This study investigates the preparation of a composite material by immobilizing Bacillus subtilis on biochar derived from chicken manure biogas residue for the removal of Cr(VI) from wastewater. The results demonstrated that the composite material (Bacillus subtilis immobilized biochar, BIB) achieved a maximum Cr(VI) removal efficiency of 94.1% in a 100 mg/L Cr(VI) solution within 4 h. The chicken manure-derived biochar not only served as an effective carrier for Bacillus subtilis but also enhanced the Cr(VI) removal efficiency through a synergistic effect with the microorganism. Functional groups such as phosphorus, carboxyl, and hydroxyl groups on the biochar surface played a key role in the sorption of Cr(VI). Bacillus subtilis primarily reduced Cr(VI) to Cr(III) by secreting cellular reductases. The combined action of biochar and Bacillus subtilis increased the Cr(VI) removal rate by 13.71% compared to biochar alone. This study presents a promising approach for Cr(VI) remediation in contaminated water and lays a theoretical foundation for the development of composite materials for Cr(VI) reduction.