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研究生:李青樺
研究生(外文):CHING-HUA Li
論文名稱:舞菇多醣體水解之分析研究
論文名稱(外文):Study on the hydrolysis of polysaccharides of Grifola frondosa
指導教授:許銘清
指導教授(外文):Hsu Ming-Ching
學位類別:碩士
校院名稱:南台科技大學
系所名稱:化學工程系
學門:工程學門
學類:化學工程學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:102
中文關鍵詞:舞菇多醣體水解
外文關鍵詞:Grifola frondosapolysaccharideshydrolysis
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本研究使用舞菇菌絲體與子實體,利用乙醇、熱水、冷鹼及熱鹼方式提取多醣,進行成分分析。總醣含量分析得知提取物的多醣含量依序為熱水提取之沉澱物(MCF-2)、冷鹼提取之沉澱物(MCF-4)、熱鹼提取之上清液(MCF-7)、熱水提取之上清液(MCF-3)、冷鹼提取之上清液(MCF-5)、熱鹼提取之沉澱物(MCF-6)分別為90.3 %、70.3 %、33.67 %、30 %、13.4 %、5.98 %。舞菇菌絲體所提取的多醣成分分子量分布範圍很廣,經SE-HPLC(Size Exclucsion-High Performance Liquid Chromatography)分析,得知各提取成分的分子量除了MCF-1(酒精提取物)和MCF-7分子量較小外,大部分的提取物分子量都大於141萬。在醛醣酸Uronic Acid含量分析,得知最高的為MCF-6與MCF-2分別為9.87 %及9.36 %。
舞菇多醣MCF-2的水解實驗,結果顯示在7 M的醋酸95 ℃下水解7 hrs時,有較佳的水解效果,其較小分子量約為11,000。
由單糖組成份的分析,得知菌絲體各提取物(MCF-1∼MCF-7)的多醣體之各單糖組成份的莫爾比例關係,顯示各成份所含主體單糖為Glucose、Mannose,而以MCF-4單糖組成比較特殊是以Glucose與Xylose為主。子實體各提取物(FCF-1∼FCF-7)的多醣體之各單糖組成份的莫爾比例關係,顯示提取物FCF-1∼7各成份所含的單糖主體是以Glucose為主。
在抗氧化性質與腫瘤抑制方面,是以MCF-2水解前後來做分析比較。結果顯示其水解後之抗氧化性質都較未水解的效果來的優異。
在腫瘤抑制的測試,得知對子宮頸癌(HeLA)細胞、乳癌(MCF-7)細胞、肝癌(Hep G2)細胞、胃癌(AGS)細胞及前列腺癌(DU-145)細胞以濃度為10 mg/mL未水解的MCF-2樣品抑制腫瘤效果較好,抑制率為25∼46 %;對纖維瘤(HT-1080)細胞而言,以濃度為10 mg/mL水解的MCF-2樣品,效果較好,抑制率為43 %。
The aim of this project is to investigate the polysaccharides of Grifola frondosa. The polysaccharides from the mycelia and the fruit bodies were extracted by ethanol first, then followed in order by hot water, cold and hot alkaline solution containing 5% urea and 10% NaOH. The extracts thus obtained from mycelia were MCF-1 (ethanol extract), MCF-2 (the precipitate of hot water extract), MCF-3(the supernatant of hot water extract), MCF-4(the precipitate of cold alkaline solution extract), MCF-5(the supernatant of cold alkaline solution extract), MCF-6(the precipitate of hot alkaline solution extract), MCF-7(the supernatant of hot alkaline solution extract). The polysaccharide contents for MCF-2, MCF-4, and MCF-6 were 90.30%, 70.30% and 5.98%, respectively, while those for MCF-3, MCF-5, and MCF-7 were 30.00%, 13.40%, and 33.67%, respectively. The gel filtration chromatograph showed wide range molecular weight distribution and were over 1,400 kDa, except MCF-1 and MCF-7. It was found that MCF-6 and MCF-2 contained higher amounts of uronic acid(9.87% and 9.36%, respectively). And it was also found that glucose and mannose were the major components of the polysaccharide extracts from mycelia with the exception of MCF-4, which major polysaccharide extracts from fruit bodies were glucose. When MCF-2 was hydrolyzed with 7 M acetic acid and hydrolysis at 95 ℃ for 7 hrs, the molecular weight of hydrolyzed polysaccharides were around 11 kDa, which however showed no promising anti-tumor and anti-oxidation activity. The anti-tumor activity assay for extracted polysaccharides indicated that the original MCF-2 at a concentration of 10 mg/mL showed the best effects against Hela cells, MCF-7, Hep G2, AGS, and Du-145 with the inhibition range from 25-46%. For HT-1080 cells, the hydrolyzed MCF-2 at the concentration of 10 mg/mL showed a best cell growth inhibition of 43%.
中文摘要……………………………………………………………………Ⅰ
英文摘要……………………………………………………………………Ⅱ
誌 謝………………………………………………………………………Ⅲ
目 錄………………………………………………………………………Ⅳ
表目錄………………………………………………………………………Ⅴ
圖目錄………………………………………………………………………Ⅵ
第一章 前言………………………………………………………………1
1-1 背景……………………………………………………………….1
1-2 研究目的……………………………………………………….…2
第二章 文獻整理…………………………………………………………3
2-1 舞菇的介紹…………………………………………………….…3
2-2 舞菇的化學成分………………………………………….………6
2-2-1、營養、食品風味成分……………………………………….6
2-2-2、目前已知舞菇之主要成份………………………….…..…7
2-3 舞菇生理功能…………………………………………….………9
2-3-1、舞菇多醣之免疫增強、抗癌、抗腫瘤多醣之特性……….9
2-3-1-1 增強免疫力……………………………………………….9
2-3-1-2 抗癌作用…………………………………………………10
2-3-1-3 抗腫瘤多醣體的特性……………………………………12
2-3-2、舞菇多醣構造………………………………………………12
2-3-2-1 活性成分β-1,3-D-glucan之結構分析………………..13
2-4 抗氧化作用………………………………………………………16
2-4-1、自氧化反應…………………………………………………18
2-4-1-1 自氧化反應機構…………………………………………18
2-4-2、抗氧化劑……………………………………………………19
2-4-3、抗氧化物……………………………………………………19
2-4-3-1 協助體內抗氧化的物質…………………………………19
2-4-4、自由基致癌、促癌作用及其發生機制……………………22
2-4-4-1自由基在化學致癌過程中的作用……………………….22
2-4-4-2自由基與促癌作用……………………………………….23
2-4-4-3自由基致癌和促癌可能發生的機制…………………….24
2-5 國內外舞菇藥理之相關研究……………………………………26
第三章 研究材料與方法…………………………….……………………30
3-1 材料與藥品………………………………………………………30
3-1-1 舞菇菌種…………………………………………………….30
3-1-2 藥品………………………………………………………….30
3-1-3 實驗儀器…………………………………………………….32
3-2 實驗方法…………………………………………………………33
3-2-1 多醣的製備……………………………………………….…33
3-2-2 總醣含量的測定……………………………………….……35
3-2-3 多醣組成份的層析………………………………….………35
3-2-4 分子量的測定…………………………………………….…36
3-2-5 醛醣酸 Uronic Acid 分析…………………………………36
3-2-6 舞菇多醣的水解………………………………………….…37
3-2-7 單糖組成份的分析………………………………….………38
3-2-8 菌株抗生活性探討……………………………………….…39
3-2-9 舞菇抗氧化性質……………………………………….……40
3-2-10 抑制腫瘤細胞成長分析……………………………………42
第四章 結果與討論……….………………………………………………45
4-1 總醣含量分析……………………………………………...……45
4-2 多醣組成份的層析…………………………………………….…47
4-3 分子量的測定………………………………………………….…51
4-4 醛醣酸 Uronic Acid 分析…………………………….…….…57
4-5 舞菇多醣的水解……………………………………………….…59
4-6 單糖組成份的分析………………………………………….……63
4-7 菌株抗生活性探討………………………………………….……85
4-8 舞菇抗氧化性質分析………………………………………...…87
4-8-1 還原能力的測定………………………………………………87
4-8-2 清除DPPH自由基能力測定…….…….………………………89
4-8-3 清除H2O2能力測定………………….…………….…………91
4-9 多醣抑制腫瘤細胞分析……………………………………….…93
第五章 結論………….……………………………………..…………..95
第六章 參考文獻………………….………………………………………97
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