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PURIFICATION AND CHARACTERIZATION OF ALKALINE PROTEINASES FROM THE VISCERA OF ANCHOVY, ENGRAVLIS JAPONICA
Author(s) -
HEU MINSOO,
PYEUN JAEHYEUNG,
KIM HYEUNGRAK,
GODBER J. SAMUEL
Publication year - 1991
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.1745-4514.1991.tb00143.x
Subject(s) - alkaline protease , sephadex , chymotrypsin , enzyme , chemistry , proteases , biochemistry , trypsin , serine , tyrosine , substrate (aquarium) , size exclusion chromatography , chromatography , biology , protease , ecology
Two electrophoretically homogeneous proteinases designated proteinase A and B were isolated from anchovy viscera. Purity was increased 17.7 and 24.6‐fold with approximately 1.9 and 1.8% yield for proteinases A and B, respectively. The maximum caseinolytic activity was found to be at pH 9.4 for proteinase A and at pH 9.6 for proteinase B at the optimum temperature of 48°C. The molecular weights of proteinase A and B were determined to be 27, 300 and 25,100 D, respectively, using Sephadex G‐100 gel filtration. The amino acid profiles of the enzymes were similar and relative proportion of amino acid residues was comparable to that in bovine pancreatic α‐chymotrypsin. Proteinase A and B were identified as α‐chymottypsin‐like serine proteases by inhibitor and substrate specificity studies. Apparent K m (K m ′) values of proteinase A and B for benzoyl‐L‐tyrosine ethyl ester were 4.6 × 10 −4 M and 1.2 × 10 −3 M, respectively.