McDermid A S, McKee A S, Marsh P D
Bacterial Metabolism Research Laboratory, Public Health Laboratory Service Centre for Applied Microbiology & Research, Salisbury, Wiltshire, England.
Infect Immun. 1988 May;56(5):1096-100. doi: 10.1128/iai.56.5.1096-1100.1988.
Since the pH of the gingival crevice increases from below neutrality in health to above pH 8 in disease, we decided to investigate the effect of environmental pH on the growth and enzyme activity of Bacteroides gingivalis W50. Cells were grown in a chemostat under hemin-excess conditions over a range of pH values; stable growth was observed only between pH 6.7 and 8.3, with the maximum yields obtained between pH 7.0 and 8.0. The enzyme profile of cells varied markedly with pH. Enzymes with a specificity for gingival connective tissue (collagenase, hyaluronidase) were produced optimally at or below neutral pH, whereas trypsinlike activity increased with the growth pH and was maximal at pH 8.0. Chymotrypsinlike activity was generally low, although its activity was highest at the extremes of growth pH, i.e., at pH 6.7 and 8.3. Inhibitor studies provided evidence that the breakdown of collagen involved the concerted action of both a collagenase and the trypsinlike enzyme. The ratio of trypsin to collagenolytic activity rose from 1:1 during growth at neutral pH and below to almost 7:1 during growth at pH 8.3. Thus B. gingivalis appears to be uniquely adapted as a periodontopathic organism in that under environmental conditions likely to prevail during the initial stages of pocket development it produces maximally those enzymes with a tissue-damaging potential. Then, as the pH of the pocket rises during the host inflammatory response, the activity of the trypsinlike enzyme increases markedly, which may enable cells to inactivate key components of the host defenses such as immunoglobulins and complement.
由于龈沟的pH值在健康状态下从低于中性增加到疾病状态下高于pH 8,我们决定研究环境pH值对牙龈类杆菌W50生长和酶活性的影响。细胞在恒化器中于血红素过量条件下在一系列pH值范围内培养;仅在pH 6.7至8.3之间观察到稳定生长,在pH 7.0至8.0之间获得最大产量。细胞的酶谱随pH值变化显著。对牙龈结缔组织具有特异性的酶(胶原酶、透明质酸酶)在中性pH或低于中性pH时产生最佳,而类胰蛋白酶活性随生长pH值增加,在pH 8.0时最大。类糜蛋白酶活性通常较低,尽管其活性在生长pH值的极端情况下最高,即在pH 6.7和8.3时。抑制剂研究提供了证据,表明胶原蛋白的分解涉及胶原酶和类胰蛋白酶的协同作用。类胰蛋白酶与胶原olytic活性的比率在中性pH及以下生长时从1:1上升到在pH 8.3生长时几乎7:1。因此,牙龈类杆菌似乎独特地适应作为一种牙周病原微生物,因为在牙周袋发育初始阶段可能普遍存在的环境条件下,它最大限度地产生那些具有组织损伤潜力的酶。然后,随着宿主炎症反应期间牙周袋pH值升高,类胰蛋白酶的活性显著增加,这可能使细胞能够使宿主防御的关键成分如免疫球蛋白和补体失活。