Abstract: This study aims to optimize the zirconia beads wall-breaking method, screen out the better broken conditions for breaking lactic acid bacteria cell wall and improve the protein yield.The weight of bacteria, the grinding buffer and time were tested and selected as important aspects to optimize the wall-breaking method, and then, the breaking effect was analyzed and evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis ( SDS-PAGE) .Under optimized conditions, six different species of strains from lactic acid bacteria were tested for cell wall-breaking treatments with the zirconia beads method, after that, the breaking effects were analyzed and evaluated by SDS-PAGE.Results showed that, on condition that the ratio of bacteria, grinding buffer and zirconia beads ( W/V/W) is 0.013 g: 300 μL: 1 g, grinding time of 15 min, the protein pattern with clear banding, high abundance; RIPA, PBS, PENP, GUS as the grinding buffer, the protein pattern with clear banding, high abundance, while 8 mol/L urea as the grinding buffer, the protein pattern with obscure banding, low abundance.After Six strains of lactic acid bacteria were broken through this method, the protein pattern with clear banding, high abundance.Therefore, zirconia beads method is a combined rapid and efficient method for breaking lactic acid bacteria cell wall.