Wine spoilage yeasts are one of the main issues in the winemaking industry, and the control of the Brettanomyces and Pichia genus is an important goal to reduce economic loses from undesired aromatic profiles. Previous studies have demonstrated that Candida intermedia LAMAP1790 produces antimicrobial peptides of molecular mass under 10 kDa with fungicide activity against Brettanomyces bruxellensis, without affecting the yeast Saccharomyces cerevisiae. So far, it has not been determined whether these peptides show biocontroller effect in this yeast or other spoilage yeasts, such as Pichia guilliermondii. In this work, we determined that the exposure of B. bruxellensis to the low-mass peptides contained in the culture supernatant of C. intermedia LAMAP1790 produces a continuous rise of reactive oxygen species (ROS) in this yeast, without presenting a significant effect on membrane damage. These observations can give an approach to the antifungal mechanism. In addition, we described a fungicide activity of these peptides fraction against two strains of P. guilliermondii in a laboratory medium. However, carrying out assays on synthetic must, peptides must show an effect on the growth of B. bruxellensis. Moreover, these results can be considered as a start to develop new strategies for the biocontrol of spoilage yeast.
In the food industry, some fungi are considered to be common spoilage microorganisms which reduce the shelf life of products. To avoid this outcome, different technologies are being developed to control their growth. Electromagnetic fields (EMF) have been used to combat bacterial growth, but there are few studies on yeasts and their possible action mechanisms. For this reason, we studied the effect of EMF between 1 to 5.9 GHz bands on the growth of Saccharomyces cerevisiae yeast and observed that all the frequencies of the band used cause the reduction of the viability of this yeast. In addition, we observed that the distance between the antenna and the sample is an important factor to consider to control the growing yeast. By using transmission electron microscopy, we found that the EMF caused a loss of continuity of the yeast cell membrane. Therefore, EMF may be used as a control method for yeast growth.
Brettanomyces bruxellensis has been described as the principal spoilage yeast in the winemaking industry. To avoid its growth, wine is supplemented with SO2, which has been questioned due to its potential harm to health. For this reason, studies are being focused on searching for, ideally, natural new antifungals. On the other hand, it is known that in wine production there are a variety of microorganisms, such as yeasts and bacteria, that are possible biological controls. Thus, it has been described that some microorganisms produce antimicrobial peptides, which might control yeast and bacteria populations. Our laboratory has described the Candida intermedia LAMAP1790 strain as a natural producer of antimicrobial compounds against food spoilage microorganisms, as is B. bruxellensis, without affecting the growth of S. cerevisiae. We have demonstrated the proteinaceous nature of the antimicrobial compound and its low molecular mass (under 10 kDa). This is the first step to the possible use of C. intermedia as a selective bio-controller of the contaminant yeast in the winemaking industry.
Dekkera bruxellensis is a spoilage yeast in wine and fuel ethanol fermentations able to produce volatile phenols from hydroxycinnamic acids by the action of the enzymes cinnamate decarboxylase (CD) and vinyphenol reductase (VR) in wine. However, there is no information about this ability in the bioethanol industry. This work evaluated CD and VR activities and 4-ethylphenol production from p-coumaric acid by three strains of D. bruxellensis and PE-2, an industrial Saccharomyces cerevisiae strain. Single and multiple-cycle batch fermentations in molasses and sugarcane juice were carried out. D. bruxellensis strains showed similar CD activity but differences in VR activity. No production of 4-ethylphenol by S. cerevisiae in any fermentation system or media was observed. The concentrations of 4-ethylphenol peaked during active growth of D. bruxellensis in single-cycle fermentation but they were lower than in multiple-cycle fermentation. Higher concentrations were observed in molasses with molar conversion (p-coumaric acid to 4-ethylphenol) ranging from 45–85%. As the first report on 4-ethylphenol production in sugarcane musts by D. bruxellensis in industry-like conditions, it opens up a new avenue to investigate its effect on the viability and fermentative capacity of S. cerevisiae as well as to understand the interaction between the yeasts in the bioethanol industry.