Effects of 5′-phosphodiesterase and yeast on the formation of Lentinus edodes taste compounds were studied in this paper. 5′-phosphodiesterases and yeast powder were added in crushed homogenate liquid of mushroom Lentinus edodes, and 5′-nucleotide yield was detected. Results showed that the taste compounds contents of fresh Lentinus edodes were strengthened by 5′-phosphodiesterase and yeast. The strengthening condition was 5% yeast, 5% 5′-phosphodiesterase, pH 4.0, reaction temperature 60℃, reaction time 60min. 5′-nucleotide yield was 30.01mg/g under the optimization condition, which was triple as the control without 5′-phosphodiesterase.