title: Immobilization of Escherichia coli novablue gamma-glutamyltranspeptidase in Ca-alginate-kappa-carrageenan beads.
abstract: The recombinant Escherichia coli gamma-glutamyltranspeptidase (EcGGT) was immobilized in Ca-alginate-kappa-carrageenan beads. Effects of alginate concentration, amount of loading enzyme, and bead size on the entrapped activity were investigated. Optimum alginate concentration for EcGGT immobilization was found to be 2% (w/v). Using a loading enzyme concentration of 1.5 mg/g alginate, maximum enzyme activity was observed. With increase in bead size from 1.9 to 3.1 mm, the immobilization efficiency was decreased significantly because of mass transfer resistance. Thermal stability of the free EcGGT was increased as a result of the immobilization. Ca-alginate-kappa-carrageenan-EcGGT beads were suitable for up to six repeated uses, losing only 45% of their initial activity. Upon 30 days of storage the preserved activity of free and immobilized enzyme were found as 4% and 68%, respectively. The synthesis of L: -theanine was performed in 50 mM Tris-HCl buffer (pH 10) containing 25 mM L: -glutamine, 40 mM ethylamine, and 1.5 mg EcGGT/g alginate at 40 degrees C for 12 h, and a conversion rate of 27% was achieved.
authors.affiliation: Department of Anesthesiology, Chang Gung Memorial Hospital, Kaohsiung, Taiwan.
authors.forename: Chih-Peng
authors.initials: CP
authors.lastname: Hung
authors.forename: Huei-Fen
authors.initials: HF
authors.lastname: Lo
authors.forename: Wen-Hwei
authors.initials: WH
authors.lastname: Hsu
authors.forename: Shih-Chun
authors.initials: SC
authors.lastname: Chen
authors.forename: Long-Liu
authors.initials: LL
authors.lastname: Lin
mesh_terms: D000464:Alginates / Q000737:chemistry*; D002118:Calcium / Q000737:chemistry*; D002351:Carrageenan / Q000737:chemistry*; D004800:Enzymes, Immobilized; D004926:Escherichia coli / Q000201:enzymology*; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D008863:Microspheres; D005723:gamma-Glutamyltransferase / Q000737:chemistry*
chemical_list: D000464:Alginates; D004800:Enzymes, Immobilized; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D002351:Carrageenan; D005723:gamma-Glutamyltransferase; D002118:Calcium