We report that catabolism of l-lactate in Bacillus subtilis depends on the previously uncharacterized yvfV-yvfW-yvbY (herein renamed lutABC) operon, which is inferred to encode three iron-sulfur-containing proteins. The operon is under the dual control of a GntR-type repressor (LutR, formerly YvfI) and the master regulator for biofilm formation SinR and is induced during growth in response to l-lactate. Operons with high similarity to lutABC are present in the genomes of a variety of gram-positive and gram-negative bacteria, raising the possibility that LutABC is a widely conserved and previously unrecognized pathway for the utilization of l-lactate or related metabolites.