Enzyme immobilization on a low-cost magnetic support: kinetic studies on immobilized and coimmobilized glucose oxidase and glucoamylase.
Glucose oxidase (GOx) and glucoamylase (GA) were immobilized and coimmobilized through their carbohydrate moieties onto polyethyleneimine-coated magnetite crosslinked with glutaraldehyde and derivatized with adipic dihydrazide. The carbohydrates were oxidized with sodium periodate, and at optimal concentration, their Vm increased up to 18% for GOx and up to 16% for GA. After immobilization, a remaining activity as high as 88% and 70% for GA with maltose and maltodextrin respectively as substrates was obtained, independently of the particle loading. On the contrary, the remaining activity of GOx strongly decreased at high particle loading. Nevertheless, half of its initial activity was recovered at low loading and was not significantly affected when GA was coimmobilized by saturating the reactive groups left on the particle. The Vm of both immobilized enzymes was improved by crosslinking their carbohydrates with adipic dihydrazide, a treatment which allows further coimmobilization of the other enzyme on a second layer.
Pieters
B R
BR
Laboratoire de Génie Enzymatique, Université Claude Bernard-Lyon, Villeurbanne, France.
Bardeletti
G
G
D004800:Enzymes, Immobilized / Q000378:metabolism*; D005087:Glucan 1,4-alpha-Glucosidase / Q000378:metabolism*; D005949:Glucose Oxidase / Q000378:metabolism*; D007202:Indicators and Reagents; D007700:Kinetics; D008280:Magnetics; D008855:Microscopy, Electron, Scanning; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000494:pharmacology; D011094:Polyethyleneimine
D004800:Enzymes, Immobilized; D007202:Indicators and Reagents; D010504:Periodic Acid; D011094:Polyethyleneimine; D005949:Glucose Oxidase; D005087:Glucan 1,4-alpha-Glucosidase