探討Aspergillus niger利用甘蔗渣生產檸檬酸之研究

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姓名

吳柏昀(Po-yun Wu) 查詢紙本館藏

畢業系所

化學工程與材料工程學系

論文名稱

探討Aspergillus niger利用甘蔗渣生產檸檬酸之研究
(Production of citric acid by Aspergillus niger using acid hydrolysates of sugarcane bagasse)

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摘要(中)

生物煉製（Biorefinery）泛指利用生質物提煉生質燃料或生物特用化學品，為近年來重點開發的技術，其中利用農林廢棄物中的木質纖維素之煉製技術更是積極開發的項目。本研究嘗試利用台灣地區常見的農業廢棄物甘蔗渣的木質纖維素作為原料，使用黑曲黴(Aspergillus niger)進行液態發酵產檸檬酸。
由於直接使用未處理之甘蔗渣做為碳源利用效果並不佳，藉由稀酸水解法可將甘蔗渣中半纖維素水解釋放出可利用之單醣。另外進行二次酸水解可以提高醣濃度，然而對微生物具有抑制現象的有害物質乙酸、5-羥甲基糠醛(HMF)以及糠醛(Furfural)等亦隨之提升，其中乙酸及5-羥甲基糠醛(HMF)已達到抑制菌體生長濃度值，為主要抑制菌體生長的有害物質。本研究採取活性碳吸附處理方式進行這類有害物質之移除，經過移除後的酸水解液可成功的進行檸檬酸發酵生產，其中檸檬酸的最大產量與組成培養基所建構對照組相較之下可達78％（酸水解液：9.3 g/L對照組：11.77 g/L），且比生長速率亦相近（酸水解液：0.075；對照組：0.083）。

摘要(英)

Biorefinery, which is one of the renewable technologies, has been developed for few years. And the use of lignocellulosic materials available in agricultural wastes as a source of raw material for citric acid is interest because of their renewable nature and abundance. The present study discusses the hydrolysate of sugar cane bagasse as carbon source for the production of citric acid by Aspergillus niger .
First, we use sugar cane bagasse in medium directly, but the results show that it was not a good method. Then we use dilute acid method for sugar cane bagasse pretreatment, which is one of the few technologies to break down the protective structure of lignin and hemicellulose, but during pretreatment process, several compounds like acetic acid and 5-hydromethylfurfural and furfural were toxic to microorganisms.
However, there need some methods to detoxify the acid hydrolysates. In this study toxic compounds were removal from hydrolysate by activated charcoal, so that the hydrolysate could be used by A. niger .After detoxic process, the maximum growth rate (μmax) and citric acid concentration of A.niger fermentation were closed to those of the control medium.

關鍵字(中)

★ 生物煉製
★ 檸檬酸
★ 酸水解液

關鍵字(英)

★ Biorefinery
★ Acid hydrolysate
★ Citric acid

論文目次

摘要 I
Abstract II
目錄 III
圖目錄 VII
表目錄 X
第一章、緒論 1
1-1 研究動機 1
1-2 研究目的 2
第二章、文獻回顧 3
2-1 檸檬酸之介紹 3
2-1-1 檸檬酸簡史 3
2-1-2 檸檬酸來源 4
2-1-3 檸檬酸應用 8
2-2 黑曲黴發酵生產檸檬酸 11
2-3 影響檸檬酸發酵營養條件 13
2-3-1 碳源 13
2-3-2 氮源 14
2-3-3 無機鹽類 14
2-3-4 微量元素 15
2-3-4 其他 15
2-4 木質纖維素 16
2-5 木質纖維素前處理 21
2-5-1 機械破碎法 22
2-5-2 氨纖維爆破法 22
2-5-3 蒸氣爆破法 (Stream explosion，STEX) 23
2-5-4 二氧化碳爆破法 (CO2 Explosion) 24
2-5-5 超音波微波處理法 24
2-6 稀酸水解木質纖維素製程 25
2-6-1 稀酸水解處理法 25
2-6-2 稀酸水解有害物質去除方法 27
2-6-2.1 氣提法(Air Stripping) 27
2-6-2.2 閃蒸法(Flash evaporation) 28
2-6-2.3 過量鹼處理法(over-liming) 28
2-6-2.4 活性碳吸附法 29
2-7 甘蔗渣的應用 31
第三章、實驗材料與方法 32
3-1 實驗材料 32
3-1-1 實驗菌株 32
3-1-2 實驗藥品 32
3-1-3 實驗儀器及其他設備 33
3-1-4 實驗裝置 35
3-2 實驗流程設計 36
3-3 實驗方法 38
3-3-1 Aspergillus niger 菌種保存及培養方法 38
3-3-2 發酵試驗操作條件 42
3-3-3 直接利用蔗渣測試效果實驗 43
3-3-4 稀酸不同水解時間前處理實驗 43
3-3-5 酸水解液有害物質影響測試 43
3-3-6 酸水解液有害物質移除效果測試 44
3-3-7 酸水解液中和鹼離子影響性測試 44
3-4 分析方法 45
3-4-1 發酵液處理流程圖 45
3-4-2 菌重分析 45
3-4-3 葡萄糖殘量分析（還原醣測定） 46
3-4-4 有害物質含量分析 47
3-4-5 檸檬酸含量分析 50
3-4-6 酸水解液總醣含量分析 51
第四章、結果與討論 53
4-1 蔗渣利用性 53
4-1-1 利用蔗渣為直接碳源測試效果 53
4-1-2 稀酸不同水解時間前處理效果 56
4-2 蔗渣酸水解液處理 57
4-2-1 酸水解液中醣類含量分析 58
4-2-2 總醣條件(Total reducing sugar)的選擇 58
4-2-2.1 總醣條件選擇後續影響 60
4-2-3 酸水解液有害物質影響性 62
4-2-3 酸水解液有害物質移除效果 68
4-2-4 酸水解液中和鹼離子影響性 69
4-3 酸水解液應用 72
4-3-1 氣泡式發酵槽實驗 72
第五章、結論與建議 75
5-1 結論 75
5-2 建議 76
第六章、參考文獻 77

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指導教授

徐敬衡(Chin-hang Shu)

審核日期

2009-7-15

推文