产3-羟基丙酸重组大肠杆菌JM109的构建及发酵条件优化

生物技术通报 ›› 2013, Vol. 0 ›› Issue (6): 200-208.

产3-羟基丙酸重组大肠杆菌JM109的构建及发酵条件优化

张晓梅^{1, 2}, 李新生¹, 许正宏²

(1.陕西理工学院资源生物重点实验室, 汉中 723001;2.江南大学药学院, 无锡 214122)

收稿日期:2013-06-20 修回日期:2013-06-20 出版日期:2013-06-20 发布日期:2013-06-20
作者简介: 张晓梅, 女, 副教授, 博士, 研究方向:生物制药;E-mail: hanqiaowu@sina.com
基金资助:
陕西省教育厅访问学者项目(12JS030), 陕西省科技厅访问学者项目(2011HBSZS003)

Construction of Recombinant E. coli JM109 Capable of Producing 3-Hydroxypropionic Acid and Its Fermentation Condition Optimization

Zhang Xiaomei^{1, 2} Li Xinsheng¹ Xu Zhenghong²

(1. Key Laboratory of Biological Resources, Shaanxi University of Technology, Hanzhog 723001;2. School of Medicine and Pharmaceutics, Jiangnan University, Wuxi 214122)

Received:2013-06-20 Revised:2013-06-20 Published:2013-06-20 Online:2013-06-20

摘要/Abstract

摘要： 将编码乙醛脱氢酶编码基因aldh连接重组质粒pEtac-dhaB, 转化大肠杆菌, 得到产3-羟基丙酸(3-HP) 重组大肠杆菌JM109(pEtac-dhaB-aldh) , 并将响应面法应用于重组大肠杆菌JM109(pEtac-dhaB-aldh) 发酵条件优化。单因素发酵试验结果表明, 影响3-HP产量的4个主要因子是初始甘油浓度, 酵母膏浓度, 维生素B₁₂浓度以及KH_{2PO₄浓度, 应用响应面分析法对各因素的最佳水平范围及其交互作用进行了研究和探讨。优化后培养基组成(g/L) : 甘油62 g/L、VB₁₂ 0.05 g/L、KH₂PO₄ 7.4 g/L及酵母膏 6.2 g/L, 在此培养基中3-HP的产量为 5.8 g/L。}

关键词: 重组大肠杆菌, 构建, 3-羟基丙酸, 响应面法, 优化

Abstract: The expression vector pEtac harboring aldh gene and dhaB gene was transformed into E.coli JM109 to construct recombinant E.coli JM109(pEtac-dhaB-aldh). And the response surface method(RSM)was firstly applied to determine and optimize the fermentation condition for the novel recombinant E.coli JM109(pEtac-dhaB-aldh)capable of producing 3- hydroxypropionic acid. A mathematical model was then developed to show the effect of each medium composition and their interactions on the production of 3- hydroxypropionic acid. The model estimated that, a maximal yield of 3-hydroxypropionic acid(5.4 g/L)could be obtained when the concentrations of glycerol, VB₁₂, yeast extract KH₂PO₄ were set at 62 g/L, 0.05 g/L, 6.2 g/L and 7.4 g/L, respectively. These predicted values were also verified by validation experiments. Compared with the values obtained by other runs in the experimental design.The yield and productivity under the optimal parameters and process can reach 5.8 g/L.

Key words: Recombinant, E. coli, Construction 3- hydroxypropionic acid, Response surface methodology, Optimization

张晓梅, 李新生, 许正宏. 产3-羟基丙酸重组大肠杆菌JM109的构建及发酵条件优化[J]. 生物技术通报, 2013, 0(6): 200-208.

Zhang Xiaomei, Li Xinsheng, Xu Zhenghong. Construction of Recombinant E. coli JM109 Capable of Producing 3-Hydroxypropionic Acid and Its Fermentation Condition Optimization[J]. Biotechnology Bulletin, 2013, 0(6): 200-208.

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