Purification and characterization of a novel protease from culture filtrates of aStreptomycessp

Autor: Joël Capdevielle, Jean-Marc Herbert, Jean-Pierre Maffrand, Anne Tuong, Jean-Marie Pereillo, Pierre Savi, Jean-Claude Guillemot, Mohamed Maftouh, Françoise Bono
Rok vydání: 1996
Předmět:
Zdroj: FEMS Microbiology Letters. 141:213-220
ISSN: 1574-6968
0378-1097
Popis: 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 °C and 50 °C. Its amino acid composition and aminoterminal sequence (17 residues) were determined. The protein exibited marked hydrolytic activity toward the substrates N -Succ-(Ala) 2 -Pro-Phe-pNA ( K m = 0.77 mM, V max = 24.2 μ mol mg −1 min −1 ) and N -Succ-(Ala) 2 -Pro-Leu-pNA ( K m = 0.92 mM, V max = 7.7 μ mol mg −1 min −1 ). It was totally inhibited by α1-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.
Databáze: OpenAIRE
Externí odkaz: