Applied Microbiology and Biotechnology

, Volume 98, Issue 13, pp 5991–6002 | Cite as

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

  • Min-Kyoung Kang
  • Jungseok Lee
  • Youngsoon Um
  • Taek Soon Lee
  • Michael Bott
  • Si Jae Park
  • Han Min Woo
Applied genetics and molecular biotechnology

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 

Supplementary material

253_2014_5714_MOESM1_ESM.pdf (72 kb)
ESM 1(PDF 72 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Min-Kyoung Kang
    • 1
  • Jungseok Lee
    • 1
    • 2
  • Youngsoon Um
    • 1
    • 4
  • Taek Soon Lee
    • 5
    • 6
  • Michael Bott
    • 7
  • Si Jae Park
    • 8
  • Han Min Woo
    • 1
    • 3
    • 4
  1. 1.Clean Energy Research CenterKorea Institute of Science and TechnologySeoulRepublic of Korea
  2. 2.Department of Chemical and Biological EngineeringKorea UniversitySeoulRepublic of Korea
  3. 3.Green School, Korea UniversitySeoulRepublic of Korea
  4. 4.Department of Clean Energy and Chemical EngineeringUniversity of Science and Technology (UST)DaejeonRepublic of Korea
  5. 5.Joint BioEnergy InstituteEmeryvilleUSA
  6. 6.Physical Biosciences DivisionLawrence Berkeley National LaboratoryBerkeleyUSA
  7. 7.Institute of Bio- and GeosciencesIBG-1: Biotechnology, Forschungszentrum JülichJülichGermany
  8. 8.Department of Environmental Engineering and EnergyMyongji UniversityYongin-siRepublic of Korea

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