Open AccessPurification and characterization of a novel protease from culture filtrates of a Streptomyces sp.
Author(s) -
Bono Françoise,
Savi Pierre,
Tuong Anne,
Maftouh Mohamed,
Pereillo JeanMarie,
Capdevielle Joel,
Guillemot JeanClaude,
Maffrand JeanPierre,
Herbert JeanMarc
Publication year - 1996
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1996.tb08387.x
Subject(s) - protease , pepstatin , phenylmethylsulfonyl fluoride , proteases , isoelectric point , biochemistry , trypsin , chemistry , serine protease , molecular mass , kunitz sti protease inhibitor , chymotrypsin , streptomyces , streptomycetaceae , enzyme , isoelectric focusing , chromatography , gel electrophoresis , actinomycetales , biology , bacteria , genetics
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.