从越南庆化地区海洋生物样品中分离和筛选放线菌的抗菌活性

Le Thi Hong Minh, Nguyen Mai Anh, Vu Thi Quyen, Vu Thi Thu Huyen, Phi Thi Dao, Nguyen Thi Thuy Linh, Doan Thi Mai Huong, Pham Van Cuong
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引用次数: 0

摘要

海洋环境条件与陆地环境条件特别不同;近年来的研究表明,海洋放线菌可能产生许多具有良好生物活性的新化合物。本研究的目的是从海洋环境中分离和筛选具有抗病原微生物活性的放线菌菌株。从沉积物、海绵、软珊瑚、棘皮动物等40个样品中分离到50株放线菌。采自越南庆化省Van Phong Bay地区。在A+培养基和发酵液中发酵的菌株用乙酸乙酯提取5次。萃取物在减压下蒸发,得到粗萃取物。定量测定提取物对3株革兰氏阴性菌(大肠埃希菌ATCC25922、铜绿假单胞菌ATCC27853、肠沙门氏菌ATCC13076)、3株革兰氏阳性菌(粪肠球菌ATCC29212、金黄色葡萄球菌ATCC25923、蜡样芽孢杆菌ATCC 14579)和酵母菌白色念珠菌ATCC10231的最小抑菌浓度。筛选结果表明:18/50菌株对至少3种被试微生物具有抑菌活性。其中,菌株G650、G654、G657、G666、G675在MIC值为8 ~ 64µg/mL时抑制革兰氏阳性试验微生物,白色念珠菌ATCC10231在MIC值为2 ~ 16µg/mL时抑制革兰氏阳性试验微生物。此外,两株菌株G657和G666对铜绿假单胞菌ATCC27853和肠链球菌ATCC13076均有抑制作用,MIC值为128µg/mL。根据所选菌株的16S rRNA基因序列进行形态学和系统发育研究表明:菌株G650属于硬链霉菌属,菌株G666与硬链霉菌属高度相似;菌株G654与沙菌Salinispora arenicola的相似性最高;菌株G657与aurantiaca小单孢子菌相似性最高;G675属黄无心藓属。
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Isolation and screening antimicrobial activity of actinomycetes from marine organisms samples of the area Khanh Hoa, Vietnam
The marine environmental conditions are particularly different from terrestrial ones; recent studies have showed that marine actinomycetes might produce many novel compounds with good biological activity. The objective of this study is to isolate and screen actinomycetes strains from the marine environment with activity against pathogenic microorganisms. Fifty strains of actinomycetes were isolated from 40 samples including: sediments, sponges, soft corals, echinoderms... collected from Van Phong Bay area of  Khanh Hoa province, Vietnam. The strains fermented in A+ medium and fermentation broths were extracted 5 times with ethyl acetate. The extracts were evaporated under reduced  pressure to yield crude extracts. Quantitative assay was used to determine MIC (minimum inhibitory concentration) of extract against 7 test strains, including three Gram-negative bacteria (Escherichia coli ATCC25922, Pseudomonas aeruginosa ATCC27853, Salmonella enterica ATCC13076), three Gram-positive bacteria (Enterococcus faecalis ATCC29212, Staphylococcus aureus ATCC25923, Bacillus cereus ATCC 14579), and the yeast Candida albicans ATCC10231. The result of screening showed that: 18/50 strains have shown the antibacterial activities against at least 3 strains of tested microorganisms. In which, strains G650, G654, G657, G666, G675 inhibited three Gram-positive test microorganisms at MIC values from 8 to 64 µg/mL and C. albicans ATCC10231 with MIC values from 2 to 16 µg/mL. In addition, the two strains G657 and G666 had inhibitory effect on  P. aeruginosa ATCC27853 và S. enterica ATCC13076 with MIC value 128 µg/mL. Morphological and phylogenetic investigations based on 16S rRNA gene sequences of the selected five strains showed that: strain G650 belonged to species Streptomyces ardesiacus and strain G666 high similarity to the genus Streptomyces; strain G654 showed the highest similarity to the Salinispora arenicola species; strain G657 highest similarity to the Micromonospora aurantiaca; whereas G675 belonged to the Nocardiopsis flavescens species.
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