Abstract: Yeast strain with local characteristics and naturally fermented pancake flavor were isolated and screened from pancake fermented batter,Y8 had rapid growth and strong acid resistance,which was identified by VITEC- 2 compact microbial identification system and prepared freeze- dried bacteria ferment whose protective agent was screened and identified. The results showed that yeast of fermented pancake batter was Saccharomyces cerevisiae,optimized formulation of cryoprotectant was glycerin 4.0%,trehalose 1.3%,skim milk12.7%. Under these conditions,the survival rate of Saccharomyces cerevisiae could reach 82%,the study provided support for continuous and industrial production of traditional foods.