Dataset Information

Analysis of acid-tolerance mechanism based on membrane microdomains in Saccharomyces cerevisiae.

ABSTRACT:

Background

Saccharomyces cerevisiae has been used in the biosynthesis of acid products such as organic acids owing to its acid tolerance. Improving the acid tolerance of S. cerevisiae is beneficial for expanding its application range. Our previous study isolated the TAMC strain that was tolerant to a pH 2.3 through adaptive laboratory evolution; however, its mechanism underlying tolerance to low pH environment remains unclear.

Results

In this study, through visual observation and order analysis of plasma membrane and membrane microdomains, we revealed that the membrane microdomains of TAMC strain play an indispensable role in acid tolerance. Transcriptomic analysis showed an increase in the expression of genes related to key components of membrane microdomains in TAMC strain. Furthermore, an obvious reduction was observed in the acid tolerance of the strain with sterol C-24 methyltransferase encoding gene ERG6 knockout for inhibiting membrane microdomain formation. Finally, colocalization analysis of H⁺-ATPase PMA1 and plasma membrane protein PMP1 showed that disruption of membrane microdomains could inhibit the formation of the H⁺-ATPase complex.

Conclusions

Membrane microdomains could provide a platform for forming H⁺-ATPase complexes to facilitate intracellular H⁺ homeostasis, and thereby improve cell acid resistance. This study proposed a novel acid tolerance mechanism, providing a new direction for the rational engineering of acid-tolerant strains.

SUBMITTER: Lv X

PROVIDER: S-EPMC10498586 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature

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Publications

Analysis of acid-tolerance mechanism based on membrane microdomains in Saccharomyces cerevisiae.

Lv Xueqin X Jin Ke K Yi Yu Y Song Lingang L Xiu Xiang X Liu Yanfeng Y Li Jianghua J Du Guocheng G Chen Jian J Liu Long L

Microbial cell factories 20230913 1

<h4>Background</h4>Saccharomyces cerevisiae has been used in the biosynthesis of acid products such as organic acids owing to its acid tolerance. Improving the acid tolerance of S. cerevisiae is beneficial for expanding its application range. Our previous study isolated the TAMC strain that was tolerant to a pH 2.3 through adaptive laboratory evolution; however, its mechanism underlying tolerance to low pH environment remains unclear.<h4>Results</h4>In this study, through visual observation and ...[more]

PMID: 37700284

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Project description:Sugarcane molasses is an ideal economical raw material for ethanol production because of its wide availability, low cost and nutrient content. However, benzoic acid compounds with toxic effects on yeast cells are commonly found in sugarcane molasses. At present, the molecular mechanism of the toxic effects of benzoic acid on Saccharomyces cerevisiae has not been elucidated. Here, the toxic effect of exogenous benzoic acid on S. cerevisiae GJ2008 cells was studied, and the genes differentially expressed in S. cerevisiae GJ2008 after 1.2 g/L benzoic acid stress were identified via Illumina RNA-Seq technology. The results indicated that benzoic acid significantly inhibited yeast cell growth, prolonged their rapid growth period, and ultimately reduced their biomass. During ethanol fermentation using 250 g/L sucrose under 1.2 g/L benzoic acid stress, several adverse effects were observed, such as high residual sugar content, low ethanol concentration and low fermentation efficiency. In addition, the cell morphology was damaged, the cell membrane permeability increased, intracellular nucleic acid and protein leakage increased, and the malondialdehyde content significantly increased. Moreover, the cells protected themselves by significantly increasing the intracellular glycerol content. Fourier transform infrared spectroscopy proved that benzoic acid could reduce the degree of unsaturation and increase cell membrane permeability by changing the yeast cell wall and cell membrane composition, leading to cell damage and even death. Transcriptomic analysis revealed that under benzoic acid stress, the expression of genes associated with sucrose and starch metabolism, thiamine metabolism, the glycolysis pathway, fructose and mannose metabolism, galactose metabolism and ABC transporters was significantly downregulated. The expression of genes related to ribosomes, lipid metabolism, ribosome biosynthesis, nucleic acid metabolism, arginine and proline metabolism, RNA polymerase, metabolism related to cofactor synthesis, and biosynthesis of valine, leucine, and isoleucine was significantly upregulated. These results indicated that benzoic acid inhibited glycolysis and reduced sugar absorption and utilization and ATP energy supply in yeast cells. In response to stress, genes related to the ribosome bioanabolic pathway were upregulated to promote protein synthesis. On the other hand, the expression of ELO1, SUR4, FEN1 and ERG1 was upregulated, which led to extension of long-chain fatty acids and accumulation of ergosterol to maintain cell membrane structure. In conclusion, this paper provides important insights into the mechanism underlying the toxicity of benzoic acid to yeast cells and for realizing high-concentration ethanol production by sugarcane molasses fermentation.

Dataset Information

Analysis of acid-tolerance mechanism based on membrane microdomains in Saccharomyces cerevisiae.

Background

Results

Conclusions

Publications

Analysis of acid-tolerance mechanism based on membrane microdomains in Saccharomyces cerevisiae.

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