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Engineering of Corynebacterium glutamicum for growth and l-lysine and lycopene production from N-acetyl-glucosamine

  • Applied microbial and cell physiology
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Abstract

Sustainable supply of feedstock has become a key issue in process development in microbial biotechnology. The workhorse of industrial amino acid production Corynebacterium glutamicum has been engineered towards utilization of alternative carbon sources. Utilization of the chitin-derived aminosugar N-acetyl-glucosamine (GlcNAc) for both cultivation and production with C. glutamicum has hitherto not been investigated. Albeit this organism harbors the enzymes N-acetylglucosamine-6-phosphatedeacetylase and glucosamine-6P deaminase of GlcNAc metabolism (encoded by nagA and nagB, respectively) growth of C. glutamicum with GlcNAc as substrate was not observed. This was attributed to the lack of a functional system for GlcNAc uptake. Of the 17 type strains of the genus Corynebacterium tested here for their ability to grow with GlcNAc, only Corynebacterium glycinophilum DSM45794 was able to utilize this substrate. Complementation studies with a GlcNAc-uptake deficient Escherichia coli strain revealed that C. glycinophilum possesses a nagE-encoded EII permease for GlcNAc uptake. Heterologous expression of the C. glycinophilum nagE in C. glutamicum indeed enabled uptake of GlcNAc. For efficient GlcNac utilization in C. glutamicum, improved expression of nagE with concurrent overexpression of the endogenous nagA and nagB genes was found to be necessary. Based on this strategy, C. glutamicum strains for the efficient production of the amino acid l-lysine as well as the carotenoid lycopene from GlcNAc as sole substrate were constructed.

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Acknowledgments

The authors thank Ute Meyer and Eva Glees (Institute of Biochemistry, University of Cologne, Germany) for excellent technical assistance, Jörn Kalinowski and Christian Rückert (Cebitec, University of Bielefeld, Germany) for granting early access to the C. glycinophilum draft genome sequence, and Jacqueline Plumbridge(Institut de Biologie Physico-Chimique, CNRS UPR9073, France) for providing the strain E. coli LR2-168 Work in the laboratories of the authors was funded in part by grants 0315589G and 0315589 F from BMBF in the CRP “Corynebacterium: improving flexibility and fitness for industrial production”.

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  1. Jung-Won Youn

    Present address: Institute of Microbiology, University of Stuttgart, Stuttgart, Germany

  2. Tomoya Maeda

    Present address: Molecular Microbiology and Biotechnology Group, Research Institute of Innovative Technology for the Earth, Kyoto, Japan

Authors and Affiliations

  1. Faculty of Biology and CeBiTec, Bielefeld University, 33501, Bielefeld, Germany

    Christian Matano, Jung-Won Youn & Volker F. Wendisch

  2. Institute of Biochemistry, University of Cologne, 50674, Cologne, Germany

    Andreas Uhde, Tomoya Maeda, Lina Clermont, Kay Marin, Reinhard Krämer & Gerd M. Seibold

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  1. Christian Matano
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Correspondence to Gerd M. Seibold.

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C. Matano and A. Uhde contributed equally to this work.

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Matano, C., Uhde, A., Youn, JW. et al. Engineering of Corynebacterium glutamicum for growth and l-lysine and lycopene production from N-acetyl-glucosamine. Appl Microbiol Biotechnol 98, 5633–5643 (2014). https://doi.org/10.1007/s00253-014-5676-9

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