Immobilization of glucose isomerase onto radiation synthesized P(AA-co-AMPS) hydrogel and its application

Abstract

Isomerization of glucose to fructose was carried out using Glucose isomerase (GI) that immobilized by entrapment into Poly(acrylic acid) P(AA) and Poly(acrylic acid-co-2-Acrylamido 2-methyl Propane sulfonic acid) P(AA-co-AMPS) polymer networks, the enzyme carriers were prepared by radiation induced copolymerization in the presence of (Methylene-bisacrylamide) (MBAA) as a crosslinking agent. The maximum gel fraction of pure P(AA) and P(AA-co-AMPS) hydrogel was found to be 95.2% and 89.6% for P(AA) and P(AA-co-AMPS), respectively at a total dose of 20 kGy. Effects of immobilization conditions such as radiation dose, MBAA concentration, comonomer composition and amount of GI were investigated. The influence of reaction conditions on the activity of immobilized GI were studied, the optimum pH value of the reaction solution is 7.5 and reaction temperature is 65 °C. The immobilized GI into P(AA-co-AMPS) and P(AA) polymer networks retained 81% and 69%, respectively of its initial activity after recycled for 15 times while it retained 87% and 71%, respectively of its initial activity after stored at 4 °C for 48 days. The Km values of free and immobilized GI onto P(AA-co-AMPS) and onto P(AA) matrices were found to be 34, 29.2 and 14.5 mg/mL, respectively while the Vmax Values calculated to be 3.87, 1.6 and 0.79 mg/mL min, respectively. GI entrapped into P(AA-co-AMPS) hydrogel show promising behavior that may be useful as the newly glucose isomerase reactor in biomedical applications.

Keywords

• Radiation
• Polymerization
• Enzyme immobilization
• Entrapment
• Glucose isomerase