title: Purification and characterization of a novel protease from culture filtrates of a Streptomyces sp.
abstract: A fibrinolytic protease has been isolated from Streptomyces sp. culture filtrate by successive chromatography on Mono S and Sephadex G50. The purified protease had a molecular mass of 33 kDa and had an isoelectric point of 6.7. It showed a sharp pH optimum at 7.8 with maximal protease activity between 35 degrees C and 50 degrees C. Its amino acid composition and amino-terminal sequence (17 residues) were determined. The protein exhibited marked hydrolytic activity toward the substrates N-Succ-(Ala)2-Pro-Phe-pNA (K(m) = 0.77 mM, Vmax = 24.2 mumol mg-1 min-1) and N-Succ-(Ala)2-Pro-Leu-pNA (K(m) = 0.92 mM, Vmax = 7.7 mumol mg-1 min-1). It was totally inhibited by alpha 1-antitrypsin, D-Phe-Pro-Arg-chloromethylketone and sodium dodecyl sulfate but was insensitive to EDTA, dithiothreitol, phenylmethylsulfonyl fluoride, soybean trypsin inhibitor, pepstatin or elastatinal. In this respect, this protease differed in its physico-chemical and biochemical properties from other extracellular proteases previously found in bacteria and fungi. The results suggest that it has properties of chymotrypsin-like serine-type proteases.
authors.affiliation: Haemobiology Research Department, Sanofi Recherche, Toulouse, France.
authors.forename: F
authors.initials: F
authors.lastname: Bono
authors.forename: P
authors.initials: P
authors.lastname: Savi
authors.forename: A
authors.initials: A
authors.lastname: Tuong
authors.forename: M
authors.initials: M
authors.lastname: Maftouh
authors.forename: J M
authors.initials: JM
authors.lastname: Pereillo
authors.forename: J
authors.initials: J
authors.lastname: Capdevielle
authors.forename: J C
authors.initials: JC
authors.lastname: Guillemot
authors.forename: J P
authors.initials: JP
authors.lastname: Maffrand
authors.forename: J M
authors.initials: JM
authors.lastname: Herbert
mesh_terms: D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D010450:Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification*; D005342:Fibrinolysis; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D011480:Protease Inhibitors; D010450:Endopeptidases