer B, Stuermer R, Friedrich T, Breuer M, Kroutil W, Faber K. Asymmetric bioreduction of activated C=C bonds using Zymomonas mobilis NCR enoate reductase and Old Yellow Enzymes OYE 1-3 from yeasts. Eur J Org Chem, 2008, 9: 1511~1516
2 Stueckler C, Hall M, Ehammer H, Pointner E, Kroutil W, Macheroux, Faber K. Stereo-complementary bioreduction of α,β-unsaturated dicarboxylic acids and dimethyl esters using enoate reductases: Enzyme-substrate-based stereocontrol. Org Lett, 2007, 9: 5409~5411
3 Schaller F, Biesgen C, Mussig C, Altmann T, Weller EW. 12-oxophytodienoate reductase 3 (OPR3) is the isoenzyme invooved in jasmonate biosynthesis. Planta, 2000, 210: 979~984
4 Shimoda K, Ito DI, Izumi S, Hirata T. Noveo reductase participation in the syn-addition of hydrogen to the C=C bond of enones in the cultured cells of Nicotiana tabacum. J Chem Soc, Perkin Trans 1, 1996, 4: 355~358
5 Williams RE, Rathbone DA, Scrutton NS, Bruce NC. Biotransformation of explosives by old yellow enzyme family of flavoproteins. Appl Environ Microbiol, 2004, 70: 3556~3574
6 Hall M, Hauer B, Stuermer R, Kroutil W, Faber K. Asymmetric whole-cell bioreduction of an α,β-unsaturated aldehyde (citral): Competing prim-alcohol dehydrogenase and C-C lyase activities. Tetrahedron Asymmetry, 2006, 17: 3058~3062
7 Joseph W, Joseph K, Eleftherios T.The effect of pH on nitrogen supply, cell lysis, and solvent production in fermentations of Clostridium acetobutylicum. Biotechnol & Bioeng, 1985, 27 (5): 681~694
8 Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual. 2nded. Beijing, China: Science Press (北京: 科学出版社), 1993
9 Huang M (黄敏), Xu H (徐辉), Cao Y (曹瑜), Zhu J (朱俊), Zhang FW (张飞伟), You FF (尤芳芳), Qiao DR (乔代蓉) Cao Y (曹毅). Characteristics of Phytase from Escherichia coli. Chin J Appl Eviron Biol (应用与环境生物学报), 2009, 15 (3): 371~375
10 汪家政, 范明. 蛋白质技术手册. 北京: 科学出版社, 2000
11 Feng ZG (冯志国), Liu HJ (刘慧娟), Lu J (陆婕), Lang J (郎君), Li YJ (李燕娇), Chen ZW (陈正望). Optimized expression of a novel glycine-rich antibacterial peptide from Drosophila in Escherichia coli. Biotechnology (生物技术), 2008, 18 (4): 18~20
12 Sorensen HP, Mortensen KK. Soluble expression of recombinant proteins in the cytoplasm of Escherichia coli. Microb Cell Factories, 2005, 4: 1~8
13 Williams RE, Bruce NC. ‘New uses for an Old Enzyme’ – the Old Yellow Enzyme family of ?avoenzymes. Microbiology, 2002, 148: 1607~1614
14 Li ZQ (李振秋), Wang HH (王花红), Wang H (王红), Li GF (李国凤), Ye HC (叶和春). Escherichia coli expression, purification and functional identification of squalene synthase from Artemisia annua L. Chin J Appl Eviron Biol (应用与环境生物学报), 2007, 13 (3): 309~312
15 Buque-Taboada EM, Straathof AJJ, Heijnen JA. Microbial reduction and in situ product crystallization coupled with biocatalyst cultivation during the synthesis of 6R-dihydrooxoisophorone. Adv Synth Catal, 2005, 347: 1147~1154
16 Surinder MS, Amulya KP. Solubilization and refolding of bacterial inclusion body proteins. J Biosci & Bioeng, 2005, 99 (4): 303~310

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Last Update: 2011-02-28