东方伊萨酵母高密度培养的研究

Studies on the High-density Cultivation of Issatchenkia orientalis

以东方伊萨酵母为试验菌株,在摇瓶中以YPD液体培养基为基础,通过单因素和响应面试验设计,得到的优化培养基为:葡萄糖质量分数4.06%,酵母粉质量分数4.06%,磷酸盐质量分数0.39%。同时对培养条件和菌体收集条件进行探讨,确定了获得东方伊萨酵母高密度菌体的最佳培养条件:接种量2%,培养基初始p H6.0,培养温度30℃,转速120 r/min。此外,比较了在间歇补料的条件下添加不同补料对菌体高密度培养的影响,结果发现在补料中适当添加氮源可促进菌体的增殖,培养结束时获得的菌体菌落数可达1.71×109CFU/mL。

In this study, an Issatchenkia orientalis has been chosen as the research object. The optimized medium composition of Issatchenkia orientalis has been conducted by means of single-factor experiments and response surface methodology. The optimal fermentation medium is： 4% glucose, 4% yeast extract powder and 0.39% K2HPO4, respectively. At the same time, the conditions for culture and collection have also been discussed in this research, at the best conditions for high-density cultivation of Issatchenkia orientalis（inoculation amount of 2%, p H 6.0, cultivation temperature of 30 ℃ and agitation speed of rotary shaker at 120 r/min）. The sugar-feeding strategy was also investigated and determined as follows： no carbon source added, only carbon source added and carbon source added with moderate nitrogen source. The result showed that supplementary food containing both carbon source and nitrogen source is more benefit to multiplication of Issatchenkia orientalis. The final colony count reach to 1.71×109CFU/mL。