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Synthetic biology platform of CoryneBrick vectors for gene expression in Corynebacterium glutamicum and its application to xylose utilization

  • Applied genetics and molecular biotechnology
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Abstract

Currently, the majority of tools in synthetic biology have been designed and constructed for model organisms such as Escherichia coli and Saccharomyces cerevisiae. In order to broaden the spectrum of organisms accessible to such tools, we established a synthetic biological platform, called CoryneBrick, for gene expression in Corynebacterium glutamicum as a set of E. coli-C. glutamicum shuttle vectors whose elements are interchangeable with BglBrick standard parts. C. glutamicum is an established industrial microorganism for the production of amino acids, proteins, and commercially promising chemicals. Using the CoryneBrick vectors, we showed various time-dependent expression profiles of a red fluorescent protein. This CoryneBrick platform was also applicable for two-plasmid expression systems with a conventional C. glutamicum expression vector. In order to demonstrate the practical application of the CoryneBrick vectors, we successfully reconstructed the xylose utilization pathway in the xylose-negative C. glutamicum wild type by fast BglBrick cloning methods using multiple genes encoding for xylose isomerase and xylulose kinase, resulting in a growth rate of 0.11 ± 0.004 h−1 and a xylose uptake rate of 3.35 mmol/gDW/h when 1 % xylose was used as sole carbon source. Thus, CoryneBrick vectors were shown to be useful engineering tools in order to exploit Corynebacterium as a synthetic platform for the production of chemicals by controllable expression of the genes of interest.

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Acknowledgments

The authors thank Prof. Anthony J. Sinskey for the kind gift of pZ8-1 and M.S. Jae Hee Jung for technical assistant. This work was supported by the National Research Foundation of Korea Grant funded by the Korean Government (Ministry of Science, ICT & Future Planning) (2014, University-Institute cooperation program) and Creative Allied Program (CAP) of the Korea Research Council of Fundamental Science and Technology (KRCF)/Korea Institute of Science and Technology (KIST) (project no. 2E24832).

Conflict of interests

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Clean Energy Research Center, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul, 136-791, Republic of Korea

    Min-Kyoung Kang, Jungseok Lee, Youngsoon Um & Han Min Woo

  2. Department of Chemical and Biological Engineering, Korea University, 145 Anam-ro, Seongbuk-gu, Seoul, 136-701, Republic of Korea

    Jungseok Lee

  3. Green School, Korea University, 145 Anam-ro, Seongbuk-gu, Seoul, 136-701, Republic of Korea

    Han Min Woo

  4. Department of Clean Energy and Chemical Engineering, University of Science and Technology (UST), 217 Gajeong-ro, Yuseong-gu, Daejeon, 305-350, Republic of Korea

    Youngsoon Um & Han Min Woo

  5. Joint BioEnergy Institute, Emeryville, CA, 94608, USA

    Taek Soon Lee

  6. Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA

    Taek Soon Lee

  7. Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich, D-52425, Jülich, Germany

    Michael Bott

  8. Department of Environmental Engineering and Energy, Myongji University, San 38-2, Nam-dong, Cheoin-gu, Yongin-si, Gyeonggido, 449-728, Republic of Korea

    Si Jae Park

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  1. Min-Kyoung Kang
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Correspondence to Han Min Woo.

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Min-Kyoung Kang and Jungseok Lee contributed equally to this work.

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Kang, MK., Lee, J., Um, Y. et al. Synthetic biology platform of CoryneBrick vectors for gene expression in Corynebacterium glutamicum and its application to xylose utilization. Appl Microbiol Biotechnol 98, 5991–6002 (2014). https://doi.org/10.1007/s00253-014-5714-7

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