An efficient ester-producing yeast (numbered as Y5) was screened from three Xiaoqu, and it was identified asWickerhamomyces anomalusby physiological biochemical tests and molecular biology method. The ester-producing test was conducted on the strain by using culture-medium, and there was obtained ethyl acetate 2.481g/L form the total of ester 2.684g/L. Though orthogonal experiments to optimize the ester-production conditions, its results showed that the appropriate culture conditions for this yeast was addition of PH value 4, alcohol 4%, fermentation temperature 30 °C and time 3d. Under this condition, comparing with the not optimized, the total ester 4.065g/L was significantly increased about 51.5% and the ethyl acetate3.677g/L was significantly increased about 48.2%. The strain has strong ability to produce ester and there is a bright prospect of applying on industry.