title: Preparation of flavocoenzyme isotopologues by biotransformation of purines.
abstract: Isotope-labeled flavins are crucial reporters for many biophysical studies of flavoproteins. A purine-deficient Escherichia coli strain engineered for expression of the ribAGH genes of Bacillus subtilis converts isotope-labeled purine supplements into the riboflavin precursor, 6,7-dimethyl-8-ribityllumazine, with yields up to 40%. The fermentation products can subsequently be converted into isotope-labeled riboflavin and the cognate flavocoenzymes, FMN and FAD, by in vitro biotransformation with better than 90% yield. Using this approach, more than 100 single or multiple (13)C-, (15)N-, (17)O-, and (18)O-labeled isotopologues of these cofactors and ligands become easily accessible, enabling advanced ligand-based spectroscopy of flavoproteins and lumazine receptor proteins at unprecedented resolution.
authors: Hamburg School of Food Science, University of Hamburg , Grindelallee 117, 20146 Hamburg, Germany.
authors: Boris
authors: B
authors: Illarionov
authors: Feng
authors: F
authors: Zhu
authors: Wolfgang
authors: W
authors: Eisenreich
authors: Adelbert
authors: A
authors: Bacher
authors: Stefan
authors: S
authors: Weber
authors: Markus
authors: M
authors: Fischer
mesh_terms: D001412:Bacillus subtilis / Q000737:chemistry*; D001711:Biotransformation; D004926:Escherichia coli / Q000737:chemistry* / Q000201:enzymology*; D005420:Flavoproteins / Q000737:chemistry*; D007553:Isotope Labeling / Q000379:methods*; D008024:Ligands; D011621:Pteridines / Q000138:chemical synthesis* / Q000737:chemistry*; D011687:Purines / Q000737:chemistry*; D012256:Riboflavin / Q000737:chemistry*; D012258:Riboflavin Synthase / Q000737:chemistry* / Q000378:metabolism
chemical_list: D005420:Flavoproteins; D008024:Ligands; D011621:Pteridines; D011687:Purines; C001061:6,7-dimethyl-8-ribityllumazine; D012258:Riboflavin Synthase; D012256:Riboflavin; C030985:purine