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OPTIMIZATION OF MEDIUM COMPOSITION FOR MONACOLIN K PRODUCTION BY MONASCUS RUBER USING RESPONSE SURFACE METHODOLOGY 黃壬章、張耀南

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OPTIMIZATION OF MEDIUM COMPOSITION FOR MONACOLIN K PRODUCTION BY MONASCUS RUBER USING RESPONSE SURFACE

METHODOLOGY 黃壬章、張耀南

E-mail: [email protected]

ABSTRACT

IN THIS STUDY, MONACOLIN K WAS PRODUCED BY MONASCUS RUBER CCRC 31535 IN FLASK CULTURE. IN PRELIMINARY STUDY, THE SUITABLE CONCENTRATION OF THE GLYCERIN CARBON SOURCE WAS

INVESTIGATED. THE OPTIMUM CONCENTRATION OF GLYCERIN WAS FOUND AT 26.4 ML/L IN THE CULTURE OF 25℃, PH 5.0 AND 25 ML OF THE RICE-GLYCERIN COMPLEX MEDIUM, AND 150 RPM IN THE SHAKER. THE AVERAGE YIELD OF MONACOLIN K WAS 0.131 MG/ML. IN ADDITION, RESPONSE SURFACE METHODOLOGY WAS USED TO OPTIMIZE THE CONCENTRATIONS OF THE RICE POWDER-GLYCERIN COMPLEX COMPOSITIONS AND TO EVALUATE THE EFFECTS OF THE COMPOSITION CONCENTRATIONS ON MONACOLIN K PRODUCTIVITY. THE ANALYSIS OF VARIANCE INDICATED THAT THE QUADRATIC TERMS OF FOUR COMPOSITIONS (RICE POWDER, PEPTONE, GLYCERIN, GLUCOSE) IN THE QUADRATIC MODEL WERE SIGNIFICANT. THE OPTIMUM COMPOSITION FOR MONACOLIN K PRODUCTION WAS FOUND TO BE 34.4 G/L RICE POWDER, 10.8 G/L PEPTONE, 26.4 ML/L GLYCERIN, 129.2 G/L GLUCOSE, 2 G/L KNO3 AND 1 G/L MGSO4.7H2O. WITH THESE COMPOUNDS, THE AVERAGE MONACOLIN K PRODUCTION WAS 0.131 MG/ML AFTER 10 DAYS OF CULTIVATION, WHILE THE PREDICTED MAXIMUM PRODUCTION WAS 0.130 MG/ML. FOR THIS KIND OF COMPLEX MEDIUM, IT WOULD BE GOOD FOR MONACOLIN K PRODUCTION BY M. RUBER CCRC 31535.

Keywords : MONASCUS RUBER, MONACOLIN K, RICE-GLYCERIN COMPLEX MEDIUM, RESPONSE SURFACE METHODOLOGY

Table of Contents

第一章 緒論--P1 第二章 文獻回顧--P8 2.1 紅麴菌簡介--P8 2.2 紅麴菌的代謝物--P9 2.2.1 紅麴色素--P9 2.2.2 膽固醇合成抑制 劑--P13 2.2.3 其他紅麴代謝物--P22 2.3 紅麴菌在保健食品醫藥上的用途--P23 2.4 紅麴菌的培養--P24 2.5 其它有關紅麴菌的 研究--P26 2.6 回應曲面法--P27 2.6.1 回應曲面法之原理--P28 2.7 二水準因子設計--P30 2.8 陡升路徑法--P33 2.9 中心混成設 計--P36 2.10 回應曲面模式適切性之統計檢驗--P37 2.11 因子影響效應之分析--P38 第三章 利用回應曲面法尋求紅麴菌生產 膽固醇合成抑制劑之培養基最適化--P40 3.1 前言--P40 3.2 材料與方法--P41 3.2.1 試驗材料--P41 3.2.2 儀器設備--P41 3.2.3 培 養方法--P42 3.2.4 膽固醇合成抑制劑(monacolin K)之HPLC定量分析--P43 3.3 甘油濃度之測試--P46 3.4 回應曲面法之實 驗設計--P46 3.4.1 部分因子之實驗設計--P48 3.4.2 陡升路徑之實驗設計--P48 3.4.3 中心混成實驗設計--P50 3.4.4 回應曲面模 式適切性之統計檢驗--P50 3.5 回應曲面實驗設計法之結果與討論--P50 3.5.1 部分因子設計實驗--P54 3.5.2 陡升路徑實 驗--P54 3.5.3 中心混成設計實驗--P57 3.5.4 回應曲面模式適切性之統計檢驗--P74 第四章 結論與展望--P77 參考文獻--P79 附 錄--P87

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