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研究生:徐鴻璿
研究生(外文):SHU, HUNG-HSUAN
論文名稱:利用Oscillatoriales sp. HSW-1 生產藻紅蛋白最佳萃取純化之研究
論文名稱(外文):Study on Optimal Extraction and Purification of Phycoerythrin Produced from Oscillatoriales sp. HSW-1
指導教授:黃思蓴
指導教授(外文):Sz-Chwun John Hwang
學位類別:碩士
校院名稱:中華大學
系所名稱:土木工程學系碩士班
學門:工程學門
學類:土木工程學類
論文種類:學術論文
論文出版年:2015
畢業學年度:103
語文別:中文
論文頁數:39
中文關鍵詞:Oscillatoriales sp. HSW-1藻紅蛋白光照色光營養源
外文關鍵詞:Oscillatoriales sp. HSW-1PhycoerythrinPhotoperiodColor lightsNutrient sources
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近年來由於綠建築的盛行,使藻類的應用又多一項綠建築的領域,除了能處理室
內空氣中的二氧化碳過高的問題,其附加產物藻膽蛋白更被發現具有良好之抗氧化能
力。藻膽蛋白成分裡含有藻紅蛋白(phycoerythrins)、藻藍蛋白(phycocyanin)及異
藻藍蛋白(allophycocyanins),而本實驗室從新竹內灣山區流域取得的藻種中,有一
株Oscillatoriales sp. HSW-1 會產生大量的藻紅蛋白,是現今藻膽蛋白所含有的蛋白中
較為稀有的部分,而其用途可當醫學用的螢光探針、製成保健食品等。本研究所使用
的Oscillatoriales sp. HSW-1 隸屬於顫藻目,外觀呈紅棕色,生存於水溫30~40℃之溫
泉水域,實驗分兩階段進行,第一階段利用光照週期、光照強度、空氣量、溫度及氮
源等變因找出本株Oscillatoriales sp. HSW-1 的最佳生長條件,實驗結果在全日照、光
照強度4000 lux、氣量1000 ml/min、溫度40℃、氮源NaNO3 的條件下的生長最快,
其最高藻乾重(DCW)為0.908 g/L。由於該藻株對於光線較為敏感,當光線超過54.6
umol m-2 s-1 (4000 lux)會受到明顯的抑制,因此對光線強度進行Haldane 抑制模式的模
擬,得到最大比生長率為21.2 (d-1),最大飽和係數(Ks)為122.3 μmol m-2 s-1,抑制係
數為1.475 μmol m-2 s-1,R2=0.8193。最第二階段則用光照週期、色光、額外添加營養
源等方式培養,觀測Oscillatoriales sp. HSW-1 藻紅蛋白之變化,在照度4000 lux,培
養溫度40℃,色光為藍光條件下,粗萃藻紅濃度達0.172 mg/ml,額外添加營養源的
部分以添加Fe 之藻紅濃度為最高,達0.190 mg/ml。
In recent years, due to the prevalence of green building, one more application area of algae was considered in here. In addition to dealing with the problem of excessive carbon dioxide in the indoor air, the additional product of Phycobiliprotein which has a good antioxidation capacity has been found from algae. Phycobiliprotein is a general terms for phycoerythrin, phycocyanin and allophycocyanins, and phycoerythrin is relatively rare as a part of protein in phycobiliprotein. The strain Oscillatoriales sp. HSW-1, which is able to produce large amounts of phycoerythrin was obtained from the watershed of Neiwan, Hsinchu. Phycoerythrin can be used as fluorescent probe on medicine purpose and made of dietary supplement food. Oscillatoriales sp. HSW-1 belonging to Oscillatoriales, normally lives in 30-40 ℃ hot spring water. The experiment was divided into two phases. In the first phase, the environmental influencing factors such as photoperiod, light intensity, air flow rate, temperature and nitrogen sources were considered. The algae, Oscillatoriales sp. HSW-1 could reach the highest dry cell weight of 0.908 g/L under the optimal growth conditions of 24-h photoperiod, 4000 lux light intensity, 1000 ml/min of air flow rate, 40 ℃, and N-source of NaNO3. In addition, due to this algal strain is very sensitive to light, it will be significantly inhibited when the light intensity is more than 54.6 μmol m-2 s-1 (4000 lux). Therefore, the algal growth kinetics was simulated using the Haldane inhibition model. The kinetic model shows the maximum specific growth rate of 21.2 (d-1), the maximum saturation coefficient (Ks) of 122.3 μmol m-2 s-1, and the inhibition coefficient of 1.475 μmol m-2 s-1 with R2 of 0.8193. In the second phase, the operating factors of the photoperiod, color of lights and an additional nutrient source were considered in cultivation of Oscillatoriales sp. HSW-1. The efficiency in production of phycoerythrin was observed. The results show that the optimal operating conditions were using blue-color light, 4000 lux, and 40 ℃, and the crude extracts of phycoerythrin could reach the concentration of 0.172 mg/ml, the amount of which could be up to 0.190 mg/ml if some extra nutrient of Fe source was added.
摘要 i
ABSTRACT ii
誌謝 iv
目錄 v
表目錄 viii
圖目錄 ix
第一章 緒論 1
1-1 研究起源 1
第二章 文獻回顧 2
2-1 選擇藻株及培養 2
2-1-1 挑選藻株 2
2-1-2 培養基選擇 2
2-1-3 光照強度對藻株生長之影響 2
2-2 藻紅蛋白生產最佳化 2
2-2-1 藻紅蛋白之萃取 2
2-2-2 藻紅蛋白計算公式 5
2-2-3 藻紅蛋白之純化 5
2-2-4 光照週期對藻紅蛋白之影響 6
2-2-5 額外添加營養源對藻紅蛋白之影響 6
2-2-6 藻紅蛋白之抗氧化能力 6
第三章 材料與方法 7
3-1 實驗架構 7
3-2 分離需要的藻株 9
3-2-1 藻株來源 9
3-2-2 藻株分離 9
3-2-3 藻株保存 10
3-2-4 藻檢量線之建立 10
3-3 藥品 11
3-4 實驗方法 12
3-4-1 一般培養 12
3-4-2 前培養 13
3-4-3 純化方式 13
第四章 結果與討論 14
4-1 乾重 14
4-2 藻生產最佳化 14
4-2-1 光照週期 14
4-2-2 光照強度 17
4-3 營養源 20
4-3-1 氮源 20
4-3-2 氣量及溫度 23
4-4 藻紅蛋白生產最佳化 27
4-4-1 藻紅蛋白萃取 27
4-4-2 光照週期 27
4-4-3 光線Model 29
4-4-4 色光 30
4-4-5 額外營養源 31
4-6 藻紅蛋白抗氧化 34
第五章 結論 36
參考文獻 37

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