Characteristics of sulfur-based autotrophic denitrification in a semi-continuous type reactor and the kinetic parameters were studied. Enriched autotrophic denitrifying culture was used for the reactor operation. Biomass growth on sulfur particles and in the liquid medium was monitored using the DAPI staining method. From the result of ion concentration changes and the biomass growth, maximum specific growth rate, ${mu}_{max}$, and the half velocity constant, $K_M$, were estimated as $0.61;d^{-1}$ and 3.66 mg/L, respectively. Growth yield coefficient, Y values for electron acceptor and donor were found as 0.49 gVSS/g N and 0.16 gVSS/g S. The biomass showed specific denitrification rate, ranging 0.86-1.13 gN/g VSS-d. A half-order equation was found to best simulate the denitrification process in the packed bed reactor operated in the semi-continuous mode.