By using cylinder-plate method， CGMCC 6624 was screened from the fresh Cupei of Shanxi vinegar. CGMCC 6624 had the most strong antibacterial ability. By the test of the effects of organic acids， hydrogen peroxide and proteinase digestion to the antibacterial property， analysized the relation on growth of CGMCC 6624 and production of antimicrobial protein， combinated the property that the antimicrobial protein could not inhibite the growed of CGMCC 6624， the antibacterial substance produced by CGMCC 6624 was determined， which including H2O2 and bacteriocine. CGMCC 6624 was identified as Bacillus subtilis by polyphasic taxonomy basing on morphological and physiological methods， 16S rDNA. Research showed CGMCC 6624 could grow in the condition of pH 2-11， temperature 30-50 ℃， alcohol 0%-8%. The optimal growth conditions of CGMCC 6624 was temperature 30 ℃ and pH 5.0. This study would supply the basis for the further utility on CGMCC 6624 and its bactericine and application in the production of Shanxi traditional brewing food.