Wild-type Corynebacterium glutamicum was metabolically engineered to convert glucose and mannose into guanosine 5′-diphosphate (GDP)-l-fucose, a precursor of fucosyl-oligosaccharides, which are involved in various biological and pathological functions. This was done by introducing the gmd and wcaG genes of Escherichia coli encoding GDP-d-mannose-4,6-dehydratase and GDP-4-keto-6-deoxy-d-mannose-3,5-epimerase-4-reductase, respectively, which are known as key enzymes in the production of GDP-l-fucose from GDP-d-mannose. Coexpression of the genes allowed the recombinant C. glutamicum cells to produce GDP-l-fucose in a minimal medium containing glucose and mannose as carbon sources. The specific product formation rate was much higher during growth on mannose than on glucose. In addition, the specific product formation rate was further increased by coexpressing the endogenous phosphomanno-mutase gene (manB) and GTP-mannose-1-phosphate guanylyl-transferase gene (manC), which are involved in the conversion of mannose-6-phosphate into GDP-d-mannose. However, the overexpression of manA encoding mannose-6-phosphate isomerase, catalyzing interconversion of mannose-6-phosphate and fructose-6-phosphate showed a negative effect on formation of the target product. Overall, coexpression of gmd, wcaG, manB and manC in C. glutamicum enabled production of GDP-l-fucose at the specific rate of 0.11 mg g cell−1 h−1. The specific GDP-l-fucose content reached 5.5 mg g cell−1, which is a 2.4-fold higher than that of the recombinant E. coli overexpressing gmd, wcaG, manB and manC under comparable conditions. Well-established metabolic engineering tools may permit optimization of the carbon and cofactor metabolisms of C. glutamicum to further improve their production capacity.
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This work was supported by the Advanced Biomass R and D Center (ABC) of Korea Grant funded by the Ministry of Education, Science and Technology (2011-0031359). This work was also supported by a grant from Marine Biotechnology Program Funded by Ministry of Land, Transport and Maritime Affairs of Korean Government.
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Chin, Y., Park, J., Park, Y. et al. Metabolic engineering of Corynebacterium glutamicum to produce GDP-l-fucose from glucose and mannose. Bioprocess Biosyst Eng 36, 749–756 (2013). https://doi.org/10.1007/s00449-013-0900-z
- Corynebacterium glutamicum
- Guanosine nucleotide