The stereochemistry of NADH utilization by the flavoenzyme monooxygenase orcinol hydroxylase.
Ribbons et al. (Ribbons, D.W., Ohta, Y., and Higgins, I.J. (1972) in Molecular Basis of Electron Transport, Miami Winter Symposic Series (Schultz, J., and Cameron, B.F., eds) Vol. 4, pp. 251-274, Academic Press, New York) presented a preliminary report that the flavoenzyme monooxygenase orcinol hydroxylase shows mixed type 4R, 4S stereospecificity with respect to dihydronicotinamide oxidation when resorcinol and m-cresol were used as substrate analogs. With the natural substrate orcinol, 4R chirality was maintained. In kinetic isotope experiments reported here, we demonstrate in fact that orcinol hydroxylase maintains 4R stereospecificity with respect to dihydronicotinamide oxidation with all three substrates, orcinol, resorcinol, and m-cresol. Deuterium and tritium kinetic isotope effects were detected under Vmax conditions with (4R)-[4-2H]-, and (4R)-[4-3H]NADH for all three substrates. No isotope effect was observed with (4S)-[4-2H]NADH and tritium labilization from assays with (4S)-[4-3H]-NADH was negligible in all cases.
D003903:Deuterium; D007700:Kinetics; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D008968:Molecular Conformation; D009243:NAD; D011485:Protein Binding; D011549:Pseudomonas / Q000201:enzymology; D012118:Resorcinols; D014316:Tritium
D012118:Resorcinols; D009243:NAD; D014316:Tritium; D003903:Deuterium; D006899:Mixed Function Oxygenases; C021911:orcinol 2-monooxygenase