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Purification and partial characterization of an extracellular serine‐proteinase of Streptomyces cyaneus isolated from Brazilian cerrado soil
Author(s) -
Petinate S. D. G.,
Branquinha M. H.,
Coelho R. R. R.,
And A. B. Vermelho,
GiovanniDeSimone S.
Publication year - 1999
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.1999.00852.x
Subject(s) - biology , streptomycetaceae , streptomyces , extracellular , actinomycetales , serine , microbiology and biotechnology , bacteria , biochemistry , enzyme , genetics
Streptomyces cyaneus , a micro‐organism isolated from Brazilian cerrado soil, produces an extracellular proteinase (SCP), which was purified 22‐fold to homogeneity from culture supernatant fluid, using a single aprotinin‐agarose affinity chromatography step. It is produced at a level corresponding to approximately 15% of total protein, but its physiological function has yet to be determined. The molecular mass of this S. cyaneus proteinase was estimated to be 120 kDa by gel filtration high performance liquid chromatography, and it migrates by SDS‐PAGE as a single band of 30 kDa. It was optimally active at 25 °C and pH 9·0, and was fully inhibited by the serine‐proteinase inhibitors PMSF and TPCK. A K m value of 1·86 × 10 −5 mmol l −1 , and V max of 2·0 × 10 −2 mmol l −1 (Abs 247 nm μg −1 min −1 ), were calculated for α‐N‐p‐ tosyl ‐ l ‐arginine‐methyl ester (TAME) as substrate.