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Properties of a Novel Glutamate Decarboxylase (GAD) from Levilactobacillus brevis B737 Isolated from Cabbage Kimchi

  • Tae Jin, Kim (Division of Applied Life Science (BK Four), Graduate School, Gyeongsang National University) ;
  • Min Jae, Kim (Division of Applied Life Science (BK Four), Graduate School, Gyeongsang National University) ;
  • Bong Sin, Kim (Environmental Agriculture Research Division, Gyeongnam Agricultural Research and Extension Services) ;
  • Ji Yeon, Yoo (Division of Applied Life Science (BK Four), Graduate School, Gyeongsang National University) ;
  • Yun Ji, Kang (Division of Applied Life Science (BK Four), Graduate School, Gyeongsang National University) ;
  • Jeong Hwan, Kim (Division of Applied Life Science (BK Four), Graduate School, Gyeongsang National University)
  • Received : 2022.06.16
  • Accepted : 2022.08.02
  • Published : 2022.09.28

Abstract

γ-Aminobutyric acid (GABA) is a multi-functional compound with broad applications for food industry. GABA producing bacteria were isolated from cabbage kimchi. Among them, B737 was the best GABA producer when culture supernatants were analyzed by TLC. B737 was identified as Levilactobacillus brevis by 16S rRNA gene sequencing. Its glutamate decarboxylase (GAD) gene was cloned by PCR and the nucleotide sequence determined. B737 GAD consisting of 485 amino acids is the largest in size among GADs reported from LAB so far. gadB from L. brevis B737 was overexpressed in Escherichia. coli BL21(DE3) using pET26b(+).pET26b(+). The recombinant GAD was purified and its size was 55 kDa by SDS-PAGE. Maximum GAD activity was observed at pH 5 and 40℃ and the activity was dependent on pyridoxal 5'-phosphate. Km and Vmax of recombinant GAD were 6.2 ± 0.06 mM and 0.34 ± 0.002 mM/min, respectively. L. brevis B737 can be used as a starter for fermented foods with high GABA contents.

Keywords

Acknowledgement

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. NRF-2020R1A2C100826711). Kim TJ, Kim MJ, Kang YJ, and Yoo JY have been supported by BK21 four program from MOE, Korea.

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