{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T14:04:03Z","timestamp":1790258643514,"version":"4.1.0"},"reference-count":46,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2025,7,2]],"date-time":"2025-07-02T00:00:00Z","timestamp":1751414400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Guizhou Provincial Science and Technology Foundation","award":["Qiankehejichu MS [2025]192"],"award-info":[{"award-number":["Qiankehejichu MS [2025]192"]}]},{"name":"Guizhou Provincial Science and Technology Foundation","award":["Qiankehejichu-ZK [2023]yiban 137"],"award-info":[{"award-number":["Qiankehejichu-ZK [2023]yiban 137"]}]},{"name":"Guizhou Provincial Science and Technology Foundation","award":["Qiankehepingtai KXJZ [2024]021"],"award-info":[{"award-number":["Qiankehepingtai KXJZ [2024]021"]}]},{"name":"Guizhou Provincial Science and Technology Foundation","award":["QKHJC-ZK [2022] YB180)"],"award-info":[{"award-number":["QKHJC-ZK [2022] YB180)"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>Yeast cells are passively exposed to ethanol stress during alcoholic fermentation, ultimately impairing cell viability and reducing fermentation efficiency. Arginine, a versatile amino acid, plays a crucial role in regulating cellular responses to various stresses. This study aimed to explore the underlying mechanism by which arginine protects Wickerhamomyces anomalus against ethanol stress. The effects of arginine supplementation (5 mM) under ethanol stress (9% v\/v) on cell survival, reactive oxygen species (ROS) production, cellular and mitochondrial membrane integrity, and nitric oxide synthesis were investigated using fluorescent staining methods. Furthermore, differentially expressed genes (DEGs) and metabolites (DEMs) were identified through transcriptomics and metabolomics analyses. The results demonstrated that exogenous arginine enhanced cell survival, reduced ROS levels, maintained cellular and mitochondrial membrane integrity, stimulated nitric oxide production, and modulated gene expression and metabolic pathways involved in pyruvate metabolism, yeast meiosis, fatty acid degradation, glycerophospholipid metabolism, and the biosynthesis of various secondary metabolites. These findings provide intriguing insights into the mechanistic role of arginine in enhancing the tolerance of W. anomalus to ethanol stress, and broaden its application in the fermentation industry for alcoholic beverages.<\/jats:p>","DOI":"10.3390\/jof11070499","type":"journal-article","created":{"date-parts":[[2025,7,3]],"date-time":"2025-07-03T04:35:52Z","timestamp":1751517352000},"page":"499","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["The Alleviating Effect of Arginine on Ethanol Stress in Wickerhamomyces anomalus"],"prefix":"10.3390","volume":"11","author":[{"given":"Yinfeng","family":"Li","sequence":"first","affiliation":[{"name":"College of Food and Pharmaceutical Engineering, Guizhou Institute of Technology, Guiyang 550000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujie","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuangyan","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Food and Pharmaceutical Engineering, Guizhou Institute of Technology, Guiyang 550000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guilan","family":"Jiang","sequence":"additional","affiliation":[{"name":"College of Food and Pharmaceutical Engineering, Guizhou Institute of Technology, Guiyang 550000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingzheng","family":"Huang","sequence":"additional","affiliation":[{"name":"College of Food and Pharmaceutical Engineering, Guizhou Institute of Technology, Guiyang 550000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaozhu","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Food and Pharmaceutical Engineering, Guizhou Institute of Technology, Guiyang 550000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1186\/s12934-015-0281-x","article-title":"Yeast as a cell factory: Current state and perspectives","volume":"14","author":"Curk","year":"2015","journal-title":"Microb. Cell Fact."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1007\/s43393-021-00058-4","article-title":"Overview of yeast environmental stress response pathways and the development of tolerant yeasts","volume":"2","author":"Lin","year":"2022","journal-title":"Syst. Microbiol. Biomanuf."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1111\/j.1365-2672.2009.04657.x","article-title":"The ethanol stress response and ethanol tolerance of Saccharomyces cerevisiae","volume":"109","author":"Stanley","year":"2010","journal-title":"J. Appl. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/j.jbiosc.2017.03.009","article-title":"Molecular mechanisms of the yeast adaptive response and tolerance to stresses encountered during ethanol fermentation","volume":"124","author":"Auesukaree","year":"2017","journal-title":"J. Biosci. Bioeng."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.procbio.2018.07.001","article-title":"Response and tolerance of yeast to changing environmental stress during ethanol fermentation","volume":"72","author":"Saini","year":"2018","journal-title":"Process Biochem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1341","DOI":"10.1007\/s00253-022-11799-0","article-title":"Ethanol stress responses in Kluyveromyces marxianus: Current knowledge and perspectives","volume":"106","year":"2022","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Cheng, Y., Du, Z., Zhu, H., Guo, X., and He, X. (2016). Protective effects of arginine on Saccharomyces cerevisiae against ethanol stress. Sci. Rep., 6.","DOI":"10.1038\/srep31311"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Li, Y., Long, H., Jiang, G., Gong, X., Yu, Z.H., Huang, M.Z., Guan, T., Guan, Y., and Liu, X. (2022). Analysis of the ethanol stress response mechanism in Wickerhamomyces anomalus based on transcriptomics and metabolomics approaches. BMC Microbiol., 22.","DOI":"10.1186\/s12866-022-02691-y"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Winter, G., Todd, C.D., Trovato, M., Forlani, G., and Funck, D. (2015). Physiological implications of arginine metabolism in plants. Front. Plant Sci., 6.","DOI":"10.3389\/fpls.2015.00534"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1080\/17429145.2010.539708","article-title":"Effect of exogenous arginine on alleviation of oxidative damage in tomato plant underwater stress","volume":"6","author":"Nasibi","year":"2011","journal-title":"J. Plant Interact."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Hussein, H.A., Alshammari, S.O., Kenawy, S.K.M., Elkady, F.M., and Badawy, A.A. (2022). Grain-priming with L-arginine improves the growth performance of Wheat (Triticum aestivum L.) plants under drought stress. Plants, 11.","DOI":"10.3390\/plants11091219"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1186\/s42269-019-0156-0","article-title":"Comparative study for the effect of arginine and sodium nitroprusside on sunflower plants grown under salinity stress conditions","volume":"43","author":"Ramadan","year":"2019","journal-title":"Bull. Natl. Res. Cent."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1128\/EC.00290-12","article-title":"Candida albicans induces arginine biosynthetic genes in response to host-derived reactive oxygen species","volume":"12","author":"Collette","year":"2013","journal-title":"Eukaryot. Cell"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1230","DOI":"10.1016\/j.procbio.2011.01.026","article-title":"Arginine: A novel compatible solute to protect Candida glabrata against hyperosmotic stress","volume":"46","author":"Xu","year":"2011","journal-title":"Process Biochem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4069","DOI":"10.1108\/BFJ-11-2020-0993","article-title":"Identification and fermentative properties of an indigenous strain of Wickerhamomyces anomalus isolated from Rosa roxburghii Tratt","volume":"123","author":"Liu","year":"2021","journal-title":"Brit. Food J."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1982","DOI":"10.4014\/jmb.1806.06014","article-title":"Ethanol induces autophagy regulated by mitochondrial ROS in Saccharomyces cerevisiae","volume":"28","author":"Jing","year":"2018","journal-title":"J. Microbiol. Biotechnol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1038\/s41564-024-01607-y","article-title":"FacZ is a GpsB-interacting protein that prevents aberrant division-site placement in Staphylococcus aureus","volume":"9","author":"Bartlett","year":"2024","journal-title":"Nat. Microbiol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Huysseune, A., Larsen, U.G., Larionova, D., Matthiesen, C.L., Petersen, S.V., Muller, M., and Witten, P.E. (2023). Bone formation in zebrafish: The significance of DAF-FM DA staining for nitric oxide detection. Biomolecules, 13.","DOI":"10.3390\/biom13121780"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Chen, Y., Wan, Y., Cai, W., Liu, N., Zeng, J., Liu, C., Peng, H., and Fu, G. (2022). Effects on cell membrane integrity of Pichia anomala by the accumulating excessive reactive oxygen species under ethanol stress. Foods, 11.","DOI":"10.2139\/ssrn.4109814"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1016\/j.plaphy.2021.08.030","article-title":"Endogenous nitric oxide and its potential sources regulate glutathione-induced cadmium stress tolerance in maize plants","volume":"167","author":"Kaya","year":"2021","journal-title":"Plant Physiol. Biochem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"115051","DOI":"10.1016\/j.foodres.2024.115051","article-title":"Effect of co-fermentation of non-Saccharomyces yeasts with Saccharomyces cerevisiae on the quality and flavor of blueberry wine","volume":"196","author":"Che","year":"2024","journal-title":"Food Res. Int."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Yao, L., Shi, X., Chen, H., Zhang, L., Cen, L., and Li, L. (2023). Major active metabolite characteristics of Dendrobium officinale rice wine fermented by Saccharomyces cerevisiae and Wickerhamomyces anomalus cofermentation. Foods, 12.","DOI":"10.3390\/foods12122370"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Fan, G., Teng, C., Xu, D., Fu, Z., Liu, P., Wu, Q., Yang, R., and Li, X. (2019). Improving ethyl acetate production in Baijiu manufacture by Wickerhamomyces anomalus and Saccharomyces cerevisiae mixed culture fermentations. Biomed Res. Int., 13.","DOI":"10.1155\/2019\/1470543"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Wang, W., Fan, G., Li, X., Fu, Z., Liang, X., and Sun, B. (2020). Application of Wickerhamomyces anomalus in simulated solid-state fermentation for Baijiu production: Changes of microbial community structure and flavor metabolism. Front. Microbiol., 11.","DOI":"10.3389\/fmicb.2020.598758"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1186\/s13568-019-0811-8","article-title":"Improvement of the aroma of lily rice wine by using aroma-producing yeast strain Wickerhamomyces anomalus HN006","volume":"9","author":"Yan","year":"2019","journal-title":"AMB Express"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Zhang, J., He, Y., Yin, L., Hu, R., Yang, J., and Zhou, J. (2023). Isolation of aroma-producing Wickerhamomyces anomalus yeast and analysis of its typical flavoring metabolites. Foods, 12.","DOI":"10.3390\/foods12152934"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"56","DOI":"10.15586\/qas.v13i2.893","article-title":"Effects of co-inoculation and sequential inoculation of Wickerhamomyces anomalus and Saccharomyces cerevisiae on the physicochemical properties and aromatic characteristics of longan (Dimocarpus longan Lour.) wine","volume":"13","author":"Liu","year":"2021","journal-title":"Qual. Assur. Saf. Crops Foods"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"136205","DOI":"10.1016\/j.chemosphere.2022.136205","article-title":"A comprehensive review on reactive oxygen species (ROS) in advanced oxidation processes (AOPs)","volume":"308","author":"Dong","year":"2022","journal-title":"Chemosphere"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Nadarajah, K.K. (2020). ROS homeostasis in abiotic stress tolerance in plants. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21155208"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"100264","DOI":"10.1016\/j.xplc.2021.100264","article-title":"Chloroplast ROS and stress signaling","volume":"3","author":"Li","year":"2021","journal-title":"Plant Commun."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"697","DOI":"10.1016\/j.jbiosc.2013.05.037","article-title":"Relationship between ethanol and oxidative stress in laboratory and brewing yeast strains","volume":"116","author":"Bleoanca","year":"2013","journal-title":"J. Biosci. Bioeng."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2853","DOI":"10.1016\/j.cell.2022.06.010","article-title":"Nitric oxide signaling in health and disease","volume":"185","author":"Lundberg","year":"2022","journal-title":"Cell"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1007\/s00709-009-0074-3","article-title":"Nitric oxide as a signaling molecule in the fission yeast Schizosaccharomyces pombe","volume":"238","author":"Kig","year":"2009","journal-title":"Protoplasma"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"111835","DOI":"10.1016\/j.plantsci.2023.111835","article-title":"Review on nitric oxide at the forefront of rapid systemic signaling in mitigation of salinity stress in plants: Crosstalk with calcium and hydrogen peroxide","volume":"336","author":"Mariyam","year":"2023","journal-title":"Plant Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"4477","DOI":"10.1093\/jxb\/erz184","article-title":"Role of nitric oxide in plant responses to heavy metal stress: Exogenous application versus endogenous production","volume":"70","author":"Sandalio","year":"2019","journal-title":"J. Exp. Bot."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2051","DOI":"10.1007\/s12298-023-01371-z","article-title":"Nitric oxide (NO) modulates low temperature-stress signaling via S-nitrosation, a NO PTM, inducing ethylene biosynthesis inhibition leading to enhanced post-harvest shelf-life of agricultural produce","volume":"29","author":"Sougrakpam","year":"2023","journal-title":"Physiol. Mol. Biol. Plants"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.niox.2024.01.002","article-title":"Harmonizing hydrogen sulfide and nitric oxide: A duo defending plants against salinity stress","volume":"144","author":"Sharma","year":"2024","journal-title":"Nitric Oxide"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1002\/jbm.a.37056","article-title":"Antibacterial activity of nitric oxide-releasing carboxymethylcellulose against periodontal pathogens","volume":"109","author":"Feura","year":"2021","journal-title":"J. Biomed. Mater. Res. Part A"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1006\/abbi.2000.1785","article-title":"Effects of nitric oxide on the copper-responsive transcription factor Ace1 in Saccharomyces cerevisiae: Cytotoxic and cytoprotective actions of nitric oxide","volume":"377","author":"Chiang","year":"2000","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"jcs253781","DOI":"10.1242\/jcs.253781","article-title":"Atg32-dependent mitophagy sustains spermidine and nitric oxide required for heat-stress tolerance in Saccharomyces cerevisiae","volume":"134","author":"Kaur","year":"2021","journal-title":"J. Cell Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.niox.2015.11.001","article-title":"Nitric oxide signaling and its role in oxidative stress response in Schizosaccharomyces pombe","volume":"52","author":"Astuti","year":"2016","journal-title":"Nitric Oxide"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"John, M.J.J., Song, Y., Hou, M., Zhou, L., Liu, X., Li, X., Fu, D., Li, Q., Cao, H., and Li, R. (2024). Multi-Omics approaches in oil palm research: A comprehensive review of metabolomics, proteomics, and transcriptomics based on low-temperature stress. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25147695"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"3840","DOI":"10.1021\/acs.jproteome.1c00139","article-title":"Integrated multi-omics analysis of mechanisms underlying yeast ethanol tolerance","volume":"20","author":"Arslan","year":"2021","journal-title":"J. Proteome Res."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"103377","DOI":"10.1016\/j.jprot.2019.103377","article-title":"Integrated transcriptomic and proteomic analysis of the ethanol stress response in Saccharomyces cerevisiae Sc131","volume":"203","author":"Li","year":"2019","journal-title":"J. Proteom."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"172017","DOI":"10.1016\/j.scitotenv.2024.172017","article-title":"L-arginine alleviates heat stress-induced mammary gland injury through modulating CASTOR1-mTORC1 axis mediated mitochondrial homeostasis","volume":"926","author":"Gai","year":"2024","journal-title":"Sci. Total Environ."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1016\/j.plaphy.2020.10.002","article-title":"What we know about plant arginases?","volume":"156","author":"Siddappa","year":"2020","journal-title":"Plant Physiol. Biochem."}],"container-title":["Journal of Fungi"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2309-608X\/11\/7\/499\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:03:28Z","timestamp":1760033008000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2309-608X\/11\/7\/499"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,2]]},"references-count":46,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2025,7]]}},"alternative-id":["jof11070499"],"URL":"https:\/\/doi.org\/10.3390\/jof11070499","relation":{},"ISSN":["2309-608X"],"issn-type":[{"value":"2309-608X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,2]]}}}