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.

This study aimed to identify and characterize lactic acid bacteria strains with antimicrobial activity against Proteus mirabilis, a gram-negative bacterium associated with Parkinson’s disease, Crohn’s disease, and nosocomial infections. Among the 987 lactic acid bacteria strains isolated from various sources, strain no. CHK903 showed high antimicrobial activity against P. mirabilis. Phylogenetic tree analysis based on the 16S rRNA gene and scanning electron microscope analysis identified the selected strain as the rod-shaped Weissella cibaria. The culture supernatants of W. cibaria CHK903 showed antimicrobial activity against some pathogens. Two antimicrobial compounds with molecular weights 189 and 365 Da were partially purified from the culture supernatants of W. cibaria CHK903 using Biogel P2 gel permeation column chromatography. The culture supernatants of W. cibaria CHK903 also showed significant antibiofilm properties, inhibiting biofilm formation by 90% and removing pre-formed biofilms by 60%. These findings suggest the potential therapeutic use of W. cibaria CHK903 as a natural antimicrobial against P. mirabilis-related infections.

Although probiotics have been shown to improve health when consumed, recent studies have reported that they can cause unwanted side effects due to bacterial-human interactions. Therefore, the importance of prebiotics that can form beneficial microbiome in the gut has been emphasized. This study isolated and identified bacteria capable of producing biopoymer as a candidate prebiotic from traditional fermented foods. The isolated and identified strain was named WCYSK01 (Wissella sp. strain YSK01). The composition of the medium for culturing this strain was prepared by dissolving 3 g K2HPO4, 0.2 g MgSO4, 0.05 g CaCl2, 0.1 g NaCl in 1 L of distilled water. The LMBP(low molecular weight biopoymers) produced when fermentation was performed with sucrose and maltose as substrates were mainly consisted of DP3 (degree of polymer; isomaltotriose), DP4 (isomaltotetraose), DP5 (isomaltopentaose), and DP6 (isomaltoheptaose). The optimization of LMBP (low molecular weight of biopolymer) production was performed using the response surface methodology. The fermentation process temperature range of 18 to 32oC, the fermentation medium pH in the range of 5.1 to 7.9. The yield of LMBP production by the strain was found to be significantly affected by q fermentation temperature and pH. The optimal fermentation conditions were found at the normal point, and the production yield was more than 75% at pH 7.5 and temperature of 23oC.

Oligosaccharide-producing lactic acid bacteria were isolated from young radish and identified as Weissella cibaria YRK005. Oligosaccharides from W. cibaria YRK005 were purified using size-exclusion high-performance liquid chromatography, and their structure was determined. The oligosaccharides consisted of glucose only, 1 H-nuclear magnetic resonance analysis revealed that the oligosaccharides consisted of 93.8% α-1,6 glycosidic linkages and 6.2% α-1,4 glycosidic linkages, and molecular weight was determined to be 1.1210 2 Da. When the oligosaccharides' prebiotic effect on probiotic strains was examined, the oligosaccharides promoted the growth of four probiotic strains, Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactococcus lactis, and Lactobacillus pentosus, compared to fructo-oligosaccharides as a reference. These results indicated that oligosaccharides from W. cibaria YRK005 could be used as potential prebiotics.

The oral care probiotic strain Weissella cibaria CMU (oraCMU) inhibits volatile sulphur compounds associated with halitosis, presumably by inhibiting the growth of associated oral pathogens. In the present study, we investigated whether oraCMU inhibits the production of these compounds by suppressing the expression of mgl . This gene encodes L-methionine-α-deamino-γ-mercaptomethane-lyase (METase) and is involved in the production of methyl mercaptan (CH3SH) by Porphyromonas gingivalis . Therefore, we specifically investigated the effects of oraCMU on the growth, CH3SH production, METase activity, and mgl expression of P. gingivalis . The minimum inhibitory concentrations of cell-free supernatant and secreted proteins from oraCMU were 125 mg/mL and 800 µg/mL, respectively. At sub-minimum inhibitory concentration levels, these metabolites inhibited CH3SH production, but they also reduced P. gingivalis viability. Only heat-killed oraCMU decreased CH3SH production without affecting P. gingivalis viability. Heat-killed oraCMU also inhibited METase activity toward L-methionine and mgl mRNA expression (p < 0.05). In summary, we demonstrated the inhibition of volatile sulphur compounds via the antimicrobial action of oraCMU and, for the first time, the inhibition of such compounds by heat-killed oraCMU, which occurred at the molecular level.

The purpose of this study is to determine if natural extracts could be used as an additive in oral health food made with Weissella cibaria CMU (oraCMU). Natural extracts of green tea, mulberry leaf, licorice, and propolis, which are reported to have antimicrobial activities, were selected and used in this study. The minimum inhibitory concentrations (MIC) of extracts on periodontal pathogens such as Fusobacterium nucleatum and Porphyromonas gingivalis and their synergy effects with oraCMU by the fractional inhibitory concentrations methods were measured. From the results obtained, all the extracts showed no effect on the growth of oraCMU. Green tea extract showed the best antibacterial activity with MIC of 1.8 mg/ml against both F. nucleatum and P. gingivalis. In addition, green tea extract had a synergistic effect with oraCMU against F. nucleatum. Therefore, these results suggested that green tea extract is available as an additive in oral health food made with oraCMU.

The objective of this study is to assess immunomodulatory effects of mixed Weissella (W.) cibaria JW15 strain with water extract of black soybean (Glycine max) and burdock (Arctium lappa) on Listeria (L.) monocytogenes infection in mice. Female 7-9 week old BALB/c mice were given a daily dose of 1 × 109 CFU of viable JW15 and JW15 mixed with black soybean (BS) and burdock (BD) in 200 μL PBS for 2 weeks. The nomal control group (NC) and positive control group (PC) were given 200 μL PBS. After 2 weeks, mice were infected with L. monocytogenes (1.0 × 105 CFU/mouse) via the tail vein. The NC was injected with 100 μL PBS without L. monocytogenes. After 2 days, mice were euthanized and their body weights were determined. In addition, their livers and spleens were weighed, and serum were analyzed for cytokine (Interleukin-1β (IL-1β) and Tumor necrosis factor-α (TNF-α)) production. The survival rate was monitored using 5 mice in each group in the same way above until the mice died. Two days after infection with L. monocytogenes, mean spleen weight per body weight (g/kg) of JW15 (5.4 ± 0.88 g/kg), JW15 + BS (6.0 ± 0.64 g/kg), and JW15 + BD (5.3 ± 0.38 g/kg) group were significantly lower than that of the PC (6.8 ± 0.57 g/kg). The level of IL-1β in the serum of JW15 + BD (113.6 ± 31.03 pg/mL) was significantly higher than that of the JW15 (67.9 ± 15.15 pg/mL). Collectively, combination W. cibaria JW15 and water extract of BD and BD have ability to induce synergistic immunomodulative effects and are suitable for consideration as a functional food for humans and functional feed additives for companion animals.

Aging is a growing concern of age-associated aberrations in immune system or immunosenescence. Probiotics contain not only these general health advantages but also other the regulation of host immune function. Among those probiotics, the immunomodulatory effects of Weissella cibaria JW15 (JW15) strain has already been reported in our previous studies. The objective of this study is to assess improved immunity of JW15 strain in aged mice. In experiment, mice were divided into five groups. Twenty eight–month-old C57BL/6J mice were given a daily dose of 1×109 CFU/ mouse (JW15-H group) and 1×108 CFU/mouse (JW15- L group) of viable JW15. The young mice group (YM) and old mice group (OM) were given PBS. After four weeks of administration, mice were euthanized. Mice blood was collected and analyzed for complete blood cell count (CBC). Then, mice spleen was weighed and analyzed for splenocyte. Finally, we measured the concentration of cytokine secreted by splenocyte and serum. The results showed splenocyte proliferation of JW15-L group by the addition of LPS and concanavalin A was significantly higher than that of OM group. Splenocyte TNF-α production was significantly increased by JW15 intake. In particular, splenocyte TNF-α production of JW15-L group by the addition of LPS (5 μg/mL) was significantly greater than that other group. Splenocyte IL-6 production was also the highest in JW15-L group. Serum IFN-γ production of JW15-L (59.13 ± 16.13 pg/mL) group was significantly higher than that of OM group (39.71 ± 2.51 pg/ mL). When compared to the OM group (32.22 ± 1.92 pg/mL), Serum TNF-α production was significantly increased by JW15-H intake (69.01± 26.51 pg/mL). In conclusion, JW15 enhanced immunomodulative effects. In particular, the effect was significantly excellent in JW15-L group (1 × 108 CFU/mouse). Therefore, Weissella cibaria JW15 would be suitable for consideration as a helpful functional food for companion animals and humans.

The objective of this study was to investigate the immunomodulatory effects mixed with Weissella cibaria JW15 strain and black soybean (Glycine max (L.) Merr.). In this experiment, JW15 was cultured in De Man Rogosa and Sharpe (MRS) broth at 37% for 17 hr, and the cells were washed twice with sterile phosphate buffered saline (PBS) (pH 7.2). And black soybean was extracted by ethanol or hot boiling water. The immuno-modulatory effects of mixed JW15 and black soybean extract were investigated by measuring the production of nitric oxide (NO), nuclear factor κB (NF-κB) and cytokine (Interleukin-1β and Tumor necrosis factor-α) in RAW 264.7 macrophage cells or RAW blue cells. The 0.1 % ethanol and hot water extract of black soybean increased NO, NF-κB, and cytokine production in a concentration dependent manner. The NF-κB activation by JW15 mixed with 0.1 % hot water extract of black soybean (0.26 ± 0.02) was significantly higher than JW15 alone (0.20 ± 0.02). Also, combination of JW15 and 0.1% hot water extract of black soybean triggered IL-1β production of 110.19 ± 4.38 pg/mL, which was significantly greater than the JW15 alone (12.06 ± 7.58 pg/mL). The results of this study indicate that combination of Weissella cibaria JW15 and black soybean extract may have an ability to activate innate immune response synergistically. According to these results, the mixture of JW15 and black soybean extract could hold great promise for use in probiotics.

본 연구에서는 김치로부터 W. cibaria 0D17와 W. confuse 0D23를 생화학적 특성 분석과 16s rRNA 염기서열 분석을 통해 분리, 동정하였으며, W. cibaria 0D17와 W. confuse 0D23 배양 상등액이 L. monocytogenes에 대한 항균 효과가 있는 것으로 나타났다. 또한, 실시간 정량 PCR을 통해 W. cibaria 0D17 및 W. confusa 0D23가 L. monocytogenes 를 공동 배양했을 때의 생육억제 효과를 분석한 결과, 37℃에서는 생육 억제 효과가 있는 것으로 확인되었다. 따라서, 김치 유래 Weissella spp. 가 갖는 L. monocytogenes에 대한 항균 활성에 대한 기초 자료로 활용가능 할 것으로 판단되며, W. cibaria 0D17 및 W. confusa 0D23가 생산하는 bacteriocin 등의 항균 물질 특성에 대한 연구가 추가 적으로 진행될 필요가 있을 것이다.

In this study, characteristics and immuno-modulatory effects of Weissella cibaria JW15 isolated from Kimchi, traditional Korean fermented food, were examined for investigation of the capacity of potentially probiotic strains. We measured acid, bile, and heat tolerance, adhesive properties to intestinal epithelial cells, and inhibitory activity against pathogens. JW15 could survive at pH 3.0 for 2 hr, but not at pH 2.0. JW15 also showed tolerance to 0.3% oxgall bile salt, and heat tolerance at 70°C and 80°C for 5 min, respectively. Adhesive ability to Caco-2 cells was similar to that of Lactobacillus rhamnosus GG (LGG), a well-known commercial probiotic. JW15 exhibited antimicrobial activities to pathogenic bacteria such as Escherichia coli, Listeria monocytogenes, Staphylococcus aureus, and Salmonella enteritidis. The immuno-modulatory effects of JW15 were compared with those of LGG, a well-known immune enhancer. For analysis, production of nitric oxide (NO), NF-κB (Nuclear factor κB), Interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) was measured. The concentration of NO induced by JW15 was higher than that by LGG at low concentration (1 × 107 cfu/mL). Low and high (5 × 107 CFU/mL) concentration of JW15 induced statistically higher production of NF-κB, IL-1β, and TNF-α than that produced by LGG, respectively. In conclusion, Weissella cibaria JW15 had ability as a probiotic strain, including acid, bile, and heat tolerance, adhesive properties to intestinal epithelial cells, and inhibitory activity against pathogens. In addition, JW15 showed better immuno-modulatory effects than LGG when NO, NF-κB, IL-1β, and TNF-α were measured. According to these results, the characteristics and immunomodulating activity of Weissella cibaria JW15 are suitable for consideration as a potential probiotic.

A new lactic acid bacteria with gluten-degrading activity which was isolated from salted sea foods (traditional Korea fermented food), identified as Weissella confusa (99%) by use of API kit and 16S rRNA sequencing, and designated as W. confusa. When the W. confusa cultured for 48 hours at 30℃ in a MRS medium containing 1% gluten, 45% of gluten was founded to be degraded. W. confusa showed 85% of survival rate at pH 3, and 94% tolerance at 0.1% oxgall, which indicates that W. confusa would survive in stomach of human. Experiments on the thermostability was confirmed that it has a stability of 70% in 50℃. W. confusa inhibited the growth of some pathogen, except for S. aureus. Results in this study suggest that using W. confusa for fermentation of grain flour containing gluten would be desirable to prepare the gluten-free foods needed for those who suffer from celia disease and gluten allergy.

오디 유산균 발효음료를 제조하기 위해서 fructose 이용능이 우수한 유산균 JS1을 선발하였고, 16S rDNA 염기서열 분석 결과 Weissella cibaria와 99.8% 일치하여 W. cibariaJP0080이라고 명명하였다. 오디 착즙액은 40%, 30%, 20%,10%로 희석하였고 fructose로 보당하여 1%로 당 농도를 맞추었다. 4가지의 오디용액에 선별한 균주인 W. cibaria JP0080을 접종하여 생육특성을 측정하였다. 오디유산균 발효음료의 생육특성으로는 2시간대부터 대수증식기로 들어가6시간에 이르기까지 급격히 성장하다가 8시간 이후부터 정지기에 들어서고, 생균수는 2시간대부터 급격히 증가하는것으로 나타냈으며, 8시간에 2.21×1010~2.14×1011 CFU/mL로 조사된 것으로 보아 증식이 빠른 것을 알 수 있었다. pH는 발효 전 4.9에서 2시간대부터 급격히 내려가 6시간대에4.03, 12시간대에 3.59까지 내려갔으며 산도는 희석 배율에 따라서 다르지만 기존의 유산발효 음료의 보고보다 상위하는 결과를 나타내었다. 제조된 오디 유산균 발효음료의 소비자 조사 결과는 맛과 전체적인 평균점수에서 40%음료가 높은 점수를 보였고, 색상, 향기, 전반적인 수응도에서 30% 음료가 높은 점수를 보였으나 40~20% 음료간에는 유의적인 차이를 보이지 않았으므로, 향후 기호도면에서 뒤지지 않으면서 경제성 면에서도 우수한 20%의 오디유산균 발효음료의 개발 가능성을 엿볼 수 있었다. 이에 기호도와 경제면에서 우수한 것으로 나타난 20% 오디 유산균 발효 음료의 기능면에서의 우수성을 알아보고자 HPLC를 이용하여 기능성 물질을 분석하였다. 그 결과 오디착즙원액을 20%로 희석하여 12시간 발효한 각 50mL에 대하여 HPLC를 이용하여 기능성 물질을 분석한 결과 Isoquer-citrin 함량은 발효 후 0.75mg/L로 발효 전보다 함량이 감소하였으며, Quercetin은 0.89mg/L에서 1.05mg/L로 증가하였다. 이러한 결과를 미루어 볼 때 20% 오디즙을 희석하여 제조한 오디 유산균 발효음료가 가장 양호한 생육과기호도를 나타내었으며, 아울러 기능성 또한 발효 후에 감소하지 않았기 때문에 향후 오디 유산균 발효음료의 개발가능성을 확인할 수 있었다.