《嗜热链球菌高密度培养及冷冻保护的研究》-毕业设计（论文）.docVIP

分类号 TS252.1 U D C 579 学校代码 10129 学 号 嗜热链球菌高密度培养及冷冻保护的研究 The Study on High Cell Density Cultivation and Freeze-drying Protection of Streptococcus thermophilus 申 请 人： 学科门类： 学科专业： 研究方向： 指导教师： 论文提交日期： 摘 要 试验以嗜热链球菌为研究对象，进行发酵酸奶优良菌株的筛选，并以嗜热链球菌ND03为模型菌株进行培养基优化、高密度发酵及冻干保护的研究。主要研究结论如下： (1)289株嗜热链球菌以商业发酵剂嗜热链球菌Yc为对照菌，依次经过发酵酸度、发酵时间、发酵特性和感官评价的分析，最终得到具有潜在酸奶发酵优势的12株优良嗜热链球菌。 (2)嗜热链球菌ND03以AST为基础培养基，依次经过碳源、氮源、碳氮总量及比例、微量元素、缓冲盐和生长因子的筛选，采用响应面设计对各成分比例进行优化后得到最适培养基。在此培养基中于37℃培养8～10h，培养液活菌数可达到1.02×1010CFU/mL，比优化前提高近13倍。 (3)在筛选出的最适培养基中对嗜热链球菌ND03静态培养条件进行优化，并在5 L液体发酵罐上对高密度发酵工艺进行优化。优化后嗜热链球菌ND03发酵时间为6h～6.5h，比优化前至少缩短2h。发酵液活菌数达到1.80×1010CFU/mL，比优化前提高76%。 (4)对嗜热链球菌ND03的保护剂添加比例、预冻温度、预冻时间进行优化，得到保护剂添加比例为1:6，最佳预冻温度为-40℃，最佳预冻时间为6h。对冻干保护剂配方中的糖类、多元醇类、聚合物类、盐类进行研究。在最佳冻干保护效果下，ND03菌粉中活菌数为1.83×1011CFU/g，冻干存活率达到84.3%，比优化前提高约20%。 (5)对筛选出的12株优良嗜热链球菌，按模型菌株ND03优化的高密度发酵工艺及冷冻干燥工艺进行高密度发酵及菌粉生产。12株嗜热链球菌中9株的发酵液活菌数达到1×1010CFU/mL，5株菌的冻干存活率达到80%，表明优化的高密度发酵工艺和冷冻干燥工艺能满足大部分嗜热链球菌高密度发酵及菌粉生产要求。 关键词：嗜热链球菌；培养基优化；高密度发酵；保护剂；冷冻干燥 The Study on High Cell Density Cultivation and Freeze-drying Protection of Streptococcus thermophilus Abstract The potential excellent strains in fermenting yogurt were screened from 289 S. thermophilus. S. thermophilus ND03 was chosen as a representative strain, and conducted with the study on medium optimization, high cell density culture and freeze-dried protection. The main conclusions are as follows: 1 By detecting the fermentation characteristics such as pH, time, sensory evaluation, 12strains were screened from 289 S. thermophilus by comparing with the fermentation characteristics of S. thermophilus Yc, which was obtained from a commercial starter. 2 Based on the medium AST, all ingredients were screened and their concentration was adjusted in order to form the optimum medium for S. thermophilus ND03. When cultivated in the optimum medium at 37℃ for 8~10h, the viable count of fermented broth can get to 1.02×1010CFU/mL, which was 14times as many as