题目指导老师谢宝全教授.DOC

國立屏東科技大學食品科學系碩士班 國立屏東科技大學食品科學系碩士班 研究生預口試之書面摘要 指導老師：郭嘉信 老師報 告 指導老師：郭嘉信 老師 報 告 者：曾暐婷 報告日期：200 發酵多醣體分液的結構特徵與其對免疫調節之影響 Studies of the immunomodulatory effects based on structural features of polysaccharides produced by Antrodia camphorata NPU-44 mycelia in solid-state fermented brown rice 摘要 牛樟芝Antrodia camphorata多醣體經證實具有免疫調節等活性，本研究由牛樟芝子實體中分離出A. camphorata NPU-44菌絲體進行固態發酵，萃取其發酵物之粗多醣體，以高效能液相層析儀通過膠體過濾管柱及折射率偵測器進行分離及檢測，結果顯示其中主要含有三種不同分子量的多醣體，分別為290 kDa (ACP-179)、8.1 kDa (ACP-50)及1.7 kDa (ACP-10)，以快速蛋白質液相層析儀分離及純化此三種牛樟芝多醣體，並檢測其對大鼠周邊血液單核球細胞(PBMC)\及脾臟免疫細胞增殖的影響，結果顯示ACP-179誘導PBMC及脾臟免疫細胞增殖的效果最佳，可達控制組的6.1倍及3.6倍，而ACP-50及ACP-10誘導PBMC免疫細胞增殖達到控制組的3及4.8倍；脾臟免疫細胞增殖則達到控制組的1.5及3倍。進一步以GC及GC-MS檢測各多醣體特徵，結果顯示ACP-179的單糖組成主要含有果糖、阿拉伯糖、甘露糖及半乳糖，其比例為14:16:6:143；ACP-50的單糖組成主要含有甘露糖及葡萄糖，其比例為42:8；ACP-10則全由葡萄糖所組成。其中ACP-179醣苷鍵結以半乳糖為1,6鍵結為主；ACP-50醣苷鍵結為甘露糖以T-鍵結及葡萄糖以1,4鍵結為主；ACP-10則是葡萄糖以1,4鍵結。最後，以核磁共振光譜儀之氫譜鑑定各多醣體結構，ACP-179具有α-D-果糖、α-D-甘露糖及β-D-半乳糖結構；ACP-50及ACP-10則具有α-D-葡萄糖結構。因此，牛樟芝菌絲體固態發酵所生產的多醣體主要以ACP-179為主，具有β-D-半乳糖結構，因此其免疫調節能力最佳；ACP-50及ACP-10可能是固態培養基中殘留碳水化合物的片段。 關鍵字：牛樟芝、菌絲體、固態發酵、多醣體、免疫調節、結構特徵 Abstract Antrodia camphorata polysaccharides (ACP) have been reported to possess potent immunomodulatory activity. In this study, A. camphorata NPU-44 was isolated from an A. camphorata fruiting body. Crude polysaccharides produced by A. camphorata NPU-44 in solid-state fermented brown rice were extracted. The crude polysaccharides were applied to an HPLC-RI equipped with a size exclusion column for determining the molecular weights of individual containing polysaccharides. The results indicated that the fermented product consisted of 3 major polysaccharides with various molecular weights of 290 kDa (ACP-179), 8.1 kDa (ACP-50) and 1.7 kDa (ACP-10). According to immunomodulatory activity study, APC-179 induced proliferations of peripheral blood mononuclear cells (PBMC) and spleen cells up to 6.1 and 3.6 folds, respective