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目的对赤子爱胜蚓中一组新型脱氧核糖核酸酶(EWDs)进行酶动力学研究。方法运用单相酶扩散法(SRED)、紫外分光光度法和林-贝氏(Lineweaver-Burk)作图法。结果EWDs的最适温度均为37℃。EWD1的最适pH为5.2,Km为1.58mg/ml,Vmax为5.36mg·ml-1·min-1;EWD2的最适pH为4.4,Km值是3.95mg/ml,Vmax值为2.87mg·ml-1·min-1;EWD3的最适pH为4.8,Km值是1.52mg/ml,Vmax值为4.89mg·ml-1·min-1。Mn2+和Ca2+是EWDs的抑制剂,Mg2+仅抑制EWD3的活性。结论蚯蚓组织中发现的此组脱氧核糖核酸酶具有特殊的酶学性质,区别于已知的脱氧核糖核酸酶类。
Objective To investigate the enzyme kinetics of a group of novel DNases (EWDs) in E. sinensis. Methods Single-phase enzyme diffusion (SRED), UV spectrophotometry, and Lineweaver-Burk mapping were used. As a result, the optimum temperature of EWDs was 37°C. The optimum pH of EWD1 was 5.2, Km was 1.58 mg/ml, Vmax was 5.36 mg·ml-1·min-1, the optimum pH of EWD2 was 4.4, Km value was 3.95 mg/ml, Vmax value was 2.87 mg· The optimum pH of ml-1·min-1; EWD3 was 4.8, the Km value was 1.52 mg/ml, and the Vmax value was 4.89 mg·ml-1·min-1. Mn2+ and Ca2+ are inhibitors of EWDs, and Mg2+ only inhibits the activity of EWD3. Conclusion This group of deoxyribonucleases found in baboon tissues has special enzymatic properties that distinguish them from known DNases.