Journal
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
Volume 34, Issue 3, Pages -Publisher
SPRINGER
DOI: 10.1007/s11274-018-2427-6
Keywords
Gamma-aminobutyric acid; Glutamate decarboxylase; Lactobacillus brevis; Expression
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Funding
- Main Research Programs of the Korea Food Research Institute (KFRI) - Ministry of Science, ICT & Future Planning [E0170602-01]
- National Research Foundation of Korea (NRF) - Ministry of Science, ICT & Future Planning [NRF-2014R1A1A1002980]
- National Research Foundation of Korea (NRF) - Ministry of Education [NRF-2016R1D1A1B03931582]
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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 degrees 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 degrees C). Kinetic parameters including K-m and V-max v alues 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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