幽门螺杆菌尿素酶的纯化和鉴定

幽门螺杆菌超声波上清经DEAE SepheroseFastFlow和PhenylSepheroseCL 4B两步柱层析得到纯化的Hp尿素酶 ,免疫印迹显示有良好的抗原性。在pH8.0和 45℃条件下酶比活约为 2 6 5U mg ,SDS PAGE显示有 6 0 0 0 0和 30 0 0 0两个亚单位。经两步柱层析纯化后 ,Hp尿素酶回收率为 30 % ,纯化倍数达 11倍。连续纯化 3批显示工艺稳定 ,为Hp尿素酶基础研究和应用奠定基础。

Purification and Characterization of Urease from Helicobacter Pylori

After the whole cells of helicobacter pylori was broken by ultrasonic wave,the urease in supernatant was purified by DEAE Sephe rose Fast Flow and Phenyl Sepherose CL 4B chromatographies.The reco very rate of urease was 30%,and the purification fold fo it reached 11.SDS PAGE proved that the purified urease consisted of 2 subunints with relative molecular weights of 60 000 and 30 000 respectively,and Western blot showed good antigenicity of it.The specific activity of the urease at 45℃ pH8.0 was about 265 U/mg.Three lots of urease wee purified,the result showed that the process was stable and laid a foundation of basic research and application of urease.;