Title |
Expression and characterization of glutamate decarboxylase from Lactobacillus brevis HYE1 isolated from kimchi
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Published in |
World Journal of Microbiology and Biotechnology, March 2018
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DOI | 10.1007/s11274-018-2427-6 |
Pubmed ID | |
Authors |
Hee Seon Lim, Dong-Ho Seo, In-Tae Cha, Hyunjin Lee, Young-Do Nam, Myung-Ji Seo |
Abstract |
A putative gene (gadlbhye1) encoding glutamate decarboxylase (GAD) was cloned from Lactobacillus brevis HYE1 isolated from kimchi, a traditional Korean fermented vegetable. The amino acid sequences of GADLbHYE1 showed 48% homology with the GadA family and 99% identity with the GadB family from L. brevis. The cloned GADLbHYE1 was functionally expressed in Escherichia coli using inducible expression vectors. The expressed recombinant GADLbHYE1 was successfully purified by Ni-NTA affinity chromatography, and had a molecular mass of 54 kDa with optimal hydrolysis activity at 55 °C and pH 4.0. Its thermal stability was determined to be higher than that of other GADs from L. brevis, based on its melting temperature (75.18 °C). Kinetic parameters including Kmand Vmaxvalues for GADLbHYE1 were 4.99 mmol/L and 0.224 mmol/L/min, respectively. In addition, the production of gamma-aminobutyric acid in E. coli BL21 harboring gadlbhye1/pET28a was increased by adding pyridoxine as a cheaper coenzyme. |
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