LuxR家族蛋白调控茂原链霉菌TGase合成的机制研究

项目名称： LuxR家族蛋白调控茂原链霉菌TGase合成的机制研究

项目编号： No.31301545

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 张莉丽

作者单位： 东北农业大学

项目金额： 22万元

中文摘要： 谷氨酰胺转氨酶（Transglutaminase，简称TGase）又名转谷氨酰胺酶，通过催化蛋白质发生交联，改善蛋白质的功能特性，受到了广泛关注。 在前期研究结果表明MgCl2胁迫促进了茂原链霉菌TGase合成，蛋白质组学研究结果表明它抑制了LuxR家族蛋白的表达。本项目将利用基因芯片技术和生物信息学分析，研究MgCl2胁迫对Streptomyces mobaraensis基因转录的影响，确定与TGase合成有关的调控LuxR家族蛋白表达的基因。利用基因突变技术，构建LuxR家族蛋白基因的阻断株和恢复突变株。利用蛋白质组学方法研究基因阻断株、恢复突变株以及野生菌株的胞内蛋白质表达的差异；利用代谢组学方法研究这三株菌的代谢路径和代谢产物的变化。另外，通过测定三株菌的生产性能,菌体形态变化，阐明LuxR家族蛋白调控TGase合成的机制。同时利用这种机制调控TGase合成，为大规模生产奠定基础。

中文关键词： 茂原链霉菌；luxR基因；TGase；合成；机制

英文摘要： Transglutaminase (shorten as TGase) is a family of enzymes that can catalyze the cross-linking of proteins to modify the functional properties of protein. It attacts a great interest. The previous results showed that MgCl2 stress improved the TGase production from Streptomyces mobaraensis and inhibited the expression of LuxR family protein. In this study, we will analyse the effects of MgCl2 stress on the gene transcription in S. mobaraensis and confirm the key gene of control LuxR family protein for regulating TGase synthesis by gene chip technology and bioinformatics analysis. Using gene silencing technology, LuxR family protein gene disruption strain and restored mutant will be constructed. Proteomics will be exploited to analyse differentially expressed proteins in disruption strains, restored mutation and wild stain. Metabolics will be used to study their changes of metabolic pathways and metabolites. In addition, changes in cell morphology of three strains will be observed. Based on above study, the mechanism of regulation TGase synthesis by LuxR family proteins will be elucidated. At the same time, the mechanism will be used to improve the TGase synthesis , which will lay the foundation for large-scale production.

英文关键词： Streptomyces mobaraensis；luxR gene；TGase；synthesis；mechanism