重组毕赤酵母发酵产PGA的条件优化

摘要

对重组毕赤酵母发酵产青霉素G酰化酶(PGA)的培养条件进行探讨.依据基础盐摇瓶发酵培养基,通过单因素试验考察了接种量、甘油浓度、pH以及诱导过程中氨水浓度、甲醇浓度等因素对PGA表达的影响.并采用Box-Behnken实验设计进行因素组合优化.结果与意义:最佳发酵条件为:甘油浓度40 g/L、氨水浓度0.1％、甲醇浓度1.0％、pH 5.6、接种量10％,在此条件下PGA酶活力达到8 680±12 U/L.本实验结果对下一步利用发酵罐进行重组毕赤酵母扩大培养有借鉴作用.%The cultivation conditions for Penicillin G acylase (PGA) production by recombinant Pichia pastoris were studied,the influences of inoculum,pH value,concentrations of glycerol,and methanol and ammonia water on the expression of PGA and the cell growth were investigated in shake flasks with Fermentation Basal Salts Medium by single factor experiments.And the experiments with the Box-Behnken experimental design were conducted to optimize the fermentation conditions of PGA production of recombinant P.pastoris.The results and implications are as follows:the optimum fermentation conditions were obtained (glycerol concentration 40 g/L,ammonia water concentration 0.1％,methanol concentration 1.0％,pH 5.6,inoculum 10％).Under these conditions,the activity of PGA was (8 680± 12) U/L.These results will be provide a reference base for the expansion cultivation of recombinant P.pastoris fermentation.