肺炎链球菌HMG-CoA合成酶动力学研究  

ASSAY OF HMG-CoA SYNTHASE ACTIVETY FROM STREPTOCOCCUS PNWUMONIAE

作　　者：贲亚琍[1,2] 崔古贞[1,2] 刘德立[1,2] 

机构地区：[1]江汉大学卫生职业技术学院医学基础部,武汉430016 [2]华中师范大学生命科学学院,武汉430079

出　　处：《广西医科大学学报》2010年第4期493-496,共4页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.30771429);教育部博士点基金项目(No.20060511002)和重点基金项目(No.106116);湖北省自然科学基金资助项目(No.2006ABA197)

摘　　要：目的:在克隆表达肺炎链球菌HMG-CoA合成酶(HMGS)的基础上,对HMGS进行动力学研究。方法:采用紫外分光光度法,以乙酰-CoA作为底物,检测乙酰乙酰-CoA在300 nm处吸收值的变化。结果:肺炎链球菌HMGS的最适pH 9.75、最适温度37℃、最适MgCl2浓度为10 mol/L,粗酶提取物的比活为0.76μmol/min.mg-1,分离纯化后的比活为3.24μmol/min.mg-1,比活提高4.26倍。结论:在37℃、pH 9.755、mol/L MgCl2的最适反应条件下,肺炎链球菌HMGS的Vmax和Km值分别为4.69μmol/min.mg-1和213μmol。Objective：The mvaS gene was cloned from S.pneumoniae and expressed in E.coli BL21.The recombinant HMG-CoA synthase（HMGS） was conducted analysis of dynamics and research of function.Methods：Changes of acetoacetyl-CoA were detected in the 300 nm absorption with acetyl-CoA as substrate by UV-Spectrometer.Results：Optimal conditions for recombinant HMGS were pH 9.75 and 10 mol/L MgCl2 at 37℃.The specific activity of the soluble crude extract was 0.76 μmol/min·mg^-1,while it increased to 3.24 μmol/min·mg^-1 after purified by HisTrapTM FF crude 1 ml column.Conclusion：The Vmax and Km were 4.69 μmol/min·mg^-1 and 213 μmol,respectively,under pH 9.75 and 5 mol/L MgCl2 at 37 ℃.

关 键 词：肺炎链球菌 HMGS 酶特性鉴定 

分 类 号：R378.12[医药卫生—病原生物学]