Immobilization of Escherichia coli novablue γ-glutamyltranspeptidase in Ca-alginate-k-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.

Characterization and immobilization of purified Aspergillus flavipesl-methioninase: continuous production of methanethiol

“…2006). A little physical interference with the enzyme catalytic residues by PVA and alginate was observed as coincide with immobilized enzymes (Hung et al. 2008; Tumturk et al.…”

“…The highest enzyme immobilization efficiency was measured by the entrapment, polyacrylamide and Ca‐alginate, assuming the low interactions with the surface catalytic residues of enzyme by the carrier (Mori et al. 1974; Hung et al. 2008; Vu and Le 2008).…”

Repeated-Batch Operation of Immobilized β-Galactosidase Inclusion Bodies-Containing Escherichia coli Cell Reactor for Lactose Hydrolysis

“…observed in other reports [12,13]. To overcome this problem, the cell viability in the alginate beads needs to be improved and new immobilization methods need to be developed.…”

Integrated study on nutrients and heavy metals removal from domestic wastewater using free and immobilized strain Scenedesmus rubescens KACC 2 (Chlorophyta, Chlorophyceae)

“…Комбинированные полимерные гранулы, содержащие 2%-ный альгинат и 0,3%-ный каррагинан, готовили путем перемешивания с 20 мМ буферного раствора трис-HCl (рН 9), а затем по каплям вносили в 50 мМ раствор того же буфера, содержащий 0,3 М CaCl 2 . Образовавшиеся гранулы оставляли в этом растворе на 2 ч (Hung et al, 2008).…”