We assessed the ability of a Pseudonocardia sp. from soil samples to bioconvert vitamin $D_3$. The optimal culture conditions for the bioconversion of vitamin $D_3$ to active $1{alpha}$,25-dihydroxyvitamin $D_3$ were investigated by varying the carbon and nitrogen sources, the metal salt concentrations, the initial pH, and the temperature. Microbial transformations were carried out with the addition of vitamin $D_3$ dissolved in ethanol. They were sampled by extraction with methanol-dichloromethane and the samples were examined by HPLC. Optimum culture conditions were found to be 0.4% yeast extract, 1% glucose, 3% starch, 1% fish meal, 0.2% NaCl, 0.01% $K_2HPO_4$, 0.2% $CaCO_3$, 0.01% NaF, and pH 7.0 at $28^{circ}C$. The optimal timing of the addition of vitamin $D_3$ for the production of calcitriol by Pseudonocardia autotrophica ID9302 was concurrent with the inoculation of seed culture broth. Maximum calcitriol productivity and the yield of bioconversion reached a value of 10.4mg/L and 10.4% respectively on the 7th day in a 75L fementer jar under the above conditions.