Applied genetics and molecular biotechnology

Applied Microbiology and Biotechnology

, Volume 98, Issue 13, pp 5991-6002

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

  • Min-Kyoung KangAffiliated withClean Energy Research Center, Korea Institute of Science and Technology
  • , Jungseok LeeAffiliated withClean Energy Research Center, Korea Institute of Science and TechnologyDepartment of Chemical and Biological Engineering, Korea University
  • , Youngsoon UmAffiliated withClean Energy Research Center, Korea Institute of Science and TechnologyDepartment of Clean Energy and Chemical Engineering, University of Science and Technology (UST)
  • , Taek Soon LeeAffiliated withJoint BioEnergy InstitutePhysical Biosciences Division, Lawrence Berkeley National Laboratory
  • , Michael BottAffiliated withInstitute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich
  • , Si Jae ParkAffiliated withDepartment of Environmental Engineering and Energy, Myongji University
  • , Han Min WooAffiliated withClean Energy Research Center, Korea Institute of Science and TechnologyGreen School, Korea UniversityDepartment of Clean Energy and Chemical Engineering, University of Science and Technology (UST) Email author 

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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.

Keywords

Corynebacterium glutamicum Synthetic biology Metabolic engineering BglBrick