Overexpression of a recombinant gamma-glutamyltranspeptidase from Escherichia coli Novablue.

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dc.contributor.authorYao, Ya-Fengen_US
dc.contributor.authorWeng, Yih-Mingen_US
dc.contributor.authorHu, Hui-Yuen_US
dc.contributor.authorLin, Long-Liuen_US
dc.date.accessioned2006-12-09en_US
dc.date.accessioned2009-05-27T09:51:45Z
dc.date.available2006-12-09en_US
dc.date.available2009-05-27T09:51:45Z
dc.date.issued2006-12-09en_US
dc.description.abstractA truncated Escherichia coli Novablue gamma-glutamyltranspeptidase (EcGGT) gene, lacking the first 48-bp coding sequence for part of the signal sequence, was amplified by polymerase chain reaction (PCR) and cloned into expression vector pQE-30 to generate pQE-EcGGT. The maximum production of His6-tagged enzyme by E. coli M15 (pQE-EcGGT) was achieved with 0.1 mM IPTG induction for 12 h at 20 degrees C. The overexpressed enzyme was purified to homogeneity by nickel-chelate chromatography to a specific transpeptidase activity of 4.25 U/mg protein and a final yield of 83%. The molecular masses of the subunits of the purified enzyme were determined to be 41 and 21 kDa respectively by SDS-PAGE, indicating the precursor EcGGT still undergoes the post-translational processing even in the truncation of signal sequence. His6-tagged EcGGT migrated relative to the molecular mass of approximately 120 kDa and its heterodimeric structure was confirmed by a native-PAGE gel.en_US
dc.description.affiliationGraduate Institute of Food Science, National Chiayi University, 300 University Road, 60083 Chiayi, Taiwan.en_US
dc.identifier.citationYao YF, Weng YM, Hu HY, Lin LL. Overexpression of a recombinant gamma-glutamyltranspeptidase from Escherichia coli Novablue. Indian Journal of Biochemistry & Biophysics. 2006 Dec; 43(6): 345-50en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/28783
dc.language.isoengen_US
dc.source.urihttps://https://www.niscair.res.in/ScienceCommunication/ResearchJournals/rejour/ijbb/ijbb0.aspen_US
dc.subject.meshEscherichia coli --enzymologyen_US
dc.subject.meshRecombinant Proteins --biosynthesisen_US
dc.subject.meshgamma-Glutamyltransferase --biosynthesisen_US
dc.titleOverexpression of a recombinant gamma-glutamyltranspeptidase from Escherichia coli Novablue.en_US
dc.typeJournal Articleen_US
dc.typeResearch Support, Non-U.S. Gov'ten_US
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