|More than hundreds of private wineries have been established since the Taiwan Tobacco and Alcohol Administration Act was enacted on April 19, 2000. It is then essential to screen local wine yeast in order to develop quality wine with unique flavor. Black Queen grape is a main ingredient for red wine brewing in Taiwan. In this study, ethanol productivity of years isolated from spontaneous fermentation of Black Queen grape must (17 and 30 °Brix) was determined. Seven yeasts identified as Saccharomyces cerevisiae by Biolog System with higher ethanol productivity were individually inoculated into Black Queen grape must to undergo fermentation at 25℃ for 14 days. The ethanol contents and volatile acidity in wine were above 15% (v/v) and below 0.7 g/L, respectively. Compared to commercial wine yeasts A, B and C, HP01 fermented red wine contained the highest total ester content, color intensity, taste grade, and overall preference value. On the other hand, HS13 fermented red wine contained the highest ethanol, higher alcohols, fatty acids and phenolic compounds content. The mtDNA restriction fragments profiles of these two wine yeast strains were different from those of other yeast strains. Due to their superiority in oenological characteristics, HP01 and HS13 can be applied in Black Queen red wine brewing to produce red wine with unique flavor.