吸水链霉菌BS-112产生的抗真菌活性物质的分离纯化与结构鉴别

【目的】分离纯化吸水链霉菌(Streptomyces hygroscopicus)BS-112产生的抗真菌活性物质,究明各活性组分的结构,测定其对黄曲霉的抑制作用,为该菌株及其产生的抗真菌活性物质的应用提供依据。【方法】通过大孔吸附树脂柱层析、硅胶柱层析及制备HPLC等方法,对该菌株产生的抗真菌活性物质进行分离纯化;利用质谱(MS)和核磁共振谱(NMR)解析各活性组分的结构;采用微量液体稀释法测定各活性组分对黄曲霉的最小抑菌浓度(MIC)和最小杀菌浓度(MFC)。【结果】从BS-112菌株发酵液中分离获得4个抗真菌活性组分,利用波谱技术确定其结构分别为Tetrins A和B、Tetramycins A和B。96孔板法测得这4个化合物对黄曲霉的MIC分别为3.13μg/mL、12.56μg/mL、1.56μg/mL、6.25μg/mL,MFC分别为6.25μg/mL、25.0μg/mL、3.13μg/mL、12.56μg/mL。【结论】BS-112菌株产生的抗真菌活性物质由Tetrins A和B、Ttramycins A和B 4个化合物组成,它们对黄曲霉均具有良好的抑制作用。

Isolation, purification and structure identification of the antifungal active substances from Streptomyces hygroscopicus BS-112

Objective]In order to provide the basis for application of Streptomyces hygroscopicus BS-112 and its antifungal active substances,we isolated and purified the active substances produced by strain BS-112,and then we examined their chemical structures and determined their inhibitory effects on Aspergillus flavus.Methods]We extracted and purified the antifungal active substances from strain BS-112 using column chromatography with X-5 macroporous resin and 200 – 300 mesh silica gel respectively,and then purified them by preparative HPLC with Venusil XBP C18 column.The chemical structures of active components were identified by means of spectroscopic analysis,including mass spectrometry(MS) and nuclear magnetic resonance spectroscopy(NMR).Their minimum inhibitory concentration and minimum fungicidal concentration against Aspergillus flavus were determined by the standard broth microdilution method.Results ] We obtained four active components from the fermentation broth of strain BS-112 and their chemical structures were the same as Tetrins A and B,Tetramycins A and B.Assay using 96-well plate illustrated that the MIC of Tetrin A,Tetrin B,Tetramycin A and Tetramycin B against Aspergillus flavus were 3.13 μg /mL,12.56 μg /mL,1.56 μg /mL and 6.25 μg / mL respectively,and their MFC were 6.25 μg /mL,25.0 μg /mL,3.13 μg /mL and 12.56 μg /mL respectively.Conclusion]The antifungal active substances of strain BS-112 were composed of four compounds,Tetrins A and B,Tetramycins A and B.The four active compounds showed strong inhibitory against Aspergillus flavus.