Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning and Expression of a Fibrinolytic Enzyme Gene, aprECJ1, from Bacillus velezensis CJ1 Isolated from Myeolchi Jeotgal

  • Yoo, Ji Yeon (Division of Applied Life Science (BK21 Four), Graduate School, Gyeongsang National University) ;
  • Yao, Zhuang (Division of Applied Life Science (BK21 Four), Graduate School, Gyeongsang National University) ;
  • Lee, Se Jin (Division of Applied Life Science (BK21 Four), Graduate School, Gyeongsang National University) ;
  • Jeon, Hye Sung (Division of Applied Life Science (BK21 Four), Graduate School, Gyeongsang National University) ;
  • Kim, Jeong Hwan (Division of Applied Life Science (BK21 Four), Graduate School, Gyeongsang National University)
  • Received : 2021.04.21
  • Accepted : 2021.05.14
  • Published : 2021.09.28
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Abstract

Bacillus velezensis CJ1, showing significant fibrinolytic activity, was isolated from Myeolchi Jeotgal, a popular Korean fermented seafood. When B. velezensis CJ1 was grown on four different culture media, the culture on the Luria-Bertani (LB) broth showed the highest fibrinolytic activity (102.94 mU/μl) at 48 h. LB was also the best medium for growth. SDS-PAGE of culture supernatant showed four major bands, 38, 35, 27, and 22 kDa in size. Fibrin zymography showed four active bands, 50, 47, 40, and 30 kDa in size. A gene homologous to aprE of the Bacillus species was cloned by PCR. DNA sequencing showed that aprECJ1 can encode a protease consisting of 382 amino acids. The translated amino acid sequence of AprECJ1 showed high identity values with those of B. velezensis strains and other Bacillus species. The aprECJ1 gene was introduced into B. subtilis WB600 using an E. coli-Bacillus shuttle vector, pHY300PLK, and overexpressed. A 27 kDa band corresponding to the mature form of AprECJ1 was produced and confirmed by SDS-PAGE and fibrin zymography. B. subtilis WB600 [pHYaprECJ1] showed 1.8-fold higher fibrinolytic activity than B. velezensis CJ1 at 48 h.

Keywords

Acknowledgement

This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2017R1D1A1B03030037) and also by a NRF grant funded by the Korea government (MSIT) (No. NRF-2020R1A2C100826711). Yoo JY, Yao Z, Lee SJ, and Jeon HS were supported by BK21 four program, MOE, Republic of Korea.

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