Rapid one-pot synthesis of riboflavin isotopomers.
Flavocoenzymes labeled with stable isotopes are important reagents for the study of flavoproteins using isotope-sensitive methods such as NMR, ENDOR, infrared, and Raman spectroscopy. We describe highly versatile one-pot methods for the preparation of riboflavin isotopomers labeled with (13)C in every desired position of the xylene moiety. The starting materials are commercially available (13)C-labeled glucose samples, which are converted into riboflavin using enzymes of the oxidative pentose phosphate pathway in combination with recombinant enzymes of the riboflavin biosynthetic pathway. The overall reaction comprises six enzyme-catalyzed reaction steps for the synthesis of the vitamin and two auxiliary enzymes for in situ recycling of cofactors. The overall yields of riboflavin based on isotope-labeled glucose are 35-50%.
Römisch
Werner
W
Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstr. 4, D-85747 Garching, Germany.
Eisenreich
Wolfgang
W
Richter
Gerald
G
Bacher
Adelbert
A
D001412:Bacillus subtilis / Q000201:enzymology; D002247:Carbon Isotopes; D004578:Electron Spin Resonance Spectroscopy; D004926:Escherichia coli / Q000201:enzymology; D005486:Flavin Mononucleotide / Q000138:chemical synthesis*; D007202:Indicators and Reagents; D019751:Intramolecular Transferases / Q000378:metabolism; D007536:Isomerism; D007553:Isotope Labeling / Q000379:methods; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D009097:Multienzyme Complexes / Q000378:metabolism; D012256:Riboflavin / Q000032:analysis* / Q000138:chemical synthesis*; D012258:Riboflavin Synthase / Q000378:metabolism; D013055:Spectrophotometry, Infrared; D013059:Spectrum Analysis, Raman
D002247:Carbon Isotopes; D007202:Indicators and Reagents; D009097:Multienzyme Complexes; D005486:Flavin Mononucleotide; C044822:6,7-dimethyl-8-ribityllumazine synthase; D012258:Riboflavin Synthase; D019751:Intramolecular Transferases; C066493:L-3,4-dihydroxy-2-butanone-4-phosphate synthase; D012256:Riboflavin