Infrared Spectroscopy During Electrocatalytic Turnover Reveals the Ni‐L Active Site State During H2 Oxidation by a NiFe Hydrogenase

A novel in situ IR spectroscopic approach is demonstrated for the characterization of hydrogenase during catalytic turnover. E. coli hydrogenase 1 (Hyd‐1) is adsorbed on a high surface‐area carbon electrode and subjected to the same electrochemical control and efficient supply of substrate as in protein film electrochemistry during spectral acquisition. The spectra reveal that the active site state known as Ni‐L, observed in other NiFe hydrogenases only under illumination or at cryogenic temperatures, can be generated reversibly in the dark at ambient temperature under both turnover and non‐turnover conditions. The observation that Ni‐L is present at all potentials during turnover under H₂ suggests that the final steps in the catalytic cycle of H₂ oxidation by Hyd‐1 involve sequential proton and electron transfer via Ni‐L. A broadly applicable IR spectroscopic technique is presented for addressing electrode‐adsorbed redox enzymes under fast catalytic turnover.     

Electrosynthesis in a microemulsion using a novel cobaloxime catalyst

[Bis[μ-[(2,3-butanedione dioximato)(2-)-O:O′]]tetrafluorodiborato(2-)-N,N′,N″,N]cobalt (COBF, 1) was used as a catalyst for the conversion of trans-1,2-dibromocyclohexane (DBCH) to cyclohexene and the photoelectrochemical cyclisation of 2-(4-bromobutyl)-2-cyclohexen-1-one (BBC) to trans-1-decalone 2 in a microemulsion. Voltammetry showed clear evidence of catalytic behaviour and bulk electrolysis showed larger turnover numbers for both reactions when compared with the same system using vitamin B_12a as catalyst. For BBC, improved turnover may result from a relatively weak carbon–cobalt bond in the alkylcobalt intermediate of 1, and from better partition of 1 into the organic phase in which reactant BBC resides.     

Flow fluorimetric investigations on reaction kinetics of a growing analytical bioreactor

The turnover of a small continuously stirred tank reactor with variable volume is measured by two independent methods simultaneously: conventional flow-through fluorimetry is used for integral measurement of the total turnover and flow-through impulse fluorimetry for measurement of the dispersed turnover of the particles themselves. The hydrolysis of resorufin-β-d-galactopyranoside by β-galactosidase immobilized on Sepharose 4B and on Eupergit C serves as fluorogenic reaction. All parameters (flow rates, stirrer speed, concentrations, gel particle density and growth functions) are controlled by a computer. The allometric turnover kinetics of the growing bioreactor are compared with those of growing organisms.     

New ligands for a general palladium-catalyzed amination of aryl and heteroaryl chlorides

The synthesis and application of monodentate N-substituted heteroarylphosphines is described. In general, the ligands are conveniently prepared by selective metallation at the 2-position of the respective N-substituted heterocycle (pyrrole, indole) by using n-butyllithium/tetramethylethylenediamine (TMEDA) followed by quenching with dialkyl- or diarylchlorophosphines. Of the different ligands prepared, the new dialkyl-2-(N-arylindolyl)phosphines (cataCXium P) perform excellently in the palladium-catalyzed amination of aryl and heteroaryl chlorides. Coupling of both activated and deactivated chloroarenes proceeds under mild conditions (room temperature to 60 degrees C). By using optimized conditions remarkable catalyst productivity (total turnover number, TON, up to 8000) and activity (turnover frequency, TOF=14000 h(-1) at 75% conversion) are observed.     

Highly efficient atom transfer radical addition reactions with a RuIII complex as a catalyst precursor

The combination of the air-stable RuIII complex [Cp*RuCl2(PPh3)] with AIBN can be used to catalyze the atom transfer radical addition reactions of polychlorinated compounds and of sulfonyl chlorides to olefins with unprecedented turnover numbers of up to 44 000.     

Unlocking Mizoroki–Heck‐Type Reactions of Aryl Cyanides Using Transfer Hydrocyanation as a Turnover‐Enabling Step

A new transfer hydrofunctionalization strategy to turnover H‐M^II‐X complexes has enabled both intra‐ and intermolecular Mizoroki–Heck (MH)‐type reactions of aryl cyanides that are challenging to realize under traditional, basic conditions. Initially, a cascade carbonickelation/MH reaction of 2‐cyanostyrenes was achieved using a key alkyne transfer hydrocyanation step. Mechanistic experiments supported the proposed catalytic cycle, including the turnover‐enabling transfer hydrocyanation step. The reactivity was then extended to the intermolecular MH reaction of benzonitriles and styrenes.     

1,2-亚苯基二硫桥[FeFe]-氢化酶模拟物选择性光催化还原CO2至CO

利用基于非贵金属的分子催化剂通过光驱动催化CO2还原生成CO是将太阳能储存为化学能和缓解CO2温室效应的有效途径之一,具有重要的科学意义和潜在的应用前景.已报道的非贵金属分子催化剂,大多数对于光驱动CO2还原表现出缓慢的催化反应速率和/或对CO产物的低选择性,反应常常伴随着质子还原产氢反应,只有很少几种非贵金属分子催化剂对光催化CO2还原生成CO表现出高催化反应速率(>100 h?1)和高选择性.研究表明,双核过渡金属配合物由于分子中邻近的两个金属中心的协同催化作用,对于CO2还原生成CO的催化活性明显高于相应的单核配合物.因此,具有两个邻近的金属离子的非贵金属双核配合物有望作为CO2选择性还原的高效分子催化剂.我们最近的研究发现,具有刚性、共轭亚苯基二硫桥结构的[FeFe]-氢化酶模拟物[(μ-bdt)Fe2(CO)6](1,bdt=苯-1,2-二巯基)能够高活性、高选择性地光化学还原CO2至CO,而与其类似的模拟物[(μ-edt)Fe2(CO)6](2,edt=乙烷-1,2-巯基)则不具有光催化还原CO2活性,表明铁铁氢化酶模拟物中硫-硫桥的结构是影响模拟物的催化性能的重要结构因素之一.可见光照射1/[Ru(bpy)3]2+/BIH(BIH=1,3-二甲基-2-苯基-2,3-二氢-1H-苯并[d]-咪唑)体系4.5 h,1催化生成CO的循环数(TON)为710,在初始1 h的转化率(TOF)为7.12 min^-1,CO的选择性达到97%,内量子效率为2.8%.有趣的是,向体系中加入TEOA时可以调节1的催化选择性,光化学反应能够在CO2还原产生CO和质子还原产生H2之间进行切换.此外,采用稳态荧光和瞬态吸收光谱研究了光催化体系中的电子转移,提出可能的光催化反应机理.该研究结果揭示了刚性硫-硫桥结构的氢化酶模拟物对光化学CO2还原至CO的特殊催化活性,拓展了铁铁氢化酶模拟物的催化多功能性.     

Photoinduced hydrogen evolution in supramolecular devices with a rhenium photosensitizer linked to FeFe-hydrogenase model complexes

Coordination of the pyridyl-attached diiron azadithiolate hexacarbonyl complexes (2 and 3) through the pyridyl nitrogen to the Re on 10-phenanthroline rhenium (5a) and 2,9-diphenyl-1,10-phenanthroline rhenium (5b) forms novel [Re-Fe] complexes 7a, 7b and 8 respectively. Under visible light illumination using triethylamine as a sacrificial electron donor and [Re-Fe] type complexes (7a, 7b or 8) as catalysts, remarkably increased efficiency was observed for photoinduced hydrogen production with a turnover number reaching 11.8 from complex 7a and 8.75 from 7b. To the best of our knowledge, these are the best values compared to other [Re-Fe] photocatalysts reported so far. In contrast to the parent molecules, the turnover number by the intermolecular combination of complexes 6a and 2 showed a value of 5.23, and that from 6b and 2 is 3.8, while no H(2) was detected from 8a and 3 under the same experimental conditions. Obviously, the intramolecular combination of rhenium(I) and [2Fe2S] as a catalyst is promising for efficient H(2) evolution, and it is better than the intermolecular multi-component system.     

Enzyme kinetics with a twist

A different approach to enzyme kinetics stressing the cyclic nature of the catalytic process is presented. The time-dependence of the substrate concentration is derived in a simple way not invoking the quasi-steady-state approximation. According to this approach the turnover rate can be written as the ratio of two parameters with a direct meaning: enzyme efficiency and average cycle duration. Real kinetic data for two enzyme-substrate pairs is used to show that the enzyme kinetic efficiency is best measured by the turnover rate.     

Isotopic turnover of a submersed macrophyte following transplant: the roles of growth and metabolism in eutrophic conditions

Stable isotopic turnover with isotopic change due to growth and metabolic tissue replacement associated with a change in environmental conditions is a critical aspect of the use of stable isotope analyses as time-integrating tracers of resource-consumer interactions. However, stable isotopic turnover in plants remains poorly understood compared with those of animals, and here we used continuous flow elemental analyzer-isotopic ratio mass spectrometry (EA-IRMS) to analyse the turnover of stable carbon and nitrogen isotopes in a submersed macrophyte (Vallisneria natans) after transplantation to hypereutrophic and mesoeutrophic treatments in a field mesocosm experiment. The direction and magnitude of the isotopic shifts of V. natans were suggested to be determined by the inorganic nutrient availability and its isotopic content in the different treatments. Based on the modelling results of turnover, the contribution of growth to the isotopic turnover was as high as those observed in various aquatic ectotherms. However, the contribution of metabolism was also considerable, especially for nitrogen in the hypereutrophic treatment, which was argued to be a response, co-occurring with growth inhabitation and biochemical disorder of V. natans, to the stress induced by the eutrophication. Our results indicated that isotope turnover in a macrophyte is a feasible technique for estimating its ecophysiological conditions in the natural environment, and that it may facilitate understanding of isotopic data in field studies of food web and habitat restoration under eutrophic conditions.     

The effects of substrate orientation on the mechanism of a phosphotriesterase

While the underlying chemistry of enzyme-catalyzed reactions may be almost identical, the actual turnover rates of different substrates can vary significantly. This is seen in the turnover rates for the catalyzed hydrolysis of organophosphates by the bacterial phosphotriesterase OpdA. We investigate the variation in turnover rates by examining the hydrolysis of three classes of substrates: phosphotriesters, phosphothionates, and phosphorothiolates. Theoretical calculations were used to analyze the reactivity of these substrates and the energy barriers to their hydrolysis. This information was then compared to information derived from enzyme kinetics and crystallographic studies, providing new insights into the mechanism of this enzyme. We demonstrate that the enzyme catalyzes the hydrolysis of organophosphates through steric constraint of the reactants, and that the equilibrium between productively and unproductively bound substrates makes a significant contribution to the turnover rate of highly reactive substrates. These results highlight the importance of correct orientation of reactants within the active sites of enzymes to enable efficient catalysis.     

Water Oxidation Catalyzed by a Dinuclear Cobalt–Polypyridine Complex

The dinuclear Co complex [(TPA)Co(μ‐OH)(μ‐O₂)Co(TPA)](ClO₄)₃ ( 1 , TPA=tris(2‐pyridylmethyl)amine) catalyzes the oxidation of water. In the presence of [Ru(bpy)₃]²⁺ and S₂O₈^2?, photoinduced oxygen evolution can be observed with a turnover frequency (TOF) of 1.4±0.1 mol(O₂) mol( 1 )^?1 s^?1 and a maximal turnover number (TON) of 58±5 mol(O₂) mol( 1 )^?1. The complex is shown to act as a molecular and homogeneous catalyst and a mechanism is proposed based on the combination of EPR data and light‐driven O₂ evolution kinetics.     

Pulling direction as a reaction coordinate for the mechanical unfolding of single molecules

The folding and unfolding kinetics of single molecules, such as proteins or nucleic acids, can be explored by mechanical pulling experiments. Determining intrinsic kinetic information, at zero stretching force, usually requires an extrapolation by fitting a theoretical model. Here, we apply a recent theoretical approach describing molecular rupture in the presence of force to unfolding kinetic data obtained from coarse-grained simulations of ubiquitin. Unfolding rates calculated from simulations over a broad range of stretching forces, for different pulling directions, reveal a remarkable "turnover" from a force-independent process at low force to a force-dependent process at high force, akin to the "roll-over" in unfolding rates sometimes seen in studies using chemical denaturant. While such a turnover in rates is unexpected in one dimension, we demonstrate that it can occur for dynamics in just two dimensions. We relate the turnover to the quality of the pulling direction as a reaction coordinate for the intrinsic folding mechanism. A novel pulling direction, designed to be the most relevant to the intrinsic folding pathway, results in the smallest turnover. Our results are in accord with protein engineering experiments and simulations which indicate that the unfolding mechanism at high force can differ from the intrinsic mechanism. The apparent similarity between extrapolated and intrinsic rates in experiments, unexpected for different unfolding barriers, can be explained if the turnover occurs at low forces.     

Direct Michael addition of alkenes via a cobalt-dinitrosyl mediated vinylic C-H functionalization reaction

We report a one-pot, direct C-H functionalization reaction of alkenes mediated by [CpCo(NO)(2)]. All intermediates in the proposed reaction sequence have been characterized. A variety of Michael acceptors can be utilized with the one-pot procedure to give the desired cyclic tetra-alkyl-substituted, γ,δ-unsaturated compounds in good yields. We also provide a preliminary result for catalytic turnover in both base and [CpCo(NO)(2)].     

Direct Spectroscopic Detection of Key Intermediates and the Turnover Process in Catalytic H2 Formation by a Biomimetic Diiron Catalyst

[FeFe(Cl₂-bdt)(CO)₆] ( 1 ; Cl₂-bdt=3,6-dichlorobenzene-1,2-dithiolate), inspired by the active site of FeFe-hydrogenase, shows a chemically reversible 2 e⁻ reduction at −1.20 V versus the ferrocene/ferrocenium couple. The rigid and aromatic bdt bridging ligand lowers the reduction potential and stabilizes the reduced forms, compared with analogous complexes with aliphatic dithiolates; thus allowing details of the catalytic process to be characterized. Herein, time-resolved IR spectroscopy is used to provide kinetic and structural information on key catalytic intermediates. This includes the doubly reduced, protonated complex 1H ⁻, which has not been previously identified experimentally. In addition, the first direct spectroscopic observation of the turnover process for a molecular H₂ evolving catalyst is reported, allowing for straightforward determination of the turnover frequency.     

1,3-Dipolar cycloaddition of organic azides to alkynes by a dicopper-substituted silicotungstate

The dicopper-substituted gamma-Keggin silicotungstate TBA 4[gamma-H2SiW10O36Cu2(mu-1,1-N3)2] (I, TBA = tetra- n-butylammonium) could act as an efficient precatalyst for the regioselective 1,3-dipolar cycloaddition of organic azides to alkynes. Various combinations of substrates (four azides and eight alkynes) were efficiently converted to the corresponding 1,2,3-triazole derivatives in excellent yields without any additives. The present system was applicable to a larger-scale cycloaddition of benzyl azide to phenylacetylene under solvent-free conditions (100 mmol scale) in which 21.5 g of the analytically pure corresponding triazole could be isolated. In this case, the turnover frequency and the turnover number reached up to 14,800 h(-1) and 91,500, respectively, and these values were the highest among those reported for the copper-mediated systems so far. In addition, I could be applied to the one-pot synthesis of 1-benzyl-4-phenyl-1H-1,2,3-triazole from benzyl chloride, sodium azide, and phenylacetylene. The catalyst effect, kinetic, mechanistic, and computational studies show that the reduced dicopper core plays an important role in the present 1,3-dipolar cycloaddition.     

Hydrogen oxidation catalysis by a nickel diphosphine complex with pendant tert-butyl amines

A bis-diphosphine nickel complex with tert-butyl functionalized pendant amines [Ni(P(Cy)(2)N(t-Bu)(2))(2)](2+) has been synthesized. It is a highly active electrocatalyst for the oxidation of hydrogen in the presence of base. The turnover rate of 50 s(-1) under 1.0 atm H(2) at a potential of -0.77 V vs. the ferrocene couple is 5 times faster than the rate reported heretofore for any other synthetic molecular H(2) oxidation catalyst.     

Determination of the stereochemistry of 2-succinyl-5-enolpyruvyl-6-hydroxy-3- cyclohexene-1-carboxylate, a key intermediate in menaquinone biosynthesis

The turnover product of the committed step of menaquinone biosynthesis was isolated and determined to be (1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate. Structural determination of this key intermediate represents a critical step to complete elucidation of the biosynthetic pathway.     

Catalytic turnover of benzylamine by a model for the lysine tyrosylquinone (LTQ) cofactor of lysyl oxidase

Lysyl oxidase differs from other copper amine oxidases in that its active quinone cofactor reflects cross-linking of a lysyl residue into the tyrosine-derived quinone nucleus found in the plasma and other copper amine oxidases. A model for the lysyl oxidase cofactor (LTQ), 3,3-dimethyl-2,3-dihydroindole-5,6-quinone (4), was synthesized and found to be stable to both hydrolysis and oxidation events that prevent simpler models from functioning as turnover catalysts. We show that 4 catalyzes the aerobic oxidative deamination of benzylamine, though turnover eventually ceases on account of oxidation of the dihydrobenzoxazole tautomer of the "product Schiff base" to form a benzoxazole, a reaction that may be physiologically relevant. The mechanism of the overall reaction profile was elucidated by a combination of optical and NMR spectroscopy and O(2) uptake studies.     

Synergistic Porosity and Charge Effects in a Supramolecular Porphyrin Cage Promote Efficient Photocatalytic CO2 Reduction**

We present a supramolecular approach to catalyzing photochemical CO₂ reduction through second-sphere porosity and charge effects. An iron porphyrin box ( PB ) bearing 24 cationic groups, FePB-2(P) , was made via post-synthetic modification of an alkyne-functionalized supramolecular synthon. FePB-2(P) promotes the photochemical CO₂ reduction reaction (CO₂RR) with 97 % selectivity for CO product, achieving turnover numbers (TON) exceeding 7000 and initial turnover frequencies (TOF_max) reaching 1400 min⁻¹. The cooperativity between porosity and charge results in a 41-fold increase in activity relative to the parent Fe tetraphenylporphyrin ( FeTPP ) catalyst, which is far greater than analogs that augment catalysis through porosity ( FePB-3(N ), 4-fold increase) or charge (Fe p-tetramethylanilinium porphyrin ( Fe-p-TMA ), 6-fold increase) alone. This work establishes that synergistic pendants in the secondary coordination sphere can be leveraged as a design element to augment catalysis at primary active sites within confined spaces.