鉴定参与酿酒酵母耐热性和耐乙醇性的关键转录因子，包括DAL80和CRZ1。

Identification of key transcription factors, including DAL80 and CRZ1, involved in heat and ethanol tolerance in Saccharomyces cerevisiae.

作者信息

Chen Rong-Rong, Wang Li, Ji Xue-Xue, Xie Cai-Yun, Tang Yue-Qin

机构信息

College of Architecture and Environment, Sichuan University, Chengdu, 610065, Sichuan, China.

Sichuan Environmental Protection Key Laboratory of Organic Wastes Valorization, Chengdu, 610065, Sichuan, China.

出版信息

Biotechnol Biofuels Bioprod. 2025 May 3;18(1):50. doi: 10.1186/s13068-025-02653-2.

DOI:10.1186/s13068-025-02653-2

PMID:40319324

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12048999/

Abstract

BACKGROUND

High temperature and ethanol are two critical stress factors that significantly challenge bioethanol production using Saccharomyces cerevisiae. In this study, the tolerance mechanisms of the multi-tolerant S. cerevisiae strain E-158 to heat stress and combined heat-ethanol stress were investigated using comparative transcriptomics.

RESULTS

Under heat stress at 44 °C, glucose transport and reactive oxygen species (ROS) scavenging were significantly upregulated, while gluconeogenesis, acetate formation, and dNDP formation showed significant downregulation. Under combined heat (43 °C) and ethanol (3% v/v) stress, glucose transport, glycolysis, acetate formation, peroxisome activity, ROS scavenging, and ribosome synthesis were significantly upregulated, while glycerol formation, cellular respiration and dNDP formation exhibited significant downregulation. Fourteen transcription factors (TFs), considered to play a key role in both stress conditions, were individually overexpressed and deleted in S. cerevisiae strain KF-7 in this study. Among these TFs, Gis1p, Crz1p, Tos8p, Yap1p, Dal80p, Uga3p, Mig1p, and Opi1p were found to contribute to enhanced heat tolerance in S. cerevisiae. Compared with KF-7, strains overexpressing DAL80 and CRZ1 demonstrated markedly improved fermentation performance under stress conditions. Under heat stress at 44 °C, glucose consumption increased by 10% and 12%, respectively, for strains KF7DAL80 and KF7CRZ1, while ethanol production increased by 12% and 15%, respectively, compared to KF-7. Under combined stress conditions of 43 °C and 3% (v/v) ethanol, glucose consumption increased by 67% and 44%, ethanol production by 116% and 77%, and ethanol yield by 29% and 22%, respectively, for KF7DAL80 and KF7CRZ1 compared to KF-7. KF7CRZ1 performs comparably to E-158, while KF7DAL80 outperforms E-158.

CONCLUSIONS

This study provides valuable theoretical insights and identifies critical TF targets, contributing to the development of robust S. cerevisiae strains for improved bioethanol production.

摘要

背景

高温和乙醇是两个关键的胁迫因素，对利用酿酒酵母生产生物乙醇构成了重大挑战。在本研究中，使用比较转录组学方法研究了多耐受性酿酒酵母菌株E-158对热胁迫和热-乙醇复合胁迫的耐受机制。

结果

在44°C热胁迫下，葡萄糖转运和活性氧（ROS）清除显著上调，而糖异生、乙酸盐形成和脱氧核苷二磷酸（dNDP）形成则显著下调。在43°C热胁迫和3%（v/v）乙醇复合胁迫下，葡萄糖转运、糖酵解、乙酸盐形成、过氧化物酶体活性、ROS清除和核糖体合成显著上调，而甘油形成、细胞呼吸和dNDP形成则显著下调。本研究在酿酒酵母菌株KF-7中分别对14个被认为在两种胁迫条件下起关键作用的转录因子（TFs）进行了过表达和缺失操作。在这些TFs中，发现Gis1p、Crz1p、Tos8p、Yap1p、Dal80p、Uga3p、Mig1p和Opi1p有助于提高酿酒酵母的耐热性。与KF-7相比，过表达DAL80和CRZ1的菌株在胁迫条件下表现出明显改善的发酵性能。在44°C热胁迫下，KF7DAL80和KF7CRZ1菌株的葡萄糖消耗分别比KF-7增加了10%和12%，而乙醇产量分别增加了12%和15%。在43°C和3%（v/v）乙醇的复合胁迫条件下，与KF-7相比，KF7DAL80和KF7CRZ1的葡萄糖消耗分别增加了67%和44%，乙醇产量分别增加了116%和77%，乙醇产率分别增加了29%和22%。KF7CRZ1的表现与E-158相当，而KF7DAL80的表现优于E-158。