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从暗黑链霉菌(Streptomyces atratus)PY-1菌株发酵液中分离鉴定抑菌活性产物,评价其对葡萄霜霉病菌(Plasmopara viticola)的抑制作用,为该菌株及其产生的抑菌活性物质的应用提供依据。PY-1菌株发酵液经二氯甲烷萃取,减压浓缩获得抑菌粗提物,稳定性测定结果显示,避光条件下,15℃~65℃、p H 6~10、4℃或常温保存6个月,抑菌活性稳定。采用硅胶柱层析、Sephadex LH-2柱层析、薄层层析和HPLC等技术,对抑菌粗提物进一步分离纯化,得到2种抑菌活性组分;采用ESI-M S、1H NM R、13C NM R等波谱分析技术,对活性组分进行结构鉴定,2种活性组分分别鉴定为5-乙酸环已基酰亚胺(5-acetoxycycloheximide)和环己酰亚胺(cycloheximide)。采用离体叶片法测定2种组分的抑菌活性,不同浓度(10~4ng·mL~(-1)、10~2ng·mL~(-1)、1 ng·mL~(-1))的5-乙酸环已基酰亚胺和环己酰亚胺对葡萄霜霉病菌的抑制作用分别为92.59%、86.30%、64.81%和97.04%、91.85%、84.07%。
The antibacterial activity product was isolated and identified from the fermentation broth of PY-1 strain of Streptomyces atratus to evaluate its inhibitory effect on Plasmopara viticola and the application of the antibacterial activity to the strain Provide evidence. The fermentation broth of PY-1 strain was extracted with dichloromethane and concentrated under reduced pressure to obtain antibacterial crude extract. The stability assay showed that the fermentation broth of PY-1 strain was stored at 15 ℃ ~ 65 ℃, p H 6 ~ 10, 4 ℃ or room temperature 6 months, antibacterial activity and stability. The antibacterial crude extracts were further isolated and purified by silica gel column chromatography, Sephadex LH-2 column chromatography, TLC and HPLC. Two kinds of antibacterial active components were obtained. The compounds were characterized by ESI-MS, R, 13C NM R and other spectral analysis techniques, the structure of the active ingredient was identified. The two active components were identified as 5-acetoxycycloheximide and cycloheximide. The antibacterial activities of the two components were determined by in vitro leaf method. The effects of different concentrations (10 ~ 4ng · mL -1, 10 ~ 2ng · mL -1, 1 ng · mL -1) The inhibitory effects of cyclohexylimidazole and cycloheximide on Plasmopara viticola were 92.59%, 86.30%, 64.81% and 97.04%, 91.85% and 84.07%, respectively.