title: Characterization of glutamate decarboxylase from Lactobacillus plantarum and its C-terminal function for the pH dependence of activity.
abstract: The gadB gene encoding glutamate decarboxylase (GAD) from Lactobacillus plantarum was cloned and expressed in Escherichia coli. The recombinant enzyme exhibited maximal activity at 40 °C and pH 5.0. The 3D model structure of L. plantarum GAD proposed that its C-terminal region (Ile454-Thr468) may play an important role in the pH dependence of catalysis. Accordingly, C-terminally truncated (Δ3 and Δ11 residues) mutants were generated and their enzyme activities compared with that of the wild-type enzyme at different pH values. Unlike the wild-type GAD, the mutants showed pronounced catalytic activity in a broad pH range of 4.0-8.0, suggesting that the C-terminal region is involved in the pH dependence of GAD activity. Therefore, this study may provide effective target regions for engineering pH dependence of GAD activity, thereby meeting industrial demands for the production of γ-aminobutyrate in a broad range of pH values.
authors.affiliation: School of Applied Biosciences, Kyungpook National University , Daegu 702-701, South Korea.
authors.forename: Sun-Mi
authors.initials: SM
authors.lastname: Shin
authors.forename: Hana
authors.initials: H
authors.lastname: Kim
authors.forename: Yunhye
authors.initials: Y
authors.lastname: Joo
authors.forename: Sang-Jae
authors.initials: SJ
authors.lastname: Lee
authors.forename: Yong-Jik
authors.initials: YJ
authors.lastname: Lee
authors.forename: Sang Jun
authors.initials: SJ
authors.lastname: Lee
authors.forename: Dong-Woo
authors.initials: DW
authors.lastname: Lee
mesh_terms: D020816:Amino Acid Motifs; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D055162:Biocatalysis; D004795:Enzyme Stability; D005968:Glutamate Decarboxylase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D048191:Lactiplantibacillus plantarum / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D005680:gamma-Aminobutyric Acid; D005968:Glutamate Decarboxylase
keywords: C-terminal region; GABA; Lactobacillus plantarum; glutamate decarboxylase; pH dependence