Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Characterization of the Recombinant Glutamate Decarboxylase of Lactobacillus brevis G144 Isolated from Galchi Jeotgal, a Korean Salted and Fermented Seafood

  • Kim, Jeong A (Institute of Agriculture and Life Science, Graduate School, Gyeongsang National University) ;
  • Park, Ji Yeong (Division of Applied Life Science BK21 4, Graduate School, Gyeongsang National University) ;
  • Kim, Jeong Hwan (Institute of Agriculture and Life Science, Graduate School, Gyeongsang National University)
  • Received : 2020.02.27
  • Accepted : 2020.04.13
  • Published : 2021.03.28
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Abstract

A γ-aminobutyric acid (GABA)-producing microorganism was isolated from galchi (hairtail fish, Trichiurus lepturus) jeotgal, a Korean salted and fermented seafood. The G144 isolate produced GABA excessively when incubated in MRS broth containing monosodium glutamate (MSG, 3%, w/v). G144 was identified as Lactobacillus brevis through 16S rRNA and recA gene sequencing. gadB and gadC encoding glutamate decarboxylase (GAD) and glutamate/GABA antiporter, respectively, were cloned and gadB was located downstream of gadC. The operon structure of gadCB was confirmed by reverse transcription (RT)-polymerase chain reaction. gadB was overexpressed in Escherichia coli and recombinant GAD was purified and its size was 54.4 kDa as evidenced by SDS-PAGE results. Maximum GAD activity was observed at pH 5.0 and 40℃ and the activity was dependent on pyridoxal 5'-phophate. The Km and Vmax of GAD were 8.6 mM and 0.01 mM/min, respectively.

Keywords

Acknowledgement

This work was supported by Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through (Agricultural Microbiome R&D Program), funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA) (918005044HD070). JA Kim and JY park were supported by BK21 Plus program, MOE, Republic of Korea.

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