Three strains, which showed obvious antibacterial activity, were isolated from lactic acid bacteria, including Lactobacillus.Acidophilus (A), Pediococcus pentosaceus(M)and Pediococcus pentosaceus(T)by the agar diffusion cup-plate method.The supernatants after eliminated organic acid and hydrogen peroxide still retained bacteriostatic activity.The effect of inhibition decreased after treatment with protease K, The above results indicated that the antibacterial substance was a kind of protein.The inhibitory spectrum from the above supernatants was broad.The bacteriocin produced by three strains was suitable for the biotype preservatives.