Supragingival human dental plaque was collected from patients with evidence of caries. The plaque was frozen and stored at -20 degrees C. Pooled plaque was homogenized in acetate buffer pH 5.0 in an ice-water bath. By incubating the homogenate at pH 5.0 with [U-14C]-sucrose the formation of glucose and fructose was followed. Incubation in acetate buffer at pH 5.0 eliminated the glycosyltransferase activities and the glycolytic pathway. Normal Michaelis-Menten kinetics were observed until about 40 mM sucrose. At higher concentration of sucrose, excess substrate inhibition occurred. Storage of the homogenate at -20 degrees C resulted in decrease of the invertase activity with time.