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  1. NTU Theses and Dissertations Repository
  2. 生物資源暨農學院
  3. 食品科技研究所
請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/60469
完整後設資料紀錄
DC 欄位值語言
dc.contributor.advisor呂廷璋
dc.contributor.authorWan-Ting Huangen
dc.contributor.author黃琬庭zh_TW
dc.date.accessioned2021-06-16T10:19:01Z-
dc.date.available2018-08-28
dc.date.copyright2013-08-28
dc.date.issued2013
dc.date.submitted2013-08-16
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/60469-
dc.description.abstract菇蕈類食品及相關保健食品,其具有 (1,6)-β-葡萄糖基之 (1,3)-β-D-葡萄聚醣為已知的生物活性調節劑 (biological response modifier),且已經發展為腫瘤抑制劑之純化藥劑的生產,此多醣為一活性成分指標。本研究選擇酵素-層析法進行分析方法的確校試驗,評估分析方法的精確性、準確性、專一性,並利用洋菇為代表性基質進行基質干擾評估,並將台灣常見的22種菇蕈類做 (1,3; 1,6)-b-D-葡萄聚醣含量及分支度分析。最後將針對另一重組酵素做其活性、專一性及轉換率試驗。實驗結果顯示所評估的分析方法於連續五天每天分析五次的精確度 (precision, % CV) 為5%,葡萄糖單體與雙體的標準線性分別為r2=0.9985與0.9991;檢測兩個年份的菇蕈類,其檢測結果為5.25 (mg/g)~207.53 (mg/g) 之間,而分支度在0.11~0.45之間,以市售的菇蕈類產品其含量範圍在9.89~54.79 (mg/g)。並以0.5 M及2 M的鹼相比較對於八種菇蕈類樣品的 (1,3; 1,6)-b-D-葡聚醣其含量及分支度比較,發現質地較軟的菇類樣品使用0.5 M的鹼溶即可達到使用2 M鹼溶的效果,但如為質地較硬的樣品則仍需使用2 M的鹼溶。對於recombinant exo-1,3-β-D-glucanase與endo-1,3-β-D-glucanase其混和使用的合適環境為40℃,及合併使用後對於非菇蕈類基質的作用,以整體而言,recombinant exo-1,3-β-D-glucanase對於β-(1,3)葡聚醣的基質較為專一。zh_TW
dc.description.abstractEdible mushrooms and their products have (1,6)-β-branched (1,3)-β-D-glucans, which play a targeting biological function as biological response modifiers, and have been developed by the pharmaceutical industry as a tumor inhibitor. This study employed enzymatic-chromatographic determination method to do validation, including precision, accuracy, specific, and matrix addition, and analysis of (1,3; 1,6)-b-D-glucan content and degree of branching for 22 species of Taiwan cultivated edible mushrooms. Then aim to recombinant exo-1,3-β-D-glucanase ( recombinant exo-1,3-ß-glucanase) to observe activity, specific, and conversion test. Samples were analyzed five times a day for five days. Results showed a precision (% CV) of 5%, with R square value 0.9985 and 0.9991 for glucose and gentiobiose, respectively. Samples of two different age mushrooms had a (1,3;1,6)-b-D-glucan content of 5.25~207.53 (mg/g), and degree of branching of 0.11~0.45. On the other hand, commercial mushroom products have 9.89-54.79 (mg/g) (1,3;1,6)-b-D-glucan content. Two alkali concentration 0.5M and 2M were used to analyze eight samples for its (1,3; 1,6)-b-D-glucan content and degree of branching. For soft textured samples, both 0.5M and 2M alkali concentration could be used and had the same effect. but for hard texture samples, 2M alkali concentration must be used for recombinant exo-1,3-β-D-glucanase mixing endo-1,3-β-D-glucanase the optimum temperature is 40℃, and action the non-(1,3; 1,6)-b-D-glucan matrix. Overall, recombinant exo-1,3-β-D-glucanase is specific of β-(1,3)-D-glucan sample, but if the specific of β-(1,3)-D-glucan sample.en
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dc.description.tableofcontents中文摘要 i
Abstract ii
總目錄 I
圖目錄 IV
表目錄 VI
第一章、 前言 1
第二章、 文獻回顧 2
第一節、 食用菇類 2
第二節、 多醣結構特徵 9
一、 菇類多醣 9
二、 酵母菌細胞壁多醣 10
第三節、 (1,3)-β-D-glucan as biological response modifiers (BRM) 11
第四節、 β-1,3-D-葡聚醣的結構及特性 14
一、 具β-(1,6)分支之(1,3)-β-D-葡聚醣的結構介紹 14
二、 (1,3; 1,6)-β-D-葡聚醣的分支度 15
三、 (1,3)-β-D-葡聚醣的分子量 15
四、 (1,3)-β-D-葡聚醣的構形特性 16
第五節、 β-1,3-D-葡聚醣的定量及檢驗方式 18
一、 螢光呈色法(aniline blue) 18
二、 Congo red呈色法 19
三、 生物活性法 20
四、 酵素法 23
五、 酵素串聯高效液相層析儀 24
第三章、 研究目的與實驗架構 26
第一節、 研究目的 26
第二節、 實驗架構 26
第一節、 實驗材料 27
一、 菇類樣品 27
第二節、 試藥與試劑 31
一、 化學藥品及試劑 31
二、 醣類標準品與酵素 31
三、 儀器 32
第三節、 實驗方法 33
一、 菇類凍乾粉末樣品製備 33
二、 總醣含量測定 ( Total sugar determined) 33
三、 還原醣含量測定 (Reducing content determined) 33
四、 以膠體過濾層析法測定分子量分布 34
五、 鹼溶性 (1,3)-β-D-glucan 含量測定 (aniline blue assay) 34
六、 (1,3; 1,6)-β-D-glucan 含量和分支度測定 35
七、 高效能陰離子交換層析系統(HPAEC-PAD) 36
八、 高效分子篩層析法(HPSEC) 37
九、 同日內與異日間之精確度與準確度( Intraday and Interday Precision and Accuracy) 37
十、 分析方法之確效 38
十一、 酵素活性測試 39
第四章、 結果與討論: 40
第一節、 鹼萃取液濃度對可食用菇蕈類 (1,3;1,6)-β-D-葡聚醣檢測之影響評估 40
一、 以膠體過濾層析法觀察鹼萃取液濃度對可溶性(1,3;1,6)-β-D-葡聚醣分子量之影響 40
二、 鹼濃度不同其 (1,3;1,6)-β-D-葡聚醣含量的變化 46
三、 鹼溶時間測定 48
第二節、 (1,3; 1,6)-β-D-葡聚醣的方法確立 48
一、 分析方法 48
二、 分析方法受非 (1,3; 1,6)-β-D-葡萄聚醣物質之影響 51
三、 準確度 53
四、 精密度 54
五、 檢量線線性及範圍的定量極限及偵測極限 55
第三節、 可食用菇類葡聚醣含量及分支度測定 56
一、 不同年份菇類樣品(1,3;1,6)-β-D-葡聚醣含量測定 56
二、 市售菇蕈樣品之 (1,3;1,6)-β-D-葡聚醣的含量及分支度 62
第四節、 重組酵素活性測試與比較 64
一、 溫度對於酵素活性之影響 64
二、 重組酵素對不同(1,3)-β-D-glucans的活性測試 65
三、 使用合併endo-與recombinant exo-1,3-β-D-glucanase 之酵素水解效果 67
第五章、 結論 75
第六章、 參考文獻: 76
dc.language.isozh-TW
dc.subjectendo-1zh_TW
dc.subjectexo-1zh_TW
dc.subject3-β-D-glucanasezh_TW
dc.subject6)-β-D-葡聚醣zh_TW
dc.subject3-β-D-glucanasezh_TW
dc.subject可食用菇類zh_TW
dc.subject3-β-D-glucanaseen
dc.subjectedible mushroomsen
dc.subjectexo-1en
dc.subject3-β-D-glucanaseen
dc.subjectand endo-1en
dc.subject6)-b-D-glucanen
dc.title"結合酵素-層析法分析可食用菇類中具分支之(1,3; 1,6)-b-D-葡萄聚醣含量"zh_TW
dc.titleBranched (1,3; 1,6)-b-D-glucan content in edible mushrooms: Enzymatic-chromatographic determinationen
dc.typeThesis
dc.date.schoolyear101-2
dc.description.degree碩士
dc.contributor.oralexamcommittee張永和,賴鳳羲,駱錫能,盧訓
dc.subject.keyword(1,3,1,6)-β-D-葡聚醣,可食用菇類,exo-1,3-β-D-glucanase,endo-1,3-β-D-glucanase,zh_TW
dc.subject.keyword(1,3,1,6)-b-D-glucan,edible mushrooms,exo-1,3-β-D-glucanase,and endo-1,3-β-D-glucanase,en
dc.relation.page101
dc.rights.note有償授權
dc.date.accepted2013-08-16
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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