产红曲色素的红曲霉菌种筛选及发酵条件优化

文档编号:SW153  文档字数:10021,页数:22

摘要：根据红曲霉菌的性质，本文采用无菌过滤法收集经研磨后的红曲米中的红曲霉孢子悬液，运用平板稀释法，实现了从红曲米中简易分离红曲霉。根据红曲霉的生物学特征，总结出分离红曲霉的原理以及分离培养基的设计与制作，分离纯化的技术方法及过程。培养结果表明：红曲霉是一种耐酸、耐热、耐乙醇的霉菌。纯化后的红曲霉在培养基上的菌落呈圆形，在30℃培养下，48h后开始生长，菌落初为白色，进而变为淡粉色、红色，直接紫红、暗红，但菌落边缘为白色。培养12天后整个菌落呈深红色。显微镜镜检观察菌丝，菌丝有横隔，多核，分支甚繁，分生孢子排列呈链状，分生孢子顶端有球形闭囊壳。红曲霉经过分纯后，针对发酵液培养基中不同碳源、氮源、pH分别进行单因素五水平的试验，确定三个较好的水平，再进行三因素三水平的正交试验以确定红曲霉产红曲色素的最佳条件。在单因素试验中，碳源以玉米粉、淀粉、麦芽浸粉较好；氮源以硝酸钠、大豆粉、蛋白胨较好；pH以3.2、4.0、 4.8较好。通过三水平三因素正交试验，确定红曲霉产红曲色素的最佳培养条件为A1B2C2，即玉米粉、硝酸钠、pH4.0为最佳组合。
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关键词：红曲米；红曲霉；分离；纯化；平板稀释法；红曲色素；色价；液态发酵条件

Abstract：According to the nature of Monascus, the paper collected by sterile filtration of red yeast rice milled in Monascus spore suspension, using plate dilution method, realized from the simple separation of red yeast rice Monascus. According to the biological characteristics of Monascus, Summed up the principle of Monascus separation and separation media design and production, separation and purification method and process technology. Culture results showed that: Monascus is an acid, heat, alcohol-resistant fungus. After purification, the medium of Monascus the colonies were round, under cultivation at 30 ℃, 48h after the growth of the early colonies were white, and then into a light pink, red, direct purple, dark red, but the edge of the colony white. The colonies were cultured for 12 days and the colonies were dark red. Microscopic examination observed mycelium, mycelium with diaphragm, multi-core, branches very complex, spores arranged in chains, conidia are spherical cleistothesium top. After hours of Monascus pure, liquid medium for the fermentation of different carbon sources, nitrogen sources, pH were the single factors and five levels of testing, to determine the level of the three good, and then for three factors and three level orthogonal test to determine the production of Monascus red pigment in the best conditions. In the single-factor test, the carbon source for the corn flour, starch, malt extract powder are better; nitrogen as nitrate, soybean powder, peptone are better; pH to 3.2,4.0, 4.8 are better. Through three levels of the three-factor orthogonal experiment to determine the  production of Monascus red
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pigment of the best medium for A1B2C2, is corn flour, sodium nitrate, pH4.0 was the best combination.
Key words: Red rice; Monascus; separation; purification; plate dilution method; red pigment; color value; liquid fermentation conditions.
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目  录
中文摘要 Ⅰ
英文摘要 Ⅲ
目录 Ⅴ
1. 绪论 1
 1.1  红曲及红曲霉简介 1
 1.2红曲霉分离原理的研究及培养条件的设计 2 
 1.3红曲霉的生物学特征 2
2. 实验部分 3
 2.1主要实验仪器、设备 3
 2.2主要实验试剂 3
 2.3培养基 3
 2.4实验方法与步骤 4
 2.5红曲霉液体培养条件与产色素关系的研究 5
3. 结果与讨论 6
 3.1红曲霉的分离纯化结果与分析 6
 3.2红曲霉液体培养条件的单因素实验结果与分析 9
 3.3正交试验最佳发酵培养条件的结果与分析 11       
4．总结与展望 12
 4.1总结 12
 4.2展望 13
致谢 15
参考文献 16

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