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Over-expression, purification, and characterization of recombinant NAD-malic enzyme from Escherichia coli K12
Wang, Jinxia; Tan, Haidong; Zhao, Zongbao (Kent)
关键词Malic Enzyme Escherichia Coli K12 Over-expression Enzyme Kinetics
刊名PROTEIN EXPRESSION AND PURIFICATION
2007-05-01
DOI10.1016/j.pep.2006.11.017
53期:1页:97-103
收录类别SCI
文章类型Article
WOS标题词Science & Technology ; Life Sciences & Biomedicine
类目[WOS]Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
研究领域[WOS]Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
关键词[WOS]POLYACRYLAMIDE-GEL ELECTROPHORESIS ; MOLECULAR-WEIGHTS ; COLI ; BINDING ; CLONING ; GENE ; SITE ; RESIDUES ; PIGEON
英文摘要NAD+-dependent malic enzyme (NAD-ME) gene from Escherichia coli K12 was inserted into an expression vector pET24b(+) and transformed into E coli BL21 (DE3). Recombinant NAD-ME was expressed upon IPTG induction, purified with affinity chromatography, and biochemically characterized. The results showed that recombinant NAD-ME could be produced mainly in a soluble form. The monomeric molecular weight of recombinant NAD-ME was about 65 kDa, whereas monomer, homotetramer, and homooctamer were formed in solution as revealed by nondenaturing polyacrylamide gel electrophoresis analysis. Finally, the K-m, values of NAD-ME for L-malate and NAD were determined as 0.420 +/- 0.174 and 0.097 +/- 0.038 mM, respectively, at pH 7.2. By using this over-expression and purification system, recombinant E coli K12 NAD-ME can now be obtained in large quantity necessary for further biochemical characterization and applications. (c) 2006 Elsevier Inc. All rights reserved.
语种英语
WOS记录号WOS:000246245000011
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被引频次:17[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/140630
专题中国科学院大连化学物理研究所
作者单位1.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China
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Wang, Jinxia,Tan, Haidong,Zhao, Zongbao . Over-expression, purification, and characterization of recombinant NAD-malic enzyme from Escherichia coli K12[J]. PROTEIN EXPRESSION AND PURIFICATION,2007,53(1):97-103.
APA Wang, Jinxia,Tan, Haidong,&Zhao, Zongbao .(2007).Over-expression, purification, and characterization of recombinant NAD-malic enzyme from Escherichia coli K12.PROTEIN EXPRESSION AND PURIFICATION,53(1),97-103.
MLA Wang, Jinxia,et al."Over-expression, purification, and characterization of recombinant NAD-malic enzyme from Escherichia coli K12".PROTEIN EXPRESSION AND PURIFICATION 53.1(2007):97-103.
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