响应面法优化短短芽胞杆菌ch2-22的发酵工艺

摘要

在摇瓶发酵条件下,优化提高短短芽胞杆菌ch2-22芽胞浓度和抑菌活性的发酵培养基和培养条件.首先在单因素试验基础上进行响应面设计对ch2-22的发酵培养基进行优化,然后使用单因素试验方法确定最佳发酵条件,得到优化发酵培养基为淀粉35.05 g/L,豆饼粉26.08 g/L,蔗糖10 g/L,鱼粉5 g/L,NaCl 1 g/L,MgSO40.3 g/L,(NH 4)2 SO43 g/L,MnSO 40.1 g/L,K 2 HPO43 g/L,酵母膏1 g/L,CaCO32 g/L.培养条件为温度32℃,转速180 r/min,装液量100 mL/500 mL,接种量4％,初始pH为7.0,发酵时间45 h.优化后的芽胞数量达到8.1×109 cfu/mL,与优化前的芽胞数量(1.6×109 cfu/mL)相比,提高了3.7倍,优化后发酵液效价达到3350 IU/mL,提高了109％,高于同类菌株的2000 IU/mL.%Fermentation medium and culture conditions to improve the spore concentration and antifungal activity of Brevibacillus brevis ch2-22 at the condition in shaking-flask were optimized. Firstly,based on single factor experiment the ferment medium for B. brevis ch-22 was optimized by response surface methodology,then confirmed the optimized ferment conditions with single factor experiment,obtained the optimized ferment medium was as followed,starch 35. 05 g/ L,soybean powder 26. 08 g/ L,sucrose 10 g/ L,fish meal 5 g/ L,NaCl 1 g/ L,MgSO40. 3g/ L, (NH4 )2SO43 g/ L,MnSO40. 1g/ L,K 2 HPO43 g/ L,yeast extract 1g/ L,CaCO32g/ L,incubated at 32 ℃,sha-king 180,filling at volume 100 mL in 500 mL,inoculum at 4％,initial pH 7. 0,culture for 45 h. The colony-forming units per milliliter were improved from 1. 6 × 109 to 8. 1 × 109 after optimization,and the spore concentration was in-creased by 3. 7 times. The broth valance reached as much as 3350 IU/ mL,increased by 109％,2000 IU/ mL higher than the strain of same kinds.