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).
- Corynebacterium glutamicum
- Metabolic engineering
- Synthetic biology