Screening of Indigenous Hanseniaspora Strains from China for Ethanol Reduction in Wine

Non-<i>Saccharomyces</i> yeasts have the potential to ameliorate wine ethanol levels, but such fit-for-purpose yeast strains are still lacking. Seventy-one indigenous non-<i>Saccharomyces</i> yeasts isolated from spontaneous fermentations of four wine regions in China (Ningxia, Xinjiang, Gansu, and Shaanxi) were screened for ethanol formation and were characterized for major metabolite profiles in synthetic grape juice fermentation to obtain non-<i>Saccharomyces</i> yeasts with low ethanol yields. Four <i>Hanseniaspora</i> strains with less volatile acidity production were primarily selected, and their ethanol yield was reduced by 22-32% compared to <i>S. cerevisiae</i>. These strains were further evaluated for oenological properties, namely ethanol and temperature tolerance, H₂S production, and killer activities against <i>S. cerevisiae</i>. Strain HuC-3-2 was then subjected to Atmospheric Room Temperature Plasma (ARTP) mutagenesis, and a mutant (HuC32-2-72) with rapid growth and optimized ethanol-reducing capability was obtained. The best-performing strains were further characterized in sequential fermentations with <i>S. cerevisiae</i> in Merlot juice, and resulted in a 1.4% <i>v</i>/<i>v</i> decrease in ethanol yield. Comprehensive analysis of yeast populations and the production of key metabolites highlighted important carbon sinks, as well as glycerol formation, partially accounting for the ethanol reduction. In addition to ethanol amelioration, the <i>Hanseniaspora</i> strains also led to alterations in many metabolites, including volatile compounds and some organic acids, which can further modulate wine aroma and flavor.