Various strains of Saccharomyces cerevisiae were isolated and characterized in an effort to screen for higher ethanol production strains from Daqu that is the starter culture of the Chinese liquor fermentation process, contains numerous enzymes and microbes. More than 200 colonies were isolated and characterized. A total of 67 yeast isolates were characterized as higher ethanol producers. The higher ethanol producers, 13-3, 13-10, 13-13 and 13-21, were selected out and prepared to protoplasts which were then mutagenized using UV-irradiation. The mutagenized protoplasts were further subjected to recursive protoplast fusions and the fusants were screened on selective medium containing ethanol of various concentrations. Furthermore, the production of ethanol was greatly increased by the genome shuffling. Many fusants showed higher ethanol yields (from 10.1% to 13.9%, v/v) than the original strain (9.5%, v/v). For two selected fusants, G13105 and G13110, showed the best fermentation characteristic. G13105 produced the highest level of ethanol, 13.9% (v/v) within 60 h fermentation, while G13110, 13.2% (v/v).