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研究生:邱子銘
研究生(外文):Zih-ming Ciou
論文名稱:幾丁聚醣奈米纖維薄膜對藻藍蛋白純度提升之研究
論文名稱(外文):The Enhancement of C-phycocyanin Purity by Chitosan Nanofiber Membrane
指導教授:張煜光張煜光引用關係
指導教授(外文):Yu-kaung Chang
口試委員:魏毓宏劉炳嵐蔡榮進
口試委員(外文):Yu-Hong WeiBing-Lan LiuJung-Chin Tsai
口試日期:2014-07-14
學位類別:碩士
校院名稱:明志科技大學
系所名稱:化學工程系生化工程碩士班
學門:工程學門
學類:化學工程學類
論文種類:學術論文
論文出版年:2014
畢業學年度:102
語文別:中文
論文頁數:77
中文關鍵詞:藻藍蛋白靜電紡絲奈米纖維薄膜吸附幾丁聚醣改質薄膜
外文關鍵詞:C-phycocyaninelectrospiningnanofiber membraneabsorptionchitosan modified membrane
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鈍頂螺旋藻胞內藻膽蛋白的含量高達60%。依據顏色和吸收特性,藻膽蛋白主要包含藻藍蛋白(CPC)和異藻藍蛋白(APC)。於此項研究中,各種官能基團的奈米纖維薄膜(例如,染料-配體、組氨酸和幾丁聚醣薄膜)分別通過不同的化學合成獲得。對於CPC和APC通過薄膜層析的突破曲線以測量評估其吸附特性。
此外,薄膜層析法用以評估CPC於澄清螺旋藻液的動態吸附行為。所有的實驗都使用薄膜膜組和ÄKTA Prime色譜系統(GE Healthcare Biosciences)進行檢測薄膜的動態吸附行為。首先,染料和組氨酸改質的薄膜被用來研究其吸附性能。不巧的是,該結果顯示,總蛋白(TP)、CPC和APC通過薄膜時同時突破。這是由於此兩種類型的薄膜缺乏選擇性。
因此,進一步進行的實驗,以幾丁聚醣改質之薄膜,並研究其吸附特性。 NaCl濃度(0.1-1.0 M),吸附之pH值(6-8),幾丁聚醣醣濃度(0.1-3.0%),CPC濃度(0.05-0.50 mg/mL)和液體流速(0.2-1.0 mL/min)的影響,於薄膜層析上進行研究該薄膜對CPC(10mL)的吸附性能。
結果顯示,於最佳操作條件下發現幾丁聚醣薄膜對蛋白質選擇性的順序為TP> APC>CPC。於此操作下,APC和TP的分子更容易被吸附在薄膜上。然而,CPC分子最容易穿透薄膜而不被吸附。
因此,將含有CPC的濾液提高了其純度。發現純化倍數和總質量通率分別為3.3倍和66%。於此情況下,最佳的操作條件為:NaCl, 0.1 M,0.3%破碎後螺旋藻液,pH為7,並且液體流速為0.2 mL/min。因此,幾丁聚醣改質之薄膜可用於除去雜蛋白質並進一步提升CPC的純度。

The amount of phycobiliprotein is as high as 60% within Spirulina platens is cells. According to the pigment and absorption characteristics, the phycobiliprotein mainly includes C-phycocyanin (CPC) and allophycocyanin (APC). In this study, various functional groups onto the nanofiber membrane (e.g., dye-ligand, Histidine, and chitosan membranes) were obtained via different chemical synthesis. The adsorption characteristics for CPC and APC by membrane chromatography were assessed by measurements of the breakthrough curves.
Moreover, the membrane chromatography was used and evaluated the dynamic adsorption behavior for CPC in clarified S. platensis. All of the experiments were carried out with a membrane module and the dynamic adsorption behavior of the membranes was monitored using an AKTA Prime chromatographic system (GE Healthcare Biosciences). Firstly, the dye and histidine modified membrane were used to investigate the adsorption performance. Unfortunately, the results showed that the total protein (TP), CPC and APC broke through the membrane at the same time. This is due to the lack of selectively of these two types of membranes.
Hence, further experiments were carried out with chitosan modified membrane to examine the adsorption characteristics. The influences of NaCl concentration (0.1-1.0 M), adsorption pH (6-8), chitosan concentration (0.1-3.0 %), CPC concentration (0.05-0.50 mg/ml), and liquid flow rate (0.2-1.0 mL/min) on the adsorption performance of membrane for CPC (10 mL) were also investigated in membrane chromatography.
The results showed that the order of selectivity of chitosan-membrane for proteins was found to be TP > APC > CPC under the optimal operating conditions. In this case, APC and TP molecules were more easily adsorbed by the membrane. However, the CPC molecules would most easily penetrate the membrane without being adsorbed. Hence, the filtrate containing CPC would enhance its purity. The purification factor and total mass flux was found to be 3.3 folds and 66%, respectively. Under the circumstances, the optimal operating conditions were 0.1 M NaCl, 0.3 % disrupted cells, pH 7, and liquid flow rate 0.2 mL/min. Therefore, the chitosan modified membrane can be used to remove undesired proteins and further to enhance the purity of CPC.
第一章 緒論 1
第一節 研究動 1
第二節 研究目的 2
第二章 文獻探討 3
第一節 藻藍蛋白 3
第二節 聚丙烯腈纖維薄膜 7
第三章 實驗材料與方法 9
第一節 實驗材料 9
第二節 研究設計 11
第三節 螺旋藻液破碎 12
第四節 PAN纖維薄膜製程 13
第五節 奈米薄膜改質 15
第六節 純化因子搜尋 21
第七節 蛋白質液相自動層析儀ÄKTA-prime簡介 28
第八節 光譜分析 29
第四章 結果與討論 30
第一節 薄膜上官能基含量之測定 30
第二節 改質薄膜於FT-IR傅立葉紅外線光譜儀分析 32
第三節 Scanning Electron Microscope (SEM)分析 34
第四節 探討不同陰離子反應性染料改質膜純化效應 36
第五節 探討不同陽離子反應性染料改質膜純化效應 38
第六節 Histidine改質纖維薄膜純化效應 40
第七節 Chitosan改質纖維薄膜純化提升探討 41
第五章 結論 80
參考文獻 81


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