一种大规模、快速的幽门螺杆菌脲酶分离纯化方法研究

The research of a large-scale and rapid method to purify urease from Helicobacter pylori

幽门螺杆菌定植于人胃黏膜并产生大量的脲酶,报道一种大规模、快速的脲酶纯化方法.脲酶用去离子水振荡及冻融法抽提,采用Q Sepharose FF阴离子交换层析,Phenyl Sepharose HP疏水相互作用和SOURCE30S阳离子交换层析等方法纯化得到脲酶,通过酚红脲酶试剂检测酶活.总蛋白回收率为2.58%,纯化后脲酶纯度大于95%,SDS-PAGE电泳显示脲酶是由相对分子质量66 000的A亚基和30 000的B亚基组成.

The bacterium Helicobacter pylori colonizes the human gastric mucosa and produces large amounts of urease.Here,a large-scale and rapid method to purify urease from Helicobacter pylori was developed.The enzyme was extracted from the bacteria by distilled water and alternate freezing and thawing,then purified by anion-exchange chomatography(Q Sepharose FF),Hydrophobic interaction(Phenyl Sepharose HP),and cation-ecxchange chomatography(SOURCE 30S).Urease enzyme activity was detected with a spectrophotometic as...