玫瑰红鹅膏主要肽类毒素的HPLC 测定及其对白色念珠菌的抑制活性

Determination of main peptide toxins from Amanita pallidorosea with HPLC and their antifungal action on Blastomyces albicans

摘要:

摘要: 【目的】检测玫瑰红鹅膏中所含肽类毒素及其含量，并对其肽类毒素的抑制白色念珠菌活性进行研究。【方法】采用HPLC 和ESI-MS 法从玫瑰红鹅膏中分离并鉴定出所含肽类毒素，并采用HPLC 法测定其子实体、菌盖及菌柄和菌托混合部分中肽类毒素的含量。同时，采用纸片法研究了玫瑰红鹅膏粗毒液和分离到的单品肽类毒素对白色念珠菌JLC31680 和JLC31681 的抑菌作用。【结果】分离并鉴定出α-鹅膏毒肽(α-AMA)、β-鹅膏毒肽(β-AMA)和二羟鬼笔毒肽( PHD) 等3 种肽类毒素。玫瑰红鹅膏子实体中α-AMA、β-AMA、PHD 的含量分别为30. 3168、6. 9932 和9. 9459 mg/g; 菌盖中含量分别为44.9573、11.0798和11.3025 mg/g; 菌柄和菌托混合部分中:α-AMA 11.6904 mg/g和PHD 7.9775 mg/g，β-AMA 未检出。粗毒液、α-AMA、β-AMA和PHD 对白色念珠菌JLC31680 均具有很好的抑制作用，抑制率分别达到11.96%、32.52%、23.29% (p＜0.01)和15. 46% (p＜0.05);粗毒液和β-AMA 对白色念珠菌JLC31681的最高抑制率分别为10.16%和11.10%(p＜0.01)，α-AMA 对白色念珠菌JLC31681最高抑菌率为6.89%(p＜0.05)。 【结论】玫瑰红鹅膏中的三种肽类毒素的含量较高，是制备肽类毒素的新资源; 其具有抑制白色念珠菌的活性，可开发利用。

Abstract:

Abstract: ［Objective］ To detect peptide toxins in Amanita pallidorasea and to study the antifungal activities of peptide toxins against Blastomyces albicans． ［Methods］We separated and identified peptide toxins and determined its contents in the fruiting body，pileus and the mixture of stipe and volva from A． pallidorasea by HPLC and ESI-MS methods．Meanwhile，we detected antifungal activities of the crude toxin and the separated peptide toxins against Blastomyces albicans JLC31680 and JLC31681by the paper disk method．［Results］We totally got three peptide toxins: α-amanitin (α-AMA)，β-amanitin (β-AMA) and phalloidin (PHD)．The contents of α-AMA，β-AMA and PHD were 30.3 mg/g，6.99 mg/g and 9.95 mg/g in fruiting body，and 45.0 mg/g，11.1 mg/g and 11.3 mg/g in pileus．The contents of α-AMA and PHD were 11.7 mg/g and 7.98 mg/g in the mixture of stipe and volva，but the β-AMA was not detected in this part． The inhibition ratio of the crude toxin and a-AMA，β-AMA and PHD to B．albicans JLC31680 were 11.96%，32.52%，23.29%(P＜0.01) and 15.46%(P＜0.05)．The inhibition ratio of the crude toxin and β-AMA to B．albicans JLC31681 was 10.16% and 11.10%(P＜0.01)，while that of α-AMA s was 6.89%(P＜0.05)．[Conclusions] A．pallidorasea is a new resource of peptide toxins with antifungal activity．