A probiotic strain dlt7a was isolated from feces of the Shennongjia golden snubnosed monkey and was identified as Enterococcus faecalis.In order to improve the viable count of dlt7a in the fermentation broth,the fermentation technology of dlt7a was optimized.The optimal culture condition with 20% (50 mL/250 mL) loading volume,2% inoculation amount,and initial pH of 6.5,was obtained by single-factorexperiment.In the study,Plackett-Burman experiment and response surface experiment were designed to identify the main nutrients affecting the viable count of dlt7a in the fermentation broth and to optimize the content of the three main nutrients including sucrose,fish meal and KH2PO4.The optimum nutrients in the media consisted of 2.55% sucrose,2.63% fish meal,2.5% soya bean,0.4% yeast extract,1.0% CaCO3,0.1% KCl,0.1% MgSO4·7H2O and 0.014% KH2PO4.E. faecalis dlt7a were then cultured by the optimized fermentation medium and condition,and the viable count reached 58.83×108 cfu/mL,which was 46.53% higher than that not optimized.