盾殼黴菌(Coniothyrium sp.)能產生多種生物活性物質，有些已開發為農作物病害防治的商業產品。本研究探討本土盾殼黴菌Coniothyrium sp. CSM生物活性物質抗癌效果。經搖瓶培養5天的發酵液以乙酸乙酯萃取，以極致效能液相色層分析(Ultra Performance Liquid Chromatography, UPLC)鑑定，確認發酵液萃取物中含有Brefeldin A (BFA)。為了提高BFA產量實驗進行菌株篩選，結果得到編號A5的菌株BFA產量最高，後續便以此菌株進行培養基修飾試驗。結果得悉增加培養基碳、氮源組成分，無法顯著提高Brefeldin A產量。培養液萃取物以HPLC分離純化成三個區分物，針對不含Brefeldin A的第二區分物，進行細胞毒性測試，得知此區分物對於抑制癌細胞亦有效果，但略遜於Brefeldin A。第二區分物濃度15μg/ml加藥處理24小時，細胞存活率為72.90 %，經過48小時處理後細胞存活率為49.19%。利用UPLC尋找第二區分物的組成最佳分離純化條件，未來將以此條件分離純化第二區分物，並探討其有效成份。 It has been well known that fungal Coniothyrium sp. can produce a variety of bioactive substances, and some of these substances have already been developed as commercial biocontrol products against plant pathogens. Besides, some Coniothyrium sp. strains are capable to produce secondary metabolites with the efficacy of anti-proliferation against cancer cells. In our lab, an endemic endophytic Coniothyrium sp. CSM has displayed the ability to produce Brefeldin A (BFA), an anti-cancer compound. The objective of this study is to improve the yield of BFA produced by this fungal strain, and to search new bioactive compounds in its fermented cultures. The experiment results briefly describe as following: the yield of BFA of this strain reached 47 mg/L after eleven day fermenting by using stir tank; a fraction of cultural extract separated by using HPLC has showed anti-proliferative effect against A549 cancer cells. The best anti-proliferative performance of this fraction was to incubate cancer cells for two day at the concentration of 15μg/ml; the fraction contains several unknown compounds, according to the analysis of UPLC. These unknown compounds are worthwhile to identify in the future.
