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PARTIAL PURIFICATION AND CHARACTERIZATION OF A THERMOSTABLE TRYPSIN FROM PYLORIC CAECA OF TAMBAQUI ( COLOSSOMA MACROPOMUM )
Author(s) -
BEZERRA RANILSON S.,
SANTOS JULIANA F.,
PAIVA PATRÍCIA M.G.,
CORREIA MARIA T.S.,
COELHO LUANA C.B.B.,
VIEIRA VERA L.A.,
CARVALHO LUIZ B.
Publication year - 2001
Publication title -
journal of food biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 47
eISSN - 1745-4514
pISSN - 0145-8884
DOI - 10.1111/j.1745-4514.2001.tb00734.x
Subject(s) - tambaqui , trypsin , sephadex , chemistry , protease , alkaline protease , chromatography , kunitz sti protease inhibitor , pepstatin , enzyme , ammonium sulfate , ammonium , salting out , biochemistry , papain , size exclusion chromatography , biology , fish <actinopterygii> , fishery , aqueous solution , organic chemistry
A 38.5 kDa alkaline protease from pyloric caeca of tambaqui (Colossoma macropomumj, a tropical freshwater fish, was partially purified in three steps: thermal treatment (45Cfor 30 min), salting‐out (ammonium sulfate at 40–80% of saturation) and gel filtration (Sephadex G‐75), The purification and yield were 51.2‐fold and 40%, respectively. The effects of pH, temperature, inhibitors, and substrates on proteolytic activities of partially purified enzyme were investigated. The optimum pH was 9.5, while the optimum temperature was 60C. This alkaline proteolytic activity remained unaltered after 30 min incubation at 55C. Active site inhibition provided additional evidence that this activity is attributed to a trypsin‐like enzyme.