三孢布拉氏霉菌发酵生产β-胡萝卜素的条件研究

论文编号:SW228  论文字数:12763,页数:24

摘  要：本文以三孢布拉氏霉菌正、负菌为生产菌种，筛选了产孢子培养基及优化了β-胡萝卜素的生物合成条件。其最佳的产孢子培养基：麦芽汁双层平板，在此培养基中，菌丝生长茂盛，产孢子多；通过对不同碳、氮源的组合筛选，确定最佳的碳、氮源及发酵终止时间，其结果为：淀粉作为碳源，黄豆粉作为氮源，发酵144 h，β-胡萝卜素合成量最大。在此基础上，利用Plackett-Burman设计分析了发酵培养基中各成分对β-胡萝卜素产量的影响，其结果为：对β-胡萝卜素产量影响的重要因素为β-紫罗酮，VB1，淀粉，植物油。最后，对该4因素以L9(34) 正交设计进一步优化，得出最佳的培养基组成为：淀粉41 g/L，黄豆粉20 g/L，植物油35 ml/L，磷酸二氢钾2 g/L，硫酸镁1 g/L，VB1 0.030 g/L，发酵40~44 h后，添加0.05%的异烟肼12 ml/L及10%的β-紫罗酮15 ml/L。β-胡萝卜素的发酵产量较比优化前发酵培养基提高了16.78%。
关键词：三孢布拉氏霉菌；β-胡萝卜素；培养基；Plackett-Burman设计；正交设计

Abstract：The produce industrially of β-carotene by Blakeslea trispora (+ −) was studied, and the process of β-carotene synthesis was optimized. The best spore produced medium was filtrated: the double layer wort plate, the mycelial growed flourishing, and produced much more spores. The fermentation medium of synthesizing β-carotene with Blakeslea trispora was optimized. The optimum carbon source, nitrogen source and the fermentation time was obtained through the compounding of different carbon source and nitrogen source. The results showed that faecula was used as carbon source, bean flour was used as nitrogen source, fermentation time was 144 h. Base on this result, the important factors which affect fermentation conditions of β-carotene were screened by Plackett-Burman design. Then by using orthogonal test, these important factors were optimized. Results showed that the best fermentation medium were faecula, 41 g/L; bean flour, 20 g/L; Vegetable oil, 35 ml/L; KH2PO4, 2 g/L; MgSO4, 1 g/L; VB1, 0.030 g/L. Isoniazid and β-ionone was appended after fermenting 40~44 h. Its yield of β-carotene increase 16.78% compared with the original fermentation medium.
Keywords: Blakeslea trispora; β-carotene; Culture medium; Plackett-Burman design;Orthogonal design