Expression and characterization of glutamate decarboxylase from Lactobacillus brevis HYE1 isolated from kimchi.
World J Microbiol Biotechnol
; 34(3): 44, 2018 Mar 02.
Article
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| MEDLINE
| ID: mdl-29500614
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 Km and Vmax values 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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MEDLINE
Asunto principal:
Levilactobacillus brevis
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Alimentos Fermentados
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Glutamato Descarboxilasa
Idioma:
En
Revista:
World J Microbiol Biotechnol
Año:
2018
Tipo del documento:
Article