식품부패 및 식중독 관련 미생물에 대한 모자반 추출물의 항균활성 = Antimicrobial Activity of Sargassum fulvellum Extract against Food Spoilage and Food Poisoning Microorganisms

다국어 초록 (Multilingual Abstract)

Antimicrobial activity of Sargassum fulvellum(SF) was investigated by disk diffusion method and MIC test, and inspected stability of processing characteristics through extreme heat and pHs. Additionally antimicrobial component was purified from SF ethanol extract. The results of antimicrobial test, SF ethanol extract showed great antimicrobial activity against B. subtilis, C. perfringens, L. plantarum, S. aureus, L. monocytogenes, S. cerevisiae and C. tropicalis by disk diffusion method at level of 4 mg/ml. On the other hand, water extract showed no effect. The MIC value of SF ethanol extract ranged from 0.05 to 0.0063% against test microbes. But SF ethanol extract showed no effect against gram negative bacteria and mold within 0.3% concentration. At the heat and pH stability test, antimicrobial activity of SF ethanol extract was not destroyed by the heating at 121？？ for 15 min, and not affected by pH 2~10. These results suggest that SF ethanol extract was greatly considered the stable substance under the change of heat and pH. Purification was carried out by liquid and liquid extraction, silica gel column chromatography, and HPLC. First, the SF ethanol extract was separated by liquid and liquid extraction, and five fractions were obtained, hexane, chloroform, ethyl acetate, butanol, and water. As a result of MIC test for each obtained fractions, the chloroform fraction showed the strongest level of antimicrobial activity. The chloroform fraction, which exhibited the antimicrobial activity at the level of 0.05~0.003%, was separated using silica gel column chromatography, and 16 of sub-fractions were obtained. Among them, CH4, which showed the highest activity, was used to perform HPLC for isolated and purification of a single substance. The fraction was separated through a ？？Bondapak ․ C18(125 ？？, 10 ？？m) column with eluted 65% THF and 3 main peaks were obtained. the antimicrobial activity of each subfraction were measured by disk diffusion method and MIC test. Among them, peak 1 and 2 showed the highest effect against S. aureus, B. subtilis, L. monocytogenes and L. Inoccua by disk diffusion method at level of 4 mg/ml. As a result of MIC test for peak 1 and 2, the peak 1 showed the strongest level of antimicrobial activity. The results confirm the possible use of Sargassum fulvellum as natural antimicrobial agents to control food spoilage and food poisoning microorganisms in food industries.

목차 (Table of Contents)

• 서 론 4
• 재료 및 방법 9
• 1. 실험 재료 9
• 1-1. 모자반 채집 9
• 1-2. 배지 9
• 1-3. 시험 균주 9
• 2. 시료 추출 10
• 3. 항균 활성
• 3-1. Disk diffusion method 12
• 3-2. Minimum Inhibitory Concentration(MIC) 측정 12
• 4. 열 및 pH 안정성
• 4-1. 열 안정성 13
• 4-2. pH 안정성 13
• 5. 모자반 유래의 항균물질 분리 및 정제
• 5-1. 액액 분배법에 의한 모자반 추출물의 분획 13
• 5-2. Silica gel column chromatography에 의한 모자반 chloroform 추출물의 분획 14
• 5-3. HPLC에 의한 항균물질 분리 14
• 결과 및 고찰 18
• 1. 모자반 물 및 에탄올 추출물의 항균활성 18
• 1-1. Disk diffusion method에 의한 항균활성 18
• 1-2. 모자반 에탄올 추출물의 최소저해농도 22
• 2. 모자반 에탄올 추출물의 열 및 pH 안정성 25
• 3. 모자반 유래의 항균물질 분리 및 정제 29
• 3-1. 모자반 분획물의 항균효과. 29
• 3-2. 모자반 chloroform 분획물의 silica gel column chromatography 항균효과 34
• 3-3. HPLC에 의해 분리된 획분의 항균효과 41
• 요 약 46