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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 呂廷璋(Ting-Jang Lu) | |
| dc.contributor.author | Hsueh-Ting Wang | en |
| dc.contributor.author | 王雪婷 | zh_TW |
| dc.date.accessioned | 2021-06-17T02:19:10Z | - |
| dc.date.available | 2018-08-24 | |
| dc.date.copyright | 2017-08-24 | |
| dc.date.issued | 2017 | |
| dc.date.submitted | 2017-08-21 | |
| dc.identifier.citation | 王鐘凰, 菇蕈中活性多醣 (1, 3; 1, 6)-β-D-葡萄聚醣的分析與功能評估. 臺灣大學食品科技研究所學位論文 2014, 1-118.
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Zhang, H.; Li, W.-J.; Nie, S.-P.; Chen, Y.; Wang, Y.-X.; Xie, M.-Y., Structural characterisation of a novel bioactive polysaccharide from Ganoderma atrum. Carbohydr. Polym. 2012, 88, 1047-1054. Zhong, H.; Voll, R. E.; Ghosh, S., Phosphorylation of NF-κB p65 by PKA stimulates transcriptional activity by promoting a novel bivalent interaction with the coactivator CBP/p300. Mol. Cell 1998, 1, 661-671. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/68376 | - |
| dc.description.abstract | 岩藻糖是誘導細胞產生免疫反應的重要抗原結合位；在本研究中以活性導向分離純化了巴西洋菇中含岩藻糖(fucose)之酸性多醣F3_FA；F3_FA具刺激巨噬細胞Raw 264.7分泌TNF-α的生物活性，經活性測試結果顯示岩藻糖在F3_FA刺激小鼠巨噬細胞RAW 264.7分泌TNF-α有著重要的角色；試驗結果並得知F3_FA在RAW 264.7細胞的接受器為TRL-4且其刺激可能為受NF-κB的調控，進一步影響下游TNF-α細胞激素的表現。巴西洋菇多醣F3_FA的化學分析結果顯示其分子量為3.45 x 105 Da.；糖組成莫耳百分比為6.4% L-fucose, 15.6% L-arabinose, 28.7% D-galactose, 14.8% D-xylose 及 24.3% 的D-galacturonic acid 並含少量的D-glucose, D-mannose, L-rhamnose 及 D-glucuronic acid ；利用甲基化衍生試驗得知F3_FA的結構中含大量1-4GalAp (24.3%)、1-4 Galp (22.1%)的鍵結結構並含有末端Fuc (6.4%)、1-5Ara (5.4%)、1-2Rha (5.1%)，推論為其結構為類似果膠多醣之結構；以果膠多醣單株抗體辨識也得到F3_FA含有(1, 4)-β-D-galactans、 (1, 5)-α-L-arabinans 及 (1, 4)-α-D-galacturonans的結構特徵；結構特徵結果在NMR圖譜中也得到呼應。本研究以四級銨鹽的選擇性沉降為分離酸性多醣、提高岩藻多醣含量的策略；以去氧糖呈色試驗估算樣品中岩藻糖的含量，結果顯示樣品中去氧糖含量由粗多醣中的4.6%經沉降區分後提升至酸性區分中含15.6%；且經比對證實F3_FA為巴西洋菇菌絲多醣非由發酵基質而來。 | zh_TW |
| dc.description.abstract | Fucose is one of important residues of recognition pattern for many immune cells. In this study, we characterized bioactive fucose-containing acidic polysaccharides from submerged fermentation of Agaricus blazei Murill. We obtained the polysaccharides through a cell-based activity-guided strategy, and used carbohydrate recognition monoclonal antibodies based Enzyme-Linked Immuno Sorbent Assay (ELISA) along with methylation and NMR analyses to investigate the structural characteristics of the polysaccharides. The polysaccharides had Mw of 3.45 × 105 Da. The major sugars were L-fucose, L-arabinose, D-galactose, D-xylose, and D-galacturonic acid in the molar ratio of 6.4, 15.6, 28.7, 14.8, and 24.3% with a small amount of D-glucose, D-mannose, L-rhamnose, and D-glucuronic acid. Results indicated that the bioactive polysaccharides consisted of a (1,4)-Galp and (1,4)-GalAp back bone; (1,2)-Xyl and (1,2)-Rha might also comprise backbone or constitute side chain; linkage (1,5)-Ara and terminal fucosyl residues were also involved in the polysaccharides. Regarding bioactivity, removal of the terminal L-fucosyl residues reduced the TNF-α cytokine stimulating activity of the polysaccharides in a RAW264.7 macrophage cell-line test, whereas NF-kapaB and TLR4 affected the polysaccharide-induced TNF-α production. Quanternary selecting precipitation as a strategy to rapid enriches fucose-containing acidic polysaccharide, deoxyl sugar contain from 4.2% upgrade to 15.6% by one step separated. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T02:19:10Z (GMT). No. of bitstreams: 1 ntu-106-D95641005-1.pdf: 2238049 bytes, checksum: 2628549d38fa0ba08619bf935ca66648 (MD5) Previous issue date: 2017 | en |
| dc.description.tableofcontents | 謝誌 i
摘要 ii Abstract iii 目錄 I 圖目錄 V 表目錄 VII 附表附圖目錄 VIII 壹、 前言 1 貳、 文獻回顧 2 第一章、 巴西洋菇 2 1.1 、巴西洋菇的化學組成 2 1.2 、巴西洋菇多醣與在生理活性的影響 2 1.2.1 、子實體多醣 2 1.2.2 、菌絲體多醣 4 第二章、 多醣結構與生理活性 5 2.1 、骨幹結構與化學修飾對多醣活性的影響 5 2.2 、構型 5 2.3 、分子量與分支度 5 第三章、 果膠多醣結構及其生理活性 7 3.1 、果膠多醣結構 7 3.1.1 、Homogalacturonan（HG） 7 3.1.2 、Rhamnogalacturonan Ⅰ (RGⅠ) 8 3.1.3 、RhamnogalacturonanⅡ (RGⅡ) 11 3.1.4 、Xylogalacturonan（XGA） 12 3.2 、果膠多醣相關的生理活性 12 3.3 、單株抗體辨識果膠多醣 13 第四章、 含岩藻糖(fucose)的多醣其生理上的重要性 17 4.1 、岩藻糖 17 4.2 、岩藻多醣的生理活性 17 4.3 、岩藻多醣的可能鍵結 17 第五章、 巨噬細胞與生理活性 19 5.1 、巨噬細胞及其功能 19 5.2 、脂多醣 (lipopolysaccharide, LPS) 誘發之生理反應與路徑 20 5.3 、核轉錄因子kappa B (nuclear transcription factor kappa B, NF-κB) 20 5.4 、活性評估指標 21 5.4.1 、誘導型環氧化酶 (cyclooxygenase-2, COX-2) 21 5.4.2 、腫瘤壞死因子-α (tumor necrosis factor alpha, TNF-α) 21 第六章、 氣相層析質譜儀於多醣分析 22 6.1 、甲基化的基本化學反應 22 6.2 、鹼試劑對甲基化反應的影響 23 6.2.1 、以金屬氧化物為甲基化鹼試劑 23 6.2.2 、以金屬氧化物的水溶液為甲基化鹼試劑 24 6.2.3 、以鹼金屬氫化物及其衍生物搭配非質子溶劑為甲基化試劑 24 6.2.4 、高鹼無水的反應環境 25 6.3 、分子的立體空間特性與反應性 27 第七章、 核磁共振光譜於多醣結構的分析 28 第八章、 四級銨鹽沉降法 30 參、 材料與方法 31 第一章、 實驗材料 31 1.1 、巴西洋菇熱水萃取 31 第二章、 多醣分析法與純化 32 2.1 、多醣分析相關方法 32 2.1.1 、碳水化合物含量測定(phenol-sulfuric acid method) 32 2.1.2 、醛醣酸之測定 32 2.1.3 、蛋白質含量測定 32 2.1.4 、去氧醣含量測定 32 2.1.5 、單糖組成測定 33 2.1.5.1 、Trifluoroacetic acid (TFA) hydrolysis 33 2.1.5.2 、Methanolysis 33 2.1.5.3 、高效能陰離子交換層析系統(High performance anion exchange chromatography with PAD detector, HPAEC-PAD) 33 2.1.6 、分子量測定 34 2.1.7 、結構特徵測定 34 2.1.7.1 、鍵結分析 (methylation analysis) 34 2.1.7.2 、 核磁共振光譜 (NMR) 37 2.1.7.3 、 果膠抗體辨識 37 2.2 、多醣純化相關方法 38 2.2.1 、 利用多醣表面電荷不同的劃分法 38 2.2.1.1 、四級銨鹽沉降法Quaternary ammonium salt precipitation: 38 2.2.1.2 、鹽度劃分法 38 2.2.1.3 、陰離子交換樹脂 38 2.2.2 、利用多醣分子量不同的劃分法 38 2.2.2.1 、膠體過濾層析法 38 第三章、 活性測試 40 3.1 、NO 測定 40 3.2 、TNF-α測定 40 3.3 、刺激RAW 264.7巨噬細胞株(含報導基因質體COX-2) 之活性評估 40 3.4 、活性路徑推論 41 第四章、 以四級銨選擇性沉降巴西洋菇多醣流程 42 第五章、 巴西洋菇生物活性多醣來源確認 43 第六章、 統計試驗 44 第七章、 實驗藥品與試劑 45 5.1化學試劑 45 5.2標準品 45 肆、 試驗架構 46 伍、 結果與討論 47 第一章、 分離純化 47 巴西洋菇菌絲發酵多醣以生物活性導向為分離純化策略。 47 1.1 、分離純化 47 1.2 、活性評估 50 第二章、 巴西洋菇生物活性多醣化學特性 54 2.1 、分子量 54 2.2 、組成與結構 55 2.2.1 、單醣組成 55 2.2.2 、單株抗體辨識 58 2.2.3 、苷鍵結分析 60 2.2.4 、NMR結構分析 62 2.3 、小結 65 第三章、 多醣F3_FA生物活性試驗 66 3.1 、具報導基因細胞殖株平台及NO釋放檢視巴西洋菇多醣F3_FA的活性表現 66 3.2 、岩藻醣對於巴西洋菇多醣F3_FA活性的影響 69 3.3 、巴西洋菇多醣F3_FA的生理活性路徑推論 71 3.4 、巴西洋菇多醣活性與岩藻糖含量關係的追蹤 74 第四章、 巴西洋菇生物活性多醣F3_FA來源確認及分離方式比較 75 4.1 、F3_FA來源確認 75 4.1.1 、結果 75 4.2 、分離純化方法的比較及確效 77 陸、 結論 84 柒、 參考文獻 85 | |
| dc.language.iso | zh-TW | |
| dc.subject | 岩藻糖 | zh_TW |
| dc.subject | 腫瘤壞死因子-α | zh_TW |
| dc.subject | 巴西洋菇 | zh_TW |
| dc.subject | 半乳聚醣 | zh_TW |
| dc.subject | 酸性多醣 | zh_TW |
| dc.subject | TNF-α stimulation | en |
| dc.subject | Agaricus blazei Murill | en |
| dc.subject | acidic polysaccharide | en |
| dc.subject | fucose | en |
| dc.subject | galactan | en |
| dc.title | 巴西洋菇菌絲發酵物中含岩藻糖之多醣特質 | zh_TW |
| dc.title | Characteristics of fucose-containing polysaccharides from submerged fermentation of Agaricus blazei Murill | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 105-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 張永和(Yung-Ho Chang),謝淑貞(Shu-Chen Hsieh),羅翊禎(Yi-Chen Lo),王惠珠(Huei-Ju Wang) | |
| dc.subject.keyword | 巴西洋菇,酸性多醣,岩藻糖,半乳聚醣,腫瘤壞死因子-α, | zh_TW |
| dc.subject.keyword | Agaricus blazei Murill,acidic polysaccharide,fucose,galactan,TNF-α stimulation, | en |
| dc.relation.page | 106 | |
| dc.identifier.doi | 10.6342/NTU201704149 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2017-08-21 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 食品科技研究所 | zh_TW |
| 顯示於系所單位： | 食品科技研究所 | |
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