NDLI: Purification and characterization of glutamate decarboxylase from Enterococcus raffinosus TCCC11660

Content Provider	Springer Nature Link
Author	Chang, Chuanyou Zhang, Jun Ma, Shenxi Wang, Lin Wang, Depei Zhang, Jian Gao, Qiang
Copyright Year	2017
Abstract	Glutamate decarboxylase (GAD) is the sole enzyme that synthesizes γ-aminobutyric acid through the irreversible decarboxylation of l-glutamate. In this study, the purification and characterization of an unreported GAD from a novel strain of Enterococcus raffinosus TCCC11660 were investigated. The native GAD from E. raffinosus TCCC11660 was purified 32.3-fold with a recovery rate of 8.3%, using ultrafiltration and ammonium sulfate precipitation, followed by ion-exchange and size-exclusion chromatography. The apparent molecular weight of purified GAD, as determined by SDS-PAGE and size-exclusion chromatography was 55 and 110 kDa, respectively, suggesting that GAD exists as a dimer of identical subunits in solution. In the best sodium citrate buffer, metal ions of Mo6+ and Mg2+ had positive effects, while Cu2+, Fe2+, Zn2+ and Co2+ showed significant adverse effects on enzyme activity. The optimum pH and temperature of GAD were determined to be 4.6 and 45 °C, while the K m and V max values for the sole l-glutamate substrate were 5.26 and 3.45 μmol L−1 min−1, respectively.
Starting Page	817
Ending Page	824
Page Count	8
File Format	PDF
ISSN	13675435
Journal	Journal of Industrial Microbiology and Biotechnology
Volume Number	44
Issue Number	6
e-ISSN	14765535
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2017-01-18
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	Glutamate decarboxylase γ-Aminobutyric acid Enterococcus raffinosus Purification Characterization Microbiology Biochemistry Inorganic Chemistry Genetic Engineering Biotechnology Bioinformatics
Content Type	Text
Resource Type	Article
Subject	Medicine Bioengineering Applied Microbiology and Biotechnology Biotechnology

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