食品科学 ›› 2008, Vol. 29 ›› Issue (6): 243-247.

• 生物工程 • 上一篇    下一篇

响应面法优化茶氨酸生物合成基因工程菌发酵条件的研究

 陆文渊, 成浩, 王丽鸳, 周健   

  1. 农业部茶叶化学工程重点实验室 中国农业科学院茶叶研究所; 农业部茶叶化学工程重点实验室; 中国农业科学院茶叶研究所;
  • 出版日期:2008-06-15 发布日期:2011-08-26

Response Surface Methodology for Optimization of Fermentation Conditions of Genetically Engineered Escherichia coli Strain for Biosynthesis of Theanine

 LU  Wen-Yuan, CHENG  Hao, WANG  Li-Yuan, ZHOU  Jian   

  1. Key Laboratory of Tea Chemical Engineering, Ministry of Agriculture, Tea Research Institute, Chinese Academy of Agricultural Sciences
  • Online:2008-06-15 Published:2011-08-26

摘要: 本实验对具有茶氨酸生物合成能力的γ-谷氨酰转肽酶(γ-GGT)基因工程菌的发酵条件进行了优化。首先采用Plackett-Burman实验设计对影响γ-GGT活性的基因工程菌发酵条件进行筛选。然后将筛选得到的初始pH、培养时间、异丙基硫代-β-D-半乳糖苷(isopropylthio-β-D-galactoside,IPTG)诱导温度3个关键影响因素进行响应面分析,通过对二次多项回归方程求解得到该基因工程菌的最佳培养条件为初始pH7.32、培养时间6.67h、IPTG31.51℃诱导。γ-GGT活性的最大预测值为4.60U/ml,实验验证值为4.64U/ml。最后通过基因工程菌催化合成茶氨酸反应,得到茶氨酸的产量为35.18g/L。

关键词: 茶氨酸, &gamma, -谷氨酰转肽酶(&gamma, -GGT), Plackett-Burman设计, 响应面法(RSM)

Abstract: This experiment studied the optimization of fermentation conditions of the genetically engineered Escherichia coli strain of γ-glutamyltranspeptidase γ-EGT for the biosynthesis of theanine. The significant effects of initial pH, cultivation time and IPTG induction temperature on the activity of γ-GGT were noted with Plackett-Burman design. It then revealed with the method of response surface methodology (RSM) that initial pH 7.32, cultivation time 6.67 h and IPTG induction temperature 31.51 ℃ are optimum for the activity of γ-GGT, with a predicted γ-GGT activity 4.60 U/ml, and the experimentalγ-GGT activity 4.64 U/ml. The yield of theanine from L-Gln and ethylamine is 35.18 g/L.

Key words: theanine; &gamma, -glytamyltranspeptidase (&gamma, -GGT); Plackett-Burman design; response surface methodology (RSM);