Vitis vinifera (variety Tempranillo) grapes were pressurized at 400 MPa by high hydrostatic pressure for 10 min and the effect on wild microbial populations, phenol extraction and wine composition was monitored. After treatment, the grapes were inoculated and fermented with Saccharomyces cerevisiae and several non-Saccharomyces yeasts: Schizosaccharomyces pombe, Torulaspora delbrueckii, Metschnikowia pulcherrima and Lachancea thermotolerans. S. pombe was used as sole fermentative yeast, but T. delbrueckii, M. pulcherrima and L. thermotolerans were used in sequential cultures with S. cerevisiae to completely ferment the sugars. The HHP treatment reduces strongly or eliminates wild microorganisms, especially yeasts, facilitating the growth and development of nonSaccharomyces yeasts. Thus, it helps to get either a better expression of enzymatic activities or metabolites production of nonSaccharomyces affecting wine quality.