title: Solvent isotope effects on alkane formation by cyanobacterial aldehyde deformylating oxygenase and their mechanistic implications.
abstract: The reaction catalyzed by cyanobacterial aldehyde deformylating oxygenase is of interest both because of its potential application to the production of biofuels and because of the highly unusual nature of the deformylation reaction it catalyzes. Whereas the proton in the product alkane derives ultimately from the solvent, the identity of the proton donor in the active site remains unclear. To investigate the proton transfer step, solvent isotope effect (SIE) studies were undertaken. The rate of alkane formation was found to be maximal at pH 6.8 and to be the same in D2O or H2O within experimental error, implying that proton transfer is not a kinetically significant step. However, when the ratio of protium to deuterium in the product alkane was measured as a function of the mole fraction of D2O, a (D2O)SIEobs of 2.19 ± 0.02 was observed. The SIE was invariant with the mole fraction of D2O, indicating the involvement of a single protic site in the reaction. We interpret this SIE as most likely arising from a reactant state equilibrium isotope effect on a proton donor with an inverse fractionation factor, for which Φ = 0.45. These observations are consistent with an iron-bound water molecule being the proton donor to the alkane in the reaction.
authors.affiliation: Department of Chemistry and ‡Department of Biological Chemistry, University of Michigan , Ann Arbor, Michigan 48109, United States.
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Waugh
authors.forename: E Neil G
authors.initials: EN
authors.lastname: Marsh
mesh_terms: D000447:Aldehydes / Q000378:metabolism; D000473:Alkanes / Q000378:metabolism*; D000458:Cyanobacteria / Q000201:enzymology* / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D007554:Isotopes; D010105:Oxygenases / Q000378:metabolism*; D012997:Solvents / Q000737:chemistry*
chemical_list: D000447:Aldehydes; D000473:Alkanes; D007554:Isotopes; D012997:Solvents; D010105:Oxygenases