响应面法优化酿酒酵母工程菌产甜蛋白monellin发酵培养基  

Optimization of fermentation medium of sweet protein monellin produced by engineering strain of Saccharomyces cerevisiae using response surface methodology

作　　者：赵沁沁[1] 刘军[1] 徐爱才[1] 欧阳文[1] 闫达中[1] 赵胜军[2] 

机构地区：[1]武汉工业学院生物与制药工程学院,湖北武汉430023 [2]武汉工业学院动物科学与营养工程学院,湖北武汉430023

出　　处：《中国酿造》2011年第9期50-55,共6页China Brewing

基　　金：湖北省教育厅重点项目(D200718002)

摘　　要：对酿酒酵母工程菌WHF9/monellin在液体培养基中产甜蛋白monellin的条件进行了优化。采用单因子试验筛选出细胞生长培养基的最适碳源为蔗糖,氮源为玉米浆干粉,且应添加微量元素溶液。在此基础上,利用Plackett-Burman试验设计筛选出影响菌体生物量的3个显著因素:蔗糖、玉米浆干粉、微量元素溶液。用最陡爬坡路径逼近最大响应区域后,利用Box-Behnken设计和响应面分析法对显著因素进行优化,得出蔗糖、玉米浆干粉、微量元素溶液的最佳浓度分别为102.2g/L,33.9g/L,4.5mL/L。菌株在优化后的生长培养基中的生物量比在YPD培养基中提高了81.38%。在诱导剂半乳糖最适浓度102.2g/L条件下,菌株在优化后的培养基中monellin的表达量达到226.7mg/L,比在YPG中表达的monellin提高了4.2倍,且表达的monellin具有生物活性。The fermentation medium of sweet protein monellin produced by engineering strain of Saccharomyces cerevisiae, WHF9/monellin, were optimized in this paper. Single factor experiments showed that the optimal carbon source was sugar, and the optimal nitrogen source was corn steep liquor powder and trace elements were added in cell growth medium. Based on these results, effects of three significant factors on biomass were selected in Plackett-Burman experiment design. The steepest ascent was used to approach the optimal region of response value. The optimal medium formula was obtained by Box-Behnken design and response surface methodology as follows： sugar 102.2g/L, corn steep liquor powder 33.9g/L and trace elements 4.5ml/L. Under these conditions, the biomass was increased by 81.38% in the cell growth medium compared with that in the YPD medium, Under the optimal concentration of galactose 102.2g/L, the production of monellin reached 226.7mg/L, which was improved by 4.2 fold compared with that of YPG medium and monellin had biological activity.

关 键 词：MONELLIN 酿酒酵母 Plackett-Burman试验设计 响应面法 优化 

分 类 号：Q93-335[生物学—微生物学]