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COVALENT IMMOBILIZATION OF BOVINE PHOSPHOLIPASE A 2
Author(s) -
NAM SEUNGHEE,
WALSH MARIE K.
Publication year - 2005
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.1745-4514.2005.00013.x
Subject(s) - glutaraldehyde , chemistry , immobilized enzyme , chromatography , enzyme , substrate (aquarium) , covalent bond , enzyme assay , hydrolysis , phospholipase a2 , sodium borohydride , biochemistry , organic chemistry , catalysis , biology , ecology
Bovine phospholipase A2(PPLA2) was immobilized onto controlled pore glass (CPG) beads using four different immobilization methods. PPLA2was immobilized directly to CPG using glutaraldehyde without and with reduction by sodium borohydride (PP1 and PP2). Whey protein isolate was directly immobilized to CPG as a spacer followed by immobilization of PPLA2without and with reduction (PP3 and PP4). Immobilized enzyme samples were characterized with respect to total amount of protein immobilized, activity with a fluorescent substrate and stability over 3 weeks. Among the methods, PP2 and PP4 showed the highest enzyme activity. All methods but PP2 showed a significant decrease in enzyme activity over 3 weeks. Enzyme immobilized by two methods (PP2 and PP4) were compared with soluble enzyme for the hydrolysis of egg phospholipids. Soluble and immobilized enzyme (PP4) resulted in similar free fatty acid values.