pckA pckA ‐deficient Porphyromonas gingivalis Porphyromonas gingivalis W83 shows reduction in hemagglutination activity and alteration in the distribution of gingipain activity

摘要

Bacterial metabolism during infection is related to bacterial persistence and virulence factors. Porphyromonas gingivalis is a key pathogen that contributes to chronic periodontitis. Our previous study showed that pckA , the gene encoding phosphoenolpyruvate carboxykinase, is a putative‐specific pathogenic gene of virulent strains of P. gingivalis . Here, a pckA ‐deficient strain (Δ PG 1676) was constructed in P. gingivalis W83. Virulence properties were compared between the mutant and wild‐type strains. Specifically, hemagglutination activity was determined by the ability to agglutinate sheep erythrocytes. Gingipain activity was detected using synthetic‐specific substrates. Gene expression levels were analyzed using RT ‐q PCR , and cell surface‐associated polysaccharides were examined by silver staining and electron microscopy. Inactivation of the pckA gene did not affect bacterial growth and lipopolysaccharide formation but led to a reduction in hemagglutination activity and downregulation in expression of the hemagglutination‐associated gene, rfa , when compared with the wild‐type strain. Additionally, the Δ PG 1676 mutant exhibited an alteration in the distribution of gingipain activity. Increased gingipain activity was detected on the cell surface, but a decrease in its activity in the culture supernatant was shown. Taken together, our results suggest that the pckA gene plays a role in modulating the virulence of P. gingivalis W83.