响应面法优化黑曲霉产葡萄糖氧化酶发酵条件

Optimization of Glucose Oxidase Fermentation with Aspergillus niger by Response Surface Methodology

DOI：10.3969/j.issn.1673-1689.2014.10.014

中文关键词: 葡萄糖氧化酶 响应面法 黑曲霉

英文关键词: glucose oxidase,response surface methodology,Aspergillus niger

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中文摘要:

为了提高葡萄糖氧化酶的产量，通过响应面法对诱变后的黑曲霉菌株的发酵条件进行优化，首先利用Plackett-Burman设计筛选出对酶活影响最显著的因素：牛肉膏蛋白胨、吐温-60和磷酸氢二铵；继而用最陡爬坡实验逼近最大影响区域；最后利用Box-Behnken设计及其响应面分析确定最优的发酵培养基（g/L）：蔗糖 87.5，牛肉膏蛋白胨3.15，NH4NO3 1.88，（NH4）2HPO4 0.34，KH2PO4 0.25，Tw-60 30.47，玉米粉12.5，培养基优化后的发酵酶活为87.5 U/mL，与优化前（45.27 U/mL）相比提高了93.28%。在7 L发酵罐中对显著影响因素吐温-60的不同添加时间进行对比，确定对数期流加吐温-60可以显著提高发酵的酶活，发酵后酶活为92.88 U/mL。

英文摘要:

For the purpose of improving the production of glucose oxidase，response surface methodology（RSM） has been employed to optimize the fermentation medium to improve glucose oxidase production by mutational Aspergillus niger. Firstly，a Plackett-Burman design was used to investigate the effects of different factors in the fermentation medium，with three most significant factors being identified as：beef extract peptone，Tw-60 and（NH4）2HPO4. Secondly，the steepest ascent procedure was employed to define the optimal response region for these three factors. Finally，the Box-Behnken method was employed to design the reaction system and the optimal fermentation medium for production of glucose oxidase consisted of the following（g/L）：sugar 87.5，beef extract peptone 3.15，NH4NO3 1.88，（NH4）2HPO4 0.34， KH2PO4 0.25，Tw-60 30.47，corn flour 12.5. Glucose oxidase production after optimization was increasing by a factor of about 93.28% from 45.27 U/mL to 87.5 U/mL. The different adding time of Tw-60 was investigated in 7 L fermentor. The result shows that adding Tw-60 to broth in logarithmic phase can improve glucose oxidase production remarkably，the total glucose oxidase activity reached 92.88 U/mL.

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