铜绿假单胞菌PAO1亚精胺脱氢酶SpdH的表达、纯化和初步晶体学研究  

Expression,Purification and Preliminary Crystallographic Analysis of Pseudomonas aeruginosa PAO1 Spermidine Dehydrogenase SpdH

作　　者：车世友 张琼林 Che Shiyou;Zhang Qionglin(College of Life Sciences,Nankai University,Tianjin 300071,China)

机构地区：[1]南开大学生命科学学院,天津300071

出　　处：《南开大学学报（自然科学版）》2020年第3期37-42,共6页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：国家自然科学基金面上项目(31570128)。

摘　　要：铜绿假单胞菌是一种具有很强环境适应性的机会致病菌,感染后通常很难治愈从而造成很严重的后果.聚胺类化学物质在很多种生物体内广泛存在,并且在细胞内发挥多种重要的功能.铜绿假单胞菌PAO1可以利用聚胺类物质作为菌体生长所需的唯一的碳源和氮源,亚精胺脱氢酶(SpdH)在铜绿假单胞菌亚精胺代谢过程中发挥重要的作用.本研究在大肠杆菌中成功表达了可溶性SpdH蛋白,并经过多种方法纯化后筛选获得了蛋白质晶体,通过X射线衍射实验收集到分辨率达到0.185 nm的衍射数据,并使用HKL2000软件对衍射数据进行了处理,这些数据为SpdH结构的解析奠定了基础.Pseudomonas aeruginosa is an important opportunistic pathogenic bacterium with strong environmental versatility,all the infections caused by Pseudomonas aeruginosa are usually lethal and the treatment can be difficult.Polyamines are widely distributed in many living things and have a great influence on many intricate cellular processes.Pseudomonas aeruginosa PAO1 can utilize polyamines as the sole source of carbon and nitrogen,spermidine dehydrogenase(SpdH)plays an important role in spermidine utilization process in Pseudomonas aeruginosa PAO1.In this research,soluble SpdH was successfully expressed in E.coli,and finally got well-diffracting protein crystals after purification and crystallization condition screen,Xray diffraction data were collected to 0.185 nm resolution and processed with HKL2000.These data established foundation to the structure determination of SpdH.

关 键 词：铜绿假单胞菌 亚精胺脱氢酶 纯化 晶体学研究 

分 类 号：Q71[生物学—分子生物学]