Most strains with capability of producing 2,3-butanediol (2,3-BDO) from glucose were isolated from different natural environment samples. The Strain 6-7 with high capability of producing 2,3-BDO was screened from the isolated strains and the production of 2,3-BDO reached 49.6 g/L. In addition to general morphological and biochemical characteristics, the strain 6-7 was identified by 16S rDNA sequence and systematic analysis. The results showed that 16S rDNA sequence of strain 6-7 had similarity of 99% with Bacillus subtilis strain BIHB332, suggesting that the strain 6-7 is one of Bacillus subtilis species. It is an environmentally friendly strain and Bacillus subtilis is generally regarded as safe microorganism used for industrial production. This strain showed the potential application in the future.