Objective: The aim of this study was to isolate and screen lactic acid bacteria with high stress resistance from homemade mulberry fruit wine， and conduct a series of identification and high-density culture studies on them. Methods: Firstly， a strain of lactic acid bacteria with excellent character was identified by the ability of acid resistance， salt tolerance and bile salt tolerance screening， and conduct physiological and biochemical identification as well as 16S rRNA sequencing on it. Then， single factor， Plackett-Burman test， and Box-Behnken test were used to optimize the medium composition and further increase the viable bacteria count of strain. Results: A strain of Weissella paramesenteroide demonstrating high stress resistance was isolated and characterized， and was assigned the designation MbWp142. The acid resistance (pH = 2) of MbWp142 was 24.4%， the salt resistance (10% NaCl) was 24.4%， and the bile salt resistance (0.9% bile salt) was 47.7%. The optimal culture conditions were determined to be 1.5 g/100 g glucose， 0.20 g/100 g ascorbic acid and 0.20 g/100 g manganese sulfate. Under the optimized conditions， the maximum number of viable bacteria in fermentation liquid could reach 1.803×1011 CFU/mL， which could reach 20 times of that before optimization (5.87×109 CFU/mL). Conclusion: In this study， excellent strains with high stress resistance were found， and the vitality and density of strains was significantly increased， which provided data support for the development of probiotics from fruit wine and the research of functional microecological preparations.