Lactic acid bacteria (LAB) with inhibition activity against food-borne spoilage yeast were screened, characterized and identified by molecular biological method. Fifty-seven LAB strains were isolated from different food products such as pickles, pickled mustards, yoghourt and hams. Overlay method showed that six LAB strains represented inhibition activity against food-borne spoilage yeast such as Candida, Pichia, Saccharomyces, Rhodotorula and Yarrowia lipolytica. Among them, strain AT6 could inhibit the growth of all eleven spoilage yeast indicators. 96 well-plate method showed that the inhibition rate of supernatant AT6 reached above 73.04%. The supernatant AT6 was not sensitive to proteinase but lose the activity at higher pH value. The inhibition activity of AT6 supernatant against Canidia albicans was maximal after 30 h incubation at 30 ℃. The genome of AT6 was extracted and the gene of 16S rRNA was amplified by PCR. The PCR products were cloned into pMD 19-T T-vector and sequenced. The sequence was BLISA and strain AT6 was identified as Weissella cibaria.