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研究生: 曾聖元
Zeng, Sheng-Yuan
論文名稱: 探討橙黃壺菌BL10產生蝦紅素之生物功能
Investigation of biological functions of astaxanthin in Aurantiochytrium strain BL10
指導教授: 陳逸民
Chen, Yi-Min
學位類別: 碩士
Master
系所名稱: 生物科學與科技學院 - 生物科技與產業科學系
Department of Biotechnology and Bioindustry Sciences
論文出版年: 2018
畢業學年度: 106
語文別: 中文
論文頁數: 61
中文關鍵詞: 橙黃壺菌蝦紅素氧化壓力
外文關鍵詞: Aurantiochytrium, astaxanthin, oxidative stress
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  • 已知一種光合微藻 - 紅球藻,能產生酯化形式(ester form)為主的3S-3′S蝦紅素來遮蔽短波長的強光,作為光保護的手段;紅法夫酵母則能透過游離態(free form)的3R-3′R蝦紅素的產生來抵禦氧化壓力。橙黃壺菌BL10已知在含G-418抗生素及照射藍光的條件下均會變紅;以薄層分析搭配分光光度計分析G-418處理後BL10細胞內的色素組成,證實其以游離態蝦紅素為主。利用高效液相層析搭配特殊的chiral column,分析該蝦紅素的構型,發現其屬於3S-3′S。當以相同的方法分析照射藍光的BL10細胞,藍光照射不同生長階段的BL10時(細胞分裂或油脂累積時期),發現在油脂累積時明顯容易誘發包括β-carotene、canthaxanthin、phoenicoxanthin、astaxanthin在內的色素。另外以氧化亞鐵處理後,BL10只會產生β-carotene;由上述結果可以推測BL10在遭遇不同的逆境時,會產生不同類型的色素來因應環境的變化,來達到保護自身的效果。最後BL10可以透過藍光照射培養的方式,使得本身不僅含有高含量的DHA,還可以含有高抗氧化能力的類胡蘿蔔素,進而增加其附加價值。

    Previous research has demonstrated that geneticin antibiotics can induce Aurantiochytrium strain BL10 to produce a red pigment. The objectives of this study were to investigate why geneticin can induce some species to produce pigmentation, whether the pigment produced by BL10 is the same as that produced by Haematococcus pluvialis, and to induce cells to produce astaxanthin and other carotenoids by stimulating them with blue light irradiation or by adding ferrous sulfate to the medium. We found that exposure to blue light and ferrous sulfate in the culture medium indeed induced the BL10 cells to turn red. Absorption spectroscopy and chiral HPLC analysis revealed that the geneticin-induced red pigment is a 3S-3'S stereoisomer of astaxanthin. Chromatography and mass spectrometry analysis further showed that four kinds of carotenoids were induced by blue light irradiation: β-carotene, canthazanthin, phoenicoxanthin, and astaxanthin. Conversely, ferrous sulfate only induced the production of β-carotene. Due to the different types and activities of reactive oxygen species (ROS) that can be produced under the aforementioned conditions, the four carotenoids produced after the induction have very different antioxidant activities. Therefore, it is likely that the types of carotenoids produced by BL10 are related to intracellular oxidation stress and the source.

    中文摘要 ii 英文摘要 iii 致謝 vii 目錄 viii 圖目錄 x 附表目錄 xii 附圖目錄 xiii 縮寫表 xiv 前言 1 1-1 蝦紅素的簡介 1 1-2 市售蝦紅素的來源與應用 4 1-3 破囊壺菌具有蝦紅素量產的潛力 5 1-4 研究目的 6 材料與方法 8 2-1 藥品與儀器 8 2-2 BL10藻細胞的凍存與活化 9 2-3 G418浸泡處理 10 2-4 藍光照射處理 13 2-5 硫酸亞鐵處理 15 2-6 統計分析 16 結果 17 3-1 G418浸泡處理 17 3-2 藍光照射處理 18 3-3 硫酸亞鐵處理 20 討論 21 參考文獻 24 圖表 32 附錄 55

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