嗜热链球菌高密度培养及冷冻保护及研究.pdfVIP

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 10 1.02×10 CFU/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 10 6h～6.5h，比优化前至少缩短 2h。发酵液活菌数达到 1.80×10 CFU/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， 11 ND03 菌粉中活菌数为 1.83×10 CFU/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 10 菌数达到 1×10 CFU/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 adjuste