猪胸膜肺炎放线杆菌尿素酶的提取及特性研究  

Purification and characterization of urease from Actinobacillus pleuropneumoniae

作　　者：王妮[1] 刁有祥[1] 刘方娜[1] 孟凡磊[1] 王辉[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018

出　　处：《西北农林科技大学学报（自然科学版）》2008年第2期34-38,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：山东农业大学科技创新基金项目(23414)

摘　　要：【目的】从猪血清7型胸膜肺炎放线杆菌中提取纯化尿素酶,并对其生物特性和理化特性进行研究。【方法】超声破碎放线杆菌粗提尿素酶,再采用硫酸铵盐析和Sephadex G-200凝胶过滤层析对该酶进行纯化,分析其活性及理化特性。【结果】最适于猪胸膜肺炎放线杆菌尿素酶保持活性的pH为8.5,温度为45℃。在该条件下其比酶活约为96.253 U/mg;SDS-PAGE电泳显示,胸膜肺炎放线杆菌尿毒酶含分子质量为11,25和60 ku的亚单位。【结论】与传统方法相比,超声破壁和Sephadex G-200凝胶过滤层析纯化法简单快速,重复性好,测定尿素酶的特性稳定,能满足实验室检测的要求。[Objective] Urease from Actinobacfllus pleuropneumoniae serotype 7 was purified to study its physical and biochemical proprties. [Method] Urease from Actinobacillus pleuropneumoniae serotype 7 was extracted,then （NH4）2SO4 fractionation and Sephadex G-200 gel filtration were used to extract and purify urease. [Result] The optimum pH and temperature for the urease to keep the activities were 8.5 and 45 ℃. The specific activity of urease under this situation was 96. 253 U/mg. The urease had three subunits, and the molecular weight for each was about 11,25 and 60 ku by SDS-PAGE. [Conclusion] Using ultrasonication and Sephadex G-200 gel filtration to extract urease was a simple and quick meature,the results of which were stable and could reach the requirement of experiment properties.

关 键 词：猪传染性胸膜肺炎 胸膜肺炎放线杆菌 尿素酶 超声破壁 酶活性 理化特性 

分 类 号：S858.286.3[农业科学—临床兽医学]