利用响应面分析法进行氨肽酶发酵培养基的优化

Optimization of Culture Medium for Methionine Aminopeptidase Fermentation by Response Surface Methodology

利用Minitab软件中的Plackett-Burman设计和响应面的Box-Behnken中心组合设计,对发酵生产甲硫氨酸氨肽酶的培养基进行优化。在培养基成分单因素优化的基础上,利用Plackett-Burman实验,确定麦芽糊精、酵母粉及硫酸镁为甲硫氨酸氨肽酶发酵生产的3个显著因素。进一步用最陡爬坡法实验将显著因素的水平逼近最优区域。最后用Box-Behnken建立这3个显著因素的二次回归模型,通过对响应面曲面进行分析,得出麦芽糊精、酵母粉和硫酸镁的最佳浓度分别为:26.00、5.3、0.204 g/L。在优化培养基中,甲硫氨酸氨肽酶生产菌的发酵水平增加约1.11倍,达到50.84 U/mL,实验值与预期值基本相符。

Optimization of culture medium for methionine aminopeptidase （MetAP） fermentation was carried out by the Plackett-Burman design and Response Surface Methodology （RSM） of Minitab software. Reference ranges of each factor were determined by single factorial experiment and three major factors significantly affected production of MetAP, which were maltodextrin ,yeast extracts and MgSO4· 7H2O, were determined by placket-burman design.The optimum fermentation conditions were optimized with Box-Behnken experiment and the largest regional of methionine aminopeptidase production was approach with the steepest ascent experiment. Estimated optimal conditions of the three factors were as the follows ： maltodextrin 26.00 g/L, yeast powder 5.3 g/L and MgSO4· 7H2O 0.204 g/L. Under the optimum fermentation medium conditions, MetAP yield was 50.84 U/mL in flask culture under the optimal culture conditions, which increased 1.11 times to that of using initial medium.