Isolation of Bacteria from Chunggukjang Prepared by Rice Straw and Identification of Protease Secreted

볏짚 청국장 발효 세균 분리 및 분비된 protease의 확인

  • Oh, Jae-Hyeon (Department of Biological Sciences, Sunchon National University) ;
  • Lee, Byeong-Jeong (Department of Biological Sciences, Sunchon National University) ;
  • Paik, Hyoung-Rok (Department of Biological Sciences, Sunchon National University) ;
  • Jung, Sang-Chul (Department of Biological Sciences, Sunchon National University) ;
  • Baik, Keun-Sik (Department of Biological Sciences, Sunchon National University) ;
  • Choi, Sang-Ki (Department of Biological Sciences, Sunchon National University)
  • Published : 2009.03.31


To isolate bacteria secreting protease, which can dissolve fibrin efficiently, we prepared chunggukjang using rice straw and isolated, preliminarily, approximately 100 bacterial stains. Their capabilities to dissolve milk protein as well as fibrin included in media were then examined and finally, five strains named J1 - J5 were selected. Among them, J-4, which is close to bacillus subtilis, showed highest activity for fibrin dissolution. Proteases secreted from the J-4 strain were partially purified from culture supernatant using DEAE-sepharose column chromatography and identified with SDS-polyacrylamide gel electrophoresis. Three proteins were subjected to analysis with MALDI-TOF and PMF (Peptide Mass Fingerprinting). 41.9 kDa protein was identified as a neutral protease. On the other hand, 45 kDa protein turned out to be bacillopeptidase F, with a molecular mass of 91.7 kDa, indicating that partially purified peptide is a degradation product.


  1. Ahn, Y. S., Y. S. Kim, and D. H. Shin. 2006. Isolation, identification, and fermentation characteristics of Bacillus sp. with high protease activity from traditional cheonggukjang. Korean J. Food Sci. Technol. 38, 82-87
  2. Chun, D. S., D. K. Kang, and H. K. Kim. 2002. Isolation and enzyme production of a neutral protease-strain, Bacillus sp. DS-1. Kor. J. Microbiol. Biotechnol. 30, 346-351
  3. Fujii, M., M. Takagi, T. Imanaka, and S. Aiba. 1983. Molecular cloning of a thermostable neutral protease gene from Bacillus stearothermophilus in vector plasmid and its expression in Bacillus stearothermophilus and Bacillus subtilis. J. Bact. 154, 831-837
  4. Godfrey, T. and S. West. 1996. Industrial Enzymology. 2nd eds., Macmillian Publishers Inc., New York
  5. Horikoshi, K. 1971. Production of alkaline enzymes by alkaalkalophilic microorganism. Agric. Biol. Chem. 35, 1407-1414
  6. Kobayashi, R., T. Yoshimoto, and D. Tsuru. 1989. Complete amino acid sequence of neutral protease from Bacillus subtilis var. amylosacchariticus. Agric. Biol. Chem. 53, 2737-2749
  7. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685
  8. Leighton, T. J., R. H. Doi, R. A. J. Warren, and R. A. Kelln. 1973. The relationship of serine protease activity to RNA polymerase modification and sporulation in Bacillus subtilis. J. Mol. Biol. 76, 103-122
  9. Paul, P., U. Faust, W. Sitting, and A. S. Dieter. 1987. Fundamentals of Biotechnology, pp. 493-498, VCH, New York
  10. Payne, W. J. 1963. Pure culture studies of the degradation of detergent compounds. Biotechnol. Bioeng. 5, 355
  11. Peng, Y., X. Yang and Y. Zhang. 2005. Microbial fibronolytic enzymes: an overview of source, production, properties, and thrombolytic activity in vivo. Appl. Microbiol. Biotechnol. 69, 126-32
  12. Rao, M. B., A. M. Tankale, M. S. Ghatage, and V. V. Deshpande. 1998. Molecular and biotechnological aspects of microbial protease. Microbiol. Mol. Biol. Rev. 62. 597-635
  13. Reed, G. 1975. Enzymes in Food Processing. pp. 123-179, Academic Press, New York
  14. Ward, O. P. 1985. Proteolytic Enzymes, pp. 789-792, In Murray, M. Y. (eds.), Comprehensive Biotechnology, Pergamon Press, New York
  15. Wu, X. C., S. Nathoo, A. S. Pang, T. Carne, and S. L. Wong. 1990. Cloning, genetic organization, and characterization of a structural gene encoding bacillopeptidase F from Bacillus subtilis. J. Biol. Chem. 265, 6845-6850

Cited by

  1. Changes in Components and Peptides during Fermentation of Cheonggookjang vol.24, pp.1, 2011,
  2. Isolation of Bacteria with Protease Activity from Cheonggukjang and Purification of Fibrinolytic Enzyme vol.23, pp.2, 2013,
  3. Protease and γ-glutamyltranspeptidase activities of bacillus spp. isolated from rice straw vol.21, pp.1, 2012,
  4. Quality Characteristics of Commercial Korean Types of Fermented Soybean Sauces in China vol.45, pp.6, 2013,