Characteristics of Trypsin-like Protease and Metalloprotease Associated with Mycelium Differentiation of Streptomyces albidoflavus SMF301

  • Kang, Sung-Gyun (Research Center for Molecular Microbiology, Seoul National University) ;
  • Kim, In-Seop (Research Center for Molecular Microbiology, Seoul National University) ;
  • Jeong, Byung-Cheol (Research Center for Molecular Microbiology, Seoul National University) ;
  • Ryu, Jae-Gon (Research Center for Molecular Microbiology, Seoul National University) ;
  • Rho, Yong-Taik (Research Center for Molecular Microbiology, Seoul National University) ;
  • Lee, Kye-Joon (Research Center for Molecular Microbiology, Seoul National University)
  • 발행 : 1995.12.01

초록

Trypsin like protease (TLP) and metalloprotease (MTP) were induced in associated with the mycelium differentiation in Streptomyces albidoflavus SMF301. TLP and MTP were purified and characterized from the culture. The molecular mass of TLP and MTP were estimated to be 32 kDa and 18 kDa, respectively. The molecular mass of TLP and MTP were estimated to be 32 kDa and 18 kDa, respectively. The optimum pH and temperature of TLP were 10 and 40.$^{\circ}C$ Those of MTP were 8 and 55 $^{\circ}C$ TLP was stable at alkaline pH (6-9) and unstable above 45.$^{\circ}C$and MTP was stable at alkaline pH and unstable above 80.$^{\circ}C$ Km and Vmax values with benzoyl-arginyl p-nitroanilide of TLP were 139 $\mu$M, and 10 nmole of nitroanilide released per min per$\mu\textrm{g}$ protein, respectively. Km, and Vmax values with a synthetic substrate, leucine p-nitroanilide, or MTP were 58.9 $\mu$M, 3.47 nmol of nitroanilide released per min per$\mu\textrm{g}$protein, respectively. TLP was inhibited competitively by leupeptin; the inhibition constant was 0.0031 $\mu$M. MTP was inhibited by EDTA, phenonthroline and bestatin.

키워드

참고문헌

  1. Protease inhibitor and biological control In Bioactive Metabolites from microorganisms, Progress in Industrial Microbiology v.27 Aoyagi,T.;M.E.Bushell(ed.);U.Gr fe(ed.)
  2. Appl. Microbiol. Biotechnol. v.34 Regulation of extracellular protease production in Streptomyces davuligerus Bascaran,V.;C.Hardisson;A.F.Brana
  3. Archiv. Biochem. Biophys. v.257 Multiple proteases from Streptomyces moderatusⅡ. Isolation and purification of five extracellular proteases Chandrasekaran,S.;S.C.Dhar
  4. Morphological and physiological differentiation in Streptomyces In Microbial Development Chater,K.F.;R.Losick(eds.);L.Sapiro(eds.)
  5. Int. J. Peptide. Protein. Res. v.8 Isolation of a trypsin-like enzyme from Streptomyces paromomycinus (paromotrypsin) by affinity adsorption through kunitz inhibitor-sepharose Chauvet,J.;J.Dostal;R.Acher
  6. J. Bacteriol. v.173 Structural gene and complete amino acid sequence of Pseudomonas aeroginosa las R gene, a transcriptional activator of elastase expression Fukusima,J.;S.Yamamoto;K.Morihara;Y.Atsumi;J.Takeuchi;S.Kawamoto;K.Okuda
  7. Can. J. Microbiol. v.34 Physiological regulation of protease activity in Streptomyces peucetius Gibb,G.D.;W.R.Strohl
  8. Antimicrob. Agents. Chemother. v.15 Sporulation and the production of serine protease and cephamycin C by Streptomyces lactamdurans Ginther,C.L.
  9. Biochim. Biophys. Acta. v.832 Interactions of derivatives of guanidinophenylalanine and guanidinophenylglycine with Streptomyces griseus trypsin Hatanaka,Y.;H.Tsunematsu;K.Mizusaki;S.Makisumi
  10. Biochim. Biophys. Acta. v.284 An anionic trypsin-like enzyme from Streptomyces erythreus: Chracterization Inoue,H.;A.Sasaki;N.Yoshida
  11. Microbiology Production dynamics of extracellular proteases accompanying morphological differentiation of Streptomyces albidoflavus SMF301 Kang,S.G.;I.S.Kim;Y.T.Rho;K.J.Lee
  12. Microbiol. v.141 Physiological roles of leupeptin and extracellular proteases in mycelium development of Streptomyces exfoliatus SMF13 Kim,I.S.;K.J.Lee
  13. Nature v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli,U.K.
  14. J. Gen. Microbiol. v.139 Characteristics of spores formed by surface and submerged cultures of Streptomyces albidoflavus SMF301 Lee,K.J.;Y.T.Rho
  15. Can. J. Microbiol. v.31 Role of substrate mycelium in colony development in Streptomyces Mendez,C.;A.F.Brana;M.B.Manzanal;C.Hardisson
  16. Methods in Enzymology v.19 Pronase Narahashi,Y.
  17. Appl. Biochem. Biotechnol. v.9 Trypsinlike enzyme from Streptomyces 771. Purification and properties of native and immobilized enzyme Palubinskas,V.I.;N.B.Yankevich;K.K.Yanulaitene;V.S.Vesa;V.G.Bendikene;A.V.Maksimenko;M.P.Torchilin;E.V.Ilyina;V.N.Smirnov;L.N.Krestyanoe;Y.E.Bartoshevich;R.C.Zabirova
  18. Actinomycete enzymes In Actinomycetes in biotechnology Peczynska-Czoch,W.;W.Mordarski;M.Goodfellow(eds.);S.T.Williams(eds.);M.Mordarski(eds.)
  19. Appl. Microbiol. Biotechnol. v.31 Streptomuces rimosus extracellular proteases 4. Trypsin-like proteinase Renko,M.;L.J.Vitale;M.Kokalj;M.Podorny
  20. Kor. J. Microbiol. v.30 Numerical identification of a Streptomyces strain producing spores in submerged culture Rho,Y.T.;H.T.Kim;K.H.Oh;H.I.Kang;A.C.Ward;M.Goodfellow;Y.C.Hah;K.J.Lee
  21. Microbiol. v.140 Kinetic studies on mycelium growth and spore formation in submerged cultures of Streptomyces albidoflavus SMF301 Rho,Y.T.;K.J.Lee
  22. Protease assay methods In Proteolytic Enzymes: A Practical Approach Sarath,G.;De La Motte RS;F.W.Wagner
  23. Biochem. J. v.109 The trypsin-like enzyme from Streptomyces griseus (Pronase) Trop,M.;Y.Birk
  24. J. Biochem. v.110 Purification and characterization of Streptomyces griseus metalloendopeptidasesⅠ and Ⅱ Tsuyuki,H.;K.Kajiware;A.Fujita;T.Kumazaki;S.Ishii
  25. J. Gen. Microbiol. v.60 Development and organization of the aerial mycelium in Streptomyces coelicolor Wildermath,H.
  26. 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
  27. FEBS Lett. v.15 An anionic trypsin-like enzymes from Streptomyces erythreus Yoshida,N.;A.Sasaki;H.Inoue