| 大孔吸附树脂联合葡聚糖凝胶Sephadex LH20分离制备鹅膏肽类毒素的研究 |
| Isolation and preparation of cyclopeptide toxins by macroporous adsorptive resins combined with sephadex LH20 column chromatography from Amanita exitialis |
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| 中文关键词:大孔吸附树脂，sephadex LH20，高效液相色谱，鹅膏毒肽，鬼笔毒肽 |
| 英文关键词:macroporous adsorptive resins, sephadex LH20, high performance liquid chromatography, amatoxins, phallotoxins |
| 基金项目:国家自然科学基金（No. 31372118，30972073） |
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| 中文摘要: |
| 鹅膏菌肽类毒素是蘑菇中毒导致死亡的最主要因素，同时也是生物学研究领域的一种重要工具试剂，但由于其结构相似，分离制备单体化合物比较困难。通过4种大孔吸附树脂对致命鹅膏Amanita exitialis子实体中主要肽类毒素组分的静态吸附与解吸附性能比较，结果表明XAD16的吸附能力最强，同时也有较强的解吸附能力，但XAD16柱层析梯度洗脱时对毒素分离效果差，表明XAD16只具有吸附富集鹅膏肽类毒素的特性但不适于分离。经XAD16柱层析富集的毒素成分，通过葡聚糖凝胶sephadex LH20柱层析分离，结果表明sephadex LH20柱层析对鹅膏肽类毒素具有较好的分离效果，经过2次sephadex LH20柱层析，获得了5个纯度达50%–80%的鹅膏肽类毒素单体化合物，进一步利用半制备高效液相色谱系统（HPLC）纯化，分离得到了7个纯度达95%以上的鹅膏肽类毒素单体，其中5个经质谱分析鉴定为：α-鹅膏毒肽（α-amanitin）、β-鹅膏毒肽（β-amanitin）、脱氧二羟毒伞素（desoxoviroidin）、羧基三羟鬼笔毒肽（phallisacin）和羧基二羟鬼笔毒肽（phallacidin）。 |
| 英文摘要: |
| The cyclopeptide toxins are mainly responsible for the death of humans in the mushroom poisoning, and these toxins are also the important tools in the biology research field. It is difficult to separate the morphon toxins from the fruiting bodies of Amanita species due to their similar structures. The static adsorption and desorption properties of four macroporous adsorptive resins on the cyclopeptide toxins of Amanita exitialis were compared and analyzed in this study. The results showed that XAD16 possessed the highest static adsorption property and relatively high desorption ability. However, gradient elution of XAD16 column chromatography could not separate the toxins. The toxins enriched by XAD16 column chromatography could be separated by repeated sephadex LH20 column chromatography and five morphon cyclopeptide toxins with purity of 50%–80% were yielded. Seven morphon cyclopeptide toxins with purity of more than 95% were obtained by semi-preparation high performance liquid chromatography purification. Five of them were identified as α-amanitin, β-amanitin, desoxoviroidin, phallisacin and phallacidin respectively by mass spectrometry analysis. |
| 胡劲松,陈作红. 大孔吸附树脂联合葡聚糖凝胶Sephadex LH20分离制备鹅膏肽类毒素的研究[J]. 菌物学报, 2014, 33(3): 549-559 |
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