Regulation of ethyl carbamate by BTN2 of Saccharomyces cerevisiae during mixed-culture Huangjiu fermentation with Pediococcus pentoses

Abstract

Ethyl carbamate (EC) is a naturally occurring potential carcinogen in fermented foods. Our prior research indicated that BTN2 knockout affected arginine metabolism in Saccharomyces cerevisiae, and the inhibitory effect was more pronounced in mixed cultures with Pediococcus pentoses (PP). Consequently, in this study, we investigated the potential mechanisms of EC regulation by BTN2 knockout strains in single- and mixed-culture fermentation systems in Huangjiu and determined their fundamental qualities. The findings revealed that the BTN2 knockout strain could reduce the EC content by decreasing the amount of EC precursors and that mixed fermentation with PP could impede the reaction between urea and ethanol, thereby exerting a more favorable EC abatement effect. The BTN2 knockout strain had a positive effect on free amino acids in Huangjiu, thus improving the flavor. Additionally, transcriptome analysis demonstrated that the knockout of BTN2 and the addition of PP affected gene expression levels, paticularly genes associated with transcription factor activity and amino acid transport in S. cerevisiae, which subsquently affected the metabolic pathways during the fermentation process.

Keywords

Ethyl carbamate / Huangjiu / Saccharomyces cerevisiae / Pediococcus pentoses / BTN2 / Mixed-culture fermentation

Author contributions

The authors confirm their contributions to the paper as follows: conceptualization: Fang R; methodology: Fang R, Hu J; writing original draft: Fang R, Xu H; writing - review & editing: Chen T, Tang X, Xiao G, Zhou A; funding acquisition: Fang R, Tang X; software: Zhou H, Hu J; formal analysis: Zhou H, Xu H; visualization: Zhou H, Lin W, Xu H; data curation: Zhou H, Lin W; investigation: Shi C, Lin W; validation: Shi C, Hu J; Zhou A; resources: Shi C, Tang X; project administration, supervision: Chen T, Xiao G. All authors reviewed the results and approved the final version of the manuscript.

Data availability

All relevant data are within the manuscript and its supplementary files.

Acknowledgments

This study was funded by the National Natural Science Foundation of China (No. 32101912), the Science and Technology Program of State Administration for Market Regulation (2022MK163), the Leading the Charge with Open Competition program in Wenzhou (ZNF2023009) and the Basic Scientific Research Project of Zhejiang University of Science and Technology (2023JLZD009).

Conflict of interest

The authors declare that they have no conflict of interest.

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