Abstract：
Abundant renewable resource lignocellulosic biomass possesses tremendous potential for green biomanufacturing, while its efficient utilization by Yarrowia lipolytica, an attractive biochemical production host, is restricted since the presence of inhibitors furfural and acetic acid in lignocellulosic hydrolysate. Given deficient understanding of inherent interactions between inhibitors and cellular metabolism, sufficiently mining relevant genes is necessary. Herein, 14 novel gene targets were discovered using clustered regularly interspaced short palindromic repeats interference library in Y. lipolytica, achieving tolerance to 0.35 % (v/v) acetic acid (the highest concentration reported in Y. lipolytica), 4.8 mM furfural, or a combination of 2.4 mM furfural and 0.15 % (v/v) acetic acid. The tolerance mechanism might involve improvement of cell division and decrease of reactive oxygen species level. Transcriptional repression of effective gene targets still enabled tolerance when xylose was a carbon source. This work forms a robust foundation for improving microbial tolerance to lignocellulose-derived inhibitors and revealing underlying mechanism.
摘要：
丰富的可再生资源木质纤维素生物质具有巨大的绿色生物制造潜力,虽然它被Yarrowialipolytica有效利用，一个有吸引力的生化生产宿主，由于在木质纤维素水解产物中存在抑制剂糠醛和乙酸而受到限制。鉴于缺乏对抑制剂和细胞代谢之间固有相互作用的理解，充分挖掘相关基因是必要的。在这里，使用Y.Lipolytica中成簇的规则间隔短回文重复干扰文库发现了14个新的基因靶标，实现对0.35％（v/v）乙酸（Y.lipolytica中报告的最高浓度）的耐受性，4.8mM糠醛，或2.4mM糠醛和0.15%(v/v)乙酸的组合。耐受机制可能涉及细胞分裂的改善和活性氧水平的降低。当木糖是碳源时，有效基因靶标的转录抑制仍然能够耐受。这项工作为提高微生物对木质纤维素衍生抑制剂的耐受性和揭示潜在机制奠定了坚实的基础。
