Production and Characterization of Cathepsin B Inhibitor from Streptomyces chromofuscus SMF28

Streptomyces chromofuscus SMF28을 이용한 Cathepsin B 저해물질의 발효생산 및 특성분석

  • Published : 1995.10.01

Abstract

The aim of the present research program was to construct an optimum fermentation system and to characterize the properties of cathepsin B inhibitor from Streptomyces chromofuscus SMF28. Glucose and casitone were proved to be good carbon source and nitrogen source, respectively. The production of inhibitor was high at lower concentration than 10 mM of inorganic phosphate. The optimum temperature and pH for the production of inhibitor were 30$\circ$C and pH 7, respectively. The production of inhibitor was related to mycelial growth and was affected by medium composition. The inhibitor in culture filtrate of S. chromofuscus SMF28 was purified by butanol extraction, silica gel chromatography, Amberlite IRC-50 (H$^{+}$ form) chromatography, preparative TLC, and preparative HPLC. From amino acid analysis and UV, IR, $^{1}$H-NMR spectroscopic analysis, the inhibitor was identified as a peptide containing valine and phenylalanine derivative.

Keywords

References

  1. Proteinases in Mammalian Cells and Tissues Barrett,A.J.(ed.);
  2. Methods in Enzymology v.80 Cathepsin B, cathepsin H, cathepsin L. Barrett,A.J.;H.Kirschke
  3. Proteinases in mammalian cells and tissues Introduction to the history and classification of tissue proteinases Barrett,A.J.(ed.)
  4. Science v.200 Muscular dystrophy : Inhibition of degeneration in vivo with protease inhibitors Stracher,A.;E.B.McGowan;S.A.Shafiq
  5. Arch. Biochem. Biophys. v.96 Increased lysosomal enzymes in genetic muscular dystrophy Tappel,A.L.;H.Zalkin;K.A.Caldwell;I.D.Desai;S.Shibko
  6. J. Biochem. v.98 Distribution of cathepsin B and H in rat tissues and peropheral blood cells Kominami,E.;T.Tsukahara;Y.Bando;N.Katunnuma
  7. Eur. J. Biochem. v.129 The slow, tight-binding inhibition of cathepsin B by leupeptin: A hysteretic effect Baici,A.;M.Gyger-Marazzi
  8. Biochemistry v.27 Inhibition of cathepsin B by peptidyl aldehydes and ketones : Slow-binding behavior of a trifluromethyl ketone Smith,R.A.;L.J.Copp;S.L.Donnelly;R.W.Spencer;A.Krantz
  9. J. Biol. Chem. v.256 Peptidyl diazomethyl ketones are specific inactivatiors of thiol proteinases Green,G.D.J.;E.Shaw
  10. FEBS. v.280 Novel epoxysuccinyl peptides-A selective inhibitor of cathepsin B, in vivo. Towatari,T.;T.Nikawa;M.Murata;C.Yokoo;M.Tamai;K.Hanada;N.Katunuma
  11. Biochem. Biophys. Res. Comm. v.155 Peptidyl O-acyl hydroxamates : Potent new inactivatior of cathepsin B. Smith, R.A.;P.J.Coles;R.W.Spencer;L.J.Copp;C.S.Hones;A.Krantz
  12. Kor. J. Appl. Microbiol. Isolation and identification of Stroptomyces chromofuscus producing cathepsin B inhibitor Lee,H.S.;I.S.Kim;H.T.Kim;S.J.Yoon;K.J.Lee
  13. Anal. Chem. v.31 Use of dinitrosalicylic acid reagent for determination of reducing sugar MIller,G.L.
  14. Anal. Biochem. v.47 A new assay for cathepsin B1 and other thiol proteinases Barrett,A.J.