To develop a functional probiotic that inhibits gingipain, a major virulence factor of Porphyromonas gingivlais (P. gingivalis), we screened over 30 probiotic strains for their ability to inhibit gingipian activity. We investigated the inhibition of expression of gingipain genes kgp, rgpA, and rgpB as well as gingipain activity, using freeze dried cell-free supernatants of Weisiella cibaria SPM402 (WC402) and Lactobacillus paracasei SMP412 (LP412), both of which demonstrated antibacterial activity against P. gingivalis. Thus, it was verified that kgp expression was reduced by approximately 0.71±0.02 folds and rgpB expression was reduced by approximately 0.71±0.14 folds at a concentration of WC402 10 mg/mL. Meanwhile, at the same concentration of 10 mg/mL of LP412, kgp expression was reduced by approximately 0.19±0.08 folds, rgpA expression was reduced by approximately 0.09±0.02 folds, and rgpB expression was reduced by approximately 0.24±0.03 folds. At a concentration of 10 mg/mL, Kgp activity was inhibited by approximately 78.65±3.58% (cell associated gingipain, CAG), 82.45±1.22% (cell-free gingipain, CFG) by WC402 and 80.71±2.11% (CAG), and 85.81±0.05% (CFG) by LP412 respectively. Rgp activity was also effectively inhibited by approximately 78.6±1.01% (CAG), 86.78±0.47% (CFG) and 82.93±1.26% (CAG), 88.81±0.36% (CFG) by WC402 and LP412 respectively. Based on these results, W. cibaria SPM402 and L. paracasei SPM412 can be regarded as functional probiotics with the ability to inhibit gingipain activity and exhibit antibacterial effects against P. gingivalis.

In this study, we have investigated to find optimal pre-treatment flocculation condition by analyzing the floc growth rate with mixing conditions and the membrane permeation flux for pre-treatment step of the membrane process. The higher mixing intensity showed a constant floc size index (FSI) values, and lower mixing intensity increased the degree of dispersion of the FSI values. Results of comparing the distribution characteristics of the FSI value and the permeation flux were more effective in increasing flux when the FSI values were 0.2 or higher. The degree of dispersion of FSI was relatively large in 40 rpm mixing condition compared to 120 rpm. In 40 rpm mixing condition, it decreased the permeation flux compared to 120 rpm because various sizes of flocs were distributed. Coagulation-UF membrane process enhanced 30%∼40% of the flux rate compare to UF alone process, and the coagulation-MF process increased up to 5% of the flux rate compare to MF alone process. Pre-treatment, that is, coagulation process, has been found to be less effects on relatively larger pore size for MF membrane. For UF membrane, the flux was a little bit same when applying only the rapid mixing process or rapid mixing with slow mixing processes together. In case of MF membrane, the flux was improved when rapid mixing process applied with slow mixing process together.