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Purification and Characterization of a Trypsin‐Like Protease from Flatfish ( P aralichthys olivaceus ) Intestine
Author(s) -
Kim Misook,
Jeong Yoonhwa
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.1745-4514.2012.00672.x
Subject(s) - trypsin , protease , benzamidine , antipain , chromatography , biochemistry , chemistry , serine protease , gel electrophoresis , hydrolysate , kunitz sti protease inhibitor , polyacrylamide gel electrophoresis , leupeptin , enzyme , hydrolysis
A trypsin‐like protease was purified from the intestine of flatfish ( P aralichthys olivaceus ) by gel filtration and anion‐exchange chromatography. The molecular weight was estimated to be 29.6 kDa by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Flatfish protease had maximal activity at 70 C and pH 7.5 using N ‐α‐benzoyl‐ dl ‐arginine‐ρ‐nitroanilide as substrate. It was stable to heat treatment up to 50 C and to pH ranges between 7.0 and 10.0. It was activated by calcium ion and completely inhibited by mercury ion and known serine‐protease inhibitors, such as phenylmethylsulfonyl fluoride, tosyl lysine chloromethyl ketone and benzamidine.