title: Characteristics and cariogenicity of a fructanase-defective Streptococcus mutants strain.
abstract: Polymers of D-fructose produced by a variety of oral bacteria are believed to function as extracellular carbohydrate reserves. Degradation of these polysaccharides in plaque following exhaustion of dietary carbohydrates is thought to contribute to the extent and duration of the acid challenge to the tooth surface and thus to the initiation and progression of dental caries. Streptococcus mutans produces a fructanase, the product of the fruA gene, which is capable of degrading beta(2,6)- and beta(2,1)-linked fructans that are commonly synthesized by dental plaque microorganisms. To evaluate the role of the FruA protein in exopolysaccharide metabolism and to assess the contribution of this enzyme to the pathogenic potential of S. mutans, a fructanase-deficient strain of S. mutans was constructed. Inactivation of a cloned fruA gene was accomplished in Escherichia coli by using a mini-Mu dE transposon, and then an isogenic mutant of S. mutans UA159 was constructed by allelic exchange. Successful inactivation of fruA was confirmed through the use of biochemical assays, Western blotting (immunoblotting) with anti-recombinant FruA antisera, and Southern hybridization. The data indicated that FruA was the only fructan hydrolase produced by S. mutans UA159. Inactivation of fruA had no significant effects on glucosyltransferase or fructosyltransferase activity. In the rat caries model using animals fed a high-sucrose diet and ad libitum, there were no significant differences in the number or severity of smooth surface, sulcal, or root caries elicited by the fruA mutant and the wild-type organism.
authors.affiliation: Department of Dental Research, University of Rochester Medical Center, New York 14642.
authors.forename: D L
authors.initials: DL
authors.lastname: Wexler
authors.forename: J E
authors.initials: JE
authors.lastname: Penders
authors.forename: W H
authors.initials: WH
authors.lastname: Bowen
authors.forename: R A
authors.initials: RA
authors.lastname: Burne
mesh_terms: D000818:Animals; D001426:Bacterial Proteins; D003731:Dental Caries / Q000209:etiology*; D005260:Female; D005798:Genes, Bacterial; D006026:Glycoside Hydrolases / Q000172:deficiency* / Q000235:genetics / Q000502:physiology; D009154:Mutation; D051381:Rats; D011919:Rats, Inbred Strains; D013295:Streptococcus mutans / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity
chemical_list: D001426:Bacterial Proteins; D006026:Glycoside Hydrolases; C021512:levanase