大孔树脂D380固定化硫酸软骨素裂解酶  

Immobilization of Chondroitinase by Macroporous Resin D380

作　　者：牛海滨[1] 蔡苏兰[1] 阎浩林[1] 刘婷婷[1] 

机构地区：[1]沈阳药科大学生命科学与生物制药学院,沈阳110016

出　　处：《中国生物制品学杂志》2010年第2期203-206,210,共5页Chinese Journal of Biologicals

摘　　要：目的以大孔树脂D380为载体,戊二醛为交联剂,进行硫酸软骨素裂解酶(ChSase)的固定化,并考察固定化酶的酶学性质。方法分别考察加酶量、吸附温度、吸附时间、吸附pH值、戊二醛交联浓度、交联时间及交联温度对ChSase固定化效果的影响,并分析该固定化酶的最适反应温度、最适反应pH值、米氏常数(Km)及其操作稳定性。结果ChSase的最佳固定化条件为:加酶量150U/g树脂,吸附温度15℃,吸附时间6h,吸附pH值7.0,戊二醛交联浓度0.01%,交联时间3h,交联温度4℃。以此条件制备的固定化酶,其酶结合效率可达79.1%。该固定化ChSase的最适反应温度为45℃;最适反应pH值为7.0;Km达1.46×10-1g/L,较游离酶高;具有较好的操作稳定性。结论以大孔树脂D380为载体固定化ChSase是可行的,所得固定化酶有较高的使用效率和稳定性,适合于工业化生产。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.

关 键 词：大孔树脂D380 硫酸软骨素裂解酶 固定化 

分 类 号：Q556.2[生物学—生物化学]