title: The activity of glucosyltransferase adsorbed onto saliva-coated hydroxyapatite.
abstract: This study aimed to determine physical and kinetic properties of glucosyltransferase (GTF) adsorbed onto hydroxyapatite (HA) surfaces. For development of a solid-phase enzyme assay, 4.0-mg samples of washed HA powder were exposed to centrifuged whole saliva (WSHA) or buffer, and subsequently exposed to a GTF solution. The activities of GTF adsorbed to HA and that remaining in solution were measured. WSHA was more effective in adsorbing GTF than was naked HA. Enzyme activity on the surface of WSHA was enhanced; more activity was detected on WSHA than was apparently removed from solution. A similar effect was observed when GTF was adsorbed to naked HA from a mixture with lysozyme or saliva; however, no enhancement was seen when GTF was adsorbed from a mixture with albumin. Compared with GTF in solution, adsorbed GTF displayed activity over a much wider range of pH values. Temperature-activity profiles indicated that GTF adsorbed to surfaces had a lower temperature optimum (40 degrees C) than did soluble enzyme (45 degrees C), and that the bound enzyme was more resistant to adverse effects of heat at elevated temperatures. The majority of glucan made by GTF adsorbed to parotid saliva-coated HA remained attached to the surface. The activity of lysozyme adsorbed to HA was reduced by adsorption of GTF to the same surface and was almost completely abolished by formation of glucans by the adsorbed GTF. These results suggest that soluble bacterial enzymes found in saliva can be incorporated into pellicle, interact with host-derived molecules on the surfaces of teeth, express enzymatic activity, and potentially influence the biological properties of pellicle.
authors.affiliation: Department of Dental Research, University of Rochester, School of Medicine and Dentistry, New York 14642, USA.
authors.forename: K M
authors.initials: KM
authors.lastname: Schilling
authors.forename: W H
authors.initials: WH
authors.lastname: Bowen
mesh_terms: D000327:Adsorption; D001426:Bacterial Proteins / Q000378:metabolism; D003741:Dental Deposits / Q000201:enzymology* / Q000382:microbiology*; D044622:Dental Pellicle; D017886:Durapatite / Q000737:chemistry*; D005936:Glucans / Q000096:biosynthesis; D005964:Glucosyltransferases / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009113:Muramidase / Q000378:metabolism; D012463:Saliva / Q000201:enzymology* / Q000382:microbiology; D013295:Streptococcus mutans / Q000201:enzymology
chemical_list: D001426:Bacterial Proteins; D005936:Glucans; D017886:Durapatite; D005964:Glucosyltransferases; D009113:Muramidase