Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Characterization of an Aminopeptidase A from Tetragenococcus halophilus CY54 Isolated from Myeolchi-Jeotgal

  • Tae Jin Kim (Division of Applied Life Science (BK21 Four), Graduate School, Gyeongsang National University) ;
  • Min Jae Kim (Division of Applied Life Science (BK21 Four), Graduate School, Gyeongsang National University) ;
  • Yun Ji Kang (Division of Applied Life Science (BK21 Four), Graduate School, Gyeongsang National University) ;
  • Ji Yeon Yoo (Division of Applied Life Science (BK21 Four), Graduate School, Gyeongsang National University) ;
  • Jeong Hwan Kim (Division of Applied Life Science (BK21 Four), Graduate School, Gyeongsang National University)
  • Received : 2022.10.03
  • Accepted : 2022.12.18
  • Published : 2023.03.28
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Abstract

In this study, a pepA gene encoding glutamyl (aspartyl)-specific aminopeptidase (PepA; E.C. 3.4.11.7) was cloned from Tetragenococcus halophilus CY54. The translated PepA from T. halophilus CY54 showed very low similarities with PepAs from Lactobacillus and Lactococcus genera. The pepA from T. halophilus CY54 was overexpressed in E. coli BL21(DE3) using pET26b(+). The recombinant PepA was purified by using an Ni- NTA column. The size of the recombinant PepA was 39.13 kDa as determined by SDS-PAGE, while its optimum pH and temperature were pH 5.0 and 60℃, respectively. In addition, the PepA was completely inactivated by 1 mM EDTA, indicating its metallopeptidase nature. The Km and Vmax of the PepA were 0.98 ± 0.006 mM and 0.1 ± 0.002 mM/min, respectively, when Glu-pNA was used as the substrate. This is the first report on PepA from Tetragenococcus species.

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 were supported by the BK21 Four Program of the MOE, Republic of Korea.

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