Evolutionary and reverse engineering in Saccharomyces cerevisiae reveals a Pdr1p mutation-dependent mechanism for 2-phenylethanol tolerance

• DOI: 10.1186/s12934-022-01996-x
• Publication Date: 2022-12-23T14:02:59Z
• Authors: Huili Xia; Yue Kang; Zilin Ma; Cuiyu Hu; Qiao Yang; Xiaoling Zhang; Shihui Yang; Jun Dai; Xiong Chen
• Keywords: 0301 basic medicine; Saccharomyces cerevisiae Proteins; 2-Phenylethanol tolerance; Research; Saccharomyces cerevisiae; Phenylethyl Alcohol; Pdr1p mutation; Microbiology; QR1-502; 03 medical and health sciences; Whole genome sequencing; Mutation; Adaptive laboratory evolution; Reactive Oxygen Species

Abstract Background 2-Phenylethanol (2-PE), a higher alcohol with rose-like odor, inhibits growth of the producer strains. However, limited knowledge regarding 2-PE tolerance mechanisms renders our current base insufficient to inform rational design. Results To improve phenotype Saccharomyces cerevisiae under high concentration, adaptive laboratory evolution (ALE) was used generate an evolved 19–2 strain. Under stress, its OD 600 and rate increased by 86% 22% than that parental strain, respectively. Through whole genome sequencing reverse engineering, transcription factor Pdr1p mutation (C862R) revealed as one main causes for tolerance. stress condition, unsaturated fatty acid/saturated acid ratio 42%, decreased cell membrane damage 81%. Using STRING website, we identified interacted some proteins, which were associated intracellular ergosterol content, reactive oxygen species (ROS), ATP-binding cassette transporter. Also, results transcriptional analysis genes encoded these proteins confirmed induced expression genes. Compared those reference content PDR1 _862 strain 72%–101%, ROS concentration 38% stress. Furthermore, also production (11% higher). Conclusions In present work, have demonstrated use ALE powerful tool yeast 2-PE. Based on physiological analysis, concluded significantly enhanced regulating proportion, ROS. It provides new insights mediated tolerance, could help in design more robust yeasts natural synthesis.