title: 2-thioriboflavin 5'-phosphate (2-thio-FMN) lactate oxidase.
abstract: The natural flavin of lactate oxidase, FMN, was removed and replaced by the synthetic flavin 2-thioriboflavin 5'-phosphate (2-thio FMN). Despite the difference in properties of the flavins, including an oxidation-reduction potential some 80 mV more positive than that of normal flavin the 2-thio-FMN enzyme behaves in practically all respects like the native enzyme. It catalyzes the oxidative decarboxylation of L-lactate with almost the same efficiency as the native enzyme and with similar kinetic constants for individual steps in the catalytic pathway. It forms covalent derivatives at the flavin N(5) and C(4a) positions in facile photochemical reactions analogous to those of the native enzyme. It also forms a flavin anion radical on photoreduction with 5-deazaflavin as catalyst and, as with the native enzyme, this radical is stabilized remarkably on formation of a complex with pyruvate. The spectral properties of the neutral flavin radical form of 2-thioflavin are also reported, as determined by photochemical reduction of 2-thio-FMN flavodoxin. Like native lactate oxidase, the 2-thio-FMN enzyme also forms a flavin N(5)-sulfite adduct in an equilibrium reaction with sulfite. These results demonstrate clearly with this enzyme that the native flavin may be removed and replaced by an artificial flavin, without altering the structural integrity of the protein.
mesh_terms: D000693:Anaerobiosis; D002384:Catalysis; D005486:Flavin Mononucleotide / Q000031:analogs & derivatives* / Q000378:metabolism; D007700:Kinetics; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D010777:Photochemistry; D011485:Protein Binding; D011773:Pyruvates / Q000378:metabolism; D019289:Pyruvic Acid; D013053:Spectrophotometry
chemical_list: D011773:Pyruvates; C027153:2-thio-FMN; D005486:Flavin Mononucleotide; D019289:Pyruvic Acid; D006899:Mixed Function Oxygenases; D010088:Oxidoreductases; C021497:lactate 2-monooxygenase