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Cloning, expression, and characterization of a thermostable glucose-6-phosphate dehydrogenase from Thermoanaerobacter tengcongensis
Li, Zilong1,2; Jiang, Ning3; Yang, Keqian2; Zheng, Jianting1
2016-03-01
发表期刊EXTREMOPHILES
ISSN1431-0651
卷号20期号:2页码:149-156
摘要Glucose-6-phosphate dehydrogenases (G6PDs) are important enzymes widely used in bioassay and biocatalysis. In this study, we reported the cloning, expression, and enzymatic characterization of G6PDs from the thermophilic bacterium Thermoanaerobacter tengcongensis MB4 (TtG6PD). SDS-PAGE showed that purified recombinant enzyme had an apparent subunit molecular weight of 60 kDa. Kinetics assay indicated that TtG6PD preferred NADP(+) (k(cat)/K-m = 2618 mM(-1) s(-1), k(cat) = 249 s(-1), K-m = 0.10 +/- 0.01 mM) as cofactor, although NAD(+) (k(cat)/K-m = 138 mM(-1) s(-1), k(cat) = 604 s(-1), K-m = 4.37 +/- 0.56 mM) could also be accepted. The K-m values of glucose-6-phosphate were 0.27 +/- 0.07 mM and 5.08 +/- 0.68 mM with NADP(+) and NAD(+) as cofactors, respectively. The enzyme displayed its optimum activity at pH 6.8-9.0 for NADP(+) and at pH 7.0-8.6 for NAD(+) while the optimal temperature was 80 degrees C for NADP(+) and 70 degrees C for NAD(+). This was the first observation that the NADP(+)-linked optimal temperature of a dual coenzyme-specific G6PD was higher than the NAD(+)-linked and growth (75 degrees C) optimal temperature, which suggested G6PD might contribute to the thermal resistance of a bacterium. The potential of TtG6PD to measure the activity of another thermophilic enzyme was demonstrated by the coupled assays for a thermophilic glucokinase.
关键词Glucose-6-phosphate Dehydrogenase Thermoanaerobacter Tengcongensis Kinetics Thermostable Dual-coenzyme Specific Coupling Enzyme
文章类型Article
WOS标题词Science & Technology ; Life Sciences & Biomedicine
DOI10.1007/s00792-016-0808-z
收录类别SCI
语种英语
关键词[WOS]GLUCOSE 6-PHOSPHATE DEHYDROGENASE ; LEUCONOSTOC-MESENTEROIDES ; THERMOTOGA-MARITIMA ; AQUIFEX-AEOLICUS ; BINDING ; ENZYME ; GENE ; G6PD
WOS研究方向Biochemistry & Molecular Biology ; Microbiology
WOS类目Biochemistry & Molecular Biology ; Microbiology
项目资助者TMO Renewables Limited ; Ministry of Science and Technology of China(2013CB734001)
WOS记录号WOS:000374832600004
引用统计
被引频次:2[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.ipe.ac.cn/handle/122111/21018
专题生化工程国家重点实验室
作者单位1.Chinese Acad Sci, Natl Key Lab Biochem Engn, Inst Proc Engn, Beijing 100190, Peoples R China
2.Chinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
3.Chinese Acad Sci, Inst Microbiol, Dept Ind Microbiol & Biotechnol, Beijing 100101, Peoples R China
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GB/T 7714
Li, Zilong,Jiang, Ning,Yang, Keqian,et al. Cloning, expression, and characterization of a thermostable glucose-6-phosphate dehydrogenase from Thermoanaerobacter tengcongensis[J]. EXTREMOPHILES,2016,20(2):149-156.
APA Li, Zilong,Jiang, Ning,Yang, Keqian,&Zheng, Jianting.(2016).Cloning, expression, and characterization of a thermostable glucose-6-phosphate dehydrogenase from Thermoanaerobacter tengcongensis.EXTREMOPHILES,20(2),149-156.
MLA Li, Zilong,et al."Cloning, expression, and characterization of a thermostable glucose-6-phosphate dehydrogenase from Thermoanaerobacter tengcongensis".EXTREMOPHILES 20.2(2016):149-156.
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