粗壮脉纹孢菌高密度培养基的优化及补料策略

Optimization of High-density Medium of Neurospora crassa and Feeding Strategy for Its High Yield

本研究以粗壮脉纹孢菌(Neurospora crassa)为实验菌株,采用单因素实验结合响应面法对粗壮脉纹孢菌的高密度培养基配方进行优化。以生物量为指标,得到最优培养基组成:蔗糖42 g/L、蛋白胨36 g/L、磷酸二氢钾2.6 g/L、硫酸锌0.038 g/L、硫酸镁0.1 g/L、硫酸锰0.03 g/L、氯化钾0.1 g/L,经验证实验发现,在最优培养基组成下菌体生物量最大可达11.295 g/L。并采用补料分批培养的方式扩大菌体生物量,进一步提高高密度培养的效果。结果表明,在最优培养基的基础上,分别于培养24、60、96 h进行补料,共补料3次,每次补加200 g/L的蔗糖溶液10 mL,可获得粗壮脉纹孢菌的最大生物量14.732 g/L,该结果为实现粗壮脉纹孢菌这一优良菌种的工业化应用提供了理论支持。

Neurospora crassa was used as the experimental strain,and the formula of high cell density medium was optimized using single factor experiment and response surface methodology.The optimum medium components were selected by the biomass of Neurospora crassa and were chosen as follows:Sucrose 42 g/L,peptone 36 g/L,KH 2PO 42.6 g/L,ZnSO 4·7H 2O 0.038 g/L,MgSO 4·7H 2O 0.1 g/L,MnSO 4·H 2O 0.03 g/L,KCl 0.1 g/L,the verification experiment found that under the optimal medium composition,the maximum biomass of the bacteria was 11.295 g/L.In order to further improve the efficiency of high cell density culture,fed-batch culture was used to expand the biomass of Neurospora crassa.Results showed that the largest biomass of Neurospora crassa(14.732 g/L)based on the optimal medium was obtained by adding 10 mL of 200 g/L sucrose solution as the fed-batch culture at 24,60 and 96 h,respectively.The results provide theoretical support for the industrial application of Neurospora crassa.