研究生: |
吳明儒 Wu, Ming-Ju |
---|---|
論文名稱: |
使用氣相層析/哨音檢測技術對酵母菌固化微管陣列薄膜發酵過程中乙醇之即時監測 The Use of a Gas Chromatography/Milli-whistle Technique for the On-line Monitoring of Ethanol Production Using Microtube Array Membrane (MTAM) Immobilized Yeast Cells |
指導教授: |
林震煌
Lin, Cheng-Huang |
學位類別: |
碩士 Master |
系所名稱: |
化學系 Department of Chemistry |
論文出版年: | 2016 |
畢業學年度: | 104 |
語文別: | 中文 |
論文頁數: | 91 |
中文關鍵詞: | 微型發音哨 、氣相層析儀 、LabVIEW 、微管陣列薄膜 、發酵 |
英文關鍵詞: | milli-whistle, gas chromatography, LabVIEW, MTAMs, fermentation |
DOI URL: | https://doi.org/10.6345/NTNU202204414 |
論文種類: | 學術論文 |
相關次數: | 點閱:169 下載:0 |
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在生質酒精的發展上,篩選發酵效果良好的菌株,是其中一項重要的環節。透過發酵反應過程中葡萄糖含量、乙醇產量、細胞活性隨時間變化的曲線,能幫助我們了解菌株的表現。但為了得到這些數據,研究人員得固守在儀器與反應裝置旁,佔用機器與研究人員的時間。另外,由於將微生物細胞固定後,能提升其代謝活性,提高其代謝產物產量,並降低抑制物質對細胞的影響。
本研究利用同軸靜電紡絲的技術,製作出包覆酵母菌的微管陣列薄膜,並以在本研究中自組裝的電磁閥進樣裝置與自組裝氣相層析/哨音檢測裝置,以五分鐘作為間隔,對於酵母菌發酵反應過程中產生的二氧化碳、乙醇進行自動化的即時線上偵測,並評估酵母菌在此薄膜應用於酒精發酵的表現。本次研究中,對包覆釀酒酵母的微管陣列薄膜一共進行11次發酵反應循環。二氧化碳訊號隨時間變化曲線的部分,由於電磁閥進樣器,一次只將反應槽少量氣體注入到分析儀器中,直到發酵反應結束為止,二氧化碳不斷累積,而當發酵反應結束後,二氧化碳不在產生,隨每次進樣的動作,反應槽內的二氧化碳逐漸減少,導致二氧化碳的變化曲線,呈現類似於高斯分佈,此曲線的頂點即是發酵反應結束地時間點,第一次至十一次發酵反應的時間分別為1.65、1.07、1.04、0.88、0.62、0.71、0.69、0.67、0.66、0.48、0.57天,此時間點的二氧化碳造成的頻率變化分別為26.54、31.70、31.13、37.35、43.50、45.94、46.72、45.18、50.80、60.88、59.25 Hz,經檢量線換算後,進樣的二氧化碳體積分別為64.20、77.03、75.62、91.07、106.36、112.42、114.36、110.53、124.50、149.55、145.50 μL。乙醇訊號隨時間變化曲線的部分,由於在定溫、定壓下,乙醇飽和蒸氣壓與溶液中乙醇濃度相關,因此可以乙醇的飽和蒸汽壓結果推估乙醇溶液的濃度,但在發酵反應結束時,乙醇無法在此時就達到飽和蒸氣壓,因此,在曲線達平衡的時間點相較於二氧化碳得到的結果有延遲的現象,此處取實驗中最後時間點收取的數據判斷乙醇的濃度。第一次到第十一次達平衡後乙醇造成的頻率變化分別為0.92、0.97、1.07、1.00、0.93、0.95、0.93、1.15、1.16、1.27、1.22 Hz,經檢量線換算後,乙醇溶液的重量百分比濃度分別5.15、5.59、6.92、6.33、5.75、5.96、5.77、6.18、6.25、6.96、6.64%,而理論產量為8.27%。由此數據能證實,透過氣相層析/哨音檢測技術能成功地對酵母菌微管陣列薄膜發酵過程進行有效的即時線上偵測,同時,也說明酵母菌微管陣列薄膜是一項在生質酒精發酵上具有潛力的技術。
另一方面,本研究同時也進行可攜式氣相層析/哨音檢測裝置的開發。開發的裝置,長36 公分、寬29公分、高17公分,總重6公斤,以氮氣作為載流氣體下,對氫氣、氦氣、氧氣、氬氣、二氧化碳,都有良好的線性結果,線性範圍都有兩個數量級,也能在管柱烘箱溫度60℃下,分離並偵測混合樣品中,氧氣、丙酮、甲醇、乙醇的訊號,顯示在此成功地開發可攜式氣相層析/哨音檢測裝置。
Hollow, poly (l-lactic acid) microtube array membranes (MTAM) were used in preparing membranes that contained immobilized yeast cells. To evaluate the performance of the developed system for continuous and fed-batch fermentation, a gas chromatography/milli-whistle device was used to on-line monitor the production of ethanol. The milli-whistle was connected to the outlet of a GC capillary, and when the fermentation gases and the GC carrier gas pass through it, a sound with a fundamental frequency is produced. The online data obtained for frequency vs. retention time can be recorded after a fast Fourier transform. In typical bioethanol fermentation, the yeast cells cannot be recycled, whereas the artificial yeast-MTAMs can be recycled. The hollow-MTAM containing immobilized yeast cells significantly enhanced bioethanol productivity and represents a novel, promising technology for bioethanol fermentation. On the other hand, a portable GC/whistle device was also developed for on-line fermentation monitoring. Herein, when nitrogen was used as carrier gas, a high linearity can be obtained from the range from 5 to 250 μL (injection volume), even various gases were used. Our data indicate that the GC/whistle device, which is economical and stable, is very useful detector for long term monitoring. Further applications can be to expected.
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