title: Optimization of polyphenol oxidase immobilization in copper alginate beads.
abstract: Polyphenol oxidase (PPO, EC 1.14.18.1) was isolated from artichoke head (Cynara scolymus L.) by using 0.1 M Tris-HCl buffer (pH 7.0), concentrated by (NH4)2SO4 precipitation, and immobilized in copper-alginate beads. Immobilization yield was determined to be 70%. The cresolase and catecholase activities of enzyme immobilized at optimum immobilization conditions were found to be 13.3 and 670 U g beads min(-1), respectively. Effects of immobilization conditions such as alginate concentration, CaCl2 concentration, amount of loading enzyme, bead size, and amount of beads on enzymatic activity were investigated. Optimum alginate and CuCl2 concentration were found to be 2 % and 3 % (w/v), respectively. Using bead (diameter 3 mm) amount of 0.25 g maximum enzyme activities were observed for both polyphenol activities. The initial concentrations of loading free enzyme were 6.5 U mL(-1) and 5815 U mL(-1) for cresolase activity and catecholase activities, respectively. Beads prepared at optimum immobilization conditions were suitable for up to 8 repeated uses.
authors.affiliation: Department of Chemistry, Faculty of Science, Trakya University, Edirne, Turkey.
authors.forename: Selin
authors.initials: S
authors.lastname: Kocaturk
authors.forename: Hulya
authors.initials: H
authors.lastname: Yagar
mesh_terms: D000464:Alginates / Q000737:chemistry; D004156:Catechol Oxidase / Q000302:isolation & purification / Q000378:metabolism; D002396:Catechols / Q000378:metabolism*; D003300:Copper / Q000737:chemistry; D031943:Cynara scolymus / Q000201:enzymology; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005419:Flavonoids / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008863:Microspheres; D014442:Monophenol Monooxygenase / Q000378:metabolism; D010316:Particle Size; D010636:Phenols / Q000378:metabolism*; D059808:Polyphenols
chemical_list: D000464:Alginates; D002396:Catechols; D004800:Enzymes, Immobilized; D005419:Flavonoids; D010636:Phenols; D059808:Polyphenols; D003300:Copper; D004156:Catechol Oxidase; D014442:Monophenol Monooxygenase