Binding and Stabilization of a Semiquinone Radical by an artificial Metalloenzyme containing a Binuclear copper (II) cofactor.

A binuclear Cu(II) cofactor was covalently bound to a lauric acid anchor. The resulting conjugate was characterized then combined with beta-lactoglobulin (ßLG) to generate a new biohybrid following the so-called "Trojan horse" strategy. This biohybrid was examined for its effectiveness in the oxidation of a catechol derivative to the corresponding quinone. The resulting biohybrid did not exhibit the sought after catecholase activity, likely due to its ability to bind and stabilize the semiquinone radical intermediate DTB-SQ. This semi-quinone radical was stabilized only in the presence of the protein and was characterized using optical and magnetic spectroscopic techniques, demonstrating stability for over 16 hours. Molecular docking studies revealed that this stabilization could occur owing to interactions of the semi-quinone with hydrophobic amino acid residues of ßLG.

Proton Domino Reactions at an Imidazole Relay Control the Oxidation of a TyrZ-His190 Artificial Mimic of Photosystem II.

A close mimic of P680 and the TyrosineZ-Histidine190 pair in photosystem II (PS II) has been synthesized using a ruthenium chromophore and imidazole-phenol ligands. The intramolecular oxidation of the ligands by the photoproduced Ru(III) species is characterized by a small driving force, very similar to PS II where the complexity of kinetics was attributed to the reversibility of electron transfer steps. Laser flash photolysis revealed biphasic kinetics for ligand oxidation. The fast phase (τ < 50 ns) corresponds to partial oxidation of the imidazole-phenol ligand, proton transfer within the hydrogen bond, and formation of a neutral phenoxyl radical. The slow phase (5 - 9 µs) corresponds to full oxidation of the ligand which is kinetically controlled by deprotonation of the distant 1-nitrogen of the imidazolium. These results show that imidazole with its two protonatable sites plays a special role as a proton relay in a 'proton domino' reaction.

Catalytic oxidation properties of an acid-resistant cross-bridged cyclen Fe(II) complex. Influence of the rigid donor backbone and protonation on the reactivity.

The catalytic properties of an iron complex bearing a pentadentate cross-bridged ligand backbone are reported. With H2O2 as an oxidant, it displays moderate conversions in epoxidation and alkane hydroxylation and satisfactory ones in aromatic hydroxylation. Upon addition of an acid to the reaction medium, a significant enhancement in aromatic and alkene oxidation is observed. Spectroscopic analyses showed that accumulation of the expected FeIII(OOH) intermediate is limited under these conditions, unless an acid is added to the mixture. This is ascribed to the inertness induced by the cross-bridged ligand backbone, which is partly reduced under acidic conditions.

Mechanistic Facets of the Competition between Cross-Coupling and Homocoupling in Supporting Ligand-Free Iron-Mediated Aryl-Aryl Bond Formations.

In the context of cross-coupling chemistry, the competition between the cross-coupling path itself and the oxidative homocoupling of the nucleophile is a classic issue. In that case, the electrophilic partner acts as a sacrificial oxidant. We investigate in this report the factors governing the cross- versus homocoupling distribution using aryl nucleophiles ArMgBr and (hetero)aryl electrophiles Ar'Cl in the presence of an iron catalyst. When electron-deficient electrophiles are used, a key transient heteroleptic [Ar2Ar'FeII]- complex is formed. DFT calculations show that an asynchronous two-electron reductive elimination follows, which governs the selective evolution of the system toward either a cross- or homocoupling product. Proficiency of the cross-coupling reductive elimination strongly depends on both π-accepting and σ-donating effects of the FeII-ligated Ar' ring. The reactivity trends discussed in this article rely on two-electron elementary steps, which are in contrast with the usually described tendencies in iron-mediated oxidative homocouplings which involve single-electron transfers. The results are probed by paramagnetic 1H NMR spectroscopy, experimental kinetics data, and DFT calculations.

Heterolytic O-O Bond Cleavage Upon Single Electron Transfer to a Nonheme Fe(III)-OOH Complex.

The one-electron reduction of the nonheme iron(III)-hydroperoxo complex, [FeIII (OOH)(L5 2 )]2+ (L5 2 =N-methyl-N,N',N'-tris(2-pyridylmethyl)ethane-1,2-diamine), carried out at -70 °C results in the release of dioxygen and in the formation of [FeII (OH)(L5 2 )]+ following a bimolecular process. This reaction can be performed either with cobaltocene as chemical reductant, or electrochemically. These experimental observations are consistent with the disproportionation of the hydroperoxo group in the putative FeII (OOH) intermediate generated upon reduction of the FeIII (OOH) starting complex. One plausible mechanistic scenario is that this disproportionation reaction follows an O-O heterolytic cleavage pathway via a FeIV -oxo species.

A Tale of Two Complexes: Electro-Assisted Oxidation of Thioanisole by an "O2 Activator/Oxidizing Species" Tandem System of Non-Heme Iron Complexes.

We report two new FeIII complexes [L1 FeIII (H2 O)](OTf)2 and [L2 FeIII (OTf)], obtained by replacing pyridines by phenolates in a known non-heme aminopyridine iron complex. While the original, starting aminopyridine [(L5 2 )FeII (MeCN)](PF6 ) complex is stable in air, the potentials of the new FeIII/II couples decrease to the point that [L2 FeII ] spontaneously reduces O2 to superoxide. We used it as an O2 activator in an electrochemical setup, as its presence allows to generate superoxide at a much more accessible potential (>500 mV gain). Our aim was to achieve substrate oxidation via the reductive activation of O2 . While L2 FeIII (OTf) proved to be a good O2 activator but a poor oxidation system, its association with another complex (TPEN)FeII (PF6 )2 generates a complementary tandem couple for electro-assisted oxidation of substrates, working at a very accessible potential: upon reduction, L2 FeIII (OTf) activates O2 to superoxide and transfers it to (TPEN)FeII (PF6 )2 leading in fine to the oxidation of thioanisole.

Proton-controlled Action of an Imidazole as Electron Relay in a Photoredox Triad.

Electron relays play a crucial role for efficient light-induced activation by a photo-redox moiety of catalysts for multi-electronic transformations. Their insertion between the two units reduces detrimental energy transfer quenching while establishing at the same time unidirectional electron flow. This rectifying function allows charge accumulation necessary for catalysis. Mapping these events in photophysical studies is an important step towards the development of efficient molecular photocatalysts. Three modular complexes comprised of a Ru-chromophore, an imidazole electron relay function, and a terpyridine unit as coordination site for a metal ion were synthesized and the light-induced electron transfer events studied by laser flash photolysis. In all cases, formation of an imidazole radical by internal electron transfer to the oxidized chromophore was observed. The effect of added base evidenced that the reaction sequence depends strongly on the possibility for deprotonation of the imidazole function in a proton-coupled electron transfer process. In the complex with MnII present as a proxy for a catalytic site, a strongly accelerated decay of the imidazole radical together with a decreased rate of back electron transfer from the external electron acceptor to the oxidized complex was observed. This transient formation of an imidazolyl radical is clear evidence for the function of the imidazole group as an electron relay. The implication of the imidazole proton and the external base for the kinetics and energetics of the electron trafficking is discussed.

Intercepting a transient non-hemic pyridine N-oxide Fe(III) species involved in OAT reactions.

In the context of bioinspired OAT catalysis, we developed a tetradentate dipyrrinpyridine ligand, a hybrid of hemic and non-hemic models. The catalytic activity of the iron(III) derivative was investigated in the presence of iodosylbenzene. Unexpectedly, MS, EPR, Mössbauer, UV-visible and FTIR spectroscopic signatures supported by DFT calculations provide convincing evidence for the involvement of a relevant FeIII-O-NPy active intermediate.

Tracking light-induced electron transfer toward O2 in a hybrid photoredox-laccase system.

Photobiocatalysis uses light to perform specific chemical transformations in a selective and efficient way. The intention is to couple a photoredox cycle with an enzyme performing multielectronic catalytic activities. Laccase, a robust multicopper oxidase, can be envisioned to use dioxygen as a clean electron sink when coupled to an oxidation photocatalyst. Here, we provide a detailed study of the coupling of a [Ru(bpy)3]2+ photosensitizer to laccase. We demonstrate that efficient laccase reduction requires an electron relay like methyl viologen. In the presence of dioxygen, electrons transiently stored in superoxide ions are scavenged by laccase to form water instead of H2O2. The net result is the photo accumulation of highly oxidizing [Ru(bpy)3]3+. This study provides ground for the use of laccase in tandem with a light-driven oxidative process and O2 as one-electron transfer relay and as four-electron substrate to be a sustainable final electron acceptor in a photocatalytic process.

Second-sphere effects on H2O2 activation by non-heme FeII complexes: role of a phenol group in the [H2O2]-dependent accumulation of FeIVO vs. FeIIIOOH.

Redox metalloenzymes achieve very selective oxidation reactions under mild conditions using O2 or H2O2 as oxidants and release harmless side-products like water. Their oxidation selectivity is intrinsically linked to the control of the oxidizing species generated during the catalytic cycle. To do so, a second coordination sphere is used in order to create a pull effect during the activation of O2 or H2O2, thus ensuring a heterolytic O-O bond cleavage. Herein, we report the synthesis and study of a new non-heme FeII complex bearing a pentaazadentate first coordination sphere and a pendant phenol group. Its reaction with H2O2 generates the classical FeIIIOOH species at high H2O2 loading. But at low H2O2 concentrations, an FeIVO species is generated instead. The formation of the latter is directly related to the presence of the 2nd sphere phenol group. Kinetic, variable temperature and labelling studies support the involvement of the attached phenol as a second coordination sphere moiety (weak acid) during H2O2 activation. Our results suggest a direct FeII → FeIVO conversion directed by the 2nd sphere phenol via the protonation of the distal O atom of the FeII/H2O2 adduct leading to a heterolytic O-O bond cleavage.

Biosynthesis of the sactipeptide Ruminococcin C by the human microbiome: Mechanistic insights into thioether bond formation by radical SAM enzymes.

Despite its major importance in human health, the metabolic potential of the human gut microbiota is still poorly understood. We have recently shown that biosynthesis of Ruminococcin C (RumC), a novel ribosomally synthesized and posttranslationally modified peptide (RiPP) produced by the commensal bacterium Ruminococcus gnavus, requires two radical SAM enzymes (RumMC1 and RumMC2) catalyzing the formation of four Cα-thioether bridges. These bridges, which are essential for RumC's antibiotic properties against human pathogens such as Clostridium perfringens, define two hairpin domains giving this sactipeptide (sulfur-to-α-carbon thioether-containing peptide) an unusual architecture among natural products. We report here the biochemical and spectroscopic characterizations of RumMC2. EPR spectroscopy and mutagenesis data support that RumMC2 is a member of the large family of SPASM domain radical SAM enzymes characterized by the presence of three [4Fe-4S] clusters. We also demonstrate that this enzyme initiates its reaction by Cα H-atom abstraction and is able to catalyze the formation of nonnatural thioether bonds in engineered peptide substrates. Unexpectedly, our data support the formation of a ketoimine rather than an α,ß-dehydro-amino acid intermediate during Cα-thioether bridge LC-MS/MS fragmentation. Finally, we explored the roles of the leader peptide and of the RiPP precursor peptide recognition element, present in myriad RiPP-modifying enzymes. Collectively, our data support a more complex role for the peptide recognition element and the core peptide for the installation of posttranslational modifications in RiPPs than previously anticipated and suggest a possible reaction intermediate for thioether bond formation.

Photoswitchable 11 nm CsCoFe Prussian Blue Analogue Nanocrystals with High Relaxation Temperature.

Photoswitchable 11 nm nanocrystals with the coordination network Cs{Co[Fe(CN)6]} were obtained using a template-free method. The nanocrystals were recovered from the colloidal solutions as solid materials surrounded by cetyltrimethylammonium (CTA) cations or embedded in the organic polymer polyvinylpyrrolidone (PVP). Complementary magnetic, spectroscopic, and structural techniques, including EPR spectroscopy, reveal a majority (∼70%) of the low-spin and photoactive diamagnetic CoIIIFeII pairs located in the core of the nanocrystals and a mixture of CoIIFeII and CoIIFeIII species present mainly within the shell of the objects. While bulk compounds with similar vacancy concentration do not exhibit noticeable photoinduced charge transfer, the observed photoactivity of the nanocrystals is ascribed to their nanometric size. The relaxation temperature of the photoinduced state shifts upward by ∼55 K when PVP is replaced by CTA. This is ascribed to the larger rigidity of the dense CsCoFe_CTA material, whose metastable state is lower than that for CsCoFe_PVP, leading to a larger relaxation energy barrier and, therefore, to a higher relaxation temperature.

Characterization in aqueous medium of an FMN semiquinone radical stabilized by the enzyme-like microenvironment of a modified polyethyleneimine.

The elusive flavin semiquinone intermediate found in flavoproteins such as cryptochromes has been obtained in aqueous solution by single electron reduction of the natural FMN cofactor using sodium ascorbate. This species was formed in the local hydrophobic microenvironment of a modified polyethyleneimine and characterized by UV-Visible, fluorescence and EPR spectroscopies.

FeIII and FeII Phosphasalen Complexes: Synthesis, Characterization, and Catalytic Application for 2-Naphthol Oxidative Coupling.

We report on the synthesis and characterization of three iron(III) phosphasalen complexes, [FeIII (Psalen)(X)] differing in the nature of the counter-anion/exogenous ligand (X- =Cl- , NO3 - , OTf- ), as well as the neutral iron(II) analogue, [FeII (Psalen)]. Phosphasalen (Psalen) differs from salen by the presence of iminophosphorane (P=N) functions in place of the imines. All the complexes were characterized by single-crystal X-ray diffraction, UV/Vis, EPR, and cyclic voltammetry. The [FeII (Psalen)] complex was shown to remain tetracoordinated even in coordinating solvent but surprisingly exhibits a magnetic moment in line with a FeII high-spin ground state. For the FeIII complexes, the higher lability of triflate anion compared to nitrate was demonstrated. As they exhibit lower reduction potentials compared to their salen analogues, these complexes were tested for the coupling of 2-naphthol using O2 from air as oxidant. In order to shed light on this reaction, the interaction between 2-naphthol and the FeIII (Psalen) complexes was studied by cyclic voltammetry as well as UV/Vis spectroscopy.

Evolution of Ate-Organoiron(II) Species towards Lower Oxidation States: Role of the Steric and Electronic Factors.

Ate-iron(II) species such as [Ar3 FeII ]- (Ar=aryl) are key intermediates in Fe-catalyzed couplings between aryl nucleophiles and organic electrophiles. They can be active species in the catalytic cycle, or lead to Fe0 and FeI oxidation states, which can themselves be catalytically active or lead to unwished organic byproducts. Analysis of the reactivity of the intermediates obtained by step-by-step displacement of the mesityl groups in high-spin [Mes3 FeII ]- by less hindered phenyl ligands was performed, and uncovered the crucial role of both steric and electronic parameters in the formation of the Fe0 and FeI oxidation states. The formation of quaternized [Ar4 FeII MgBr(THF)]- intermediates allows the bielectronic reductive elimination energy required for the formation of Fe0 to be reduced. Similarly, the small steric pressure of the aryl groups in [Ar3 FeII ]- enables the formation of aryl-bridged [{FeII (Ar)2 }2 (µ-Ar)2 ]2- species, which afford the FeI oxidation state by bimetallic reductive elimination. These results are supported by 1 H NMR, EPR, and 57 Fe Mössbauer spectroscopies, as well as by DFT calculations.

Hydroxylation of Aromatics by H2 O2 Catalyzed by Mononuclear Non-heme Iron Complexes: Role of Triazole Hemilability in Substrate-Induced Bifurcation of the H2 O2 Activation Mechanism.

Rieske dioxygenases are metalloenzymes capable of achieving cis-dihydroxylation of aromatics under mild conditions using O2 and a source of electrons. The intermediate responsible for this reactivity is proposed to be a cis-FeV (O)(OH) moiety. Molecular models allow the generation of a FeIII (OOH) species with H2 O2 , to yield a FeV (O)(OH) species with tetradentate ligands, or {FeIV (O); OH. } pairs with pentadentate ones. We have designed a new pentadentate ligand, mtL4 2 , bearing a labile triazole, to generate an "in-between" situation. Two iron complexes, [(mtL4 2 )FeCl](PF6 ) and [(mtL4 2 )Fe(OTf)2 ]), were obtained and their reactivity towards aromatic substrates was studied in the presence of H2 O2 . Spectroscopic and kinetic studies reflect that triazole is bound at the FeII state, but decoordinates in the FeIII (OOH). The resulting [(mtL4 2 )FeIII (OOH)(MeCN)]2+ then lies on a bifurcated decay pathway (end-on homolytic vs. side-on heterolytic) depending on the addition of aromatic substrate: in the absence of substrate, it is proposed to follow a side-on pathway leading to a putative (N4 )FeV (O)(OH), while in the presence of aromatics it switches to an end-on homolytic pathway yielding a {(N5 )FeIV (O); OH. } reactive species, through recoordination of triazole. This switch significantly impacts the reaction regioselectivity.

Base-controlled mechanistic divergence between iron(iv)-oxo and iron(iii)-hydroperoxo in the H2O2 activation by a nonheme iron(ii) complex.

Activation of hydrogen peroxide by FeII salts (Fenton systems) leads to a myriad of oxidizing agents whose nature, FeIVO, or hydroxyl radicals and FeIII species, is dictated by the reaction conditions, in particular the pH value. Using the non heme FeII complex [FeII(L52)(CH3CN)]2+ (1) (where L52 is the pentadentate ligand N-methyl-N,N',N'-tris(2-pyridylmethyl)ethane-1,2-diamine) we have observed the simultaneous formation of two reaction intermediates, [FeIV(O)(L52)]2+ and [FeIII(OOH)(L52)]2+, in its reaction with excess hydrogen peroxide in the presence of sub-stoichiometric amounts of triethylamine. Kinetic and spectroscopic monitoring of the reaction mixture and of independently prepared [FeIV(O)(L52)]2+ in the presence of the different constituents of the reaction mixture allows drawing a mechanistic scheme. These two reactive species are formed simultaneously following two independent and competitive pathways. [FeIV(O)(L52)]2+ is obtained via heterolytic O-O cleavage of the oxidant assisted by the base in a peroxidase-like mechanism whereas [FeIII(OOH)(L52)]2+ is generated upon homolytic O-O cleavage of hydrogen peroxide. The relative contribution of these two pathways can be tuned by adjusting the amount of base used.

Electronic and spin delocalization in a switchable trinuclear triphenylene trisemiquinone bridged Ni3 complex.

A trinuclear triphenylene trisemiquinone complex containing paramagnetic NiII is obtained under ambient conditions from the reaction of deprotonated tricatecholate hexahydroxytriphenylene (H6HHTP) with NiII capped with a trispyrazolyl borate tridentate ligand. The magnetic and EPR data are consistent with delocalization of the electronic spin over the three NiII species. The two-electron reduced complex shows an EPR spectrum corresponding to a S = 1/2 species due to a large antiferromagnetic coupling between the radical and only one of the NiII ions highlighting the localization of the electronic spin. No EPR signal is observed for the one- and three-electron reduced species consistent with the closed shell of the bridging ligand.

Non-Heme FeII Diastereomeric Complexes Bearing a Hexadentate Ligand: Unexpected Consequences for the Spin State and Catalytic Oxidation Properties.

The reactivity and selectivity of non-heme FeII complexes as oxidation catalysts can be substantially modified by alteration of the ligand backbone or introduction of various substituents. In comparison with the hexadentate ligand N,N,N',N'-tetrakis(pyridin-2-ylmethyl)ethane-1,2-diamine (TPEN), N,N'-bis[1-(pyridin-2-yl)ethyl]-N,N'-bis(pyridin-2-ylmethyl)ethane-1,2-diamine (2Me L6 2 ) has a methyl group on two of the four picolyl positions. FeII complexation by 2Me L6 2 yields two diastereomeric complexes with very similar structures, which only differ in the axial/equatorial positions occupied by the methylated pyridyl groups. In solution, these two isomers exhibit different magnetic behaviors. Whereas one isomer exhibits temperature-dependent spin-state conversion between the S=0 and S=2 states, the other is more reluctant towards this spin-state equilibrium and is essentially diamagnetic at room temperature. Their catalytic properties for the oxidation of anisole by H2 O2 are very different and correlate with their magnetic properties, which reflect their lability/inertness. These different properties most likely depend on the different steric constraints of the methylated pyridyl groups in the two complexes.

Selective Formation of an FeIV O or an FeIII OOH Intermediate From Iron(II) and H2 O2 : Controlled Heterolytic versus Homolytic Oxygen-Oxygen Bond Cleavage by the Second Coordination Sphere.

We demonstrate that the devised incorporation of an alkylamine group into the second coordination sphere of an FeII complex allows to switch its reactivity with H2 O2 from the usual formation of FeIII species towards the selective generation of an FeIV -oxo intermediate. The FeIV -oxo species was characterized by UV/Vis absorption and Mössbauer spectroscopy. Variable-temperature kinetic analyses point towards a mechanism in which the heterolytic cleavage of the O-O bond is triggered by a proton transfer from the proximal to the distal oxygen atom in the FeII -H2 O2 complex with the assistance of the pendant amine. DFT studies reveal that this heterolytic cleavage is actually initiated by an homolytic O-O cleavage immediately followed by a proton-coupled electron transfer (PCET) that leads to the formation of the FeIV -oxo and release of water through a concerted mechanism.