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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 謝志誠(Jyh-Cherng Shieh) | |
dc.contributor.author | Che-Wei Yang | en |
dc.contributor.author | 楊哲維 | zh_TW |
dc.date.accessioned | 2021-06-14T17:00:13Z | - |
dc.date.available | 2013-08-06 | |
dc.date.copyright | 2008-08-06 | |
dc.date.issued | 2008 | |
dc.date.submitted | 2008-07-28 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/40785 | - |
dc.description.abstract | 本研究以向日葵葉柄作為原料本研究以台灣台南田間產的農業廢棄物之向日葵葉柄作為原料,其原料之組成分析為32.09% 纖維素、15.13% 半纖維素、18.83%木質素、21.98% 其他成分、7.29% 灰分、3.02% 醋酸與1.66%之水分,在前處理階段先以0.5%與4% ( w/v )之氫氧化鈉(NaOH)溶液,於1大氣壓下、溫度95℃等條件下,對向日葵葉柄進行前處理90分鐘,判斷其較佳之鹼處理濃度之後,以4%氫氧化鈉(NaOH)溶液為鹼處理濃度,提升其處理溫度為103℃與延長浸泡時間為3小時,此製程本文中以4.0% NaOHIII表示,接著將 4.0% NaOHIII之前處理物進行二階段過氧化氫處理,並分為有無添加氫氧化鈉之方式,使其pH值分別為7.5與11,再利用NREL Methods進行此五種前處理製程之前處理物進行組成分析,除此之外再利用單一酵素之纖維水解酶Cellulclast® 1.5L劑量為21 FPU/g,對此五種前處理物進行酵素水解72小時,並由其組成分析以及水解效果來判斷最佳之鹼前處理製程。在酵素水解階段使用最佳前處理製程4.0% NaOHIII+H2O2V之前處理物於後續酵素水解進行最佳水解製程之探討,並使用混合酵素纖維水解酶Cellulclast® 1.5L搭配葡萄糖苷酶Novozyme® 188,於溫度50℃、pH值4.5與90 rpm等條件下,對酵素活性比例、酵素劑量、基質濃度、批次式與饋料式水解等方式進行酵素水解72小時,由水解液中纖維雙醣、葡萄糖、木糖、還原醣濃度、水解產率與葡萄糖產率等數據來判斷最佳酵素水解製程。醱酵階段中,使用市售酵母發粉,以1 g酵母發粉配置事先滅菌處理之營養液 ( broth ),以1 g︰100 mL比例完成酵母接種物,於恆溫相中控制溫度37℃與120 rpm等條件下活化至少6小時,接著取90mL酵素水解製程之水解液,選擇葡萄糖濃度較高之水解液,也就是批次式基質15%與20%之水解液,以及兩種饋料式基質15%之水解液,搭配10mL之已活化之酵母接種物於溫度37℃與120 rpm等條件下,進行醱酵24小時,探討計算其葡萄糖、木糖、醋酸、乙醇濃度以及其理論值,最後獲致下列結論:
研究結果顯示:最佳前處理製程為以4.0% NaOHIII加上二階段過氧化氫以及添加氫氧化鈉之處理,本文中4.0% NaOHIII+H2O2V表示,其組成分析成分為50.74%纖維素、8.87%半纖維素、19.90%木質素、16.33%其他成分、1.66%水分與2.50%之灰分,前處理後利用濾袋固液分離後之前處理液則利用高效能液相層析儀檢測含有葡萄糖濃度0.43 mg/mL、木糖濃度 0.30 mg/mL與醋酸2.72 mg/mL之濃度,若將前處理液作醱酵處理,由於水解液中可醱酵醣濃度過低,加上有醋酸等會抑制醱酵之因素,顯示鹼前處理液並不適用做醱酵處理。在水解酵素階段,最佳酵素活性比例為FPU/g:CBU/g=10:2,而在此最佳酵素活性比例下之最佳劑量為21 FPU/g 、4.2 CBU/g之混合酵素,可於基質濃度5%、溫度50℃、90 rpm酵素水解72小時下,獲得13.03 mg/mL之葡萄糖濃度、60.96%之水解產率。除此之外若提高水解基質濃度,其葡萄糖產率以及水解產率反而呈現下降狀態,因此在高基質濃度下水解需採用饋料式之酵素水解方式,以同樣為基質濃度15%為例,以饋料式之酵素水解其最終葡萄糖濃度與水解產率分別為31.86 mg/mL與45.66%,而相對的批次式酵素水解基質濃度15%者之葡萄糖濃度與水解產率分別為28.15 mg/mL與42.25%,顯現在高基質濃度下,饋料式酵素水解為較理想之製程。在醱酵階段,將四種高葡萄糖濃度之水解液,以未添加葡萄糖、排毒、滅菌等方式進行醱酵24小時,則可在24小時內消耗大部分葡萄糖,而最終乙醇濃度為13.14~15.40 mg/mL,其理論值約為89.82~94.79%。 | zh_TW |
dc.description.abstract | In this study, sunflower stalks which contained 32.09% cellulose, 15.13% semicellulose, 18.83% lignin, 21.98% others, 7.29% ash , 3.02% acetic acid and 1.66% moisture, was first pretreated by 0.5% and 4.0% NaOH solution at 95℃for 90 mins under atmosphere pressure. In the next step, we change the pretreatment parameters such as temperature, immerse time and second hydrogen peroxide pretreatment. In order to find the best pretreatment process, dried solid fraction was hydrolyzed by enzymes of cellulase from Trichoderma reesei C2730 ( Celluclast 1.5L ) under conditions of pH 4.5 and 50℃ for 72 hours with shaking water bath. In hydrolysis step, different mixing enzyme proportion, loadings and substrate ratios were studied to find out the optimum hydrolysis parameters. And then the hydrolysate was fermented with Saccharomyces cerevisiae under conditions at 37℃ and 120 rpm for 24 hours. The glucose, xylose, acetic acid and ethanol concentration were investigated in fermentation step.
As result, 0.12~0.43 mg/mL of glucose and 0.16~0.30 mg/mL of xylose were observed in liquid fraction. In hydrolysis step, the enzyme loading of 21 FPU/g Celluclast 1.5L plus 4.2 CBU/g Novozyme 188 represented the best balance between economy and efficiency. 260.60 mg/mL of glucose yield and 60.96% of conversion ratio were obtained under this enzyme loading with 5% substrate ratio and rising the substrate ratio did help improving the concentration of glucose and reducing sugar in the hydrolysate. In fermentation step, the four kind of hydrolysates without sterilization and detoxification were fermented for 24 hours and comsuming the most part of glucose. The ethanol concentration was measured around 13.14~15.40 mg/mL, corresponding to 89.82~94.79% of theoretical ethanol yield. | en |
dc.description.provenance | Made available in DSpace on 2021-06-14T17:00:13Z (GMT). No. of bitstreams: 1 ntu-97-R95631021-1.pdf: 809723 bytes, checksum: 6e85dac86391914da7a6796c6c71a6fb (MD5) Previous issue date: 2008 | en |
dc.description.tableofcontents | 目錄
摘要………………………………………………………………………..i Abstract…………………………………………………………………..iii 圖目錄………………………………………………………………….....ix 表目錄………………………………………………………………….....xi 第一章 前言…………………………………………………….………1 第二章 文獻探討………………………………………………….……4 2.1 纖維乙醇之料源…………………………………………………….….……....4 2.2 纖維乙醇之製程………………………………………………….……….…....8 2.2.1 稀酸水解製程.................................................................................................8 2.2.2 濃酸水解製程……………………………………………………….………9 2.3 酵素水解製程…………………………………………………………………11 2.3.1 前處理……………………………………………………………………...13 2.3.2 酵素水解…………………………………………………………………...15 2.3.2.1 纖維水解酶..............................................................................................15 2.3.2.2 纖維水解酶之製備……………………..……………………………....16 2.3.2.3 纖維水解酶之活性..................................................................................17 2.3.2.4 纖維水解酶作用機制..............................................................................19 2.3.2.5 影響酵素水解之因素..............................................................................19 2.3.2.6 表面活性劑之使用..................................................................................21 2.3.3 醱酵................................................................................................................21 2.4 前處理與酵素水解回顧…………………………………………………...…..23 2.5分開酵素水解與醱酵製程回顧…………………………………………….….30 2.6 同步醣化與醱酵製程回顧……………………………………………….…....35 第三章 材料研究方法…………………………………………………41 3.1 實驗材料與設備……………………………………………………………….41 3.2 實驗方法與內容……………………………………………………………….44 3.2.1 前處理………………………………………………………………...…….45 3.2.1.1 鹼前處理濃度分析……………………………………………………...45 3.2.1.2 浸泡式鹼前處理………………………………………………………...46 3.2.1.3 二階段過氧化氫前處理………………………………………………...46 3.2.2 酵素水解………………………………………………………………..…..47 3.2.2.1選擇最佳之FPU與CBU比值.….…..……………….………….….…..47 3.2.2.2 選擇最佳之酵素劑量……………………………………..……….…….48 3.2.2.3 基質濃度對水解之影響…………………………………………………49 3.2.2.4 基質濃度15%下,饋料批次式進行酵素水解(Fed-Batch)…..…..….49 3.2.2.5 基質濃度15%下,饋料批次式進行酵素水解(Fed-Batch)II……......50 3.2.3 酵母菌醱酵.....................................................................................................50 3.2.3.1 酵母菌之活化…………………………………………………………....51 3.2.3.2 酵母菌接種物之製備………………………………………………...….51 3.2.3.3 醱酵……………………………………………………………….…...…52 3.2.4 物料組成成分分析…………………………………….….…………………52 3.2.5 酸不可溶木質素與灰分量測方法………………………..…………………53 3.2.6 酵素活性之檢測方法…………………………………………………...…...54 3.2.6.1 FPU之分析方法…………………………………………………………..54 3.2.6.2 Cellobiase之分析方法………………………………….………………....55 3.2.7 葡萄糖、木糖、其他成分與乙醇等濃度分析…………………..…………56 3.2.7.1 檢量線之建立……………………………………………………….……57 3.2.7.2 葡萄糖、木糖與其他標準樣品之濃度分析………………………….....58 3.2.7.3 葡萄糖檢量線之建立……………………………………………...…..…59 3.2.7.4 木糖檢量線之建立……………………………………………………….60 3.2.8 乙醇濃度分析………………………………………..……………………....62 3.2.8.1 乙醇檢量線之建立……………………………………………………….62 第四章 結果與討論…………………………………………………….64 4.1 向日葵葉柄之組成成分………………………………………………………..64 4.2 鹼前處理………………………………………………………………………..66 4.2.1前處理物之成分………………………………………………………….….66 4.2.2前處理液之成分……………………………………………………………..68 4.2.3 鹼前處理進一步酵素水解分析…………………………………………….69 4.3 酵素水解………………………………………………………………………..71 4.3.1 最佳之FPU與CBU比值…….……………….…………………………….71 4.3.2 最佳之酵素劑量.……………………………………………………………75 4.3.3基質濃度對水解之影響( Batch ) …………………………………………..77 4.3.4 基質濃度15%饋料批次式酵素水解( Fed-Batch )…………………..……78 4.3.5基質濃度15%饋料批次式進行酵素水解( Fed-Batch II )………………...79 4.4 醱酵…………………………………………………………………………….80 第五章 結論……………………………………………………………85 參考文獻…………………………………………………………………91 圖目錄 圖2-1 纖維素之化學結構 6 圖2-2 微纖維之結構 7 圖2-3 稀酸水解製程 9 圖2-4 濃酸水解製程 10 圖2-5 水解與醱酵分離之酵素水解製程 12 圖2-6 醣化與醱酵同步之酵素水解製程 13 圖2-7木質纖維素之前處理目的 13 圖2-8 纖維水解酶作用機制 20 圖3-1 實驗流程圖 45 圖3-2 1.0% ( w/v ) 之葡萄糖標準溶液層析圖 59 圖3-3 葡萄糖檢量線 60 圖3-4 1.0% ( w/v ) 之木糖標準溶液層析圖 61 圖3-5 木糖檢量線 61 圖3-6 1.0% ( v/v ) 之乙醇標準溶液層析圖 63 圖3-7 乙醇檢量線 63 圖4-1 向日葵葉柄之組成成分 64 圖4-2原料組成分析層析圖 65 圖4-3不同酵素季量比例對酵素水解之葡萄糖濃度影響 73 圖4-4 酵素劑量之還原醣濃度與水解產率關係圖 74 圖4-5 不同酵素劑量與水解產率之成本關係圖 76 圖4-6 批次式基質濃度15%水解液之醱酵24小時 81 圖4-7 批次式基質濃度20%水解液之醱酵24小時 82 圖4-8 饋料式( Fed-Batch )水解液之醱酵24小時 83 圖4-9 饋料式( Fed-Batch II )水解液之醱酵24小時 84 表目錄 表2-1 常見之農業與一般廢棄物之纖維素、半纖維素與木質素組成比例 6 表2-2 纖維水解酶之活性測定 18 表3-1 不同酵素劑量比例之吸光值、還原糖濃度及FPU/mL 55 表3-2 不同酵素劑量比例之稀釋倍率與CBU/mL 56 表4-1 其他研究者採用之向日葵葉柄成分比較 65 表4-2 前處理後之前處理物組成分析 67 表4-3 不同鹼處理濃度與其他研究者向日葵前處理後之成分比較 68 表4-4 前處理後之前處理物組成分析( 乾重 ) 68 表4-5 前處理物之前處理液組成濃度分析 69 表4-6 不同前處理條件下,前處理經72小時酵素水解 70 表4-7 不同FPU與CBU比值之水解72小時葡萄糖、還原醣濃度與水解產率 74 表4-8 在最佳酵素活性比例之下之最佳酵素劑量酵素水解72小時 ……76 表4-9 最佳酵素劑量對於不同基質濃度之酵素水解72小時 78 表4-10基質濃度15%下饋料批次式酵素水解72小時 79 表4-11基質濃度15%下饋料批次式II酵素水解72小時 80 表4-12批次式基質濃度15%者後續醱酵24小時 81 表4-12批次式基質濃度20%者後續醱酵24小時 82 表4-13饋料式水解( Fed-Batch ) 後續醱酵24小時 83 表4-14饋料式水解( Fed-Batch II ) 後續醱酵24小時 84 表5-1原料轉換葡萄糖與酒精 89 表5-2 4.0% NaOHII+H2O2V前處理物轉換葡萄糖與酒精 90 | |
dc.language.iso | zh-TW | |
dc.title | 鹼前處理向日葵葉柄產製生質乙醇之研究 | zh_TW |
dc.title | Alkaline Pretreatment of Sunflower Stalks on the Bioethanol Production | en |
dc.type | Thesis | |
dc.date.schoolyear | 96-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 李允中(Yeun-Chung Lee),周楚洋(Chu-Yang Chou),陳力騏(Richie Chen) | |
dc.subject.keyword | 鹼前處理,向日葵葉柄,生質乙醇,酵素水解,醱酵, | zh_TW |
dc.subject.keyword | Alkaline pretreatment,Sunflower stalks,Bioethanol,Enzymatic hydrolysis,Fermentation, | en |
dc.relation.page | 103 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2008-07-30 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 生物產業機電工程學研究所 | zh_TW |
顯示於系所單位: | 生物機電工程學系 |
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