马克思克鲁维酵母有高温下快速生长，在工业发酵中降低能耗减少污染等优势。我们在该酵母中实现了共利用葡萄糖和木糖生产乙醇、丙酮酸及木糖醇等生物基化学品，但木质纤维素生物质预处理产生的抑制物会降低该酵母的发酵效率。目前对酵母抑制物的耐受以直接相关的蛋白的研究为主，与抑制物耐受特有的转录调控因子的研究很少。我们通过该酵母在抑制物存在与否时的转录组分析发现一种与抑制物尤其是糠醛类的耐受有关的转录因子（FRR），并证实该基因的敲除或过表达可以降低或提高此酵母对抑制物的耐受。本项目将以此转录因子为研究对象，以其对糠醛类耐受作用为重点，拟采用ChIP-Seq、RNA-seq、EMSA及报告基因分析等技术，运用基因敲除及过表达等基因工程技术手段确定FRR调控的目标基因，并对调控的基因在抑制物耐受中的功能进行鉴定，解析FRR作为关键节点的糠醛类耐受机理，为提高该酵母对抑制物的耐受提供工程改造的重要信息。

Kluyveromyces marxianus is a thermo-tolerant yeast which can grow at temperature higher than 42℃ and keeps relatively high ethanol fermentation efficiency. Fermentation at elevated temperature has some advantages such as reduced cool cost and contamination in large scale fermentation. We have constructed a strain which can assimilate glucose and xylose simultaneously to produce ethanol, pyruvate, xylitol, but the inhibitors produced during the pretreatment of lignocellulosic biomass reduced the fermentation efficiency. Current researches of inhibitor tolerance mainly focus on directly related enzymes and proteins. The reports of inhibitor-specific transcriptional regulator are seldom. We found a potential transcriptional factor FRR through the RNA-seq of K. marxianus with or without inhibitor. Disruption of the gene of this protein led decreased inhibitor tolerance, and overexpression of FRR could imporve the inhibitor tolerance. In this application, we will use ChIP-seq, EMSA, RNA-seq and reporter system to screen and identify the DNA binding sites and target genes of this protein; use gene disruption and overexpression to figure out the function of target genes; analyze the protein and the target genes function in inhibitor (mainly furfural and HMF) tolerance; summary the interaction of the FRR and those genes encoded proteins. The results of this research will provide important information for higher inhibitor tolerant strain construction in biochemical production with lignocellulosic biomass.