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High crude violacein production from glucose by Escherichia coli engineered with interactive control of tryptophan pathway and violacein biosynthetic pathway

文献类型： 外文期刊

作者： Fang, Ming-Yue 1 ; Zhang, Chong 1 ; Yang, Song 2 ; Cui, Jin-Yu 2 ; Jiang, Pei-Xia 3 ; Lou, Kai 4 ; Wachi, Masaaki 5 ; Xin 1 ;

作者机构： 1.Tsinghua Univ, Dept Chem Engn, Beijing 10084, Peoples R China

2.Qingdao Agr Univ, Sch Life Sci, Qingdao 266109, Peoples R China

3.Chinese Acad Sci, Inst Microbiol, Beijing 10084, Peoples R China

4.Xinjiang Acad Agr Sci, Inst Microbiol, Urumqi 830000, Peoples R China

5.Tokyo Inst Technol, Dept Bioengn, Yokohama, Kanagawa 2268503, Japan

关键词： Violacein;Tryptophan;Glucose;Pathway optimization;Escherichia coli

期刊名称：MICROBIAL CELL FACTORIES （ 影响因子：5.328； 五年影响因子：5.588 ）

ISSN： 1475-2859

年卷期： 2015 年 14 卷

页码：

收录情况： SCI

摘要： Background: As bacteria-originated crude violacein, a natural indolocarbazole product, consists of violacein and deoxyviolacein, and can potentially be a new type of natural antibiotics, the reconstruction of an effective metabolic pathway for crude violacein (violacein and deoxyviolacein mixture) synthesis directly from glucose in Escherichia coli was of importance for developing industrial production process. Results: Strains with a multivariate module for varied tryptophan productivities were firstly generated by combinatorial knockout of trpR/tnaA/pheA genes and overexpression of two key genes trpE(fbr)/trpD from the upstream tryptophan metabolic pathway. Then, the gene cluster of violacein biosynthetic pathway was introduced downstream of the generated tryptophan pathway. After combination of these two pathways, maximum crude violacein production directly from glucose by E. coli B2/pED + pVio was realized with a titer of 0.6 +/- 0.01 g L-1 in flask culture, which was four fold higher than that of the control without the tryptophan pathway up-regulation. In a 5-L bioreactor batch fermentation with glucose as the carbon source, the recombinant E. coli B2/pED + pVio exhibited a crude violacein titer of 1.75 g L-1 and a productivity of 36 mg L-1 h(-1), which was the highest titer and productivity reported so far under the similar culture conditions without tryptophan addition. Conclusion: Metabolic pathway analysis using C-13 labeling illustrated that the up-regulated tryptophan supply enhanced tryptophan metabolism from glucose, whereas the introduction of violacein pathway drew more carbon flux from glucose to tryptophan, thereby contributing to the effective production of crude violacein in the engineered E. coli cell factory.

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