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The study of lactic acid bacteria with independent intellectual property resource library, and on this basis, screening and development for our industry, the dairy industry and the development of lactobacillus probiotic lactic acid bacteria, starter cultures and probiotics is particularly necessary.Optimisation of this experimental test is to promote the growth of Lactobacillus acidophilus, a high density culture. Single factor experiments, Plackett-Burman experiment, Box-Behnken experiment and response surface analysis were used to find the levels of vitamin C, sorbitol, glutamic acid, leucine, arginine, lysine, methionine, xylose, Phe, Pro needed to promote a better factor of Lactobacillus acidophilus.The culture medium contains the basic nutrients for eosinophilic Lactobacillus, but it cannot reach the content of amino acid in cell growth because its protein hydrolysis ability is limited, so in order to improve the purity and activity of eosinophilic Lactobacillus, the culture medium requires added amino acids on demand. The predicted value of the Plackett-Burman experiment is 97.85%, while it can achieve 96.95%. The most important factor and the best composite bacteria culture medium included vitamin C, glutamic acid and lysine, sorbitol, leucine, arginine, methionine, phenylalanine, proline and xylose; the optimum enrichment medium ratio is glutamic acid 0.0276 g/L, lysine 0.0243 g/L, vitamin C 0.0308 g/L.The best ratio of enrichment medium was: glutamate, lysine, vitamin C. Culture temperature was 37°C, initial fermentation pH was 6.5, 4% inoculation, harvest time was 20 hours after fermentation.Supported by a project grant from Longdong University doctoral research (Grant No. XYBE1605). Central finance forestry science and technology extension demonstration project  ZYTG8 Qingyang science and technology support program (NK2011-41).