L.plantarumFCJX 102 andL. acidophiluFCJX 104 producing acetaldehyde dehydrogenase were selected from five kinds of lactic acid bacteria including Lactobacillus brevis, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus acidophilus and Lactobacillus rhamnosus. On the basis of single factor experiment, the fermentation conditions of hawthorn puerarin juice producing aldehyde dehydrogenase were studied by single factor and orthogonal optimization, hawthorn pueraria ratio, strain ratio and fermentation time as impact factors. Experimental results showed that when the rate of hawthorn and puerarinin the medium was 1:2 and the amount ratio ofL. plantarumFCJX 102 andL. acidophilu FCJX104 was 2:1, and cultured at 37 for 16,the highest enzyme activity of acetaldehyde dehydrogenase was obtained after optimization was 16.083U / g.