본 연구는 한국전통식품인 김치에서 분리한 W. cibaria SPM402, L. paracasei SPM412의 포괄적인 항 치주염 효 과를 확인하였다. WC402 10 mg/mL농도에서 P. gingivalis 의 생물막 형성이 37.30±8.23%, LP412 10 mg/mL에서 51.36±5.95% 억제되었고, F. nucleatum의 생물막 형성의 경우 WC402 10 mg/mL에서 76.77±2.77%, LP412 10 mg/ mL에서 95.99± 0.73% 억제되었다. LP412 10 mg/mL에서 P. gingivalis 부착소인 fimA의 RQ값이 0.08±0.05로 약 12 배 감소함을 확인하였고, F. nucleatum의 부착소인 radD 의 RQ값은 0.08±0.008으로 radD는 거의 90배 이상 억제 되었다. 사람 잇몸 상피세포주인 YD-38에 Pg OMV에 의한 염증반응을 유도 후 WC402 15 mg/mL 처리 결과 IL-1β유 전자 발현이 약 150배 가량 억제되었고, LP412 0.1 mg/mL 처리 결과 IL-1β유전자 발현이 약 3.6배 가량 억제됨을 확 인하였다. YD-38세포주에 F. nucleatum에 의한 염증반응 을 유도 후 1 mg/mL의 WC402를 처리한 결과 IL-8유전자 발현이 약 3배 정도, 1 mg/mL의 LP412를 처리한 결과 IL- 8유전자 발현이 약 5.6배 정도 억제되었다. 이상의 결과를 볼 때 W. cibaria SPM402, L. paracasei SPM412는 구강 병원성 세균의 생물막 형성 관련 병인인자 발현 억제, 직 접적인 생물막 형성 억제 및 병원성 세균에 의해 유도된 염증반응을 효과적으로 억제하는 기능을 보유한 균주로 구강질환에 대한 치료제나 구강 건강에 도움이 되는 구 강 건강기능성 식품으로 사용될 가능성이 있음을 확인하 였다.
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.
A galactose fermentation bacterium producing lactose from red seaweed, which was known well to com-promise the galactose as main reducing sugar, was isolated from button mushroom bed in Buyeo-Gun, Chungchugnam-do province. The lactic acid bacteria MONGB-2 was identified as Lactobacillus paracasei subsp. tolerans by analysisof 16S rRNA gene sequence. When the production of lactic acid and acetic acid by L. paracasei MONGB-2 was inves-tigated by HPLC analysis with various carbohydrates, the strain MONGB-2 efficiently convert the glucose and galactoseto lactic acid with the yield of 18.86 g/L and 18.23 g/L, respectively and the ratio of lactic acid to total organic acidswas 1.0 and 0.91g/g for both substrates. However, in the case of acetic acid fermentation, other carbohydrates besidesgalactose and red seaweed hydrolysate could not be totally utilized as carbon sources for acetic acid production by thestrain. The lactic acid production from glucose and galactose in the fermentation time courses was gradually enhancedupto 60 h fermentation and the maximal concentration reached to be 16-18 g/L from both substrates after 48 h of fer-mentation. The initial concentration of glucose and galactose were completely consumed within 36 h of fermentation, ofwhich the growth of cell also was maximum level. In addition, the bioconversion of lactic acid from the red seaweedhydrolysate by L. paracasei MONGB-2 appeared to be about 20% levels of the initial substrates concentration and thisresults were entirely lower than those of galactose and glucose showed about 60% of conversion. The apparent resultsshowed that L. paracasei MONGB-2 could produce the lactic acid with glucose as well as galactose by the homof-ermentation through EMP pathway
본 연구는 여성의 손으로부터 분리한 두 유산균의 co-culture에 의한 향장 활성에 대한 것이다. Lactobacillus rhamnosus (L. rhamnosus)와 Lactobacillus paracasei (L. paracasei)를 함께 배양하는 것은 최초이며, co-culture를 이용한 유산균은 tyrosinase 저해율이 L. paracasei와 L. rhamnosus에 비해 20.68%로 가장 높은 수치를 보였다. 또한, co-culture 유산균은 melanin 생성 저해율에서도 63.5%의 가장 높은 활성을 보였으며, 주름개선활성에 영향을 미치는 MMP-1의 생성량이 3726.3 pg/mL로 확인되었다. 반면 단일 유산균 배양시 얻어지는 MMP-1 생성량은 각각 L. rhamnosus가 13613.5 pg/mL, L. paracasei는 13012.0 pg/mL이 확인되었다. Collagen 생성량을 확인한 결과 co-culture 유산균이 380.7 ng/mL로 323.4 ng/mL의 양이 확인된 L. paracasei와 304.1 ng/mL의 양이 확인된 L. rhamnosus보다 많은 양이 생성됨을 확인하였다. 상기의 결과는 유산균의 co-culture가 향장 활성 증진을 시킬 수 있다는 것을 나타내고, 이는 향장 활성에 영향을 미치는 항산화 활성을 측정한 결과 단일 유산균 배양을 통한 결과보다 co-culture 유산균의 항산화 활성이 더 높은 것으로 확인할 수 있었다.