Fusobacterium nucleatum is a Gram-negative anaerobic rod-shaped bacterium frequently isolated from human dental plaque. It is capable of the desulfuration of cysteine and methionine, resulting in the formation of sulfide and thiol volatiles, respectively. Intact cells, as well as cell-free extracts produced by French pressure cell lysis of F. nucleatum, hydrolyzed radiolabeled cysteine to produce sulfide, pyruvic acid, and ammonia. The hydrolysis products of radiolabeled methionine were a volatile thiol, ketobutyrate, and ammonia. Both activities were associated with the cytoplasmic component, not the membrane. The desulfuration mechanisms are heat-labile, inhibited by the presence of excess substrate, and rates are dependent upon substrate concentration. These dissimilar pathways by F. nucleatum can account in part for the presence of sulfur-containing volatile products that occur in the mouth.