利用被孢霉发酵生产花生四烯酸工艺配方优化研究

Optimization of the culture medium of mortierella alpina in the production of arachidonic acid by fermentation

利用高山被孢霉Mortierella alpina I49-N18发酵生产花生四烯酸。优化筛选出最适于高山被孢霉斜面培养、种子摇瓶培养以及摇瓶发酵培养的配方,稳定的培养基配方为后续发酵生产提供保障。通过培养基配方单因子试验分别对碳源、氮源、无机盐、氨基酸、植物油等原料的合适添加量进行初步确定,然后设计正交实验进行验证。考察初始葡萄糖浓度在高山被孢霉发酵生产花生四烯酸过程中对脂肪酸组分的影响,分析脂肪酸主要组分在发酵过程中的变化趋势。考察结果显示发酵培养基中补充添加800 1000mg/kg磷酸盐,添加600 1000mg/kg谷氨酸钠盐、添加0.04%0.12%植物油可以有效提高发酵生产花生四烯酸的产量。通过发酵工艺放大,在200m3发酵罐规模的发酵生产中得到应用,发酵产量得到大幅度提升,培养7d发酵生产花生四烯酸(ARA)的产量达到10g/L以上。

Mortierella alpina I49-N18 was used to produce Arachidonic acid.The culture to best suitable for Mortierella alpina slant culturing was optimized.Culturing and fermentation was done by flask shaking.Through the single factorial experiment,the raw material,the source for carbon and nitrogen,the content of inorganic salt,amino acid,vegetable oil were all tested on the influence of the culture.The orthogonal test was then carried out to validate.The effect of start glucose concentration on the constitution of fatty acids during the fermentation with Mortierella alpine was studied by the analyzing the main fatty acids during fermentation.The experiment result showed that adding 8001000mg/kg phosphate,6001000mg/kg monosodium glutamate,0.04%0.12% vegetable oil into the fermentation medium increased the yield of fermentation products.The above formular was applied in 200m3 fermentor,the fermentation yield increased significantly.The concentration of arachidonic acid（ARA）reached 10g/l after 7-day＇s fermentation.