棘豆蠕孢菌发酵液中次级活性产物分离纯化初探  

Preliminary Study on the Isolation and Purification of Active Secondary Metabolites in the Fermentation Broth of Embellisia oxytropis

作　　者：时东方[1] 李丽[1] 郑梅竹[1] 李广洲 张红晶[3] 王琦[3] 

机构地区：[1]长春师范学院中心实验室,吉林长春130032 [2]本溪矿柱林总场卧龙林场,辽宁本溪117000 [3]吉林农业大学,吉林长春130118

出　　处：《安徽农业科学》2009年第33期16213-16215,共3页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金项目(30970554)

摘　　要：[目的]为寻找具有合成与宿主植物相同或相似活性成分能力的内生真菌奠定理论基础。[方法]利用布氏漏斗抽滤将菌丝体和发酵液分开,通过预处理除去发酵液中的杂蛋白、色素、高价金属离子和多糖,然后用大孔树脂分离发酵产物并利用硅胶柱层析进行纯化,最后通过红外光谱检测确定样品与宿主植物成分为同一类生物碱。[结果]样品在改良碘化铋钾的作用下呈橘红色斑点,在Ehrli-ch’s显色剂作用下呈紫红色斑点,两者的Rf值均为0.78。水洗脱段对3种沉淀试剂均呈阴性反应;醇水混合段对碘化铋钾呈阴性反应,对苦味酸和鞣酸呈阳性反应;醇洗脱段对3种沉淀试剂均呈阳性反应。鉴定结果表明所分离的样品为羟基取代吲哚里西啶类生物碱。[结论]内生真菌棘豆蠕孢菌具有合成与其宿主植物甘肃棘豆相同或相似活性成分的能力。[ Objective ] The study aimed to lay a theoretical foundation for seeking for endophytic fungi with capability of synthesizing the same or similar active components which was synthesized in their host plants. [ Method] The mycelia and fermentation broth were isolated by pumping filtration with Buchner funnel, the impure proteins, pigments, muhivalent metal ions and polysaccharides were scavenged from fermentation broth through pretreatments. Then the fermentation products were isolated with macroporous resin and purified by silica gel chromatography. Finally, the sample and the component from host plants were ascertained to belong to the same class of alkaloid by infrared spectrum detection. [ Result] The improved bismuth potassium iodide made sample show as a tangerine spot and the Ehrlich＇s ehromogenic reagent made it show as a purplish red spot. Both of their Rf values were 0.78. The reactions of the water elution products and the 3 kinds of precipitants were negative. The reaction of alcohol-water elution products and bismuth potassium iodide was negative and that of alcohol-water elution products and picric acid and tannic acid were positive. The reactions of alcohol elution products and the 3 kinds of precipitants were positive. It was shown in the identification results that the isolated sample was hydroxyl substituted indolizidine alkaloids. [ Conclusion ] The endophytic fungus Embellisia oxytropis had the capability of synthesizing the same or similar active components which was synthesized in its host plant Gansu whin.

关 键 词：棘豆蠕孢菌 分离纯化 生物碱 

分 类 号：S132[农业科学—农业基础科学]