利用响应面分析法优化深绿木霉Tr16液体发酵产孢培养基研究

Optimization the Culture Medium of Liquid Fermentation for Spore Production of Trichoderma atroviride Strain Tr16 by Utilizing Response Surface Analysis Method(RSAM)

采用响应面分析法(RSAM)对深绿木霉(Trichoderma atroviride)菌株Tr16液体发酵产孢培养基进行优化。通过Plackett-Burman(PB)方法,先筛选出培养基中影响产孢量的3个主要因素:MgSO 4、H 8MoN 2O 4和酵母提取物,再运用“最陡爬坡路径法”和RSAM,确定主要因子之间的交互影响并筛选出最佳液体发酵培养基配方。结果表明,最佳培养基配方:葡萄糖5 g/L、KH 2PO 48 g/L、MgSO 41.29 g/L、丙三醇10 mL/L、蔗糖10 g/L、H 8MoN 2O 40.99 mL/L、酵母提取物4.83 g/L、可溶性淀粉5 g/L。经过3次平行测试,Tr16的平均孢子产量为6.19×108 CFU/mL,与预测最大孢子产量接近,比未优化的培养基孢子产量提高了70.8%。

Response surface analysis method(RSAM)was used to optimize the liquid fermentation culture medium for spore production of Trichoderma atroviride strain Tr16.Through Plackett-Burman method(PB),three main factors(MgSO 4,H 8MoN 2O 4 and yeast extract)affecting sporulation quantity in the medium were firstly screened out.Then,the steepest climbing path method(SCPM)and RSAM were used to determine the interaction among the main factors and the best formula of liquid fermentation medium was screened and determined.The results showed that the optimal formula of the medium was glucose 5 g/L,KH 2PO 48 g/L,MgSO 41.29 g/L,glycerol 10 mL/L,sucrose 10 g/L,H 8MoN 2O 40.99 mL/L,yeast extract 4.83 g/L,soluble starch 5 g/L.After three parallel tests,the average spore yield of Tr16 was 6.19×108CFU/mL,which was close to the predicted maximum spore yield and was 70.8%higher than that of the unoptimized medium.