{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T23:06:35Z","timestamp":1788822395813,"version":"build-2803163510"},"reference-count":46,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,5,29]],"date-time":"2025-05-29T00:00:00Z","timestamp":1748476800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Junta Andalucia (Spain) Program: Specialized Transfer Actions Universities\u2013CEI-RIS3-FEDER","award":["PYC20 RE 068 UCO"],"award-info":[{"award-number":["PYC20 RE 068 UCO"]}]},{"name":"Junta Andalucia (Spain) Program: Specialized Transfer Actions Universities\u2013CEI-RIS3-FEDER","award":["PID 2021-127766OB-I00"],"award-info":[{"award-number":["PID 2021-127766OB-I00"]}]},{"name":"Singular Project AGROMIS-ceiA3, Subsector: Traditional Wines of Andalusia","award":["PYC20 RE 068 UCO"],"award-info":[{"award-number":["PYC20 RE 068 UCO"]}]},{"name":"Singular Project AGROMIS-ceiA3, Subsector: Traditional Wines of Andalusia","award":["PID 2021-127766OB-I00"],"award-info":[{"award-number":["PID 2021-127766OB-I00"]}]},{"name":"Call 2020: Grants for R+D+I projects in the field of Innovation Ecosystems of the International Centers of Excellence (PY2020 ECOSIS INNOV CEIS)","award":["PYC20 RE 068 UCO"],"award-info":[{"award-number":["PYC20 RE 068 UCO"]}]},{"name":"Call 2020: Grants for R+D+I projects in the field of Innovation Ecosystems of the International Centers of Excellence (PY2020 ECOSIS INNOV CEIS)","award":["PID 2021-127766OB-I00"],"award-info":[{"award-number":["PID 2021-127766OB-I00"]}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science, Innovation, and Universities (MICIU\/European Union FEDER)","doi-asserted-by":"publisher","award":["PYC20 RE 068 UCO"],"award-info":[{"award-number":["PYC20 RE 068 UCO"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science, Innovation, and Universities (MICIU\/European Union FEDER)","doi-asserted-by":"publisher","award":["PID 2021-127766OB-I00"],"award-info":[{"award-number":["PID 2021-127766OB-I00"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>This study evaluated the oenological potential of two non-Saccharomyces yeast strains, Hanseniaspora opuntiae TR-5 and Zygosaccharomyces bailii L-25, isolated from extreme winemaking environments in southern Spain. Out of 156 yeast isolates screened from high-sugar musts and flor yeast biofilms, strains were selected based on their \u03b2-glucosidase activity, killer phenotype, and ethanol production, traits associated with aroma release and microbial competition. Fermentation trials on sugar-rich synthetic medium showed that both H. opuntiae and Z. bailii achieved ethanol yields of 10% v\/v and residual sugars at 4 g\/L. Co-culture and sequential inoculation, with H. opuntiae introduced first and Z. bailii added on day four, resulted in complete alcoholic fermentation and a reduction in undesirable acetoin levels compared to single-strain fermentations. These findings highlight the practical potential of using selected non-Saccharomyces strains in sequential fermentations to improve aroma complexity, fermentation reliability, and sensory quality in wines, even in the absence of Saccharomyces cerevisiae. The application of these strains offers a novel approach for precision oenology and varietal expression in challenging musts.<\/jats:p>","DOI":"10.3390\/microorganisms13061260","type":"journal-article","created":{"date-parts":[[2025,5,29]],"date-time":"2025-05-29T07:46:26Z","timestamp":1748504786000},"page":"1260","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Selection of Non-Saccharomyces Yeasts from Extreme Oenological Environments for Potential Use in Winemaking"],"prefix":"10.3390","volume":"13","author":[{"given":"Mar\u00eda Trinidad","family":"Alcal\u00e1-Jim\u00e9nez","sequence":"first","affiliation":[{"name":"Department of Agricultural Chemistry, Edaphology and Microbiology, Agrifood Campus of International, Excellence CeiA3, Universidad de C\u00f3rdoba, 14014 C\u00f3rdoba, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan Carlos","family":"Garc\u00eda-Garc\u00eda","sequence":"additional","affiliation":[{"name":"Department of Agricultural Chemistry, Edaphology and Microbiology, Agrifood Campus of International, Excellence CeiA3, Universidad de C\u00f3rdoba, 14014 C\u00f3rdoba, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5694-7653","authenticated-orcid":false,"given":"Juan Carlos","family":"Mauricio","sequence":"additional","affiliation":[{"name":"Department of Agricultural Chemistry, Edaphology and Microbiology, Agrifood Campus of International, Excellence CeiA3, Universidad de C\u00f3rdoba, 14014 C\u00f3rdoba, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7646-9345","authenticated-orcid":false,"given":"Juan","family":"Moreno","sequence":"additional","affiliation":[{"name":"Department of Agricultural Chemistry, Edaphology and Microbiology, Agrifood Campus of International, Excellence CeiA3, Universidad de C\u00f3rdoba, 14014 C\u00f3rdoba, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0136-8793","authenticated-orcid":false,"given":"Rafael","family":"Peinado","sequence":"additional","affiliation":[{"name":"Department of Agricultural Chemistry, Edaphology and Microbiology, Agrifood Campus of International, Excellence CeiA3, Universidad de C\u00f3rdoba, 14014 C\u00f3rdoba, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2746-2753","authenticated-orcid":false,"given":"Teresa","family":"Garc\u00eda-Mart\u00ednez","sequence":"additional","affiliation":[{"name":"Department of Agricultural Chemistry, Edaphology and Microbiology, Agrifood Campus of International, Excellence CeiA3, Universidad de C\u00f3rdoba, 14014 C\u00f3rdoba, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"117209","DOI":"10.1016\/j.lwt.2024.117209","article-title":"Microbial diversity during wine fermentation in Beijing region and its influence on wine flavor","volume":"215","author":"Wu","year":"2025","journal-title":"LWT"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Gao, J., Wang, M., Huang, W., You, Y., and Zhan, J. (2022). Indigenous Saccharomyces cerevisiae could better adapt to the physicochemical conditions and natural microbial ecology of prince grape must compared with commercial Saccharomyces cerevisiae FX10. Molecules, 27.","DOI":"10.3390\/molecules27206892"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Yin, X., Liu, B., Maimaitiyiming, R., Wang, L., Zhao, L., Zhang, H., Chen, K., and Aihaiti, A. (2024). The Effect of Non-Saccharomyces Cerevisiae Torulaspora delbrueckii on the aroma composition of munage grape base-wine and the mechanism of the effect. Fermentation, 10.","DOI":"10.3390\/fermentation10050266"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1111\/j.1567-1364.2009.00579.x","article-title":"Controlled mixed culture fermentation: A new perspective on the use of non-Saccharomyces yeasts in winemaking","volume":"10","author":"Ciani","year":"2010","journal-title":"FEMS Yeast Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.fm.2013.10.001","article-title":"Biodiversity study of wine yeasts belonging to the \u201cterroir\u201d of Montepulciano d\u2019Abruzzo \u201cColline Teramane\u201d revealed Saccharomyces cerevisiae strains exhibiting atypical and unique 5.8S-ITS restriction patterns","volume":"39","author":"Tofalo","year":"2014","journal-title":"Food Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"141248","DOI":"10.1016\/j.foodchem.2024.141248","article-title":"Volatile aroma compound-based decoding and prediction of sweet berry aromas in dry red wine","volume":"463","author":"Li","year":"2025","journal-title":"Food Chem."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Lombardi, S.J., Pannella, G., Iorizzo, M., Moreno-Arribas, M.V., Tremonte, P., Succi, M., Sorrentino, E., Macciola, V., Di Renzo, M., and Coppola, R. (2018). Sequential inoculum of Hanseniaspora Guilliermondii and Saccharomyces Cerevisiae for winemaking campanino on an industrial Scale. World J. Microbiol. Biotechnol., 34.","DOI":"10.1007\/s11274-018-2540-6"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2625","DOI":"10.1002\/fsn3.1116","article-title":"Volatile organic compounds of Hanseniaspora Uvarum increase strawberry fruit flavor and defense during cold storage","volume":"7","author":"Wang","year":"2019","journal-title":"Food Sci. Nutr."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1016\/j.lwt.2019.04.045","article-title":"Impact of co-inoculation of Saccharomyces Cerevisiae, Hanseniaspora Uvarum and Oenococcus Oeni autochthonous strains in controlled multi starter grape must Fermentations","volume":"109","author":"Capozzi","year":"2019","journal-title":"LWT"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/S0168-1605(03)00245-9","article-title":"Yeast interactions and wine flavour","volume":"86","author":"Fleet","year":"2003","journal-title":"Int. J. Food Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/S0168-1605(03)00289-7","article-title":"Yeast associated with spontaneous fermentations of white wines from the \u201cTxakoli de Bizkaia\u201d region (Basque Country, North Spain)","volume":"86","author":"Rementeria","year":"2003","journal-title":"Int. J. Food Microbiol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Morata, A., Escott, C., Loira, I., Manuel Del Fresno, J.M., Vaquero, C., Antonia Ba\u00f1uelos, M., Palomero, F., L\u00f3pez, C., and Gonz\u00e1lez, C. (2022). pH Control and Aroma Improvement Using the Non-Saccharomyces Lachancea thermotolerans and Hanseniaspora spp. Yeasts to improve wine freshness in warm areas. Grapes Wine, Intechopen.","DOI":"10.5772\/intechopen.100538"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"422","DOI":"10.1099\/mic.0.2007\/011445-0","article-title":"Ethanol tolerance of sugar transport, and the rectification of stuck wine fermentations","volume":"154","author":"Santos","year":"2008","journal-title":"Microbiology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3","DOI":"10.3390\/fermentation1010003","article-title":"Evaluation of Zygosaccharomyces bailii to metabolize residual sugar present in partially-fermented red wines","volume":"1","author":"Zuehlke","year":"2015","journal-title":"Fermentation"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Petruzzi, L., Capozzi, V., Berbegal, C., Corbo, M.R., Bevilacqua, A., Spano, G., and Sinigaglia, M. (2017). Microbial resources and enological significance: Opportunities and benefits. Front. Microbiol, 8.","DOI":"10.3389\/fmicb.2017.00995"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.micres.2015.02.002","article-title":"Evaluation of Zygosaccharomyces bailii BCV 08 as a co-starter in wine fermentation for the improvement of ethyl esters production","volume":"173","author":"Garavaglia","year":"2015","journal-title":"Microbial. Res."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Solieri, L. (2021). The revenge of Zygosaccharomyces yeasts in food biotechnology and applied microbiology. World J. Microbiol. Biotechnol., 37.","DOI":"10.1007\/s11274-021-03066-7"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Alcal\u00e1-Jim\u00e9nez, M.T., Garc\u00eda-Mart\u00ednez, T., Mauricio, J.C., Moreno, J., and Peinado, R.A. (2025). Influence of terroir on microbial diversity and wine volatilome. Appl. Sci., 15.","DOI":"10.3390\/app15063237"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4686","DOI":"10.1128\/AEM.70.8.4686-4691.2004","article-title":"Genetic instability of heterozygous hybrid populations of natural wine yeasts","volume":"70","author":"Vinagre","year":"2004","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_20","unstructured":"Kaiser, C., Michaelis, S., and Mitchell, A. (1994). Methods in Yeast Genetics, Cold Spring Harbor Laboratory Press."},{"key":"ref_21","unstructured":"Lodder, J., and Kreger-van Rij, N.J.W. (1952). The Yeasts: A Taxonomic Study, Elsevier. [1st ed.]."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"02018","DOI":"10.1051\/bioconf\/20235602018","article-title":"Impact of non-Saccharomyces autochthonous yeasts on ethanol reduction and chemical profile of Chilean Sauvignon Blanc wine","volume":"Volume 56","author":"Urtubia","year":"2023","journal-title":"Proceedings of the BIO Web of Conferences"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Spedding, G. (2016). Alcohol and its measurement. Brewing Materials and Processes, Academic Press.","DOI":"10.1016\/B978-0-12-799954-8.00007-1"},{"key":"ref_24","unstructured":"(2023). International Code of Oenological Practices, OIV International Organisation of Vine and Wine."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1093\/jaoac\/68.2.188","article-title":"Determination of ethanol in wine by titrimetric and spectrophotometric dichromate methods: Collaborative study","volume":"68","author":"Pilone","year":"1985","journal-title":"J. AOAC Int."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"6389","DOI":"10.1021\/jf049369o","article-title":"Gas chromatographic quantification of major volatile compounds and polyols in wine by direct injection","volume":"52","author":"Peinado","year":"2004","journal-title":"J. Agric. Food Chem."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Miranda, A., Pereira, V., Jardim, H., Malfeito-Ferreira, M., and Marques, J.C. (2023). Impact of Non-Saccharomyces yeast fermentation in madeira wine chemical composition. Processes, 11.","DOI":"10.3390\/pr11020482"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Ruiz-Mu\u00f1oz, M., Hern\u00e1ndez-Fern\u00e1ndez, M., Cordero-Bueso, G., Mart\u00ednez-Verdugo, S., P\u00e9rez, F., and Cantoral, J.M. (2022). Non-Saccharomyces are also forming the veil of flor insherry wines. Fermentation, 8.","DOI":"10.3390\/fermentation8090456"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1016\/j.lwt.2017.09.026","article-title":"Production and characterization of a \u03b2-glucosidase from Issatchenkia terricola and its use for hydrolysis of aromatic precursors in Cabernet Sauvignon wine","volume":"87","author":"Cavello","year":"2018","journal-title":"LWT"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/1574-6968.12348","article-title":"\u03b2-Glucosidase activities of lactic acid bacteria: Mechanisms, impact on fermented food and human health","volume":"352","author":"Michlmayr","year":"2014","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1080\/10408398.2020.1813684","article-title":"Glycosidically bound aroma precursors in fruits: A comprehensive review","volume":"62","author":"Liang","year":"2022","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1381","DOI":"10.1007\/s11947-013-1161-1","article-title":"Immobilization of \u03b2-glucosidase and its application for enhancement of aroma precursors in muscat wine","volume":"7","author":"Ramirez","year":"2014","journal-title":"Food Bioprocess Technol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/j.tifs.2022.06.015","article-title":"Microbial interactions in winemaking: Ecological aspects and effect on wine quality","volume":"127","author":"Englezos","year":"2022","journal-title":"Trends Food Sci. Technol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1066","DOI":"10.1128\/AEM.66.3.1066-1076.2000","article-title":"Prevention of yeast spoilage in feed and food by the yeast mycocin HMK","volume":"66","author":"Lowes","year":"2000","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1023\/A:1010396232420","article-title":"Stress effect of ethanol on fermentation kinetics by stationary-phase cells of Saccharomyces cerevisiae","volume":"23","author":"Blondin","year":"2001","journal-title":"Biotechnol. Lett."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2705","DOI":"10.1007\/s00217-019-03384-z","article-title":"Using Torulaspora delbrueckii, Saccharomyces cerevisiae and Saccharomyces bayanus wine yeasts as starter cultures for fermentation and quality improvement of mead","volume":"245","author":"Sottil","year":"2019","journal-title":"Eur. Food Res. Technol."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Del Fresno, J.M., Loira, I., Escott, C., Carrau, F., Gonz\u00e1lez, C., Cuerda, R., and Morata, A. (2021). Application of Hanseniaspora vineae yeast in the production of ros\u00e9 wines from a blend of Tempranillo and Albillo grapes. Fermentation, 7.","DOI":"10.3390\/fermentation7030141"},{"key":"ref_38","unstructured":"Moreno, J., and Peinado, R. (2012). Enological Chemistry, Academic Press."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1464","DOI":"10.1016\/j.foodchem.2005.11.045","article-title":"Volatile components of zalema white wines","volume":"100","author":"Cacho","year":"2007","journal-title":"Food Chem."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Martin, V., Valera, M.J., Medina, K., Boido, E., and Carrau, F. (2018). Oenological impact of the Hanseniaspora\/Kloeckera yeast genus on wines\u2014A review. Fermentation, 4.","DOI":"10.3390\/fermentation4030076"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Guittin, C., Ma\u00e7na, F., Picou, C., Perez, M., Barreau, A., Poitou, X., Sablayrolles, J.-M., Mouret, J.-R., and Farines, V. (2023). New online monitoring approaches to describe and understand the kinetics of acetaldehyde concentration during wine alcoholic fermentation: Access to production balances. Fermentation, 9.","DOI":"10.3390\/fermentation9030299"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"105660","DOI":"10.1016\/j.jfca.2023.105660","article-title":"Mechanisms and effects of non-Saccharomyces yeast fermentation on the aromatic profile of wine","volume":"124","author":"Wang","year":"2023","journal-title":"J. Food Compos. Anal."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"111649","DOI":"10.1016\/j.foodres.2022.111649","article-title":"Impact of Starmerella bacillaris and Zygosaccharomyces bailii on ethanol reduction and Saccharomyces cerevisiae metabolism during mixed wine fermentations","volume":"159","author":"Capece","year":"2022","journal-title":"Food Res. Int."},{"key":"ref_44","unstructured":"Epifanio, S. (2005). La Influencia de la Tecnolog\u00eda de la Vinificaci\u00f3n en la Microbiolog\u00eda y el Desarrollo de la Fermentaci\u00f3n Alcoh\u00f3lica. [Ph.D. Thesis, Universidad de la Rioja]."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1016\/S0168-1605(03)00255-1","article-title":"Acetate ester formation in wine by mixed cultures in laboratory fermentations","volume":"86","author":"Rojas","year":"2003","journal-title":"Int. J. Food Microbiol."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Filippousi, M.E., Chalvantzi, I., Mallouchos, A., Marmaras, I., Banilas, G., and Nisiotou, A. (2024). The use of Hanseniaspora opuntiae to improve \u2018Sideritis\u2019 wine quality, a late-ripening Greek grape variety. Foods, 13.","DOI":"10.3390\/foods13071061"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/6\/1260\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:43:03Z","timestamp":1760031783000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/13\/6\/1260"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,5,29]]},"references-count":46,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2025,6]]}},"alternative-id":["microorganisms13061260"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms13061260","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,29]]}}}