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Metabolic engineering of Corynebacterium glutamicum to produce GDP-l-fucose from glucose and mannose

Title
Metabolic engineering of Corynebacterium glutamicum to produce GDP-l-fucose from glucose and mannose
Authors
Chin Y.-W.Park J.-B.Park Y.-C.Kim K.H.Seo J.-H.
Ewha Authors
박진병
SCOPUS Author ID
박진병scopus
Issue Date
2013
Journal Title
Bioprocess and Biosystems Engineering
ISSN
1615-7591JCR Link
Citation
vol. 36, no. 6, pp. 749 - 756
Indexed
SCI; SCIE; SCOPUS WOS scopus
Abstract
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. © 2013 Springer-Verlag Berlin Heidelberg.
DOI
10.1007/s00449-013-0900-z
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엘텍공과대학 > 식품공학전공 > Journal papers
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