响应面法优化微小毛霉固态发酵生产凝乳酶工艺研究

Optimization of solid state fermentation using response surface methodology for milk-clotting enzyme production by Mucor pusillus

采用麦麸培养基进行固态发酵,以研究发酵条件对产酶活力的影响,在此基础上应用Plackett-Burman、最陡爬坡实验和Box-Behnken实验设计,对微小毛霉的固态发酵培养基进化以达到最佳产酶量。实验结果表明,最佳发酵条件为:发酵时间5d,发酵温度30℃,固液比值0.75,接种量为3份,测得凝乳酶酶活为828U/mL。并且当麸皮质量为15.50g、(NH4)2SO4质量为0.15g、KH2PO4质量为0.08g时,微小毛霉产凝乳酶活力达到1200U/mL。较未优化前活力提高了66.67%。

Using Plackett-Burman design combined with the path of steepest ascent method and Box-Behnken design,the optimum fermnetation properies of mucor pusillus including fermentation time 5d,fermentation temperature 30℃,solid-to-liquid ratio 0.75 and application with 4 samples were determined.Under such conditions,with increasing the amount of bran,（NH4）2SO4 and KH2PO4 in the culture to 15.50g,0.15g and 0.08g respectively,the highest enzyme milk-clotting activity 1200U/mL was achieved,66.67% higher than the milk-clotting activity before optimization.