海洋枝顶孢霉BH0531发酵液杀线虫组分的分离及活性测定

实验采用生物活性跟踪法，通过超滤、Sephadex凝胶层析、离子交换树脂、硅胶板薄层层析，以及特异性鉴别染色等方法对海洋枝顶孢霉BH0531发酵液的杀线虫活性组分进行分级分离和初步的性质分析。结果表明，Sephadex凝胶柱层析（G-25，G-10）可有效分离获得杀线虫活性组分，3 d的线虫校正死亡率达98.33%；以正丁醇:水:无水乙醇=1∶1∶1(V/V)为展开剂的薄层层析，可将活性组分有效分离，Rf = 0.31，斑点洗脱物的线虫校正死亡率达到94.00%；定性染色结果显示，茚三酮反应阳性，证实有α-氨基存在，茚三酮吡啶呈红色，显现出寡肽性质；双甲酮染色呈亮黄色，表明具有酮糖性质。结果表明，BH0531杀线虫活性代谢物应为含有酮糖基团的短肽类化合物。

Separation and activity determination of nematicidal components from marine-derived Acremonium sp. BH0531 fermentation broth

The nematicidal active components from marine-derived Acremonium sp. BH0531 fermentation broth were fractionated and preliminarily identified through the methods of ultra-filtration, Sephadex gel chromatography, anion-exchange resin, thin layer chromatography (TLC), as well as specific differential staining under the guidance of bio-active assay. The results showed that the nematicidal active components were separated and purified effectively by Sephadex G-25 and Sephadex G-10 gel chromatography, the adjusted mortality of the nematodes at 3 d was up to 98.33%. The TLC results indicated that the active components can be effectively separated using 1-butanol, H2O, anhydrous ethanol (1∶1∶1) (V/V) as developing solvent, Rf = 0.31. The spot of silica gel plate, which adjusted mortality of the nematodes was 94.00%. The qualitative staining results indicated that the inhydrin color reaction was positive, which proved the presence of α-amino group. The inhydrin pyridine color reaction was red, showing oligopeptide properties. The dimedone staining was bright yellow, showing the properties of ketose. These results suggested that the marine-derived fungus Acremonium sp. BH0531 fermentation broth with nematicidal activity should contain oligopeptides compounds with ketose groups.