세균 배양액과 반응한 Lactobacillus plantarum의 생장속도 및 항균 활성에 대한 연구
Growth Rate and Antimicrobial Activity of Lactobacillus plantarum Reacting with Bacterial Culture Supernatant
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인간의 위장관은 매우 복잡한 장내균총으로 이루어져 있으며, 장내균총은 인간의 생리 기능에 있어 매우 중요한 역할을 담당하고 있다. 특히 유아기의 장내균총의 불균형은 면역연관 질병을 일으키는 요인이며 치료 목적으로 probiotics가 유용하게 활용되고 있다. Probiotics의 대표 균종은 L.plantarum으로서 영유아기에 병원균보다 가장 먼저 정착해야 면역 질환을 예방할 수 있다. 그러나 아직 Lactobacillus가 장의 우점종으로 정착하기 위한 환경적 요인과 항생물질 분비 조절에 대한 이해는 부족한 실정이다. 이에 본 연구는 세균의 배양액과 반응한 L. plantarum은 생장 속도와 항균활성이 증가되는 것을 관찰하였다. 본 연구는 균의 특성과 반응을 살펴보기 위해 시험 균을 M9 minimal media에 배양한 후 상층액을 감압건조 방법으로 농축하여 실험을 진행하였다. Colony forming unit assay 방법을 통해, 다양한 잠재적 병원성 세균에 대한 L. plantarum 의 생장억제 기능을 확인하였다. 세균을 L. plantarum에 처리하여 함께 배양한 후, L. plantarum의 생장속도를 흡광도(OD600)로 측정한 결과, 처리하지 않은 대조군(OD600= 0.48) 과 비교하여 생장속도(E. coli 처리 군: OD600 = 0.848, S.typhimurium 처리 군: OD600 = 0.848)가 증가되었다. 반면L. plantarum 농축액 자체는 L. plantarum의 생육증가를 유도하지 않았다. 이러한 결과는 Lactrobacillus species의 보편적 현상으로 관찰되었다. 뿐만 아니라 E. coli와 S.typhimurium의 배양액과 반응한 L. plantarum은 항균활성이 증가되었으며, disc diffusion assay를 통해 다양한 세균에 대한 생육저해결과가 관찰되었다. 마지막으로, L.plantarum의 생장속도와 항균활성을 증가시킨 세균 배양액의 특성을 분석해본 결과, 병원균은 분해효소(a-amylase, lipase, lysozyme, proteinase K), pH 2−8와 열(95℃)에 내성이 있음이 관찰되었으므로, 이 물질은 다양한 조건에서도 매우 안정한 small molecule일 가능성을 제시한다.
Abstract
In this study, L. plantarum, when reacting with the culture media of potential pathogenic bacteria, exhibited an increase in growth rate and antimicrobial activity. In order to examine the characteristics and the nature of the reaction with the bacteria, this study carried out experiments involving culturing the test bacteria in M9 minimal media. Subsequently, the supernatant was incrassated by the decompression-drying method. Through colony forming unit assay, it was confirmed that L. plantarum had the function of growth inhibition to various bacteria. After culturing L. plantarum with bacterial media, the growth rate of L. plantarum was measured by absorbance (OD600), the results showed that the growth rate (E. coli treatment group: OD600 = 0.848, S. typhimurium treatment group: OD600 = 0.848) increased, as compared with the non-treated control group (OD600 = 0.48). In contrast, the concentrate itself did not induce the growth of L. plantarum. These results were observed as a universal phenomenon of the Lactobacillus species. Moreover, the increase in antimicrobial activity was observed in L. plantarum, which reacted with the culture media of E. coli and S. typhimurium, through a disc diffusion assay, and the result of growth inhibition against various bacteria was induced. Finally, based on the analysis results of the characteristics of bacteria culture media, which increased the growth rate of L. plantarum and antibacterial activity, the bacterial media had a tolerance for catabolic enzymes, pH 2−8 and heat. Therefore, this substance can be said to be a small molecule which is highly stable under various conditions.
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