2,3-Butanediol (2,3-BD), especially when it exists in the chirally pure form, is a valuable chemical feedstock with numerous applications in various industries. However, in most cases, the 2,3-BD naturally produced by the microorganism is a mixture of two of the three isomers (d-2,3-BD, l-2,3-BD, and meso-2,3-BD), which has restricted applications and thus a much lower value than the chirally pure one. In this study, we report a new isolate Bacillus spp. FJ-4 strain, which can produce highly chirally pure d-2,3-BD (>99.9%) to high levels, with very high productivity and yield. The composition of the fermentation medium for the strain was sequentially optimized using statistical experimental methodologies. During a fed-batch experiment, 100.0 g/L of d-2,3-BD was produced from 226.8 g/L glucose, with a yield of 0.44 g/g (88.2% of the theoretical maximum). Acetoin was the primary byproduct, and no other byproduct such as lactate, acetate, or ethanol was detected, which would be advantageous for the downstream purification process. Moreover, no special conditions such as limited oxygen level (which generally limits the cell growth and thus final d-2,3-BD production) are required for the fermentation. These results indicated that Bacillus spp. FJ-4 is a robust workhorse for efficient d-2,3-BD production from low-value carbon sources.