Cloning, expression, and characterization of a thermostable glucose-6-phosphate dehydrogenase from Thermoanaerobacter tengcongensis
文献类型：期刊论文
| 作者 | Li, Zilong1,2; Jiang, Ning3; Yang, Keqian2; Zheng, Jianting1 |
| 刊名 | EXTREMOPHILES
 |
| 出版日期 | 2016-03-01 |
| 卷号 | 20期号:2页码:149-156 |
| 关键词 | Glucose-6-phosphate dehydrogenase Thermoanaerobacter tengcongensis Kinetics Thermostable Dual-coenzyme specific Coupling enzyme |
| ISSN号 | 1431-0651 |
| 英文摘要 | 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. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biochemistry & Molecular Biology ; Microbiology |
| 研究领域[WOS] | Biochemistry & Molecular Biology ; Microbiology |
| 关键词[WOS] | GLUCOSE 6-PHOSPHATE DEHYDROGENASE ; LEUCONOSTOC-MESENTEROIDES ; THERMOTOGA-MARITIMA ; AQUIFEX-AEOLICUS ; BINDING ; ENZYME ; GENE ; G6PD |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000374832600004 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/21018]  |
| 专题 | 过程工程研究所_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 |
| 推荐引用方式 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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来源：过程工程研究所
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