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研究生:何慧琳
研究生(外文):Hui-Lin HO
論文名稱:利用市售纖維素酶和半纖維素酶降解麥麩生成木寡糖與甘露寡糖
論文名稱(外文):Use of commercial cellulase and hemicellulase on degrading wheat bran to produce xylo-oligosaccharide and mannooligosaccharide
指導教授:陳錦樹陳錦樹引用關係
口試委員:黃至盛林澤群
口試日期:2015-01-30
學位類別:碩士
校院名稱:國立中興大學
系所名稱:食品暨應用生物科技學系所
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2015
畢業學年度:103
語文別:中文
論文頁數:56
中文關鍵詞:麥麩纖維素酶半纖維素酶木寡糖甘露寡糖
外文關鍵詞:wheat brancellulasehemicellulasexylooligosaccharidemannooligosaccharide
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麥麩為麵粉製造過程之副產物,其主要由糊粉層、種皮和果皮部分
所構成。麥麩含有豐富的蛋白質、碳水化合物、脂肪、礦物質,也含膳食纖維及酚酸等生理活性物質。麥麩之粗纖維包含纖維素、半纖維素和木質素,其中半纖維素經過酵素或稀酸降解後得到寡糖。寡糖具有多元的保健功效,可選擇性促進腸道益生菌之生長,達到改變腸道菌相而改善宿主健康。
本研究使用麥麩為基質,以商業酵素纖維素酶和半纖維素酶進行酵素水解,利用HPLC法分析酵素水解液中寡糖組成,探討不同酵素混合比例纖維素酶:半纖維素酶(用量比,w/w)和水解時間(0-12小時)對寡糖生成種類及量之影響,以確立水解之較適條件。
將5 g麥麩加入50 mL水經高溫滅菌後,於50℃下,添加單一酵素纖維素酶0.5 g,結果於水解開始時即有寡糖成分出現,以木三糖最多,10.81 mg/mL,其次為木二糖(2.14 mg/mL)。同樣地,甘露寡糖中,以甘露三糖及甘露四糖為主,分別為0.64 mg/mL及0.63 mg/mL。而添加單ㄧ半纖維素酶組中,於水解開始時,亦以木三糖(8.68 mg/mL)為主,其次為木五糖(3.59 mg/mL),而甘露寡糖則以甘露三糖(0.27 mg/mL)及甘露四糖(0.18 mg/mL)為主。至於混合酵素組,發現當纖維素酶及半纖維素酶比例為0.4:0.1(w/w)時,木三糖在水解開始時即含有13.78 mg/mL,木二糖則在水解6小時後,達4.97 mg/mL。上述結果顯示將纖維素酶及半纖維素酶以單一或混合方式添加於麥麩中,發現在短時間下即有寡糖生成推測來自高溫滅菌使半纖維素組成降解所致,所得水解產物預期可提高益生效果。

Wheat bran is a by-product of milling process during the production of flour. Wheat bran is composed of aleurone layer, testa and pericarp, and is rich in proteins, carbohydrates, fats, minerals, dietary fibers and other bioactive substances such as phenolic acids. Crude fiber in wheat bran includes cellulose, hemicellulose and lignin. After enzymatic or acidic treatment, the hemicellulosic component will be degraded into oligosaccharides. These oligosaccharides could provide multiple health effects, including selectively stimulating the growth of gastrointestinal probiotics and promotion of the host’s health. In this study, five grams of wheat bran were suspended in 50 mL water, followed by autoclaving. Hydrolysis reactions were subsequently performed by adding commercial cellulase and/or hemicellulose. The composition of oligosaccharides in the hydrolysates was analyzed using HPLC method. Effects of the mixing ratio (w/w) of two enzymes and hydrolysis time (0-12 h) on the kinds and amount of oligosaccharides were investigated. The results showed that oligosaccharides especially xylotriose (10.81 g/mL) and xylobiose (2.14 mg/mL) were detected in the hydrolysates right after the cellulase alone (0.5 g) was added. In addition, mannotriose at 0.64 mg/mL and mannotetraose at 0.63 mg/mL were detected, too. As for those using hemicellulase alone, not only xylotriose (8.68 mg/mL) and xylopentose (3.59 mg/mL) were found right after the addition of the enzyme, but also mannotriose at 0.27 mg/mL and mannotetraose at 0.18 mg/mL were detected. Finally, when combinations of cellulase and hemicellulase at ratio of 0.4 to 0.1 (w/w) were used, xylotriose at 13.78 mg/mL was detected immediately after the enzymes were added, followed by the appearance of xylobiose at 4.97 mg/mL after hydrolysis for 6 h. The above results indicated possible thermal destruction of wheat bran occurred during the autoclaving, both xylo- and manno-oligosaccharides were produced shortly whenever cellulase or hemicellulase alone or combinations of both enzymes were used, the hydrolysis reactions led eventually to production of hydrolysates with probiotic effect.

摘要 i
Abstract ii
第一章 前言 1
第二章 文獻回顧 2
一、小麥之介紹 2
二、麥麩之介紹 5
三、半纖維素之介紹 8
(一)植物木質纖維素成分 8
(二)半纖維素之總類 8
四、半纖維素酶之介紹 10
(一)聚木糖酶之介紹 10
(二)聚甘露糖酶之介紹 11
五、寡糖(oligosaccharides)之介紹 13
(一)木寡糖(xylooligosaccharides, XOS)之介紹 13
(二)甘露寡糖(Mannooligosaccharides, MOS)之介紹 14
六、益生質(Prebiotics)之介紹 15
第三章 材料與方法 17
一、實驗材料 17
(一) 實驗原料 17
(二) 商業酵素 17
(三) 化學藥劑 17
二、儀器設備 18
三、套裝軟體 18
四、實驗方法 19
(一)實驗架構 19
(二)麥麩前處理 20
(三)麥麩基本成分分析 20
(四)分析方法 23
第四章 結果與討論 24
一、麥麩基本成分分析 24
二、較適條件探討 24
(一)最適水解時間 24
(二)最適酵素濃度 24
三、木三糖之比較分析 28
第五章 結論 46
第六章 未來展望 47
第七章 參考文獻 48

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