Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Characterization of $Co^{2+}-Activated$ Extracellular Metalloprotease from Bacillus sp. JH108

  • Jung, Hyun-Joo (Department of Food and Microbial Technology, Faculty of Natural Sciences, Seoul Women's University) ;
  • Kim, Haek-Won (Department of Biology, Faculty of Natural Sciences, Seoul Women's University) ;
  • Kim, Jong-Il (Department of Food and Microbial Technology, Faculty of Natural Sciences, Seoul Women's University)
  • Published : 1999.12.01
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Abstract

An extracellular protease was purified to homogeneity from the culture supernatant of psychrotrophic bacteria Bacillus sp. JH 108 using procedures including ammonium sulfate fractionation, anion exchange chromatography, gel filtration chromatography, and cation exchange chromatography. The enzyme exhibited a molecular weight of 36 kDa, an optimum pH of 8 to 9, and optimum temperature of $60^{\circ}C$. The enzyme preferentially hydrolyzed leucine at the N-terminus of peptides and thus can be classified as an aminopeptidase. It was strongly inhibited by metal chelating agents such as EDTA and l, l0-phenanthroline. The activity lost by EDTA was restored with $Zn^{2+}{\;}or{\;}Co^{2+}$. These divalent cations also stimulated the native enzyme. This suggests that the enzyme is a metalloprotease acting as a leucine aminopeptidase.

Keywords

References

  1. Biotechnol. Appl. Biochem. v.19 Purification and characterization of an aminopeptidase from Lactobacillus casei subsp. rhamnosus S93 Arora, G.;B. H. Lee
  2. Eur. J. Biochem. v.212 Specificity of Streptomyces friseus aminopeptidase and modulation of activity by divalent metal ion binding and substitution Ben-Meir, D.;A. Spungin;R. Ashkenazi;S. Blumberg
  3. Proteolytic enzymes Beynon, R. J.;J. S. Bond
  4. 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.
  5. Proc. Natl. Acad. Sci. USA v.87 Molecular structure of Leucine aminopeptidase at 2.7- A resolution Burley, S. K.;P. R. David;A. Taylor;W. N. Lipscomb
  6. J. Biochem. v.105 Purification and properties of an aminopeptidase from Alaska pollack, Theragra chalcoramma, roe Chiou, T. K.;T. Matsui;S. Konosu
  7. Appl. Environ. Microbiol. v.56 Purification and characterization of Acidolysin, an acidic metalloprotease produced by Clostridium actobutylicum ATCC824 Croux, C.;V. Paquet;G. Goma;P. Soucalille
  8. J. Gen. Microbiol. v.132 Isolation and characterization of a Vibrio aginolyticus mutant that overproduces extracellular proteases Deane, S. M.;E. T. Robb;D. R. Wood
  9. Biochim. Biophys. Acta v.251 Bacillus cereus neutral protease Feder, J. L.;L. Keay;L. Garett;N. Circulis;M. H. Moseley;B. S. Wildi
  10. Advances in Biochemical Engineering and Biotechnology Microbial protienases Fiechter, A.
  11. J. Bacteriol. v.171 Structural gene and complete amino acid sequence of Pseudomonas gene and complete amino acid sequence of Pseudomonas aeruginosa IFO 3455 elastase Fukushima, J.;S. Yamamoto;Y. Morihara;H. Atsumi, Takeuchi;S. Kawamoto;K. Okuda
  12. Biochemistry v.13 Human liver aminopeptidase. Role of metal ions in mechanism of action Garner, C. W.;F. J. Behal
  13. FEMS Microbiol. Rev. v.18 Bacterial aminopeptidase: Properties and functions Gonzales, T.;J. Robert-Baudroy
  14. Biochem. Biophys. Res. Commun. v.173 Leukotrience A4 hydrolase: An epoxide hydrolase with peptidase activity Haeggstrom J. Z.;A. Wetterhilm;B. L. Valee;B. Samuelsson
  15. Biochim. Biophys. Acta v.1204 Purification and characterization of an aminopeptidase from tuna Pyloric caeca Hajjou, M.;Y. Le Gal
  16. J. Microbiol. Biotechnol. v.8 Characterization of alkaline serine proteases secreted from the coryneform bacterium TU-119 Kang, S. C.;S. Park;M. C. Choi
  17. J. Kor. Microbiol. v.33 Characteristics of Trypsin-like protease and metalloprotease associated with mycelium differentiation of Streptomyces albidoflavus SMF 301 Kang, S. G.;B. C. Jeong;K. J. Lee
  18. J. Biochem. v.92 Purification and characterization of an aminopeptidase from porcine Kawata, S.;T. Imamura;K. Ninomiya;S. Makisumi
  19. J. Biochem. v.121 Purification and characterization of an extracellular metalloprotease from Pseudomonas fluorscens Kim, H.;Y. Tamanoue;G. H. Jeohn;A. Iwamatsu;A. Yokota;Y. T. Kim;T. Takahashi;K. Takahashi
  20. Nature v.277 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli, U. K.
  21. FEMS Microbiol. Lett. v.79 Characterization of a metalloprotease from psychrophilic Xanthomonas maltoplhila Margesin, R.;F. Schinner
  22. Biochim. Biophys. Acta v.613 Purification and properties of human intestine alanine aminopeptidase McClellan, J. B.;C. W. Garner
  23. Encyclopedia of Microbiology v.2 Low-temperatur environments Morita, R. Y.
  24. Eur. J. Biochem. v.44 Purification and properties of an extracelluar proteinase of psychrophilic Escherichia freundii Nakazima, M.;K. Mizusawa;F. Yoshida
  25. Agr. Biol. Chem. v.55 Purification and properties of aminopeptidase C from chicken skeletal muscle Nishimura, T.;Y. Kato;A. Okitani;H. Kato
  26. Methods Enzymol. v.45 Aeromonas aminopeptidase Prescott, J. M.;S. H. Wikes
  27. Appl. Environ. Microbiol. v.58 Extracellular protease from the Antarctic yeast Candida humicola Ray, M. K.;G. Uma Devi;G. Seshu Kumar;S. Shivaji
  28. J. Microbiol. Biotechnol. v.10 Extracellular protease from Bacillus coagulans, a psychrotropic, antarctic bacterium - production, purification and characterization Sai Ram, M.;L. Singh;S. I. Alam;M. K. Aggarwal
  29. Clin. Chim. Acta v.104 Multiple molecular forms of human pancreas alanine aminopeptidase Shidorowicz, W.;G. C. Jackson;F. J. Behal
  30. Anal. Biochem. v.150 Measurement of protein using bicinchoninic acid Smith, P. K.;R. J. Krohn;G. T. Hermanson;A. K. Mallia;F. H. Gartner;M. D. Provenzano;E. K. Ferjimoto;N. M. Goeke;B. J. Olson;D. C. Klenk
  31. Appl. Microbiol. Biotechnol. v.27 Bacillus stearothermophilus KP1236 neutral protease with strong thermostability comparable to thermolysin Takii, Y.;H. Taguchi;H. Shimoto;Y. Suzuki
  32. J. Biol. Chem. v.25 The relative binding of cobalt substitution on specific activity Thompson, G. A.;F. H. Carpenter
  33. Neoplasma v.37 A chloride activated alanine aminopeptidase from melanoma cell line Tsushima, H.;V. K. Hopsu-Havu
  34. Biomed. Biochim. Acta v.49 Low molecular weight alanine aminopeptidase of human serum Tsushima, H.;H. Sumi;R. Ikeda;E. Yoshida;H. Mihara;V. K. Hopsu-Havu
  35. Biochemistry v.20 Metal binding stoichiometry and mechanism of metal ion modulation of the activity of procine kindney Leucine aminopeptidase Van Wart, H. E.;S. H. Lin
  36. Arch. Microbiol. v.165 An intracellular aminopeptidase from Streptomyces rimosus that prefers basic amino acids Vitale, L.;I. Skrtic;M. Abramic
  37. Biochemistry v.20 Purification and characterization of an enkephalin aminopeptidase from rat brain Wagner, G. W.;M. A. Tavianni;K. M. Harrmann;J. E. Dixon
  38. J. Biol. Chem. v.244 The reliability of molecular weight determinators by the dodecylsulfate-polyacrylamid gel clectrophoresis Weber, K.;M. Osborn
  39. J. Biol. Chem. v.273 Purification, characterization and cloning of cytosolic Aspartyl aminopeptidase Wilk, S.;E. Wilk;R. P. Magnusson
  40. J. Bacteriol. v.160 Cloning of the neutral protease gene of Bacillus subtilis and the use of the cloned gene to create an in vitro derived deletion mutation Yang, M. Y.;E. Ferrari;D. J. Henner
  41. J. Bateriol. v.166 Specific excretion of Serratia marcescens protease through the outer membrane of Escherichia coli Yanagida, N.;T. Uozumi;T. Beppu
  42. Eur. J. Biochem. v.211 Purification and characterization of an aminopeptidase A from Staphylococcus chromogenes Yoshpe-Besancon, I.;D. Auriol;F. Paul;P. Monsan;J. C. Gripon;B. Ribadeau-Dumas
  43. Eur. J. Biochem. v.211 Purification and characterization of an aminopeptidase A from Staphylococcus chromogenes Yoshpe-Besancon, I.;D.Auriol;F.Paul;P.Monsan;J.C.Gripon;B. Ribadeau-Dumas