Production of activated TDP-deoxysugars in recombinant microorganisms

A method of producing a thymidine diphosphate (TDP)-deoxysugar, comprising:

• culturing a recombinant microorganism comprising Escherichia coli strain zucTKDG in culture medium comprising glucose, fructose, gluconate, glycerol, or combinations thereof, under conditions suitable for conversion of glucose, fructose, gluconate, glycerol, or combinations thereof by the recombinant microorganism to a TDP-deoxysugar;
• and
• isolating the TDP-deoxysugar,
• wherein the recombinant microorganism:
• (a) comprises a functional deletion of rmIC (dTDP-4-dehydrorhamnose 3,5-epimerase), rmlD (dTDP-4-dehydrorhamnose reductase), wecE (TDP-4-oxo-6-deoxy-D-glucose transaminase), and wecD (TDP-fucosamine acetyltransferase);
• (b) expresses a recombinant rmIA (glucose-1-phosphate thymidylyltransferase);
• and
• (c) expresses:
• (i) a TDP-deoxysugar biosynthetic protein consisting of Fcf1 to produce TDP-D-fucose;
• (ii) TDP-deoxysugar biosynthetic proteins consisting of Fcf1 and Fcf2 to produce TDP-D-fucofuranose;
• (iii) TDP-deoxysugar biosynthetic proteins consisting of KijB1 or Olev, KijD10 or OleW, and ChIC5 to produce TDP-D-olivose;
• (iv) a TDP-deoxysugar biosynthetic protein consisting of RmIC to produce TDP-L-rhamnose;
• (v) TDP-deoxysugar biosynthetic proteins consisting of RmIC, NovU, and NovS to produce TDP-L-noviose;
• (vi) TDP-deoxysugar biosynthetic proteins consisting of NovW or GerF and GerKI to produce TDP-6-deoxy-D-allose;
• (vii) TDP-deoxysugar biosynthetic proteins consisting of KijB1 or OleV, MegDII or EvaB, DvaC or EvaC, EvaD or HedK, and EvaE to produce TDP-L-eremosamine;
• (viii) TDP-deoxysugar biosynthetic proteins consisting of KijB1 or OleV, KijD10 or OleW, SpnQ or UrdQ, and SpnR to produce TDP-N,N-didemethyl-D-forosamine;
• (ix) TDP-deoxysugar biosynthetic proteins consisting of DesI, DesII, DesV, or MegDII, and DesVI or MegDIII to produce TDP-D-desosamine;
• (x) a TDP-deoxysugar biosynthetic protein consisting of DesI to produce TDP-4-amino-D-quinovose;
• or (xi) TDP-deoxysugar biosynthetic proteins consisting of DesI or DesII and DesV or MegDII to produce TDP-N,N-didemethyl-D-desosamine.