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Purification and Partial Characterization of Trypsin from the Viscera of Tropical Sierra ( Scomberomorus Sierra ) from the Gulf of California
Author(s) -
ValdezMelchor R.G.,
EzquerraBrauer J.M.,
CincoMoroyoqui F.J.,
CastilloYáñez F.J.,
CardenasLopez J.L.
Publication year - 2013
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.1745-4514.2012.00667.x
Subject(s) - trypsin , kunitz sti protease inhibitor , protease , biochemistry , chemistry , enzyme , serine protease , affinity chromatography , chromatography , sepharose , trypsin inhibitor , hydrolysis
Trypsin from the viscera of sierra ( S comberomorus sierra ) was purified by affinity chromatography on S epharose‐4 B coupled to soybean trypsin inhibitor and characterized with respect to its purity, sensitivity to temperature, pH and inhibition. Trypsin was purified from sierra viscera with 11.9‐fold and 29.7% yield. The enzyme had a molecular weight of 25.4 kDa estimated by SDS‐PAGE and two possible trypsin isoforms were observed in activity gels. Trypsin activity was strongly inhibited by soybean trypsin inhibitor and porcine trypsin inhibitor, showing a partial inhibition by a serine protease inhibitor. The optimal activity of the enzyme was observed at pH 9 and 60 C with n‐α‐benzoyl‐dl‐arginine‐p‐nitroanilide as a substrate. The enzyme maintained more than 50% of its activity in temperatures up to 50 C and within the pH range of 8–10 for a period of up to 2 h.