Method for production of GDP-L-fucose by overexpression of NADPH-regenerating and guanosine nucleotide biosynthetic enzymes in recombinant Escherichia coli

摘要

The present invention in addition to overexpression of the relevant enzymes which mediate the route of synthesis of GDP- fucose, glucose-6-phosphate-dehydro through dehydratase and / or guanosine-inosine kinase to add specific to the recombinant E. coli that over-expression used herein relates to a method for producing the GDP- fucose, ; Representative NADPH reproduced enzyme glucose-6-phosphate dehydro through Kinase (glucose-6-phosphate dehydrogenase; G6PDH) present in the E. coli using the ManB, ManC, Gmd and recombinant E. coli expressing a simultaneous WcaG, pH- stat ( pH-stat) were produced up to 235.2 ± 3.3mg / L of GDP- fucose in fed-batch culture method, which is a result of a 21% improvement compared to the expression of the recombinant E. coli have not added G6PDH. ; In addition, guanosine-inosine kinase (guanosine-inosine kinase; Gsk) were added to the production of recombinant E. coli using the expression, up to 305.5 ± 5.3mg / L of GDP- fucose in the pH- stat fed-batch culture method, This is 58% better results than the expression of the recombinant E. coli did not add gsk. ; G6PDH, GDP-L-fucose, Gsk, GTP, NADPH, E. coli, fed-batch culture, fucose