人胰型及唾液型淀粉酶的分离纯化与鉴定

Purification and characterization of human pancreatic and salivary amylases

目的 为获得高比活性、高纯度的人胰型及唾液型淀粉酶。方法 用硫酸铵分级沉淀、Sephadex G-50凝胶层析及DEAE-纤维素离子交换层析法分别从人胰腺及唾液中分离纯化人胰型及唾液型淀粉酶,并用聚丙烯酰胺凝胶电泳法进行纯度、酶活性及分子量鉴定。结果 获得了高比活性及高纯度的人胰型及唾液型淀粉酶,胰型及唾液型淀粉酶的表现分子量分别为55KD和57KD。结论 本方法是获得高纯度及高比活性人胰型及唾液型淀粉酶的有效途径。

Objective To obtain human pancreatic and salivary amylase with high specific activity and high purity. Methods Pancreatic and salivary amylases were separated and purified from human pancreas and saliva with ammonium sulfate fractionation, Sephadex G - 50 gel chro-matography, and DEAE - cellulose ion - exchange chromatography. Polyacrylamide gel elec-trophoresis was also applied to determine the purity, activity, and molecular weights of pancreatic and salivary amylases. Results Pancreatic and salivary amylases with high specific activity and high purity were obtained.The apparent molecular weights of pancreatic and salivary amylases were 55KD and 57KD,respectively.Conclusion This protocol is an effective approach to obtain pancreatic and salivary amylases with high specific activity and high purity.