臺灣博碩士論文加值系統

近年飲茶人口的增加，使得世界各國茶葉用量逐年上升，因此茶葉廢棄物成為需被正視的環境問題。而茶葉廢棄物其主要成分為木質纖維素，在先前文獻中發現，預處理對於木質纖維素生物質轉化過程是相當重要的步驟，藉由改變木質纖維素生物質的結構以利酵素進行糖化作用，另外，亦有文獻利用高壓加工技術提高酵素水解效率，故本研究將兩種茶葉廢棄物 - 高山烏龍茶渣及紅烏龍茶渣，先利用不同溫度及鹼濃度進行預處理，再配合纖維素酶、半纖維素酶與不同條件的高壓加工技術將其分解成小分子醣類，並使用高效液相層析分析其成分與含量，凝膠滲透層析分析其分子量，薄層層析推測茶渣水解液中醣類組成，藉由掃描式電子顯微鏡、X 射線繞射測定和傅立葉轉換紅外線光譜了解茶渣的表徵變化，以此了解 NaOH 預處理和酵素水解對處理後茶渣木質纖維素結構之影響。結果顯示，在預處理實驗中，為了得到較多的寡醣、較少的單糖，因此 40°C、1% NaOH 為最佳預處理條件，在不同加工處理方法中，相比於酵素水解組別，高壓加工技術使水解液中寡醣占比顯著降低，且提高單醣含量，因此選用酵素處理作為實驗組別。而在微觀結構分析的實驗結果中，隨著鹼處理而後酵素水解，可以看到茶渣表面出現孔洞及有序纖維素結構，在結晶度分析的結果，木質纖維素結構的崩解，使結晶度有上升的趨勢。在傅立葉轉換紅外線光譜分析結果中，可以得到木質素的鍵結斷裂、纖維素鍵結的顯露之結果。而在高效液相層析實驗中可以得知高山烏龍及紅烏龍水解液中皆含有木三糖、麥芽糖以及葡萄糖之成分，而在凝膠滲透層析和薄層層析則推測出水解液中含有四糖五糖之寡醣成分。綜合上述，透過本研究可將茶葉廢棄物分解為小分子寡醣類，以達到解決大量茶葉廢棄物的環境問題。

In recent years, the tea consumption in countries around the world has been increasing year by year since more and more people drink tea. Therefore, the environmental issues of tea leaves have to be faced. The main component of tea leaves is lignocellulose. It has been found in previous literature that the pretreatment plays an important role in lignocellulosic biomass conversion process. For example, it is required to prepare an altered structure of lignocellulosic biomass for the propose of accessibility on enzymatic saccharification. In addition, the high pressure processing can also be used to improve enzymes hydrolyzing. In this study, high-mountain oolong tea leaves and red oolong tea leaves were pretreated with different temperatures and alkali concentrations, and then combined with cellulase, hemicellulase and different conditions of high-pressure processing, finally, tea leaves were hydrolyzed into small molecule carbohydrate. The composition and content of the hydrolyzed solution were analyzed by high performance liquid chromatography, the molecular weight of the substance in the solution was analyzed by gel permeation chromatography, the carbohydrate components of the solution was analyzed by thin layer chromatography and the characterization changes of high-mountain oolong tea leaves and red oolong tea leaves were carried out by scanning Electron Microscope, X-ray diffraction and FT-IR Spectrometer, so as to study the influence of NaOH pretreatment and enzyme hydrolysis on the altered lignocellulose structure. In the pretreatment step, for preparing the composition of the tea leaves hydrolysate with more proportion of oligosaccharides and less proportion of monosaccharides, the pretreatment conditions are 40°C and 1% NaOH. Among the different processing methods, to compare with the enzyme hydrolysis group only, HPP significantly reduced the proportion of oligosaccharides and significantly increased the proportion of monosaccharides in the hydrolyzated components. Therefore, enzyme hydrolysis only was finally chosen as the experimental group. In the experimental results of the microstructural characteristics, the alkali treatment and enzyme hydrolysis increased the pores on the surface of tea leaves and ordered cellulose structure. In the results of the crystallinity index, the removal of lignocellulose structure increased the crystallinity slightly. In the results of FT-IR spectrometer analysis, the chemical bonding of the lignin were broken or the chemical bonding of the cellulose were exposed. In the experiments of high performance liquid chromatography, it can be known that the composition of the hydrolysates from high-mountain oolong tea leaves and red oolong tea leaves contained xylotriose, maltose, and glucose. By gel permeation chromatography and thin layer chromatography, it can be inferred that the composition of the hydrolysates contained tetrasaccharides and pentasaccharides. Based on these results, it is known that the tea leaves can be decomposed into small molecular oligosaccharides, so as to solve the environmental problem of a large amount of tea leaves.

中文摘要 I
Abstract II
縮寫表 IV
致謝 VI
壹、 前言 1
貳、 文獻回顧 3
一、 茶渣 3
二、 木質纖維素 (Lignocellulose) 5
三、 木質纖維素之組成分 8
(一) 纖維素 (Cellulose) 8
(二) 半纖維素 (Hemicellulose) 10
(三) 木質素 (Lignin) 12
四、 木質纖維素生物質之預處理方法 13
五、 鹼預處理 (Alkali pretreatment) 16
六、 益生質 (Prebiotics) 18
七、 膳食纖維 (Dietary fiber) 19
八、 寡醣 (Oligosaccharides) 20
(一) 纖維寡醣 (Cello-oligosaccharides, COS) 22
(二) 木寡醣 (Xylo-oligosaccharides, XOS) 23
九、 酵素協同作用 24
(一) 纖維素酶 (Cellulase) 24
(二) 半纖維素酶 (Hemicellulase) 25
十、 高壓加工技術 (High pressure processing, HPP) 27
參、 實驗架構與目的 28
一、 實驗架構 28
二、 實驗目的 29
肆、 實驗材料與方法 30
一、 實驗材料 30
二、 化學藥品及酵素 30
三、 儀器設備 31
四、 實驗方法 32
(一) 茶渣預處理 32
(二) 不同之 HPP 條件偕同酵素水解 32
(三) 高效液相層析分析 (High performance liquid chromatography, HPLC) 33
(四) 掃描式電子顯微鏡 (Scanning Electron Microscope, SEM) 33
(五) X 射線繞射測定 (X-ray diffraction, XRD) 33
(六) 傅立葉轉換紅外線光譜儀分析 (FT-IR Spectrometer) 34
(七) 一般成分分析 34
(八) 凝膠滲透層析分析 (Gel Permeation Chromatography, GPC) 36
(九) 酒精純化 36
(十) 薄層層析分析 (Thin layer chromatography, TLC) 36
(十一) 統計分析 36
伍、 結果與討論 37
一、 預處理條件之探討 37
(一) 高山烏龍茶渣 37
(二) 紅烏龍茶渣 41
二、 不同加工處理之比較 46
(一) 高山烏龍茶渣 46
(二) 紅烏龍茶渣 49
三、 一般成分分析 53
四、 木質纖維素之表徵分析 54
(一) 微觀結構分析 (Microstructural characteristics) 54
(二) 結晶度分析 56
(三) 傅立葉轉換紅外線光譜儀分析 (FT-IR Spectrometer) 60
五、 茶渣水解產物之醣類成分分析 64
(一) 高效液相層析分析 (High performance liquid chromatography, HPLC) 64
(二) 凝膠滲透層析分析 (Gel Permeation Chromatography, GPC) 66
(三) 薄層層析分析 (Thin layer chromatography, TLC) 69
陸、 結論 71
柒、 參考文獻 72
捌、 附圖 86
一、 高效液相層析分析 - 標準曲線 86
二、 凝膠滲透層析分析 - 多醣標準品檢量線 89
三、 核磁共振光譜 (Nuclear Magnetic Resonance, NMR) 90










表目錄
表一、2017 至 2020 年茶葉年生產量 4
表二、木質纖維素生物質中纖維素、半纖維素、木質素、木聚醣之占比 11
表三、在 1% NaOH 下不同處理溫度對高山烏龍茶渣之各類醣類占比 39
表四、在 2% NaOH 下不同處理溫度對高山烏龍茶渣之各類醣類占比 40
表五、在 1% NaOH 下不同處理溫度對紅烏龍茶渣之各類醣類占比 43
表六、在 2% NaOH 下不同處理溫度對紅烏龍茶渣之各類醣類占比 44
表七、高山烏龍茶渣在不同高壓條件偕同酵素水解所產生之各類醣類占比 48
表八、紅烏龍茶渣在不同高壓條件偕同酵素水解所產生之各類醣類占比 51
表九、茶渣之一般成分分析 53
表十、不同加工處理高山烏龍及紅烏龍茶渣之結晶度 59
表十一、高山烏龍茶渣水解液中醣類之分子量及其占比 67
表十二、紅烏龍茶渣水解液中醣類之分子量及其占比 68











圖目錄
圖一、木質纖維素組成 6
圖二、木質素 - 碳水化合物鍵的結構 7
圖三、纖維二糖單元 9
圖四、木質纖維素生物質之各種預處理過程概述 15
圖五、木質纖維素生物質之寡醣類 21
圖六、不同加工處理高山烏龍茶渣及紅烏龍茶渣之 SEM 圖譜 55
圖七、不同加工處理高山烏龍茶渣之 XRD 圖譜 57
圖八、不同加工處理紅烏龍茶渣之 XRD 圖譜 58
圖九、不同加工處理高山烏龍茶渣之 FT-IR 光譜圖 62
圖十、不同加工處理紅烏龍茶渣之 FT-IR 光譜圖 63
圖十一、標準品混入於紅烏龍茶渣水解液之 HPLC 圖譜 65
圖十二、高山烏龍茶渣水解液之 GPC 圖譜 67
圖十三、紅烏龍茶渣水解液之 GPC 圖譜 68
圖十四、高山烏龍茶渣水解液之 TLC 分析 69
圖十五、紅烏龍茶渣水解液之 TLC 分析 70








附表目錄
附表一、高山烏龍茶渣水解液對應麥芽糖、葡萄糖之光譜位置 92

附圖目錄
附圖一、木三糖標準曲線 86
附圖二、麥芽糖標準曲線 87
附圖三、葡萄糖標準曲線 88
附圖四、多醣標準品檢量線 89
附圖五、高山烏龍茶渣水解液之氫光譜 90
附圖六、高山烏龍茶渣水解液之碳光譜 91
附圖七、麥芽糖及葡萄糖之化學結構 92

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