食品科学

• 生物工程 • 上一篇    下一篇

高产靛蓝色素大肠杆菌基因工程菌的构建及其发酵条件优化

程 雷1,尹 胜1,孙宝国1,陈 敏2,王成涛1,*,隆歆然1,王子豪1   

  1. 1.北京工商大学 北京市食品添加剂工程技术研究中心,食品营养与人类健康北京高精尖创新中心,北京 100048;
    2.中国农业大学食品科学与营养工程学院,北京 100083
  • 出版日期:2016-12-15 发布日期:2016-12-21

Construction of Genetically Engineered E. coli and Optimization of Its Fermentation Conditions for High-Level Production of Indigo

CHENG Lei1, YIN Sheng1, SUN Baoguo1, CHEN Min2, WANG Chengtao1,*, LONG Xinran1, WANG Zihao1   

  1. 1. Beijing Engineering and Technology Research Center of Food Additives, Beijing Innovation Centre of Food Nutrition and Human Health, Beijing Technology and Business University, Beijing 100048, China; 2. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China
  • Online:2016-12-15 Published:2016-12-21

摘要: 通过基因重组技术构建重组苯乙烯单加氧酶基因的高产靛蓝色素大肠杆菌基因工程菌Escherichia coli AB,研究优化工程菌E. coli AB生物合成靛蓝色素的发酵条件。单因素试验分别考察不同发酵温度、底物质量浓度、起始pH值和接种量4 个因素对于靛蓝色素合成的影响,选取发酵温度、底物浓度和起始pH值3 个对靛蓝色素产量影响显著的因素进行响应面设计试验优化。经模型分析获得优化发酵条件为:底物质量浓度0.95 mg/mL,发酵温度29 ℃,起始pH 6.9。在该条件下,工程菌E. coli AB催化合成靛蓝色素产量为67.65 mg/mL。优化发酵条件的实验结果与预测值相符,说明回归模型切实可行。

关键词: 靛蓝色素, 苯乙烯单加氧酶, 工程菌, 发酵条件, 响应面试验

Abstract: A genetically engineered strain of Escherichia coli for the expression of the styrene monooxygenase gene from
Pseudomonas putida was constructed by recombinant DNA technique, and the fermentation conditions of the recombinant
strain were optimized for enhanced indigo production. Firstly, the effects of four factors including substrate concentration,
fermentation temperature, initial pH and inoculum amount on indigo production were investigated. Then, the response surface
methodology was applied to optimize the variables showing significant influence on indigo production, substrate concentration,
fermentation temperature and initial pH. Results revealed that the optimized fermentation conditions were determined as substrate
concentration of 0.95 mg/mL, fermentation temperature of 29 ℃ and initial pH 6.9. The predicted indigo production was
67.65 mg/mL, which was in close agreement with the predicted value, indicating that the model was feasible.

Key words: indigo, styrene monooxygenase, genetically engineered strain, fermentation conditions, response surface methodology

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