Chondroitin sulfatases in oral bacteria.

Abstract

A possible function of bacteria in the etiology of periodontal disease may involve production of mucopolysaccharidases. The gel phase of ground substance of connective tissue contains acid mucopolysaccharides including chondroitin and chondroitin sulfates A, B, and C. These substrates, especially chondroitin sulfates, may be attacked by mucopolysaccharidases. By attacking the intercellular cementing substances of tissue, these enzymes could be a factor in the inflammation and destruction of the gingiva; they could increase tissue permeability to toxic substances in the oral cavity or destroy connective tissue. Several investigators have demonstrated the presence of such enzymes among a small number of poorly classified bacterial strains isolated from the oral cavity (PEPPER ET AL, J Dent Res 37:623, 1958; SCHULTZ-HAUDT ET AL, J Dent Res 33:624, 1954; SCHULTZ-HAUDT ET AL, J Dent Res 35:299, 1956.) The purpose of this study was to investigate more carefully the correlation between oral bacteria implicated in the production of periodontal disease and their ability to produce chondroitin sulfatase. Twenty bacterial strains were chosen. Eighteen were considered representative of the human oral microflora. Of these, 13 (Streptococcus mutans, strains BHT, 6715, 134, GS-5, FA-1, 20, HS-6; S faecalis, strains 547, 539; S sanguis, strain ATCC 10556, Actinomyces viscous, strain ATCC 74187, and Actinomyces spp strains I, J) have been implicated in formation of dental plaque, induction of periodontal disease, or etiology of dental caries. Three oral staphylococci were included (Staphylococcus aureus, strains HL-1, HL-2 and Staph epidermidis). Two strains of Proteus vulgaris, strain ATCC 6896 which produces chondroitin sulfatase and strain ATCC 13315 which does not produce the enzyme, were chosen to verify the methods of enzyme preparation and assay. The other oral strains were Actinomyces naeslundii, ATCC 12104 and Rothia dentocariosa, ATCC 14191. All strains were grown in appropriate media of minimal carbohydrate content with 0.1% chondroitin sulfate added as an enzyme inducer. Cells were separated by centrifugation at 37,000 X g and washed in 0.1 M phosphate buffer (pH 7.2), then resuspended in buffer and broken by homogenization with plastic beads. The broken cells were separated from the beads by passing