Modulation of guanosine 5'-diphosphate-D-mannose metabolism in recombinant Escherichia coli for production of guanosine 5'-diphosphate-L-fucose.
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 BL21star(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.
Lee
Won-Heong
WH
Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, Republic of Korea.
Han
Nam-Soo
NS
Park
Yong-Cheol
YC
Seo
Jin-Ho
JH
D002244:Carbon / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005285:Fermentation / Q000187:drug effects; D005798:Genes, Bacterial; D005818:Genetic Engineering; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D006154:Guanosine Diphosphate Fucose / Q000096:biosynthesis*; D006155:Guanosine Diphosphate Mannose / Q000378:metabolism*; D008358:Mannose / Q000378:metabolism / Q000494:pharmacology
D029968:Escherichia coli Proteins; D006154:Guanosine Diphosphate Fucose; D006155:Guanosine Diphosphate Mannose; D002244:Carbon; D005947:Glucose; D008358:Mannose