黑殭菌素製備與分析研究探討

Preparation and analysis of Destruxins from Metarhizium anisopliae

許惠婷(Hui-ting Shu)

指導教授 ： 曾耀銘 ； 劉炳嵐

朝陽科技大學/理工學院/生化科技研究所/碩士(2012年)

摘要

黑殭菌屬(Metarhizium)是昆蟲病原真菌的一屬，常在世界各地農業害蟲及疾病媒介控制方案中，被作為化學殺蟲劑替代品。廣泛使用真菌的二次代謝產物包括黑殭菌作為生物殺蟲劑，已經增強其基本生物學的興趣。黑殭菌已被廣泛用於合成環狀六縮酚酸胜肽—稱為黑殭菌素，乃是由a-羥基酸及五個胺基酸殘基組成。事實上，環縮酚酸胜肽加上天然環胜肽，合併成具有一個以上酯鍵連結為特色之天然產物大家族，這一類天然產物，因其表現出的多樣化生物活性而在科學界引起廣大興趣，包括抗癌、抗生素、抗真菌、免疫抑制劑或抗發炎活性，與其有趣的作用機制及引人注意的分子結構一致。本實驗中以5 L攪拌式發酵槽發酵培養黑殭菌素，再以乙酸乙酯萃取出其成分，並且利用毛細管電泳分析與定量黑殭菌素，半製備型HPLC分離純化。實驗結果顯示，毛細管電泳分析條件為40 mM Sodium broate、pH 9.8、15 kV、215 nm、57 cm×50 μm×37.5 μm即有相當好的分析效果。而質譜方面，destruxin B分子量為593，經由ESI-MS正離子模式可得到m/s 594，ESI-MS/MS分析可得到分子量481之片段離子。

關鍵字

毛細管電泳 ； 黑殭菌素 ； 黑殭菌

並列摘要

Metarhizium is a genus of entomopathogenic fungi used world-wide as alternatives to chemical insecticides in agricultural pest and disease-vector control programs. The wide utilization of M. anisopliae as bioinsecticide has increased interest in its basic biology, including its fungal secondary metabolites.M. anisopliae has been widely used for synthesis of cyclic hexadepsipeptides,known as destruxins (dtxs) which consist of an a-hydroxyl acid and five amino acid residues.together with natural cyclic peptides, emerge as a broad family of natural products characterized by the occurrence of at least one ester linkage. The great interest that this class of natural products has elicited in the scientific community is explained by the diverse range of biological activities that they exhibit, including antitumoral, antibiotic, antifungi, immunosuppressant, or anti-inflammatory activities, coincidence with intriguing mechanisms of action and attractive molecular architectures. The destruxin used in this study were isolated and purified from submerged culture.Qualitative and quantitative analysis was performed on a Capillary Electrophoresis system.The optimum conditions were pH 9.8、40 mM Sodium broate、10％ Acetonitrile, at voltage15 kV, and detected wavelength 214 nm. The ESI mass spectrum shows an abundant protonated molecule (dxt B), [M+H]+ m/z 594, and MS2 analysis of the m/z 481 product ion of MS of m/z 594.

並列關鍵字

Destruxins ； Capillary Electrophoresis ； Metarhizium anisopliae

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