产丁醇梭菌耐受高浓度丁醇的生理及代谢调节机制研究

项目名称： 产丁醇梭菌耐受高浓度丁醇的生理及代谢调节机制研究

项目编号： No.21466014

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 有机化学

项目作者： 李汉广

作者单位： 江西农业大学

项目金额： 50万元

中文摘要： 作为一种极具潜力的生物燃料，生物丁醇已经成为世界能源领域的研究热点。然而，丁醇对细胞的毒害作用导致其产量、生产强度和底物转化率较低是目前丙酮丁醇发酵过程中存在的一个关键科学问题。本项目拟采用诱变新技术及结合可视化筛选模型获得高丁醇耐受菌；通过宏蛋白质组学和转录组学技术，找出原始菌与突变菌在高浓度丁醇胁迫下差异表达蛋白及其编码基因在转录水平上的变化；采用代谢通量分析技术，分析同一条件下原始菌与突变菌代谢通量及代谢途径酶含量变化；以及系统分析突变菌与原始菌在高浓度丁醇胁迫下细胞形态、细胞内微环境及细胞膜脂变化等。从多层次、多角度获得突变菌在高浓度丁醇胁迫下发生变化的信息组，以期较为全面的揭示产丁醇梭菌耐受高浓度丁醇的生理及代谢调节机制，为提高产丁醇梭菌合成丁醇能力提供理论指导与技术支持。本研究结果将为提高该微生物的工业适应性和其它微生物耐受有机溶剂机理提供科学的探索。

中文关键词： 丙酮丁醇发酵；丁醇耐受性；丙酮丁醇梭菌PW12；宏蛋白质组学；转录组学

英文摘要： As a great potential biofuel, acetone, butanol and ethanol (ABE) fermentation by clostridial species has regained much attention in both industrial and academic worlds recently. However, butanol toxicity to the current producing microorganisms limits its accumulation in the fermentation broth, and for this reason the biobutanol production processes is of low yield, productivity and conversion efficiency. Therefore, this presents a key scientific challenge in ABE fermentation process. We propose to use an effective mutation screening method to obtain high butanol tolerance strain. The expression of different proteins and the corresponding changes of gene at the transcription level will be identified under high butanol concentration conditions between wild-type strain and mutant strain by metaproteomics and transcriptomics. Metabolic flux changes and enzyme levels will also be investigated under the same conditions by metabolic flux analysis. Furthermore, we will study the changes of cell morphology, cellular microenvironment and membrane lipid under high butanol concentration stress between the wild-type strain and mutant strain. After successful completion, the mechanisms of physiological and metabolic regulation will be fully revealed and understood in mutant strain. In addition, our results will provide the theoretical foundation and technical support to further enhance butanol production by clostridial, and improve the industrial adaptability and capabilities of this microorganism, thus providing scientific know-how of solvent tolerance mechanisms.

英文关键词： Acetone butanol and ethanol fermentation;Butanol tolerance;Clostridium acetobutylicum PW12;Metaproteomics;Transcriptomics