Xia Huili, Song Na, Liu Daoqi, Zhou Rong, Shangguan Lingling, Chen Xiong, Dai Jun
College of Biological and Food Engineering, Huanghuai University, Zhumadian, 463000, Henan, China.
Cooperative Innovation Center of Industrial Fermentation (Ministry of Education & Hubei Province), Key Laboratory of Fermentation Engineering (Ministry of Education), National "111" Center for Cellular Regulation and Molecular Pharmaceutics, Hubei Key Laboratory of Industrial Microbiology, School of Life and Health Sciences, Hubei University of Technology, Wuhan, 430068, Hubei, People's Republic of China.
Biotechnol Biofuels Bioprod. 2024 Aug 1;17(1):109. doi: 10.1186/s13068-024-02559-5.
The 2-phenylethanol (2-PE) tolerance phenotype is crucial to the production of 2-PE, and Pdr1p mutation can significantly increase the tolerance of 2-PE in Saccharomyces cerevisiae. However, its underlying molecular mechanisms are still unclear, hindering the rational design of superior 2-PE tolerance performance.
Here, the physiology and biochemistry of the PDR1_862 and 5D strains were analyzed. At 3.5 g/L 2-PE, the ethanol concentration of PDR1_862 decreased by 21%, and the 2-PE production of PDR1_862 increased by 16% than those of 5D strain. Transcriptome analysis showed that at 2-PE stress, Pdr1p mutation increased the expression of genes involved in the Ehrlich pathway. In addition, Pdr1p mutation attenuated sulfur metabolism and enhanced the one-carbon pool by folate to resist 2-PE stress. These metabolic pathways were closely associated with amino acids metabolism. Furthermore, at 3.5 g/L 2-PE, the free amino acids content of PDR1_862 decreased by 31% than that of 5D strain, among the free amino acids, cysteine was key amino acid for the enhancement of 2-PE stress tolerance conferred by Pdr1p mutation.
The above results indicated that Pdr1p mutation enhanced the Ehrlich pathway to improve 2-PE production of S. cerevisiae, and Pdr1p mutation altered the intracellular amino acids contents, in which cysteine might be a biomarker in response to Pdr1p mutation under 2-PE stress. The findings help to elucidate the molecular mechanisms for 2-PE stress tolerance by Pdr1p mutation in S. cerevisiae, identify key metabolic pathway responsible for 2-PE stress tolerance.
2-苯乙醇(2-PE)耐受性表型对于2-PE的生产至关重要,Pdr1p突变可显著提高酿酒酵母对2-PE的耐受性。然而,其潜在的分子机制仍不清楚,这阻碍了具有优异2-PE耐受性能的合理设计。
在此,对PDR1_862和5D菌株的生理生化进行了分析。在2-PE浓度为3.5 g/L时,PDR1_862的乙醇浓度比5D菌株降低了21%,PDR1_862的2-PE产量比5D菌株提高了16%。转录组分析表明,在2-PE胁迫下,Pdr1p突变增加了艾氏途径相关基因的表达。此外,Pdr1p突变减弱了硫代谢并增强了叶酸一碳池以抵抗2-PE胁迫。这些代谢途径与氨基酸代谢密切相关。此外,在2-PE浓度为3.5 g/L时,PDR1_862的游离氨基酸含量比5D菌株降低了31%,在游离氨基酸中,半胱氨酸是Pdr1p突变赋予2-PE胁迫耐受性增强的关键氨基酸。
上述结果表明,Pdr1p突变增强了艾氏途径以提高酿酒酵母的2-PE产量,并且Pdr1p突变改变了细胞内氨基酸含量,其中半胱氨酸可能是2-PE胁迫下响应Pdr1p突变的生物标志物。这些发现有助于阐明酿酒酵母中Pdr1p突变对2-PE胁迫耐受性的分子机制,确定负责2-PE胁迫耐受性的关键代谢途径。