During the processing of fermented beef, Pichia diba from Xinyang cured meat and Baryces hensonensis from Sichuan cured meat were used as the fermentation strains. In order to study the effects of fermentation with a single strain on the quality of fermented beef, the local special products and blank groups without starter were used as controls, the water content, water activity (aw), pH value, protein content, nitrosamine content, texture, sensory score and other characteristic indicators of fermented beef were analyzed. The results showed that the two types of fermented beef produced with a single strain, Pichia diba or Baryces hensonensis, had the moderate hardness, with their elasticity and chewiness being significantly higher than those of traditional products. There was no significant difference in overall acceptability compared with traditional cured meat, and the water content, pH, and nitrosamine content were significantly lower than those of traditional cured meat products. Comprehensive analysis reveals that both yeasts can be used as the starters for the production of fermented meat products to improve product quality and safety.