Mechanism of the Sharpless Epoxidation Reaction: A DFT Study.

The Sharpless reaction is an enantioselective epoxidation of prochiral allylic alcohols that employs a Ti(IV) catalyst formed from titanium tetra(isopropoxide), Ti(O-i-Pr)4, diethyl tartrate (DET), and the oxidizing agent tert-butyl hydroperoxide. The M06-2X DFT functional with the 6-311+G(d,p) basis set has been employed to model the structures and energetics of the Sharpless epoxidation reaction. The monomeric tetracoordinate titanium(IV) diethyltartrate is thermodynamically strongly favored to dimerize, producing a pentacoordinate catalyst, [Ti(DET)(O-i-Pr)2]2, that is a more reactive chiral epoxidation catalyst. The rapid ligand exchange reactions needed to generate the "loaded" catalyst and to repeat the overall catalytic cycle have been examined and are found to have activation energies that are much lower than the epoxidation barriers. The transition structures and activation energies for the enantioselective epoxidation of allyl alcohol, trans-methyl-allyl alcohol and trans-tert-butyl-allyl alcohol with the "loaded" Sharpless catalyst, [Ti2(DET)2(O-i-Pr)2-(OAllyl)-(OOt-Bu)], are presented. The effect of the C═O···Ti interactions on the activation energies and the significance of the O-C-C═C dihedral angle on the enantioselectivity of the epoxidation reaction are discussed.

Mechanism of Orbital Interactions in the Sharpless Epoxidation with Ti(IV) Peroxides: A DFT Study.

The M06-2X DFT functional has been employed to examine monomeric titanium(IV) hydroperoxo catalysts that model the individual steps in the dimeric titanium(IV)-catalyzed Sharpless reaction. This is the first example of a transition structure for titanium(IV) tert-butyl hydroperoxide-catalyzed epoxidation that describes the molecular motion required for oxygen atom transfer. These epoxidation catalysts have been examined for both bimolecular reactions with E-2-butene and the intramolecular epoxidation of allyl alcohol. The transition structure for the bimolecular peroxyacetic acid epoxidation of E-2-butene has been shown to be spiro in nature, and likewise, the intramolecular epoxidation of allyl alcohol is also nearly spiro. The significance of the O-C-C═C dihedral angle of allyl alcohol is examined for the Ti(IV) tert-butyl hydroperoxide epoxidation mechanism. Evidence is presented that supports a hexacoordinate titanium peroxo environment that exists in the dimeric form of the Sharpless catalyst. The mechanism for a 1,3-rearrangement of the alkoxide ligand in a titanium hydroperoxide to the Ti center in concert with oxygen atom transfer of the proximal oxygen to the C═C bond of the substrate is presented. The dimerization of Ti(IV)-(R,R)-diethyl tartrate-diisopropoxide and its hydrolysis have been calculated. The mechanism for rapid ligand exchange with alkyl hydroperoxides involving the Ti(O-i-Pr)4 precursor is examined to show how the active epoxidation catalyst is produced.

The Bond Dissociation Energy of the N-O Bond.

Bond dissociation energy (BDE) has been calculated for a series of compounds that contain N-O bonds. These structures encompass model N,N,O-trisubstituted hydroxylamines that include O-methoxy, O-acyl, and O-phenyl hydroxylamines. The calculations used three accurate composite methods, CBS-QB3, CBS-APNO, and G4 methods and the computationally more affordable M06-2X/6-311+G(3df,2p) density functional theory (DFT) functional. The calculated N-O single-bond BDEs are 5-15 kcal/mol higher than a generic N-O BDE of 48 kcal/mol quoted in the literature and in textbooks. The M06-2X DFT functional provides BDEs that are in excellent agreement with the higher-level composite methods. We also provided a comparison of the N-O BDE for pyridine-N-oxide to simple trialkylamine oxides. Based on an experimental BDE of 63.3 ± 0.5 kcal/mol for pyridine-N-oxide, our best estimate gives 56.7 ± 0.9 kcal/mol N-O BDE for trimethylamine-N-oxide and 59.0 ± 0.8 kcal/mol for triethylamine-N-oxide.

Bond Dissociation Energy of Peroxides Revisited.

When dealing with organic peroxides in both laboratory and industrial applications, the relative strengths of the O-O bond are of vital importance, both from a safety and mechanistic perspective. Although it is well known that such oxidizing agents are highly reactive, reliable O-O bond dissociation energies (BDEs) have only recently been established. In an earlier report, we suggested a median O-O BDE value of ca. 45 kcal/mol for a variety of well-known peroxides based upon G2 ab initio calculations. In the present report, we have calculated the O-O BDE of twenty peroxides of varying structures at a more accurate CBS-APNO level. We have also compared these bond energies to the corresponding BDEs calculated with several DFT functionals and shown that the M06-2X functional produces O-O bond energies that compare very favorably with G4 and CBS-APNO values.

Structure and Mechanism for Alkane Oxidation and Alkene Epoxidation with Hydroperoxides, α-Hydroxy Hydroperoxides, and Peroxyacids: A Theoretical Study.

DFT calculations (B3LYP/6-311+G(d,p)) have been employed to reexamine the mechanism of the oxidation of saturated hydrocarbons and the epoxidation of alkenes with a series of hydroperoxides, α-hydroxy perhydrates, and peroxyacids. Hydrocarbon oxidation and alkene epoxidation with the hydroperoxide group involve a diradicaloid process initiated by a homolytic O-O bond cleavage involving a somersault rearrangement (1,2-hydrogen shift) induced abstraction of a hydrogen atom followed by a final product forming an "oxygen rebound" step. The epoxidation of alkenes and the oxidation of saturated hydrocarbons with peroxyacids is a concerted process involving a 1,4-hydrogen shift in the transition state. Accurate (G4) O-O bond dissociation energies (BDEs) for the series of peroxides are included. Although the O-O BDEs for the series of peroxides examined differ by only 3 kcal/mol, the activation energies reported differ by more than an order of magnitude. Both SCRF-PCM and natural bond order (NBO) analyses have been included.

The DMDO Hydroxylation of Hydrocarbons via the Oxygen Rebound Mechanism.

Both DFT and G4 molecular orbital calculations have been employed to reexamine the mechanism of dimethyldioxirane (DMDO) oxidation of saturated hydrocarbons and the epoxidation of alkenes. The UM062X DFT functional provided the most accurate bond O-O dissociation energies for a series of typical peroxides. A diradicaloid process initiated by an O-O homolytic bond cleavage involving abstraction of hydrogen from the C-H bond followed by a final product forming "oxygen rebound" step best describes the DMDO oxidation of saturated hydrocarbons. In contrast, this study showed that the DMDO epoxidation of alkenes is a concerted process best described with the B3LYP DFT functional.

Mechanism of N-hydroxylation catalyzed by flavin-dependent monooxygenases.

Aspergillus fumigatus siderophore (SidA), a member of class B flavin-dependent monooxygenases, was selected as a model system to investigate the hydroxylation mechanism of heteroatom-containing molecules by this group of enzymes. SidA selectively hydroxylates ornithine to produce N(5)-hydroxyornithine. However, SidA is also able to hydroxylate lysine with lower efficiency. In this study, the hydroxylation mechanism and substrate selectivity of SidA were systematically studied using DFT calculations. The data show that the hydroxylation reaction is initiated by homolytic cleavage of the O-O bond in the C(4a)-hydroperoxyflavin intermediate, resulting in the formation of an internal hydrogen-bonded hydroxyl radical (HO(•)). As the HO(•) moves to the ornithine N(5) atom, it rotates and donates a hydrogen atom to form the C(4a)-hydroxyflavin. Oxygen atom transfer yields an aminoxide, which is subsequently converted to hydroxylamine via water-mediated proton shuttling, with the water molecule originating from dehydration of the C(4a)-hydroxyflavin. The selectivity of SidA for ornithine is predicted to be the result of the lower energy barrier for oxidation of ornithine relative to that of lysine (16 vs 24 kcal/mol, respectively), which is due to the weaker stabilizing hydrogen bond between the incipient HO(•) and O3' of the ribose ring of NADP(+) in the transition state for lysine.

Mechanistic aspects regarding the elimination of H2O2 from C(4a)-hydroperoxyflavin. The role of a proton shuttle required for H2O2 elimination.

DFT calculations presented for C(4a)-hydroperoxyflavin (C(4a)-FLHOOH) at the B3LYP/6-311+G(d,p) level suggest a new mechanism for the elimination of H2O2. The calculated activation barrier for a concerted four-centered elimination (ΔE(‡) = 32.86 kcal/mol) strongly suggests that in the absence of interactions with the local environment a spontaneous elimination is not feasible. A proton shuttle from the N5 hydrogen to the proximal oxygen of the OOH moiety involving three water molecules has an activation barrier that is reduced to 17.11 kcal/mol. Calculations that utilize CH3OH to model the role of a local Thr or Ser residue shows that an alcohol functionality hydrogen bonded to the N5 H-atom can catalyze the elimination of H2O2 with a free energy of activation of 21.5 kcal/mol. Interaction of amines and amide residues (CH3NH2 and CH3(C═O)NH2) with the N5 locus of C(4a)-hydroperoxyflavin markedly reduce the activation barrier for H2O2 elimination relative to the concerted pathway. Proton transfer from a COOH group (ΔG(‡) = 8.36 kcal/mol) or the NH2 group of a positively charged Arg model (ΔG(‡) = 9.99 kcal/mol) to the proximal oxygen of the OOH moiety of C(4a)-FLHOOH in the TS for H2O2 elimination strongly enhances elimination of H2O2.

The role of acid catalysis in the Baeyer-Villiger reaction. A theoretical study.

Quantum mechanical calculations at the B3LYP/6-311+G(d,p) level have examined the overall mechanism of the Baeyer-Villiger (BV) reaction with peroxyacetic acid. A series of reactions that include both the addition step and the subsequent alkyl group migration step included ketones, acetone, t-butyl methyl ketone, acetophenone, cyclohexyl methyl ketone, and cyclohexyl phenyl ketone. The combined data suggested that the first step for addition of the peroxyacetic acid oxidation catalyst to the ketone carbonyl to produce the Criegee or tetrahedral intermediate is rate-limiting and has activation barriers that range from 38 to 41 kcal/mol without the aid of a catalyst. The rate of addition is markedly reduced by the catalytic action of a COOH functionality acting as a donor-acceptor group affecting both its proton transfer to the ketone C═O oxygen in concert with transfer of the OOH proton to the carboxylic acid carbonyl. The second or alkyl group migration step has a much reduced activation barrier, and its rate is not markedly influenced by acid catalysis. The rate of both steps in the BV reaction is greatly influenced by the catalytic action of very strong acids.

Role of the somersault rearrangement in the oxidation step for flavin monooxygenases (FMO). A comparison between FMO and conventional xenobiotic oxidation with hydroperoxides.

Model quantum mechanical calculations presented for C-4a-flavin hydroperoxide (FlHOOH) at the B3LYP/6-311+G(d,p) level suggest a new mechanism for flavoprotein monooxygenase (FMO) oxidation involving a concerted homolytic O-O bond cleavage in concert with hydroxyl radical transfer from the flavin hydroperoxide rather than an S(N)2-like displacement by the substrate on the C-4a-hydroperoxide OOH group. Homolytic O-O bond cleavage in a somersault-like rearrangement of hydroperoxide C-4a-flavinhydroperoxide (1) (FLHO-OH → FLHO···HO) produces an internally hydrogen-bonded HO(•) radical intermediate with a classical activation barrier of 27.0 kcal/mol. Model hydroperoxide 1 is used to describe the transition state for the key oxidation step in the paradigm aromatic hydroxylase, p-hydroxybenzoate hydroxylase (PHBH). A comparison of the electron distribution in the transition structures for the PHBH hydroxylation of p-hydroxybenzoic acid (ΔE(‡) = 23.0 kcal/mol) with that of oxidation of trimethylamine (ΔE(‡) = 22.3 kcal/mol) and dimethyl sulfide (ΔE‡ = 14.1 kcal/mol) also suggests a mechanism involving a somersault mechanism in concert with transfer of an HO(•) radical to the nucleophilic heteroatom center with a hydrogen transfer back to the FLH-O residue after the barrier is crossed to produce the final product, FLH-OH. In each case the hydroxylation barrier was less than that of the O-O rearrangement barrier in the absence of a substrate supporting an overall concerted process. All three transition structures bear a resemblance to the TS for the comparable hydroxylation of isobutane (ΔE(‡) = 29.2 kcal/mol) and for simple Fenton oxidation by aqueous iron(III) hydroperoxides. To our surprise the oxidation of N- and S-nucleophiles with conventional oxidants such as alkyl hydroperoxides and peracids also proceeds by HO(•) radical transfer in a manner quite similar to that for tricyclic hydroperoxide 1. Stabilization of the developing oxyradical produced by somersault rearrangement for concerted enzymatic oxidation with tricyclic hydroperoxide 1 results in a reduced overall activation barrier.

The rate-limiting step in P450 hydroxylation of hydrocarbons a direct comparison of the "somersault" versus the "consensus" mechanism involving compound I.

Model theoretical quantum mechanical (QM) calculations are described for the P-450 hydroxylation of methane, isobutane, and camphor that compare the concerted somersault H-abstraction mechanism with the oxidation step involving Cpd I. Special emphasis has been placed on maintaining a balanced basis set in the oxidation step. QM calculations, employing the 6-311+G(d,p) basis set on the Fe atom and all of the key surrounding atoms involved in the C-H abstraction step, reaffirm a mechanism involving rearrangement of the iron hydroperoxide group (FeO-OH --> FeO...HO(*)) in concert with hydrogen abstraction from the C-H bond of the substrate by the incipient bound hydroxyl radical HO(*). The barrier for the somersault rearrangement of model Cpd 0 (FeO-OH) is calculated to be 21.4 kcal/mol in the absence of substrate. The overall activation energy for the oxidation of camphor involving the somersault motion of the FeO-OH group of P450 model porphyrin iron(III) hydroperoxide [Por(SH)Fe(III)-OOH(-)] --> [Por(SH)Fe(III)-O....HO(-)] in concert with hydrogen abstraction is DeltaE(++) = 12.4 kcal/mol. The corresponding abstraction of the hydrogen atom from the C-H bond of camphor by Cpd I has an activation barrier of 17.6 kcal/mol. Arguments are presented that the somersault rearrangement is induced by steric compression at the active site. Kinetic isotope effect data are discussed that provides compelling evidence for a rate-limiting step involving C-H bond cleavage.

Transient inverted metastable iron hydroperoxides in fenton chemistry. A nonenzymatic model for cytochrome p450 hydroxylation.

Quantum mechanical calculations (DFT) have provided a mechanism for the oxidative C-H bond cleavage step in Fenton-like hydrocarbon hydroxylation. A transition structure for hydrocarbon oxidation by aqueous solvated cationic iron(III) hydroperoxides ((H(2)O)(n)Fe(III)OOH) is presented that involves a novel rearrangement of the hydroperoxide group (FeO-OH --> FeO...HO) in concert with hydrogen abstraction by the incipient HO* radical with activation barriers ranging from 17 to 18 kcal/mol. In every hydroperoxide examined, the activation barrier for FeO-OH isomerization, in the absence of the hydrocarbon, is significantly greater than the overall concerted activation barrier for C-H bond cleavage in support of the concept of O-O bond isomerization in concert with hydrogen abstraction. The transition structure for the oxidation step in simple anionic iron(III) hydroperoxides has been shown to bear a remarkable resemblance to model porphyrin calculations on cytochrome P450 hydroxylation.

Ring strain energy in the cyclooctyl system. The effect of strain energy on [3 + 2] cycloaddition reactions with azides.

Ring strain energies (SEs) and enthalpies of hydrogenation (DeltaH(hyd)) of a series of E- and Z-alkenes, cyclic alkynes and allenes (C(5)-C(9)) are computed at the G3 level of theory. The SE for cycloheptyne, cyclohexyne, and cyclopentyne are calculated to be 25.4, 40.1, and 48.4 kcal/mol, respectively. The SE for E-cycloheptene and E-cyclohexene are calculated to be 25.2 and 49.3 kcal/mol (G3). The SE of cyclooctyne is 2.0 kcal/mol greater than that of E-cyclooctene (17.9 kcal/mol) but only 7.7 kcal/mol greater than that of cyclooctane. The SE of 3,3-difluorocyclooctyne (DIFO) is predicted to be slightly reduced (DeltaSE = 2.6 kcal/mol) relative to the parent cyclooctyne to 17.3 kcal/mol. The SE and DeltaH(hyd) are correlated with activation barriers for the [3 + 2] cycloaddition of a series of azides to E- and Z-cycloalkenes and alkynes at the G3 level of theory. The energy barrier for the cycloaddition of methyl azide to cyclooctyne is 9.2 kcal/mol lower than addition to 4-octyne and 3.1 kcal/mol lower for reaction with E-cyclooctene. The activation energies for [3 + 2] cycloaddition of benzyl azide and acetamido azide ((2)HN(C=O)CH(2)-N(3)) to DIFO are 2.3 and 5.3 kcal/mol lower in energy than cycloaddition to cyclooctyne [B3LYP/6-311+G(3df,2p)].

The Curtius rearrangement of cyclopropyl and cyclopropenoyl azides. A combined theoretical and experimental mechanistic study.

A combined experimental and theoretical study addresses the concertedness of the thermal Curtius rearrangement. The kinetics of the Curtius rearrangements of methyl 1-azidocarbonyl cycloprop-2-ene-1-carboxylate and methyl 1-azidocarbonyl cyclopropane-1-carboxylate were studied by (1)H NMR spectroscopy, and there is close agreement between calculated and experimental enthalpies and entropies of activation. Density functional theory (DFT) calculations (B3LYP/6-311+G(d,p)) on these same acyl azides suggest gas phase barriers of 27.8 and 25.1 kcal/mol. By comparison, gas phase activation barriers for the rearrangement of acetyl, pivaloyl, and phenyl azides are 27.6, 27.4, and 30.0 kcal/mol, respectively. The barrier for the concerted Curtius reaction of acetyl azide at the CCSD(T)/6-311+G(d,p) level exhibited a comparable activation energy of 26.3 kcal/mol. Intrinsic reaction coordinate (IRC) analyses suggest that all of the rearrangements occur by a concerted pathway with the concomitant loss of N2. The lower activation energy for the rearrangement of methyl 1-azidocarbonyl cycloprop-2-ene-1-carboxylate relative to methyl 1-azidocarbonyl cyclopropane-1-carboxylate was attributed to a weaker bond between the carbonyl carbon and the three-membered ring in the former compound. Calculations on the rearrangement of cycloprop-2-ene-1-oyl azides do not support pi-stabilization of the transition state by the cyclopropene double bond. A comparison of reaction pathways at the CBS-QB3 level for the Curtius rearrangement versus the loss of N2 to form a nitrene intermediate provides strong evidence that the concerted Curtius rearrangement is the dominant process.

Mechanism of thiolate-disulfide interchange reactions in biochemistry.

Both density functional theory (DFT) (B3LYP) and CCSD ab initio calculations were employed in a theoretical investigation of the mechanism of thiolate-disulfide exchange reactions. The reaction pathway for degenerate thiolate-disulfide exchange reactions with dimethyl disulfide has been shown to proceed through a SN2-like transition structure that is very close in energy to the corresponding trisulfur anionic intermediate ([delta-S-S-Sdelta(-)]). When relatively small substituents are involved, the level of theory must be increased to CCSD to make this rather subtle mechanistic distinction. With the more sterically hindered exchange reaction involving t-butyl mercaptide and di-t-butyl disulfide, the potential energy surface exhibits a distinct preference for the S(N)2 displacement pathway with an activation barrier of 9.8 kcal/mol. When corrections for solvent polarity are included (COSMO), an S(N)2 mechanism is also implicated in both polar and nonpolar solvents. DFT studies on thiolate-disulfide exchange, when the substituent is a model peptide, strongly support the intermediacy of a trisulfur intermediate that lies 10.7 kcal/mol below isolated reactants. A well depth of this magnitude should provide a sufficient lifetime of the intermediate to accommodate the requisite conformational adjustments that accompanies formation of the new disulfide bond.

Mechanism of SN2 disulfide bond cleavage by phosphorus nucleophiles. Implications for biochemical disulfide reducing agents.

The B3LYP variant of DFT has been used to study the mechanism of S-S bond scission in dimethyl disulfide by a phosphorus nucleophile, trimethylphospine (TMP). The reaction is highly endothermic in the gas phase and requires significant external stabilization of the charged products. DFT calculations (B3LYP) were performed with explicit (water molecules added) and implicit solvent corrections (COSMO model). The transition structures for this SN2 displacement reaction in a number of model systems have been located and fully characterized. The reaction barriers calculated with different approaches for different systems are quite close (around 11 kcal/mol). Remarkably, the calculations suggest that the reaction is almost barrierless with respect to the preorganized reaction complex and that most of the activation energy is required to rearrange the disulfide and TMP to its most effective orientation for the SMe group transfer way. Different reactivities of different phosphorus nucleophiles were suggested to be the result of steric effects, as manifested largely by varying amounts of hindrance to solvation of the initial product phosphonium ion. These data indicate that the gas-phase addition of a phosphine to the disulfide moiety will most likely form a phosphonium cation-thiolate anion salt, in the presence of four or more water molecules, that provide sufficient H-bonding stabilization to allow displacement of the thiolate anion, a normal uncomplicated SN2 transition state is to be expected.

The effect of carbonyl substitution on the strain energy of small ring compounds and their six-member ring reference compounds.

High level ab initio calculations have been applied to the estimation of ring strain energies (SE) of a series of three- and six-member ring compounds. The SE of cyclohexane has been estimated to be 2.2 kcal/mol at the CBS-APNO level of theory. The SE of cyclopropane has been increased to 28.6 kcal/mol after correction for the one-half of the SE of cyclohexane. The SEs of a series of carbonyl-containing three-member ring compounds have been estimated at the CBS-Q level by their combination with cyclopropane to produce a six-member ring reference compound. The SEs of cyclopropanone (5), the simplest alpha-lactone (6) [oxiranone], and alpha-lactam (7) [aziridinone] have been predicted to be 49, 47, and 55 kcal/mol, respectively, after correction for the SE of the corresponding six-member ring reference compound. The SEs of cyclohexanone, delta-valerolactone, and delta-valerolactam have been estimated to be 4.3, 11.3, and 5.1 kcal/mol, respectively. Marked increases in the SE of silacyclopropane and siladioxirane have been established, while significant decreases in the SEs of phosphorus, sulfur, dioxa- and diaza-containing three-member ring compounds were observed. The ring strain energies of the hydrocarbons (but not heterocycles) exhibit a strong correlation with their C-H bond dissociation energies.

The "somersault" mechanism for the p-450 hydroxylation of hydrocarbons. The intervention of transient inverted metastable hydroperoxides.

A series of model theoretical calculations are described that suggest a new mechanism for the oxidation step in enzymatic cytochrome P450 hydroxylation of saturated hydrocarbons. A new class of metastable metal hydroperoxides is described that involves the rearrangement of the ground-state metal hydroperoxide to its inverted isomeric form with a hydroxyl radical hydrogen bonded to the metal oxide (MO-OH --> MO....HO). The activation energy for this somersault motion of the FeO-OH group is 20.3 kcal/mol for the P450 model porphyrin iron(III) hydroperoxide [Por(SH)Fe(III)-OOH(-)] to produce the isomeric ferryl oxygen hydrogen bonded to an *OH radical [Por(SH)Fe(III)-O....HO(-)]. This isomeric metastable hydroperoxide, the proposed primary oxidant in the P450 hydroxylation reaction, is calculated to be 17.8 kcal/mol higher in energy than the ground-state iron(III) hydroperoxide Cpd 0. The first step of the proposed mechanism for isobutane oxidation is abstraction of a hydrogen atom from the C-H bond of isobutane by the hydrogen-bonded hydroxyl radical to produce a water molecule strongly hydrogen bonded to anionic Cpd II. The hydroxylation step involves a concerted but nonsynchronous transfer of a hydrogen atom from this newly formed, bound, water molecule to the ferryl oxygen with a concomitant rebound of the incipient *OH radical to the carbon radical of isobutane to produce the C-O bond of the final product, tert-butyl alcohol. The TS for the oxygen rebound step is 2 kcal/mol lower in energy than the hydrogen abstraction TS (DeltaE() = 19.5 kcal/mol). The overall proposed new mechanism is consistent with a lot of the ancillary experimental data for this enzymatic hydroxylation reaction.

Chemical behavior of the biradicaloid (HO...ONO) singlet states of peroxynitrous acid. The oxidation of hydrocarbons, sulfides, and selenides.

Various high levels of theory have been applied to the characterization of two higher lying biradicaloid metastable singlet states of peroxynitrous acid. A singlet minimum (cis-2) was located that had an elongated O-O distance (2.17 A) and was only 12.2 kcal/mol [UB3LYP/6-311+G(3df,2p)+ZPVE] higher in energy than its ground-state precursor. A trans-metastable singlet (trans-2) was 10.9 kcal/mol higher in energy than ground-state HO-ONO. CASSCF(12,10)/6-311+G(d,p) calculations predict the optimized geometries of these cis- and trans-metastable singlets to be close to those obtained with DFT. Optimization of cis- and trans-2 within the COSMO solvent model suggests that both exist as energy minima in polar media. Both cis- and trans-2 exist as hydrogen bonded complexes with several water molecules. These collective data suggest that solvated forms of cis-2.3H(2)O and trans-2.3H(2)O represent the elusive higher lying biradicaloid minima that were recently (J. Am. Chem. Soc. 2003, 125, 16204) advocated as the metastable forms of peroxynitrous acid (HOONO). The involvement of metastable trans-2 in the gas phase oxidation of methane and isobutane is firmly established to take place on the unrestricted [UB3LYP/6-311+G(d,p)] potential energy surface (PES) with classical activations barriers for the hydrogen abstraction step that are 15.7 and 5.9 kcal/mol lower than the corresponding activation energies for producing products methanol and tert-butyl alcohol formed on the restricted PES. The oxidation of dimethyl sulfide and dimethyl selenide, two-electron oxidations, proceeds by an S(N)2-like attack of the heteroatom lone pair on the O-O bond of ground-state peroxynitrous acid. No involvement of metastable forms of HO-ONO was discernible.