Fiehn N E
Scand J Dent Res. 1986 Apr;94(2):132-40. doi: 10.1111/j.1600-0722.1986.tb01377.x.
The purpose of the present investigation was to detect strains of small-sized oral spirochetes isolated from subgingival plaque for protease, peptidase, lipase, glycosidase, phosphatase, hyaluronidase and chondroitinsulfatase activities. The analyses were routinely carried out with cultures in the early stationary phase of growth after 4 days incubation. Both culture media and harvested spirochete cells were examined for the different enzyme activities. The enzymes were assayed by use of the API ZYM system, by p-nitroanilide derivatized peptides, and by hydrolyzing of mucopolysaccharides incorporated in solid bacterial medium. Relatively strong activities of trypsin-like enzymes, mainly bound to the cells, were observed in all strains. Similarly all strains showed acid phosphatases bound to the cells, too. Extracellular hyaluronidase- and chondroitinsulfatase activities were detected qualitatively in all strains after 7 days growth. The activities of the two mucopolysaccharide degrading enzymes almost disappeared after 10 subcultivations. Weak lipase (butyrate), higher lipase (caprylate), and weak phosphoamidase activities were observed in all cell pellets. No glycosidase activities were found. The observations are discussed by regarding the spirochetal enzymes as potential virulence factors for the development of marginal periodontitis.
本研究的目的是检测从龈下菌斑分离出的小型口腔螺旋体菌株的蛋白酶、肽酶、脂肪酶、糖苷酶、磷酸酶、透明质酸酶和硫酸软骨素酶活性。分析在培养4天后处于生长早期稳定期的培养物中常规进行。对培养基和收获的螺旋体细胞都进行了不同酶活性的检测。通过使用API ZYM系统、对硝基苯胺衍生肽以及水解掺入固体细菌培养基中的粘多糖来测定酶活性。在所有菌株中均观察到相对较强的类胰蛋白酶活性，主要与细胞结合。同样，所有菌株也都显示出与细胞结合的酸性磷酸酶。在生长7天后，在所有菌株中定性检测到细胞外透明质酸酶和硫酸软骨素酶活性。经过10次传代培养后，这两种粘多糖降解酶的活性几乎消失。在所有细胞沉淀中均观察到弱脂肪酶（丁酸盐）、高级脂肪酶（辛酸盐）和弱磷酸酰胺酶活性。未发现糖苷酶活性。通过将螺旋体酶视为边缘性牙周炎发展的潜在毒力因子来讨论这些观察结果。
