探討培養溫度對Amycolatopsis thermoflava轉化阿魏酸生產香草醛的影響

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李奕徵(Yi-Cheng Lee) 查詢紙本館藏

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化學工程與材料工程學系

論文名稱

探討培養溫度對Amycolatopsis thermoflava轉化阿魏酸生產香草醛的影響

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摘要(中)

香草醛(Vanillin, 4-Hydroxy-3-methoxybenzaldehyde)，又名香蘭素，其為香草莢氣味的主要來源，香草莢是世界上最受歡迎且使用最廣泛的香料之一，獨特的香氣使得它在許多領域被廣泛應用。目前市場上供應的香草醛主要分為化學合成香草醛和天然香草醛，化學合成香草醛在生產過程中，會對環境產生嚴重汙染，而天然香草醛因昂貴且量少，無法滿足市場需求。因此，近年來開始積極研究利用微生物發酵生產香草醛的方法，以取代化學合成香草醛及滿足消費者對天然香草醛的需求。阿魏酸是生物合成香草醛的理想原料，因其便宜易取得，且在農產廢棄物中含量豐富。利用Amycolatopsis thermoflava轉化阿魏酸生產香草醛，可帶來豐厚商業利益，也可解決農產廢棄物所帶來的環境問題，故兼具了商業應用及環境保護雙重效益。
在微生物發酵過程中，環境溫度會極大地影響微生物的生理狀態及發酵行為，故本論文的目的是探討培養溫度對Amycolatopsis thermoflava轉化阿魏酸生產香草醛的影響。單一溫度實驗結果顯示，在發酵初期培養溫度為45℃可得最大菌重濃度，而培養溫度為35℃可得最大香草醛濃度458 mg/L，相較於培養溫度45℃的操作提升209%。藉由兩階段溫度操作提升香草醛產量，第一階段溫度設定45℃適合菌重增長，然後第二階段調整至35℃以利香草醛生成，在此操作下獲得最大香草醛產量為681 mg/L，相較於單一培養溫度35℃的操作提升49%。透過調控培養溫度可有效提升香草醛之產量，且能更加完善Amycolatopsis thermoflava轉化阿魏酸生產香草醛的研究。

摘要(英)

Vanillin (3-methoxy-4-hydroxybenzaldehyde) is one of the major constituents of ‘‘natural vanilla’’ flavor responsible for its characteristic aroma and is the most widely used flavor ingredient in the cosmetics and food industries. The price of ‘‘natural’’ vanillin is very high compared to the chemically synthesized form, mainly due to the limited availability of vanilla pods, which cannot meet the demand of the growing global market for natural vanillin, search for attractive alternative resource is important. Fermentation products are from microorganisms, which are generally considered as natural products. However, only vanillins biotechnologically produced from ferulic acid are considered as food-grade additives by most food safety control authorities worldwide. Furthermore, ferulic acid is rich in agricultural wastes. The microbial fermentation produces high value-added products from inexpensive feedstock such as agricultural wastes, so the cost of production is relatively low, and is more environmental friendly. It is consistent with the concept of sustainable development.
In microbial fermentation procedures, environmental temperature will greatly affect not only the physiological status but also the behavior of fermentation. Ergo, the aim of this study was to investigate the effects of culture temperature on the production of vanillin from ferulic acid by Amycolatopsis thermoflava. The results of experiments showed that the maximum biomass concentration when the culture temperature is 45℃ at the initial stage of fermentation. The optimal vanillin production temperature was 35℃. In this optimal operation, the yield and productivity of vanillin were the highest among other operations, the production of vanillin was 458 mg/L, which was about two hundred and nine percent higher than that obtained from the operation of which culture temperature was fixed at 45℃. Therefore, using the two-stage culture temperature operation, the first stage culture temperature was set at 45℃ to cause more biomass, and then the second stage culture temperature was shifted to 35℃ to enhance the production of vanillin. In this operation, the production of vanillin was 681 mg/L, which was about forty nine percent higher than that obtained from the operation of which culture temperature was fixed at 35℃. As a result, adjusting the culture temperature can make the product formation approaching the ideal type, thereby improving and enhancing the fermentation performance.

關鍵字(中)

★ Amycolatopsis thermoflava
★ 阿魏酸
★ 香草醛
★ 溫度

關鍵字(英)

★ Amycolatopsis thermoflava
★ Ferulic acid
★ Vanillin
★ Temperature

論文目次

中文摘要.........i
ABSTRACT.........iii
誌謝.........v
目錄.........vi
圖目錄.........x
表目錄.........xiii
一、緒論.........1
1-1 研究背景.........1
1-2 研究動機.........4
1-3 研究目的.........6
二、文獻回顧.........7
2-1 香莢蘭介紹.........7
2-2 香草醛介紹.........10
2-2-1 香草醛基本性質.........10
2-2-2 香草醛市場價值.........11
2-2-3 香草醛生產方法.........13
2-3 香草酸介紹.........17
2-4 阿魏酸介紹.........18
2-5 實驗菌株介紹.........19
2-6 影響發酵表現的物理化學因子.........21
2-6-1 碳源.........21
2-6-2 氮源.........22
2-6-3 無機鹽類與微量元素.........22
2-6-4 酸鹼值.........22
2-6-5 攪拌速率.........23
2-6-6 溫度.........23
2-7 農產廢棄物提取阿魏酸.........24
三、材料與方法.........27
3-1實驗規劃.........27
3-2 實驗材料.........28
3-2-1 實驗菌株.........28
3-2-2 實驗藥品.........29
3-2-3 實驗儀器與設備.........30
3-3 實驗方法.........32
3-3-1 菌種保存.........32
3-3-2 菌種固態培養.........32
3-3-3 液態種瓶培養.........33
3-3-4 香草醛生產的培養溫度實驗.........34
3-3-5 香草醛生產的兩階段培養溫度實驗.........35
3-3-6 鹼處理提取阿魏酸.........37
3-4 分析方法.........39
3-3-1 菌重濃度分析.........39
3-4-2 pH值分析.........40
3-4-3 還原糖濃度分析.........40
3-4-4 阿魏酸、香草醛和香草酸濃度分析.........42
四、結果與討論.........45
4-1 不同培養溫度對香草醛生產的影響.........45
4-1-1 不同培養溫度對Amycolatopsis thermoflava生長的影響...45
4-1-2 不同溫度對轉化阿魏酸生產香草醛和香草酸的影響.........48
4-1-3 不同培養溫度的發酵動力曲線.........50
4-2 兩階段溫度對香草醛生產的影響.........55
4-2-1 兩階段溫度對Amycolatopsis thermoflava生長的影響...55
4-2-2 兩階段溫度對轉化阿魏酸生產香草醛和香草酸的影響......58
4-2-3 兩階段溫度的發酵動力曲線.........60
4-3 不同的鹼處理條件對阿魏酸提取之影響.........65
4-3-1 不同提取時間對阿魏酸提取量的影響.........65
4-3-2 不同提取溫度對阿魏酸提取量的影響.........66
4-3-3 不同氫氧化鈉濃度對阿魏酸提取量的影響.........67
4-3-4 不同固液比對阿魏酸提取量的影響.........68
4-3-5 不同原料對阿魏酸提取量的影響.........69
4-3-6 鹼性溶液重複提取阿魏酸之影響.........70
五、結論與建議.........71
5-1 結論.........71
5-2 建議.........72
參考文獻.........74

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指導教授

徐敬衡(Chin-Hang Shu)

審核日期

2019-8-13

推文