(Chemo)enzymatic synthesis of dTDP-activated 2,6-dideoxysugars as building blocks of polyketide antibiotics.
The flexible substrate spectrum of the recombinant enzymes from the biosynthetic pathway of dTDP-beta-L-rhamnose in Salmonella enterica, serovar typhimurium (LT2), was exploited for the chemoenzymatic synthesis of deoxythymidine diphosphate- (dTDP-) activated 2,6-dideoxyhexoses. The enzymatic synthesis strategy yielded dTDP-2-deoxy-alpha-D-glucose and dTDP-2,6-dideoxy-4-keto-alpha-D-glucose (13) in a 40-60 mg scale. The nucleotide deoxysugar 13 was further used for the enzymatic synthesis of dTDP-2,6-dideoxy-beta-L-arabino-hexose (dTDP-beta-L-olivose) (15) in a 30-mg scale. The chemical reduction of 13 gave dTDP-2,6-dideoxy-alpha-D-arabino-hexose (dTDP-alpha-D-olivose) (1) as the main isomer after product isolation in a 10-mg scale. With 13 as an important key intermediate, the in vitro characterization of enzymes involved in the biosynthesis of dTDP-activated 2,6-dideoxy-, 2,3,6-trideoxy-D- and L-hexoses can now be addressed. Most importantly, compounds 1 and 15 are donor substrates for the in vitro characterization of glycosyltransferases involved in the biosynthesis of polyketides and other antibiotic/antitumor drugs. Their synthetic access may contribute to the evaluation of the glycosylation potential of bacterial glycosyltransferases to generate hybrid antibiotics.
Amann
S
S
Institute of Enzyme Technology, Heinrich-Heine-University, Düsseldorf Research Center Jülich, D-52426 Jülich, Germany.
Dräger
G
G
Rupprath
C
C
Kirschning
A
A
Elling
L
L
D000818:Animals; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry; D002237:Carbohydrate Dehydrogenases / Q000378:metabolism; D002238:Carbohydrate Epimerases / Q000378:metabolism; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003837:Deoxy Sugars / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006836:Hydro-Lyases / Q000378:metabolism; D008274:Magnesium / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D009713:Nucleotidyltransferases / Q000378:metabolism; D011817:Rabbits; D019779:Salmonella enterica / Q000201:enzymology*; D013379:Substrate Specificity; D013942:Thymine Nucleotides / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D001426:Bacterial Proteins
D000900:Anti-Bacterial Agents; D002237:Carbohydrate Dehydrogenases; D002238:Carbohydrate Epimerases; D003837:Deoxy Sugars; D006836:Hydro-Lyases; D008274:Magnesium; D009713:Nucleotidyltransferases; D013942:Thymine Nucleotides; C439753:dTDP-2,6-dideoxy-4-ketoglucose; C439751:dTDP-2,6-dideoxyglucose; C439752:dTDP-olivose; C027923:thymidine 5'-diphosphate; C107144:dTDP-4-dehydrorhamnose reductase; C020587:glucose-1-phosphate thymidylyltransferase; C022771:dTDPglucose 4,6-dehydratase; C020092:dTDP-4-ketorhamnose 3,5-epimerase; D001426:Bacterial Proteins; C024157:thymidine 5'-triphosphate