Natural kefir grains are rich in beneficial bacteria, and analysis of their microbial diversity is a necessary condition for developing and applying kefir grains. In this study, six commercially available natural kefir grains were used as raw materials to explore their microbial diversity by metagenomics. The results showed that there were 14794 genes in 6 kinds of natural kefir grains, and the number of unique genes of X1, X2, X3, X4, X5, X6 were 111, 11, 0, 1899, 552, 1, respectively. From the relative abundance table of boundary, phylum, class, order, family, genus and species, the microbial diversity at each level was analyzed.The two dominant genera at the genus level are