Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Isolation of Bacillus subtilis SJ4 from Saeu (Shrimp) Jeotgal, a Korean Fermented Seafood, and Its Fibrinolytic Activity

  • Yao, Zhuang (Division of Applied Life Science (BK21 plus), Graduate School, Gyeongsang National University) ;
  • Meng, Yu (Division of Applied Life Science (BK21 plus), Graduate School, Gyeongsang National University) ;
  • Le, Huong Giang (Division of Applied Life Science (BK21 plus), Graduate School, Gyeongsang National University) ;
  • Kim, Jeong A (Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Kim, Jeong Hwan (Division of Applied Life Science (BK21 plus), Graduate School, Gyeongsang National University)
  • Received : 2019.06.10
  • Accepted : 2019.07.10
  • Published : 2019.12.28
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Abstract

A Bacillus strain, SJ4, exhibiting strong fibrinolytic activity was isolated from saeu (shrimp, Acetes chinensis) jeotgal, a Korean traditional fermented food and was identified as B. subtilis. The B. subtilis SJ4 strain can grow at a NaCl concentration of up to 15% (w/v). The fibrinolytic activity of B. subtilis SJ4 (152.0 U/ml) cultured in Luria-Bertani (LB) broth for 48 h at 37℃ with aeration was higher than that of B. subtilis SJ4 cultured in TSB (124.5 U/ml) under same culture conditions. The major proteins in the LB culture supernatant of B. subtilis SJ4 were analyzed by SDS-PAGE, which revealed three major bands (23, 25, and 28 kDa). The band (23 kDa) with strong fibrinolytic activity, analyzed on fibrin zymogram, was observed at 60-96 h of cultivation. The aprESJ4 gene encoding the major fibrinolytic enzyme, AprESJ4, was cloned by PCR. The aprESJ4 gene sequence exhibited high similarities with the fibrinolytic gene sequences of other Bacillus species. The amino acid sequence of AprESJ4 exhibited 98.9 and 98.4% similarity with subtilisin NAT and AprE2 of B. subtilis, respectively. Hence, B. subtilis SJ4 can be a potential starter culture for jeotgal products.

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References

  1. Schallmey M, Singh A, Ward OP. 2004. Developments in the use of Bacillus species for industrial production. Can. J. Microbiol. 50: 1-17. https://doi.org/10.1139/w03-076
  2. Cui W, Han L, Suo F, Liu Z, Zhou L, Zhou Z. 2018. Exploitation of Bacillus subtilis as a robust workhorse for production of heterologous proteins and beyond. World J. Microbiol. Biotechnol. 34: 145. https://doi.org/10.1007/s11274-018-2531-7
  3. Stein T. 2005. Bacillus subtilis antibiotics: structure, syntheses and specific functions. Mol. Microbiol. 56: 845-857. https://doi.org/10.1111/j.1365-2958.2005.04587.x
  4. Cutting SM. 2011. Bacillus probiotics. Food Microbiol. 28: 214-220. https://doi.org/10.1016/j.fm.2010.03.007
  5. Elshaghabee FMF, Rokana N, Gulhane RD, Sharma C, Panwar H. 2017. Bacillus as potential probiotics: status, concerns, and future perspectives. Front. Microbiol. 8: 1490. https://doi.org/10.3389/fmicb.2017.01490
  6. Mondol MA, Shin HJ, Islam MT. 2013. Diversity of secondary metabolites from marine Bacillus species: chemistry and biological activity. Mar. Drugs 11: 2846-2872. https://doi.org/10.3390/md11082846
  7. Weng Y, Yao J, Sparks S, Wang KY. 2017. Nattokinase: an oral antithrombotic agents for the prevention of cardiovascular disease. Int. J. Mol. Sci. 18. pii: E523.
  8. Dabbagh F, Negahdaripour M, Berenjian A, Behfar A, Mohammadi F, Zamani M, et al. 2014. Nattokinase: production and application. Appl. Microbiol. Biotechnol. 98: 9199-9206. https://doi.org/10.1007/s00253-014-6135-3
  9. Yao Z, Liu X, Shim JM, Lee KW, Kim HJ, Kim JH. 2017. Properties of a fibrinolytic enzyme secreted by Bacillus amyloliquefaciens RSB34, isolated from doenjang. J. Microbiol. Biotechnol. 27: 9-18. https://doi.org/10.4014/jmb.1608.08034
  10. Peng Y, Huang Q, Zhang R, Zhang YZ. 2003. Purification and characterization of a fibrinolytic enzyme produced by Bacillus amyloliquefaciens DC-4 screened from douchi, a traditional Chinese soybean food. Comp. Biochem. Physiol. B. Biochem. Mol. Biol. 134: 45-52. https://doi.org/10.1016/S1096-4959(02)00183-5
  11. Jo HD, Kwon GH, Park JY, Cha J, Song YS, Kim JH. 2011. Cloning and overexpression of aprE3-17 encoding the major fibrinolytic protease of Bacillus licheniformis CH3-17. Biotechnol. Bioprocess Eng. 16: 352-359. https://doi.org/10.1007/s12257-010-0328-0
  12. Kwon GH, Lee HA, Park JY, Kim JS, Lim JK, Park CS, et al. 2009. Development of a RAPD-PCR method for identification of Bacillus species isolated from cheonggukjang. Int. J. Food Microbiol. 129: 282-287. https://doi.org/10.1016/j.ijfoodmicro.2008.12.013
  13. Kim GM, Lee AR, Lee KW, Park JY, Chun J, Cha J, et al. 2009. Characterization of a 27 kDa fibrinolytic enzyme from Bacillus amyloliquefaciens CH51 isolated from cheonggukjang. J. Microbiol. Biotechnol. 19: 997-1004. https://doi.org/10.4014/jmb.0811.600
  14. Lee AR, Kim GM, Park JY, Jo HD, Cha J, Song YS, et al. 2010. Characterization of a 27 kDa fibrinolytic enzyme from Bacillus amyloliquefaciens CH86-1 isolated from cheonggukjang. J. Korean Soc. Appl. Biol. Chem. 53: 56-61. https://doi.org/10.3839/jabc.2010.010
  15. Yao Z, Kim JA, Kim JH. 2018. Gene cloning, expression, and properties of a fibrinolytic enzyme secreted by Bacillus pumilus BS15 isolated from gul (oyster) jeotgal. Biotechnol. Bioprocess Eng. 23: 293-301. https://doi.org/10.1007/s12257-018-0029-7
  16. Graudal N, Jürgens G. 2018. Conflicting evidence on health effects associated with salt reduction calls for a redesign of the salt dietary guidelines. Prog. Cardiovasc. Dis. 61: 20-26. https://doi.org/10.1016/j.pcad.2018.04.008
  17. Jeong SJ, Kwon GH, Chun J, Kim JS, Park CS, Kwon DY, et al. 2007. Cloning of fibrinolytic enzyme gene from Bacillus subtilis isolated from Cheonggukjang and its expression in protease-deficient Bacillus subtilis strains J. Microbiol. Biotechnol. 17: 1018-1023.
  18. Nakamura T, Yamagata Y, Ichishima E. 1992. Nucleotide sequence of the subtilisin NAT gene, aprN, of Bacillus subtilis (Natto). Biosci. Biotechnol. Biochem. 56: 1869-1871. https://doi.org/10.1271/bbb.56.1869
  19. Jang JS, Kang DO, Chun MJ, Byun SM. 1992. Molecular cloning of a subtilisin J gene from Bacillus stearothermophilus and its expression in Bacillus subtilis. Biochem. Biophys. Res. Commun. 184: 277-282. https://doi.org/10.1016/0006-291X(92)91189-W
  20. Peng Y, Yang X, Zhang Y. 2005. Microbial fibrinolytic enzymes: An overview of source, production, properties, and thrombolytic activity in vivo. Appl. Microbiol. Biotechnol. 69: 126-132. https://doi.org/10.1007/s00253-005-0159-7
  21. Prieto ML, O'Sullivan L, Tan SP, McLoughlin P, Hughes H, Gutierrez M, et al. 2014. In vitro assessment of marine Bacillus for use as livestock probiotics. Mar. Drugs 12: 2422-2445. https://doi.org/10.3390/md12052422
  22. Wang G. 2006. Diversity and biotechnological potential of the sponge-associated microbial consortia. J. Ind. Microbial Biotechnol. 33: 545-551. https://doi.org/10.1007/s10295-006-0123-2
  23. Herzog B, Overy DP, Haltli B, Kerr RG. 2016. Discovery of keratinases using bacteria isolated from marine environments. Syst. Appl. Microbiol. 39: 49-57. https://doi.org/10.1016/j.syapm.2015.10.004