响应曲面法优化多粘类芽孢杆菌HT16产生抗菌蛋白的培养基

Optimization of culture medium for antifungal protein production by paenibacillus polymaxa HT16 using response surface methodology

通过单因子试验对多粘类芽孢杆菌HT16菌株产抗菌蛋白的发酵培养基碳氮源进行优化,确定以硫酸铵、黄豆饼粉为氮源;以蔗糖为碳源。采用Plackett-Burman法筛选出发酵培养基中对产抗真菌蛋白有显著影响的3个组分,分别为土豆、硫酸铵、蔗糖三个因素,以抑菌圈直径大小作为产抗菌蛋白能力大小的判断依据,通过最陡爬坡试验和Box-Behnken试验进一步对3个主要因素进行优化,并对优化结果进行验证,验证结果表明,预测值和实际值有良好的拟合性,此优化模型可靠。最后确定优化发酵培养基组成:30%土豆,0.4%(NH4)2SO4,1.5%蔗糖,0.05%K2HPO4,0.05%MgSO4·7H2O,0.1%黄豆饼粉,pH6.5。该优化发酵培养基,其抗菌活性增加了1倍,且芽孢数提高了2.5倍。

The fermentation medium for antifungal protein production by paenibacillus polymaxa HT]6 wasoptimized.According to single factor experiments, ammonium sulfate and bean cake powder, sucrose were themost suitable nitrogen sources and carbon sources, respectively.The main factors （potato, ammonium sulfate,sucrose）were determined by Plackett-Burman design.The activity of antifungal protein produced was judgedaccording to the size of the inhibition zone diameter.With the help of Box-Behnken design,the maximum responsevalue could be identified.The results showed good fitting between the estimated value and the real value throughverification,and the model was reliable.The optimization of fermentation medium was as follows：30% potato,0.4%ammonium sulfate, 1.5% sources, 0.05% K2HPO4,0.05% MgSO4 · 7H2O,0.1% bean cake powder, pH6.5. Theinhibition zone diameter of the optimized culture medium after fermentation with p.polymaxa HT16 was impoved 1 times compared to that of the original medium.The number of the spores was improved 2.5 times.