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Article

Step-Wise Ethanol Adaptation Drives Cell-Wall Remodeling and ROM2/KNR4 Activation in Brettanomyces bruxellensis

by
Leslie Hernandez-Cabello
1,
Nachla Rojas-Torres
1,
Liliana Godoy
2,*,
Camila G-Poblete
1,
Yarabi Concha
1,
Verónica Plaza
3,
Luis Castillo
3,
Héctor M. Mora-Montes
4 and
María Angélica Ganga
1,*
1
Departamento en Ciencia y Tecnología de los Alimentos, Universidad de Santiago de Chile, Alameda 3363, Santiago 9170022, Chile
2
Departamento de Fruticultura y Enología, Facultad de Agronomía y Sistemas Naturales, Pontificia Universidad Católica de Chile, Santiago 8320165, Chile
3
Laboratorio de Biología Molecular y Bioquímica, Departamento de Biología, Universidad de La Serena, La Serena 1700000, Chile
4
Departamento de Biología, División de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta s/n, Col. Noria Alta, C.P., Guanajuato 36050, Mexico
*
Authors to whom correspondence should be addressed.
Microorganisms 2025, 13(7), 1489; https://doi.org/10.3390/microorganisms13071489
Submission received: 14 May 2025 / Revised: 16 June 2025 / Accepted: 20 June 2025 / Published: 26 June 2025
(This article belongs to the Special Issue Wine Fermentation Microorganisms)
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Abstract

Brettanomyces bruxellensis has been described as the main spoilage microorganism in wines due to its ability to produce volatile phenols, which negatively impact the final product’s organoleptic properties. This yeast can grow and survive in environments that are too nutritionally poor and stressful for other microorganisms, and one of the stressful conditions it can endure is the high alcohol content in wine. In this study, cell wall morphology and the expression of some genes related to its composition were characterized under increasing ethanol concentrations to establish a possible ethanol resistance mechanism. B. bruxellensis LAMAP2480 showed greater resistance to β-1,3-glucanase activity when grown in media supplemented with 5% or 10% ethanol compared with the control assay (without ethanol). Transmission electron microscopy showed no significant differences in cell wall thickness during the different adaptation stages. However, the amount of wall polysaccharides and chitin briefly increased at 1% ethanol but returned to baseline at 5% and 10%. The amount of wall-associated protein increased progressively with each increment in ethanol concentration. In addition, overexpression of the ROM2 and KNR4/SMI1 genes was observed at 10% ethanol. These results suggest that the integrity of the cell wall might play an important role in the adaptation of B. bruxellensis to an ethanol-containing medium.
Keywords: Brettanomyces bruxellensis; cell wall integrity; spoilage wine yeast; ethanol adaptation Brettanomyces bruxellensis; cell wall integrity; spoilage wine yeast; ethanol adaptation

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MDPI and ACS Style

Hernandez-Cabello, L.; Rojas-Torres, N.; Godoy, L.; G-Poblete, C.; Concha, Y.; Plaza, V.; Castillo, L.; Mora-Montes, H.M.; Ganga, M.A. Step-Wise Ethanol Adaptation Drives Cell-Wall Remodeling and ROM2/KNR4 Activation in Brettanomyces bruxellensis. Microorganisms 2025, 13, 1489. https://doi.org/10.3390/microorganisms13071489

AMA Style

Hernandez-Cabello L, Rojas-Torres N, Godoy L, G-Poblete C, Concha Y, Plaza V, Castillo L, Mora-Montes HM, Ganga MA. Step-Wise Ethanol Adaptation Drives Cell-Wall Remodeling and ROM2/KNR4 Activation in Brettanomyces bruxellensis. Microorganisms. 2025; 13(7):1489. https://doi.org/10.3390/microorganisms13071489

Chicago/Turabian Style

Hernandez-Cabello, Leslie, Nachla Rojas-Torres, Liliana Godoy, Camila G-Poblete, Yarabi Concha, Verónica Plaza, Luis Castillo, Héctor M. Mora-Montes, and María Angélica Ganga. 2025. "Step-Wise Ethanol Adaptation Drives Cell-Wall Remodeling and ROM2/KNR4 Activation in Brettanomyces bruxellensis" Microorganisms 13, no. 7: 1489. https://doi.org/10.3390/microorganisms13071489

APA Style

Hernandez-Cabello, L., Rojas-Torres, N., Godoy, L., G-Poblete, C., Concha, Y., Plaza, V., Castillo, L., Mora-Montes, H. M., & Ganga, M. A. (2025). Step-Wise Ethanol Adaptation Drives Cell-Wall Remodeling and ROM2/KNR4 Activation in Brettanomyces bruxellensis. Microorganisms, 13(7), 1489. https://doi.org/10.3390/microorganisms13071489

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