利用二階段醱酵法生產嗜鹽菌Haloferax mediterranei 類胡蘿蔔素

PRODUCTION OF CAROTENOIDS FROM HALOPHILIC HALOFERAX MEDITERRANEI USING TWO-STAGE FERMENTATION

劉耀中(Yao-Chung Liu)

指導教授 ： 林銘澤

大同大學/工程學院/生物工程學系所/碩士(2011年)

摘要

嗜鹽菌Haloferax mediterranei 之類胡蘿蔔素具有抗氧化、保護細胞傷害等功效。H. mediterranei 於不同碳源、氮源及鹽類和不同濃度下，對其菌體生長及類胡蘿蔔素的生產有很大的影響。增加葡萄糖濃度對菌體生長速率無明顯變化，但高碳源濃度能維持較長久的菌體生長，因此葡萄糖濃度越高，其最大菌體量越高；以醋酸為碳源時，其菌體生長速率皆遠低於葡萄糖為碳源者；而低濃度丁酸 (≤0.5%) 不會抑制菌體生長，但丁酸濃度高於1%時，隨濃度增加會抑制菌體生長以及最大菌體量。另外，隨時間增加總類胡蘿蔔素產量亦隨之提高，而以0.5%丁酸為碳源者產量最高。
利用二階段批次醱酵培養以誘導類胡蘿蔔素的生產。首先，以2%葡萄糖的ATCC1176 培養基進行第一階段醱酵培養，經36 小時後，分別饋入以葡萄糖、丁酸或醋酸為碳源且不含NaCl 之等體積培養基進行第二階段培養，發現NaCl 濃度減半後，加入0.1%醋酸會抑制菌體生長，其生長較為緩慢，且葡萄糖使用率亦下降。而0.1%丁酸與葡萄糖其菌體生長及葡萄糖代謝速率相近，但丁酸培養基則可快速誘導類胡蘿蔔素的產生，而葡萄糖培養基誘導生產類胡蘿蔔的素速率最慢。

關鍵字

二階段醱酵法 ； 類胡蘿蔔素 ； 嗜鹽菌

並列摘要

Carotenoids of Haloferax mediterranei have anti-oxidant effect and protection against cell damage. The proliferation of H. mediterranei and generation of carotenoids are greatly influenced by different carbon sources and salt concentrations. Increasing the concentrations of glucose, the growth rate didn’t change obviously, but elongated the time for the cell growth and had higher biomass. The cell growth rates were lower by acetate as carbon source than ones by glucose. Lower concentrations of butyrate (≤0.5%) didn’t inhibit the growth, but the cell growth rates and maximum biomass were decreased when the media with butyrate were more than 1%. Additionally, the total production of carotenoids were increased along the culture time. The maximum production was in 0.5% butyrate as carbon source.
The two-stage fed-batch fermentation in the presence of different carbon sources and without NaCl was used for the growth of H. mediterranei and enhancing the production of carotenoids. At first, ATCC1176 medium with 2% glucose was used for first stage of fermentation. After 36 hours, the equal volumes of media with 0.2% different carbon sources and without NaCl were fed, respectively, for second stage of fermentation. The concentrations of NaCl in media were decreasing 50% to induce the productions of carotenoids. The growth rate and the utilization efficiency of residual glucose was inhibited by the addition of 0.1% acetate, but were similar between glucose and butyrate. However, the butyrate boosted the production of carotenoids, and the enhanced production of carotenoids was lowest for glucose.

並列關鍵字

HALOFERAX MEDITERRANEI ； CAROTENOIDS ； TWO-STAGE

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