Isolation of collagenase inhibitor from soil bacterium and its application on periodontal disease.

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pervious study ha shown that Bacteroides gingivalid produces collagenolytic enzyme which is qenerally believed one of perfiodontopathogens. This enzyme is strongly inhibited by the substance(s) prosuced by a strain of soil bacterium. The purpose of this study was to clatilify the characteristics of this enzyme inhibitor and to search, by the experiment using human gingival fibroblast cell culture system, whether this material can be applied for the prevention of periodontal disease.The soil bacterium used for the isolation source of the collagenolytic enzyme inhibitor was identified as one of streptomyces by its morphological and biological characteristics. The inhibitow ras isolated and purified by procedures including an ultrafiitration using minimodule membrane, gel filtration on Sephadex G-15, followed by reverse-phase high pressure liwquid chromatography. Through these procedures, four active fractions as the collagenolytic enzyme inhibitors were finally obrained and subjected to amino acis analyss. The inhibitors were of low molecular weight peptides and consisted of only one to two amino acids including undetermined ones. This substrance also inhibited protease(s) isolated from B. gingivalis, but did not the activity of trypsin from bovine pancreas. The inhibitors had the strong preventive effect on the cytopathogenecity of the collagenolytic enzyme from B. gingivalis Human gingival fibroblast (HGF) cells, which had been grown to confluence, were seberely damaged by the further incubation with the collagenolytic enzyme. However, the presence of the inhibitor isolated gfor this study prevented from morphological damages of HGF cells. These results suggest that this collagenolytic enzyme inhibitor isolated from Streptomyces maintains normal turmnover of gingival cells ans preventsfrom the developemtn of periodontal disease.