摘要
目的 将血管紧张素转换酶 (ACE)固定化并测定其动力学性质。方法 以甲壳糖为载体 ,碳二亚胺 (EDC)为交联剂 ,与分离纯化的ACE混合 ,使ACE固定化 ;并以马尿酰甘氨酰甘氨酸 (HGG)为底物 ,分别测定了其游离酶和固定化酶的动力学性质。结果 固定化酶活力 110u g湿重 ,固定化率为 2 2 % ,与游离酶比较 ,固定化酶稳定且具良好的耐热性 ,在 5 0～ 80℃固定化酶较游离酶稳定。在pH7.1的底物缓冲体系中 ,37℃ ,游离酶Km =0 .0 2 6mol L ;在间歇振摇下固定化酶的表观Km =0 .0 4 0mol L。游离酶最适pH为 8.0 ,固定化酶最适pH为 9.0。结论 用甲壳糖固定ACE方法简便可行 ,且固定后的甲壳糖性能稳定 。
Objective Fixing ACE with chitosan,and detecting its kinetic characterization.Methods Isolated and purified ACE was mixed with vector,chitosan,and crosslinking agent,carbodiimide(EDC),and ACE was immobilized.The kinetics characterization of free enzyme and immobilized ACE were detected using the substrate hippuricylglycylglycine (HGG).Results One gram damp immobilized ACE's activity was 110u.Immobilized rate of ACE was 22%.Compared with free enzyme immobilized ACE was stable and had a good heat resistance,even at 50～80℃.Km value of free enzyme was 0.026mol/L at 37℃ in pH7.1 substrate buffer system; apparent Km of immobilized ACE was 0.040mol/L with intermission vibration.The optimum pH of free enzyme was 8.0,and the optimum pH of immobilized ACE was 9.0.Conclusion The method of immobilized ACE is feasible,and the characteristic of immobilized chitoson is steady.This method has very exrensive applied foreground.
出处
《潍坊医学院学报》
2004年第3期169-170,F003,共3页
Acta Academiae Medicinae Weifang
关键词
甲壳糖
血管紧张素转换酶
固定化
ACE
动力学性质
Chitosan
Angiotensin converting enzyme
Immobilized ACE
Kinetics characterization
