臺灣博碩士論文加值系統

雨生紅球藻富含天然蝦青素,具有高抗氧化性,可去除自由基, 提高免疫力、抗老及抗癌的效果,是迄今為止人類發現自然界最強的 抗氧化劑,遠超過其他的種類的抗氧化劑。本研究分兩階段模式培養 雨生紅球藻,在第一階段策略培養前先進行小量批次培養,發現定時 更換新鮮培養液能有效幫助雨生紅球藻生長。第一階段策略培養中, 設計一套田口實驗方法尋找養殖雨生紅球藻最佳培養條件,實驗結果 以培養基 JM 的濃度為基礎,其中氮、碳、磷濃度比為 1:2:5 時,並 在紅光 LED 照射下,光照強度為 1600 lux,培養 10 天,可得最佳生 長速率為 0.262 d-1。在第二階段策略培養中,實驗結果顯示,以缺乏 主要氮源之培養基 JM 培養並採藍光 LED 照射,光照強度為 8000 lux 下可加速雨生紅球藻由綠轉紅,可得到蝦青素含量約 3.2 mg L-1。使 用直立式光合反應器培養,採未循環的方式培養,培養 7 天後,藻密 度達 50×104 cell mL-1,生長速率可達到 0.291 d-1。

Haematococcus pluvialis contain with abundant natural astaxanthin, which is full of antioxidant capacity that can quench free radicals, improve immune system, and prevent aging and cancer. Up to now, natural astaxanthin has been discovered as a most powerful antioxidant in natural resources. In this study, it’s divided into two steps to cultivate Haematococcus pluvialis. Before the first step, we found that replacing fresh medium regularly can nurture Haematococcus pluvialis effectively. In the first step of culture, use Taiguchi method to search for the optimal experimental condition for Haematococcus pluvialis production. The result shows that the optimum growth rate is 0.262 d-1, with the medium (JM) in N:C:P=1:2:5, irradiation of red light LED at illumination of 1600 lux, culturing for 10 days. In the second step of culture, it shows that the color in Haematococcus pluvialis will be accelerated from green to red, and get the concentration of Astaxanthin about 3.2 mg L-1, with the medium (JM), which is without main Nitrogen, irradiation of blue light LED at illumination of 8000 lux. Using vertical tubular photobioreactor to cultive Haematococcus pluvialis for 7 days in un-circulated condition, the concentration of algae reached 50×104 cell mL-1. As a result, the growth rate of algae was 0.291 d-1.

致 謝 I
摘要 II
Abstract III
目錄 IV
圖目錄 IX
表目錄 XII
第一章 緒論 1
1-1 研究動機 1
1-2 研究目的 4
第二章 文獻回顧 5
2-1 藻類 5
2-2 現代藻類的應用 5
2-2-1 能源 5
2-2-2 保健與營養食品 7
2-2-2-1 藻體 8
2-2-2-2 萃取物 10
2-3 培養方式 13
2-3-1 自營生長 13
2-3-2 異營生長 14
2-3-3 混營生長 15
2-3-4 二階段策略生長 15
2-4 雨生紅球藻(Haematococcus pluvialis) 16
2-5 蝦青素(Astaxanthin) 17
2-5-1 由水產生物提取蝦青素 18
2-5-2 由酵母菌提取蝦青素 19
2-5-3 由藻類提取蝦青素 19
2-6 影響雨生紅球藻生長因子 20
2-6-1 pH值 20
2-6-2 光源 21
2-6-3 溫度 22
2-6-4 營養鹽與微量元素 23
2-7 二階段策略培養雨生紅球藻 24
2-8 培養反應器 25
2-8-1 開放式培養系統 25
2-8-2 封閉式培養系統 27
2-8-3 現階段培養雨生紅球藻之系統 36
第三章 實驗方法與實驗流程 40
3-1 實驗目的及方法 40
3-2 實驗規劃 41
3-3 培養基組成 41
3-4 培養反應器 43
3-5 種源與其來源 47
3-5-1 種原的純化與保存 47
3-5-2 小量活性培養及放大 49
3-6 實驗流程 51
3-6-1 光合混營之培養 51
3-6-1-1 雨生紅球藻細胞大量化 51
3-6-1-2 雨生紅球藻迅速累積蝦青素 52
3-6-2 光合反應器培養 52
3-6-3 藻體收集及乾燥 53
3-6-4 培養條件 54
3-7 實驗分析 55
3-7-1 細胞數測定(Cells Number) 55
3-7-2 藻體密度測定(Dry Weight) 55
3-7-3 光學密度測定(Optical Density) 55
3-7-4 pH測定 56
3-7-5 光照強度測定 56
3-7-6 生長速率測定 56
3-7-7 蝦青素濃度測定 57
3-7-7-1 蝦青素之特徵峰 58
3-7-7-2 蝦青素之檢量線 58
3-8 田口實驗設計 60
第四章 實驗結果與討論 63
4-1 種原處理及逐步放大 63
4-1-1 固態保種 63
4-1-2 小量活性培養及放大 65
4-2 雨生紅球藻細胞大量化 67
4-2-1 營養鹽濃度對一階段策略培養之影響 67
4-2-2 光源對一階段策略培養之影響 69
4-2-3 採用最佳條件培養雨生紅球藻 71
4-3 雨生紅球藻迅速累積蝦青素 74
4-3-1 不同光照強度累積蝦青素之關係 74
4-3-2 藻細胞密度變化與蝦青素累積之關係 78
4-3-3 pH變化與蝦青素累積之關係 79
4-4 光合反應器 80
4-4-1 單一管培養雨生紅球藻 81
4-4-2 光合反應器操作之可行性 84
第五章 結論與未來展望 89
附錄 91
附錄A: 實驗化學藥品與儀器 91
附錄A-1: 實驗化學藥品 91
附錄A-2: 實驗儀器 93
附錄A-3: 藥品儀器供應商資料 94
附錄B: 光源的波長及能量密度分析 96
附錄B-1: 紅光LED 96
附錄B-2: 綠光LED 97
附錄B-3: 藍光LED 98
附錄B-5 T5日光燈管 99
參考文獻 100

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