Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning of aprE86-1 Gene Encoding a 27-kDa Mature Fibrinolytic Enzyme from Bacillus amyloliquefaciens CH86-1

  • Lee, Ae-Ran (Division of Applied Life Science (BK21 Program), Graduate School, Gyeongsang National University) ;
  • Kim, Gyoung-Min (Division of Applied Life Science (BK21 Program), Graduate School, Gyeongsang National University) ;
  • Kwon, Gun-Hee (Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Lee, Kang-Wook (Division of Applied Life Science (BK21 Program), Graduate School, Gyeongsang National University) ;
  • Park, Jae-Yong (Institute of Agriculture and Life Science, Gyeongsang National University) ;
  • Chun, Ji-Yeon (Department of Food Science and Technology, Sunchon National University) ;
  • Cha, Jae-Ho (Department of Microbiology, Pusan National University) ;
  • Song, Young-Sun (School of Food and Life Science, Inje University) ;
  • Kim, Jeong-Hwan (Division of Applied Life Science (BK21 Program), Graduate School, Gyeongsang National University)
  • Published : 2010.02.28
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Abstract

A gene encoding the major secreted fibrinolytic protein of Bacillus amyloliquefaciens CH86-1 was cloned from genomic DNAs. DNA sequencing showed that the gene, aprE86-1, could direct the synthesis of a mature protein 275 amino acids in length after processing. When aprE86-1 was introduced into B. subtilis, a mature 27-kDa protein was produced as expected. The fibrinolytic activity of the B. subtilis transformant (TF) was higher than that of B. amyloliquefaciens CH86-1, showing the possibility of increasing the fibrinolytic activity of Bacillus strains through genetic engineering.

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References

  1. Bradford, M. M. 1976. Rapid and sensitive methods for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254. https://doi.org/10.1016/0003-2697(76)90527-3
  2. Choi, N. S., K. H. Yoo, K. S. Yoon, K. T. Chang, P. J. Maeng, and S. H. Kim. 2005. Identification of recombinant subtilisins. J. Microbiol. Biotechnol. 15: 35-39.
  3. Choi, N. S., D. M. Chung, C. H. Ryu, K. S. Yoon, P. J. Maeng, and S. H. Kim. 2006. Identification of three extracellular proteases from Bacillus subtilis KCTC 3014. J. Microbiol. Biotechnol. 16: 457-464.
  4. Jeong, S. J., G. H. Kwon, J. Chun, J. S. Kim, C. S. Park, D. Y. Kwon, and J. H. Kim. 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.
  5. Kim, G. M., A. R. Lee, K. W. Lee, J. Y. Park, J. Chun, J. Cha, Y. S. Song, and J. H. Kim. 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
  6. Kwon, G. H., W. J. Jeong, A. R. Lee, J. Y. Park, J. Cha, Y. S. Song, and J. H. Kim. 2008. Heterologous gene expression of aprE2 encoding a 29 kDa fibrinolytic enzyme from Bacillus subtilis in Bacillus licheniformis ATCC 10716. Food Sci. Biotechnol. 17: 1372-1375.
  7. Kwon, G. H., H. A. Lee, J. Y. Park, J. S. Kim, J. Lim, C. S. Park, D. Y. Kwon, Y. S. Kim, and J. H. Kim. 2009. Development of a RAPD-PCR method for identification of Bacillus species from Cheonggukjang. Int. J. Food Microbiol. 129: 282-287. https://doi.org/10.1016/j.ijfoodmicro.2008.12.013
  8. Lee, A. R., G. M. Kim, J. Y. Park , H. D. Jo, J. Cha, Y. S. Song, J. Chun, and J. H. Kim. 2010. Characterization of a 27 kDa Fibrinolytic Enzyme from Bacillus amyloliquefaciens CH86-1 Isolated from Cheonggukjang J. Korean Soc. Appl. Biol. Chem. 53(1): in-Press.
  9. Park, S. S., S. L. Wong, L. F. Wang, and R. H. Doi. 1989. Bacillus subtilis subtilisin gene (aprE) is expressed from a ${\sigma}^A({\sigma}^{43})$ promoter in vitro and in vivo. J. Bacteriol. 171: 2657-2665.
  10. Wu, X. C., W. Lee, L. Tran, and S. L. Wong. 1991. Engineering a Bacillus subtilis expression-secretion system with a strain deficient in six extracellular proteases. J. Bacteriol. 173: 4952-4958.

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