1,3-丙二醇（1,3-PD）在食品、化工、医药等领域具有非常广泛的应用，而通过微生物发酵粗甘油生产1,3-PD是近年来的热点研究问题。酪酸梭菌是生物法生产1,3-PD的理想发酵菌种之一，但该菌仍然存在1,3-PD耐受性低的问题。本课题以自主筛选的益生菌—酪酸梭菌作为出发菌株，通过Genome shuffling技术，定向进化并筛选1,3-PD的高耐受性突变株。在此基础上，采用反向代谢工程技术，通过全基因组测序，获得与1,3-PD耐受性相关的差异基因；然后，利用RNA-Seq、2-DE和iTRAQ技术分别从转录和蛋白质水平，分析样品RNA和蛋白的变化差异情况，综合分析并确定与1,3-PD耐受性相关的基因与蛋白；最后，对其中的关键基因和蛋白进行功能验证，综合解析酪酸梭菌的1,3-PD耐受性机制。该课题的开展可为粗甘油的综合利用及酪酸梭菌的1,3-PD高效生产提供理论和实践上的参考。

1,3-propanediol has a wide range of application in many fields, such as food, chemical, pharmaceutical, and cosmetics. It is a hot ressearch issue that 1,3-propanediol is obtained from the trasformation of crude glycerol through microbial fermentation in these fields. Although Clostridium butyricum is one of the most ideal fermentation strains which can produced 1,3-propanediol throuth biological method, there is still a shortcoming with low tolerance of 1,3-propanediol in this strain. In present project, in order to obtain a mutant strain with high 1,3-propanediol tolerance, genome shuffling technology is used in the directed evolution of C. butyricum which was a original probiotic strain screened from baby feces. On this base, difference genes associated with 1,3-propanediol tolerance will be obtained using reverse metabolic engineering technology through the whole genome sequencing. Subsequently, RNA and protein’s defferences will also be analyzed using the RNA-Seq, 2-DE and iTRAQ technology on transcription and protein levels, respectively. Then, genes and proteins associated with 1,3-propanediol tolerance will be determined by systems analysis. Furthermore, molecular mechanism about 1,3-propanediol tolerance of C. butyricum will be comprehensive analyzed base on the function validation of key genes and proteins. This research will enhance the comprehensive utilization of crude glycerol and the biosynthesis of 1,3-propanediol by C. butyricum on the theoretical and technical levels.