Journal
BIORESOURCE TECHNOLOGY
Volume 100, Issue 24, Pages 6143-6148Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.biortech.2009.07.035
Keywords
Recombinant Escherichia coli; GDP-L-fucose; GDP-D-mannose; Fed-batch fermentation
Funding
- Seoul RBD Program [11000]
- WCU (World Class University) program through the Korea Science
- Ministry of Education, Science and Technology [08-0238]
- Korean Ministry of Commerce, Industry and Energy [10028380-2006-11]
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Guanosine 5'-diphosphate (GDP)-L-fucose, an activated form of a nucleotide sugar, plays an important role in a wide range of biological functions. In this study, the enhancement of GDP-L-fucose production was attempted by supplementation of mannose, which is a potentially better carbon source to be converted into GDP-L-fucose than glucose, and combinatorial overexpression of the genes involved in the biosynthesis of GDP-D-mannose, a precursor of GDP-L-fucose. Supply of a mannose and glucose led to a 1.3-fold-increase in GDP-L-fucose concentration (52.5 +/- 0.8 mg l(-1)) in a fed-batch fermentation of recombinant E. coli BL21 star(DE3) overexpressing the gmd and wcaG genes, compared with the case using glucose as a sole carbon source. A maximum GDP-L-fucose concentration of 170.3 +/- 2.3 mg l(-1), corresponding to a 4.4-fold enhancement compared with the control strain overexpressing gmd and wcaG genes only, was achieved in a glucose-limited fed-batch fermentation of a recombinant E. coli BL21star(DE3) strain overexpressing manB, manC, gmd and wcaG genes. Further improvement of GDP-L-fucose production was not obtained by additional overexpression of the manA gene. (C) 2009 Elsevier Ltd. All rights reserved.
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