基于毛细管电泳技术的微生物胞内目标代谢物在线分析

项目名称： 基于毛细管电泳技术的微生物胞内目标代谢物在线分析

项目编号： No.21475019

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 数理科学和化学

项目作者： 杨丽

作者单位： 东北师范大学

项目金额： 80万元

中文摘要： 微生物胞内代谢物的定量水平关系着微生物代谢途径和代谢流量的掌握。基于高效分离技术发展快速、准确、高通量的在线分析方法对胞内代谢物进行精确测定，有助于微生物代谢途径的分析和调控，对发酵工业、生物制药、环境微生物等研究领域具有重要意义。项目提出以毛细管电泳分离技术为基础，建立集高通量细胞进样、快速在线细胞溶膜、以及高效分离检测于一体的新型微生物胞内代谢物在线分析方法，着重以微生物培养液中重要的碳源物质-单糖，和氮源物质-氨基酸作为目标代谢物分析，针对生物制品合成中若干重要的微生物，检测不同培养环境和细胞生长时刻的胞内代谢产物，并与胞外培养液检测结果对比，分析胞内外相关代谢物的动态变化。项目旨在建立一种简单、快速、高效的微生物胞内代谢物在线检测方法，加深对微生物代谢途径和代谢流的认识，探索对目标产物产率的优化和控制，进一步拓展毛细管电泳技术在生物、复杂及痕量物质分析中的应用。

中文关键词： 毛细管电泳；微生物；目标代谢物；在线分析

英文摘要： Quantitative measurement of intracellular microbial metabolites relates to understanding the network and flux of microbial metabolism. Based on high performance separation technique, to develop rapid, accurate and high throughput online analysis method for rapid and accurate determination of intracellular microbial metabolites is of great importance in controlling metabolic pathways, optimizing fermentation process, developing pharmaceuticals, investigating environmental microbiology, and so on. The proposal presents a novel method for online analysis of intracellular microbial metabolites based on capillary electrophoresis, which combines high throughput cell injection, fast online cell lyses, and high efficient separation and detection techniques. The study will focus on two kinds of target metabolites, i.e., monosaccharides and amino acids, which are important carbon source and nitrogen source in culture solution respectively. We will online measure the intracellular metabolites of several important microorganisms in synthesis of bio-products, with different culture environments and at different cellular growth time. By comparing the intercellular analysis results with those of extracellular measurements, we will analyze the dynamic change of both intracellular and extracellular metabolites. The purpose of the proposal aims at developing a simple, fast and efficient method for online analysis of intracellular microbial metabolites, adding our understanding of the microbial metabolic pathways and flux, thus exploring optimization and control of the yield of target bio-products. Our study will further expand the application of capillary electrophoresis in analysis of biological, complex and trace amount samples.

英文关键词： Capillary electrophoresis;Microbe;Targeted metabolite;Online analysis