Abstract: In search for a glycosidase that can enhance flavor in fruit wine. Four strains of β-D-glucosidase were screened from 63 strains of Lactobacillus with Esculin color-developing plate method. The p-nitro-phenyl-β-D-glucopyranoside(PNPG)were used as a substrate and method of differential centrifugation graded precipitation was utilized to the initial location of β-D-glucosidase.In addition,traditional identification methods,such as morphology,physiological and biochemical tests,and sequence analysis of 16S rDNA were used to identify the four strains. The result indicated that the β-D-glucosidases generated by four strains were all endoenzyme;FL12 and Hsb were Lactobacillus plantarum,and CM3 was Lactobacillus pentosus and T61 was Lactobacillus brevis.