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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
Source PublicationEXTREMOPHILES
ISSN1431-0651
Volume20Issue:2Pages:149-156
AbstractGlucose-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.
KeywordGlucose-6-phosphate Dehydrogenase Thermoanaerobacter Tengcongensis Kinetics Thermostable Dual-coenzyme Specific Coupling Enzyme
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
DOI10.1007/s00792-016-0808-z
Indexed BySCI
Language英语
WOS KeywordGLUCOSE 6-PHOSPHATE DEHYDROGENASE ; LEUCONOSTOC-MESENTEROIDES ; THERMOTOGA-MARITIMA ; AQUIFEX-AEOLICUS ; BINDING ; ENZYME ; GENE ; G6PD
WOS Research AreaBiochemistry & Molecular Biology ; Microbiology
WOS SubjectBiochemistry & Molecular Biology ; Microbiology
Funding OrganizationTMO Renewables Limited ; Ministry of Science and Technology of China(2013CB734001)
WOS IDWOS:000374832600004
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/21018
Collection生化工程国家重点实验室
Affiliation1.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
Recommended Citation
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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