Impact of stress combination on Brettanomyces bruxellensis growth and volatile phenol production
Abstract
Brettanomyces bruxellensis is a major wine spoilage yeast responsible for the production of volatile phenols, including 4-ethylphenol (EP) and 4-ethylguaiacol (EG), from grape-derived hydroxycinnamic acids. This study investigated how oenological stress factors, particularly temperature, pH, and p-coumaric acid concentration, modulate B. bruxellensis growth, viability, p-coumaric acid consumption, and conversion into 4-vinylphenol (VP) and EP. Three B. bruxellensis strains, 2OT13_02, L0463 and 1961-Mx-M1 E2, representing distinct genetic backgrounds, were grown in synthetic wine at two temperatures with varying p-coumaric acid concentrations. Complementarily, strain 2OT13_02 was exposed to pH 3.3 and pH 5.6 across a large range of p-coumaric acid concentrations. Cell counts and viability were monitored by flow cytometry, while LC-UV-MS/MS quantified p-coumaric acid, VP and EP. Elevated temperature strongly impaired growth and viability in a strain-dependent manner. At 28 °C, 1961-Mx-M1 E2 showed near-zero growth and mortality close to 100 %, whereas 2OT13_02 maintained the highest metabolic activity. At 20 °C, all strains consumed almost all p-coumaric acid, but their conversion profiles differed, with 2OT13_02 and 1961-Mx-M1 E2 producing mainly EP and L0463 accumulating both VP and EP. Increasing temperature reduced EP formation and favoured VP accumulation, indicating a limitation of VP reduction under stress. Wine pH also reshaped the pathway: pH 5.6 reduced p-coumaric acid consumption but prevented the metabolic block observed at high p-coumaric acid concentrations and pH 3.3. These results show that volatile phenol production by B. bruxellensis is not solely determined by precursor availability, but by the interaction between strain background, acid stress, and temperature. Taking into account these stress combinations may help winemakers better anticipate Brettanomyces spoilage risks and optimise non-chemical control strategies.
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BibTeX
@article{bodin2026impact,
title = {{Impact of stress combination on Brettanomyces bruxellensis growth and volatile phenol production}},
author = {Paul Bodin and Somaya Sachot and Aurelie Roland and Carole Camarasa and Fabienne Remize},
journal = {OENO One},
year = {2026},
month = oct,
doi = {10.20870/oeno-one.2026.60.4.10269},
url = {https://hal.inrae.fr/hal-05794209v1},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Impact of stress combination on Brettanomyces bruxellensis growth and volatile phenol production
AU - Paul Bodin
AU - Somaya Sachot
AU - Aurelie Roland
AU - Carole Camarasa
AU - Fabienne Remize
PY - 2026
DA - 2026/10/12
JO - OENO One
DO - 10.20870/oeno-one.2026.60.4.10269
UR - https://hal.inrae.fr/hal-05794209v1
AB - Brettanomyces bruxellensis is a major wine spoilage yeast responsible for the production of volatile phenols, including 4-ethylphenol (EP) and 4-ethylguaiacol (EG), from grape-derived hydroxycinnamic acids. This study investigated how oenological stress factors, particularly temperature, pH, and p-coumaric acid concentration, modulate B. bruxellensis growth, viability, p-coumaric acid consumption, and conversion into 4-vinylphenol (VP) and EP. Three B. bruxellensis strains, 2OT13_02, L0463 and 1961-Mx-M1 E2, representing distinct genetic backgrounds, were grown in synthetic wine at two temperatures with varying p-coumaric acid concentrations. Complementarily, strain 2OT13_02 was exposed to pH 3.3 and pH 5.6 across a large range of p-coumaric acid concentrations. Cell counts and viability were monitored by flow cytometry, while LC-UV-MS/MS quantified p-coumaric acid, VP and EP. Elevated temperature strongly impaired growth and viability in a strain-dependent manner. At 28 °C, 1961-Mx-M1 E2 showed near-zero growth and mortality close to 100 %, whereas 2OT13_02 maintained the highest metabolic activity. At 20 °C, all strains consumed almost all p-coumaric acid, but their conversion profiles differed, with 2OT13_02 and 1961-Mx-M1 E2 producing mainly EP and L0463 accumulating both VP and EP. Increasing temperature reduced EP formation and favoured VP accumulation, indicating a limitation of VP reduction under stress. Wine pH also reshaped the pathway: pH 5.6 reduced p-coumaric acid consumption but prevented the metabolic block observed at high p-coumaric acid concentrations and pH 3.3. These results show that volatile phenol production by B. bruxellensis is not solely determined by precursor availability, but by the interaction between strain background, acid stress, and temperature. Taking into account these stress combinations may help winemakers better anticipate Brettanomyces spoilage risks and optimise non-chemical control strategies.
SN - 2494-1271
ER -