重组酵母产中性米曲蛋白酶的工艺研究

摘要

With experiments of single factor, the optimal process conditions of recombined Pichia pastoris for producing neutral Aspergillus oryzae protease were researched. The results showed the most optimal process conditions for Pichia pastoris expressing neutral Aspergillus oryzae protease were the induced time 132 h, induced temperature 26℃, induced inoculum concentration OD600 value 6.0, induced incubating shaking speed 240 r/min, inducer (methanol) addition 0.8% (volume ratio), YNB primary concentration 7g/L and pH 7.0. Under the most optimal conditions, the activity of neutral Aspergillus oryzae protease in the broth can bring to 538 u/mL and was twice higher than the primary productivity.%　　通过单因素试验探索重组酵母产中性米曲蛋白酶的最佳条件。结果显示，重组酵母菌产蛋白酶的最佳表达条件是：诱导时间132 h，诱导温度26℃，诱导接种量为OD600值6.0，诱导培养振荡速度为240 r/min，诱导剂（甲醇）添加量0.8%（体积比），YNB添加量7 g/L，pH 7.0。在最佳培养条件下，发酵液中的最高酶活达到538 u/mL。与初始的发酵条件相比，中性米曲蛋白酶的产量提高1倍。