본 연구에서는 꽃송이버섯 건물과 powder를 각각 Yeast Extract broth와 MRS broth에서 Lactoballius 4종(L. plantarum, L. acidophilus, L. helveticus, and L. delbrueckii)과 함께 발효시켰다. 발효액의 final pH는 pH 3.9±0.02로 유산균의 일반적인 final pH 값과 일치함을 통해 유산균의 발효를 확인하였고, 유산균에 의한 꽃송이버섯 발효의 효과를 β-glucan 함량을 통하여 확인하였다. Yeast Extract broth 발효액은 YSB-2LP 1.65 g/100 g으로 β-glucan을 거의 함유하고 있지 않았지만 MRS broth 발효액은 Yeast Extract broth 발효액의 10배에 달하는 MPB-1LP 10.84 g/100 g을 함유하고 있었다. MPB-1LP는 꽃송이버섯 powder와 L.actobacillus plantarum으로 발효한 발효액으로 pH stress로 인해 만들어진 L. plantarum의 β-D-glucosidase에 의해 분해된 고분자량의 수용성 β-glucan이라 사료된다. 꽃송이버섯을 powder 상태로 L. plantarum과 함께 발효시켰을 때 꽃송이버섯의 수용성 β-glucan 추출 효율을 높일 수 있으며, 본 연구의 결과는 꽃송이버섯 유산균 발효물의 면역촉진 작용 연구를 위한 기초연구자료로 활용될 수 있을 것이다.

According to the previous study, Lactobacillus pentosus A67 and L. plantarum subsp. plantarum C2 were isolated and screened. In this study, culture broths of strains A67 and C2 were performed various physiological activity assays including antimicrobial, antioxidant, cytotoxicity, cell proliferation activity. The remarkable anti-microbial activities using the culture broths of the two strains were showed by disc diffusion method, MIC tests. In the 2,2-diphenyl-1-picryl hydrazyl (DPPH) and superoxide dismutase (SOD) assays to verify antioxidant activity, culture broths of strains A67 and C2 significantly scavenged free radical maximum by 91.33% and 81.95% respectively compared with arbutin. Also, they showed equivalent activity same as 297.22 units and 255.56 units of SOD respectively. In RAW264.7 cells, culture broths of strain A67 and C2 showed no cytotoxicity. Moreover, they showed significant cell proliferative effects by 173.5% and 143.5% compared to control group respectively. Overall, this study showed that the culture broths of strain A67 and C2 had antimicrobial and antioxidant activities, which could synergically increase the physiological activities. And they had no cytotoxicity and cell proliferative effects. Therefore, it is expected that L. pentosus A67 and L. plantarum C2 could be applied to a wide variety of functional materials including food and medicine as new probiotics.

Lactobacillus pentosus A67 and L. planarum subsp. plantarum C2 was tested the probiotics characterization such as acid, bile acid and antibiotics resistance. Isolates were isolated from the traditional Korean food, collected from traditional markets and homes in Seoul and Gyeonggi Province, and were selected the isolates A67 and C2, which were the most antibacterial to E. coli. According to the analysis of 16S rRNA base sequences, the results showed that L. pentosus ATCC 8041 (similarity 100%), and L. plantarum subsp. plantarum ATCC 14917 (similarity 100%). L. pentosus A67 showed a survival rate of about 98.35% and the L. plantarum C2 showed a survival rate of about 95.62% at pH 2.5, respectively. L. pentosus A67 showed a survival rate of about 94.56% and the L. plantarum C2 showed a survival rate of about 88.81% on the 0.3% bile salt, respectively. L. pentosus A67 and L. plantarum C2 were survived by antibiotics such as the quinolone, aminoglycoside, macrolide, glycopeptide, tetracycline, cephalosporin and phenicillin families. On basis of these results, L. pentosus A67 and L. plantarum C2 are the candidates for useful probiotics.