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Rational engineering of multiple module pathways for the production of l-phenylalanine in Corynebacterium glutamicum

  • Metabolic Engineering and Synthetic Biology
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Microbial production of l-phenylalanine (l-Phe) from renewable sources has attracted much attention recently. In the present study, Corynebacterium glutamicum 13032 was rationally engineered to produce l-Phe from inexpensive glucose. First, all the l-Phe biosynthesis pathway genes were investigated and the results demonstrated that in addition to AroF and PheA, the native PpsA, TktA, AroE and AroA, and the heterologous AroL and TyrB were also the key enzymes for L-Phe biosynthesis. Through combinational expression of these key enzymes, the l-Phe production was increased to 6.33 ± 0.13 g l−1 which was about 1.48-fold of that of the parent strain C. glutamicum (pXM-pheA fbr-aroF fbr) (fbr, feedback-inhibition resistance). Furthermore, the production of l-Phe was improved to 9.14 ± 0.21 g l−1 by modifying the glucose and l-Phe transport systems and blocking the acetate and lactate biosynthesis pathways. Eventually, the titer of l-Phe was enhanced to 15.76 ± 0.23 g l−1 with a fed-batch fermentation strategy. To the best of our knowledge, this was the highest value reported in rationally engineered C. glutamicum 13032 strains. The results obtained will also contribute to rational engineering of C. glutamicum for production of other valuable aromatic compounds.

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Acknowledgments

This work was financially supported by the Key Program of the National Natural Science Foundation of China (973 Program, 2014CB745103, 2013CB733602), the National Natural Science Foundation of China (31200020), the Jiangsu Planned Projects for Postdoctoral Research Funds (1301010B), the 111 Project and the Priority Academic Program Development of Jiangsu Higher Education Institutions and the Self-determined Research Program of Jiangnan University (JUSRP51303A).

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Authors and Affiliations

  1. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China

    Chuanzhi Zhang, Junli Zhang & Zhen Kang

  2. Synergetic Innovation Center of Food Safety and Nutrition, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China

    Chuanzhi Zhang, Junli Zhang, Zhen Kang, Guocheng Du & Jian Chen

  3. School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China

    Chuanzhi Zhang, Junli Zhang, Zhen Kang, Guocheng Du & Jian Chen

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  1. Chuanzhi Zhang
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  2. Junli Zhang
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Correspondence to Zhen Kang.

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Zhang, C., Zhang, J., Kang, Z. et al. Rational engineering of multiple module pathways for the production of l-phenylalanine in Corynebacterium glutamicum . J Ind Microbiol Biotechnol 42, 787–797 (2015). https://doi.org/10.1007/s10295-015-1593-x

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