| 一种大规模、快速的幽门螺杆菌脲酶分离纯化方法研究 |
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| 引用本文： | 任金琪,刘建东,周曾芬,马岚. 一种大规模、快速的幽门螺杆菌脲酶分离纯化方法研究[J]. 云南大学学报(自然科学版), 2006, 0(Z1) |
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| 作者姓名： | 任金琪 刘建东 周曾芬 马岚 |
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| 作者单位： | [1]云南大学单克隆抗体工程技术中心 [2]昆明医学院第一附属医院消化内科 [3]云南大学单克隆抗体工程技术中心 云南昆明 [4]云南昆明 |
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| 摘 要： | 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.
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| 关 键 词： | 幽门螺杆菌 脲酶 纯化 |
The research of a large-scale and rapid method to purify urease from Helicobacter pylori |
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| REN Jin-qi,LIU Jian-dong,ZHOU Zeng-fen,MA Lan. The research of a large-scale and rapid method to purify urease from Helicobacter pylori[J]. Journal of Yunnan University(Natural Sciences), 2006, 0(Z1) |
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| Authors: | REN Jin-qi LIU Jian-dong ZHOU Zeng-fen MA Lan |
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| Affiliation: | REN Jin-qi~1,LIU Jian-dong~2,ZHOU Zeng-fen~2,MA Lan~1 |
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| Abstract: | 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 assay based on phenol red.The overall protein recovery was 2.58%,and purity of urease was over 95%.The urease showed the typical subunit of M_r 66 000 and M_r 30 000 when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. |
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| Keywords: | Helicobacter pylori urease purification |
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