Enzymatic characterization of a Type Ⅱ isocitrate dehydrogenase from pathogenic Leptospira interrogans serovar Lai strain 56601
作者单位：Institute of Molecular Biology and BiotechnologyAnhui Normal University
会议名称：《第八届全国医学生物化学与分子生物学第五届全国临床应用生物化学与分子生物学2013华东六省一市生物化学与分子生物学联合学术研讨会》
会议日期：2013年
学科分类：1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 1001[医学-基础医学(可授医学、理学学位)] 100103[医学-病原生物学] 10[医学]
关 键 词：Leptospira interrogans isocitrate dehydrogenase kinetics coenzyme specificity diagnosic candidate
摘 要：Leptospira interrogans,a Gram-negative pathogen,could cause infections in a wide variety of mammalian hosts,but due to their fastidious cultivation requirements and the lack of genetic systems,the pathogenic factor is still not *** dehydrogenase(IDH) is a key enzyme in the tricarboxylation(TCA) cycle,which could have an important impact on the growth and pathogenesis of the *** the present study,we first report the cloning,heterologous expression,and detailed characterization of the EDH gene from *** serovar Lai strain 56601(LiIDH).The molecular weight of LilDH was determined to be 87 kDa by filtration chromatography,suggesting LilDH is a typical *** optimum activity of LilDH was found at 60 ℃,and its optimum pH was 7.0(Mn) and 8.0(Mg).Heat-inactivation studies showed that heat treatment for 20 min at 50 ℃ caused a 50%loss of enzyme *** was completely divalent cation dependent as other typical dimeric IDHs and Mg was its best *** recombinant LilDH specificities(k/K values for NADPand NAD) in the presence of Mg and Mn were 6,269-fold and 1,000-fold greater for NADPthan NAD,*** current work is expected to shed light on the functions of metabolic enzymes in *** and provide useful information for LilDH to be considered as a possible candidate for serological diagnostics and detection of *** infection.