Immobilization of Chondroitinase by Macroporous Resin D380

Objective To immobilize chondroitinase(ChSase)using macroporous resin D380 as carrier and glutaraldehyde as cross linking agent,and investigate the enzymological property of immobilized chondroitinase.Methods The effects of quantity of enzyme added,the temperature,time and pH value for adsorption as well as the glutaraldehyde concentration,time and temperature for cross linking on immobilization of ChSase were investigated,based on which the optimal working temperature and pH value,Km and operational stability of immobilized ChSase were analyzed.Results The optimal quantity of enzyme added and optimal temperature,time and pH value for adsorption were 150 U/g resin,15℃,6 h and 7.0,while the optimal glutaraldehyde concentration,time and temperature for cross linking were 0.01%,3 h and 4℃,respectively.The combining efficiency of ChSase immobilized under the optimal condition was 79.1%.The optimal working temperature and pH value of immobilized ChSase were 45℃and 7.0 respectively.The immobilized ChSase showed good operational stability,and its Km was 1.46×10-1 g/L,which was significantly higher than that of free ChSase.Conclusion It was feasible to immobilize ChSase using macroporous resin D380 as carrier,and the immobilized ChSase showed high combining efficiency and stability,which was suitable for industrial production.