Identification of Total Extracellular Fibrinase from Bacillus sp. DJ Using One-or Two-Dimensional Fibrin Zymography for Proteomic Approach

  • CHOI, NACK-SHICK (Proteome Research Laboratory, Korea research Institute of Bioscience and Biotechnology) ;
  • JIN-YOUNG LEE (Proteome Research Laboratory, Korea research Institute of Bioscience and Biotechnology) ;
  • KAB-SEOG YOON (Proteome Research Laboratory, Korea research Institute of Bioscience and Biotechnology) ;
  • KYOUNG-YOEN HAN (Proteome Research Laboratory, Korea research Institute of Bioscience and Biotechnology) ;
  • SEUNG-HO KIM (Proteome Research Laboratory, Korea research Institute of Bioscience and Biotechnology)
  • Published : 2001.12.01

Abstract

An extracellular fibrinolytic-enzyme-producing bacterium was isolated from Doen-Jang, a Korean traditional fermented flood, and identified as Bacillus sp. DJ based on its morphology and cellular fatty acid composition. The total extracellular fibrinase (EF) from Bacillus sp. DJ was analyzed using three fibrin zymographic techniques, SDS-fibrin zymography (SDS-FZ), isoelectrofocucing-fibrin zymographs(IEF-FZ), and a two-dimensional SDS-fibrin zymographic analysis (2D SDS-FZ). As a result, the EP map of Bacillus sp. DJ was established. The results suggest that the 2D SDS-FZ method will be a useful tool for the proteomic approach for many other bacterial pretenses.

Keywords

References

  1. Anal. Biochem. v.72 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Bradford, M. M.
  2. Biotechnol. Techniq. v.13 Application of fibrin zymography ofor determination the optimum culture time for protease activity Choi, N. S.;S. H. Kim
  3. Anal. biochem. v.281 Two fibrin zymography methods for analysis of plasminogen activators on gels Choi, N. S.;S. H. Kim
  4. J. Biol. Chem. v.226 A simple method for the isolation and purification of total lipids from animals tissures Folch, J.;M. Lees;G. H. Stanley
  5. Anal. Biochem. v.102 Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates Heussen, C.;E. B. Dowdle
  6. Anal. Biochem. v.270 Electrophoretic analysis of protease inhibitors in fibrin zymography Kim, S. H.;N. S. Choi
  7. Biosci. Biotechnol. Biochem. v.64 Purification and characterization of subtilisin DJ-4 screted by Bacillus sp. strain DJ-4 screened from Doen-Jang Kim, S. H.;N. S. Choi
  8. Anal. Biochem. v.263 Fibrin zymography: A direct analysis of fibrinolytic enzyme on gels Kim, S. H.;N. S. Choi;W. Y. Lee
  9. Kor. J. Microbiol. v.34 Isolation of Bacillus strains secreting fibrinolytic enzymes from Doen-Jang Kim, S. H.;N. S. Choi;W. Y. Lee;D. H. Kim
  10. Appl. Environ. Microbiol. v.62 Purification and characterization of a fibrinolytic enzyme produced from Bacillus sp. strain CK 11-4 screened from Chungkook-Jang Kim, W.;K. Choi;Y. Kim;H. Park;J. Choi;Y. Lee;H. Oh;I. Kwon;S. Lee
  11. Anal. Biochem. v.218 Quantitative zymography: Detection of picogram quantities of gelatinases Kleiner, D. E.;W. G. Stetler-Stevenson
  12. Anal. Biochem. v.249 Zymography: a single-step staining method for quantitation of proteolytic activity on substrate gels Leber, T. M.;F. R. Balkwill
  13. Arch. Biochem. Biophys. v.319 Casein zymography: A method to study mu-calpain, m-calpain, and their inhibitor agents Raser, K. J.;A. Posner;K. K. W. Wang
  14. Int. J. Syst. Bacteriol. v.44 Bacillus mojavensis sp. nov., distinguishable from Bacillus subtilis by sexual isolation, divergence in DNA sequence, and differences in fatty acid composition Roberts, M. S.;L. K. Nakamura;F. N. Cohan
  15. Anal. Biochem. v.167 Rapid isoelectric focusing in a vertical polyacrylamide minigel system Robertson, E. F.;K. Dannelly;P. J. Malloy;H. C. Reeves
  16. J. Micobiol. Biotehcnol. v.9 The roles of tryptophan and histidine residues in the catalytic activities of β-cyclodextrin glucanotransferase from Bacuillus firmus var. alkalophilus Shin, H. D.;C. Kim;Y. H. Lee
  17. Acta Haematol. v.84 Enhancement of the fibrinolytic activity in plasma by oral administration of nattokinase Sumi, H. H. Hamada;K. Nakanishi;H. Hiratani
  18. Experientia v.43 A novel fibrinolytic enzyme (nattokinase) in the vegetable cheese Natto: A typical and popular soybean food in the Japanese diet Sumi, H.;H. Hamada;H. Tsushima;H. Mihara;H. Muraki