Genetic Improvement of <i>Torulaspora delbrueckii</i> for Wine Fermentation: Eliminating Recessive Growth-Retarding Alleles and Obtaining New Mutants Resistant to SO₂, Ethanol, and High CO₂ Pressure

Abstract

The use of Torulaspora delbrueckii has been repeatedly proposed to improve a wine’s organoleptic quality. This yeast has lower efficiency in completing wine fermentation than Saccharomyces cerevisiae since it has less fermentation capability and greater sensitivity to SO2, ethanol, and CO2 pressure. Therefore, the completion of fermentation is not guaranteed when must or wine is single-inoculated with T. delbrueckii. To solve this problem, new strains of T. delbrueckii with enhanced resistance to winemaking conditions were obtained. A genetic study of four wine T. delbrueckii strains was carried out. Spore clones free of possible recessive growth-retarding alleles were obtained from these yeasts. These spore clones were used to successively isolate mutants resistant to SO2, then those resistant to ethanol, and finally those resistant to high CO2 pressure. Most of these mutants showed better capability for base wine fermentation than the parental strain, and some of them approached the fermentation capability of S. cerevisiae. The genetic stability of the new mutants was good enough to be used in industrial-level production in commercial wineries. Moreover, their ability to ferment sparkling wine could be further improved by the continuous addition of oxygen in the culture adaptation stage prior to base wine inoculation.

Keywords

• <i>Torulaspora delbrueckii</i>
• wine fermentation
• spore clone
• sparkling wine
• ethanol resistance
• SO₂ resistance