项目名称: 谷氨酸棒杆菌中3-羟基苯甲酸/龙胆酸代谢途径的调控研究
项目编号: No.31271333
项目类型: 面上项目
立项/批准年度: 2013
项目学科: 生物科学
项目作者: 周宁一
作者单位: 中国科学院武汉病毒研究所
项目金额: 80万元
中文摘要: 谷氨酸棒杆菌 ATCC 13032通过龙胆酸分解代谢3-羟基苯甲酸。本项目研究该菌中3-羟基苯甲酸/龙胆酸代谢途径在转录水平上的调控。在其基因组中,参与3-羟基苯甲酸代谢的基因可能位于genIKL、genR、genTX等三个操纵子上。其中,genR基因编码一个IclR家族调控蛋白,其他基因编码3-羟基苯甲酸代谢所需的酶;genR基因缺失后,谷氨酸棒杆菌中的龙胆酸和3-羟基苯甲酸代谢功能丧失,这表明该调控蛋白正调控整个代谢途径。另外,生物信息学显示,CRP-FNR家族调控蛋白GlxR可能抑制这条代谢途径。本研究拟通过DNase I 足迹法、EMSA等方法研究调控蛋白GenR和GlxR与每个启动子的结合情况,以及解析启动子的结构,揭示它们对每个操纵子的调控及自调控,从而首次在分子水平阐释IclR家族调控蛋白和CRP-FNR家族调控蛋白同时参与3-羟基苯甲酸/龙胆酸代谢的调控机理。
中文关键词: 谷氨酸棒杆菌;3-羟基苯甲酸;龙胆酸;分解代谢;转录调控
英文摘要: Corynebacterium glutamicum ATCC13032 catabolize 3-hydroxybenzoate via gentisate. This project examines the regulation of 3-hydroxybenzoate/gentisate catabolism in this strain at transcriptional level. Three operons (genIKL, genR, genTX) likely participate in the 3-hydroxybenzoate metabolism in this strain. genR encodes an IclR family regulator and other genes code for 3-hydroxybenzoate metabolic enzymes. The genR-deleted strain lost the ability to grow on gentisate and 3-hydroxybenzoate, indicating that the GenR positively regulates their metabolism. In addition, bioinformatics analysis shows that a CRP-FNR family regulator GlxR may inhibit the transcription of the operons in this metabolic pathway. In this study, the interaction between regulatory proteins GenR/GlxR and each promoter of the operons will be performed by the methods of DNase I footprinting, EMSA. The structure of the promoter will be resolved, which will further reveal the regulation of each operon and their auto-regulation. This study will be the first one to interpret the involvement of an IclR family regulator and a CRP-FNR regulator in the regulation of 3-hydroxybenzoate metabolism.
英文关键词: Corynebacterium glutamicum;3-hydroxybenzoate;gentisate;catabolism;transcriptional regulation