Study Of Electron Transfer Reactions Research Articles

Enhancing electrochemiluminescence intensity through emission layer control

Electrochemiluminescence (ECL), an intriguing luminescent phenomenon induced by electrochemical stimulation, has evolved from studying electron transfer reactions to a powerful analytical method and imaging technique. ECL can be generated through annihilation or co-reactant methods, with recent advancements integrating it into imaging devices for diverse applications. This review traces the evolution of ECL from its early applications to recent developments in imaging technology. Notably, the utilization of charge-coupled devices (CCD) and electron multiplying charge-coupled devices (EMCCD) in ECL microscopy has revolutionized imaging capabilities, making it a cost-effective option for point-of-care testing. The review explores the heterogeneous ECL mechanism, emphasizing its limitations and challenges in visualizing objects away from the electrode surface. The synergy between ECL and microscopy is highlighted, showcasing its diverse applications and contributions to understanding ECL mechanisms and improving its use in biological contexts. Finally, the review encapsulates innovative approaches in material design, surface modification, and electrode architecture, providing a comprehensive overview of strategies to control the active layer of the ECL and advance ECL microscopy.

Single-Entity Bioelectrochemistry

Interfacing biological catalytic entities, such as whole cells or enzymes, with electrodes, has a wide range of applications, ranging from fundamental studies of the entities themselves to power generation, bioremediation, chemical synthesis, and biosensing. In all cases, the operation of these bioelectrocatalytic systems is based on electron transfer phenomena within protein molecules, whether isolated or residing inside cells or parts of cells, as well as between proteins and electronics. Typically, the electron transfer and catalytic properties of proteins are studied using ensemble averaging methods, which obscure any heterogeneity and dynamics of individual molecules. This information, however, is especially important for understanding electron transfer and catalytic processes within a living cell where only one or a few protein molecules are present. While single-entity electrochemical measurements are particularly useful for studying electron transfer reactions at the nanoscale level, they have limited applications for investigating biological entities due to the experimental challenges associated with direct measurements of small electrical currents. 1,2 Here we will discuss our recent progress in investigating biological electron transfer reactions at the single-entity level, from bacterial cells to single redox proteins, using nano-impact (collision) single-entity electrochemistry. 3 We demonstrate how advances in electrode fabrication methods and detection setups enable current detection in the fA range. When combined with the developed data processing algorithm based on unsupervised learning and template matching techniques, this enabled unprecedented insight into electron transfer processes at the nanoscale. We will talk about insights provided by the method in the study of several important bioelectrocatalytic systems: single E.coli cells with overexpressed Fe-Fe hydrogenases, single bilirubin oxidase molecules, and single catalase molecules. In addition, we will discuss models that we developed for understanding the single bioentity electrode interactions for diffusion-controlled enzymes when the product of the catalytic reaction is detected on the electrode, as well as systems based on direct electron transfer between the bioentity and the electrode. Overall, we show how single-entity bioelectrochemistry can shed light on biological electron transfer and catalysis processes and discuss the challenges it currently faces. Zhang, J.-H.; Zhou, Y.-G. Nano-Impact Electrochemistry: Analysis of Single Bioentities. TrAC Trends Anal. Chem. 2019, 115768.Davis, C.; Wang, S. X.; Sepunaru, L. What Can Electrochemistry Tell Us About Individual Enzymes? Curr. Opin. Electrochem. 2020, 100643.Sekretaryova, A. N.; Vagin, M. Yu.; Turner, A. P. F.; Eriksson, M. Electrocatalytic Currents from Single Enzyme Molecules. J. Am. Chem. Soc. 2016, 138 (8), 2504–250

Cyclic voltammetric studies of electron transfer reactions associated with bioactive iron–KA complexation at different biological pH

This study explores electrochemical interactions of a pharmaceutical drug, kojic acid (5-hydroxy-2-hydroxymethyl-4 H-pyran-4-one, KA), with bioactive iron Fe(III) at different pH systems by using cyclic voltammetric technique. For this study, buffers relevant to physiological pH of blood and digestive system have been selected. Bioactive iron was found to form an electroactive complex [Fe(III)L3] in the presence of KA at all studied pH, whereas electrochemical behavior of KA was also changed by complex formation. This electrochemical investigation reveals that under the experimental conditions, [Fe(III)L3] complex undergoes a diffusion controlled, one electron redox process that is quasi-reversible in nature. It was also found that [Fe(III)L3] complex undergoes coupled chemical reaction according to EC (an electron transfer reaction followed by a chemical reaction) mechanism, and this interaction is pH dependent. Modified Nicholson and Shain, Gileadi, and Kochi methods were used to determine standard heterogeneous electron transfer rate constant ( k0) of [Fe(III)L3] complex. It was found to be in the range of 10−3 to 10−5 cm/s.

Extended Mulliken–Hush Method with Applications to the Theoretical Study of Electron Transfer

A novel adiabatic-to-diabatic (ATD) transformation strategy, namely, the extended Mulliken-Hush (XMH) method, is proposed to evaluate diabatic properties including electronic couplings, potential energy surfaces, and their crossings. The XMH method is developed by adopting our recently proposed ATD transformation formula of a general vectorial physical observable, in which a useful ATD transformation is further determined by using an auxiliary dipole between localized frontier orbitals as a simple approximation of the diabatic transition dipole. The XMH method is simple and practical that provides a flexible way to construct diabatic states. To some extent, it can be regarded as an extension of the generalized Mulliken-Hush (GMH) method since the latter takes a stronger approximation, in which the diabatic transition dipole is assumed to be vanishing. Test calculations on the HeH2+ system show that the electronic couplings predicted by the XMH method are closer to the ones calculated by the valence bond block-diagonalization approach than the GMH ones since the XMH method takes into account both the magnitude and direction of the diabatic transition dipole, which is consistent with the properties of this molecule. In the study of electron transfer in the two kinds of donor-bridge-acceptor systems, the XMH method maintains the simplicity of the GMH method and gives reasonable results even when the latter fails, wherein the diabatic transition dipole is nearly perpendicular to the difference of the initial and final adiabatic dipoles. More importantly, the XMH method can be easily combined with high-level electronic structure methods, in which the properties of the ground and excited states may be more accurately calculated, and hence, one may expect that further development of the XMH method would result in a general computational model for studying electron transfer reactions.

Assessing the correctness of pressure correction to solvation theories in the study of electron transfer reactions.

Liquid state theories have emerged as a numerically efficient alternative to costly molecular dynamics simulations of electron transfer reactions in solution. In a recent paper [Jeanmairet et al., Chem. Sci. 10, 2130-2143 (2019)], we introduced the framework to compute the energy gap, free energy profile, and reorganization free energy using molecular density functional theory. However, this technique, as other molecular liquid state theories, overestimates the bulk pressure of the fluid. Because of the very high pressure, the predicted free energy is dramatically exaggerated. Several attempts were made to fix this issue, either based on simple a posteriori correction or by introducing bridge terms. By studying two model half reactions in water, Cl → Cl+ and Cl → Cl-, we assess the correctness of these two types of corrections to study electron transfer reactions. We found that a posteriori correction, because it violates the Variational principle, leads to an inconsistency in the definition of the reorganization free energy and should not be used to study electron transfer reactions. The bridge approach, because it is theoretically well grounded, is perfectly suitable for this type of systems.

Active Intermediates in Copper Nitrite Reductase Reactions Probed by a Cryotrapping-Electron Paramagnetic Resonance Approach.

Redox active metalloenzymes catalyse a range of biochemical processes essential for life. However, due to their complex reaction mechanisms, and often, their poor optical signals, detailed mechanistic understandings of them are limited. Here, we develop a cryoreduction approach coupled to electron paramagnetic resonance measurements to study electron transfer between the copper centers in the copper nitrite reductase (CuNiR) family of enzymes. Unlike alternative methods used to study electron transfer reactions, the cryoreduction approach presented here allows observation of the redox state of both metal centers, a direct read‐out of electron transfer, determines the presence of the substrate/product in the active site and shows the importance of protein motion in inter‐copper electron transfer catalyzed by CuNiRs. Cryoreduction‐EPR is broadly applicable for the study of electron transfer in other redox enzymes and paves the way to explore transient states in multiple redox‐center containing proteins (homo and hetero metal ions).

Active Intermediates in Copper Nitrite Reductase Reactions Probed by a Cryotrapping‐Electron Paramagnetic Resonance Approach

AbstractRedox active metalloenzymes catalyse a range of biochemical processes essential for life. However, due to their complex reaction mechanisms, and often, their poor optical signals, detailed mechanistic understandings of them are limited. Here, we develop a cryoreduction approach coupled to electron paramagnetic resonance measurements to study electron transfer between the copper centers in the copper nitrite reductase (CuNiR) family of enzymes. Unlike alternative methods used to study electron transfer reactions, the cryoreduction approach presented here allows observation of the redox state of both metal centers, a direct read‐out of electron transfer, determines the presence of the substrate/product in the active site and shows the importance of protein motion in inter‐copper electron transfer catalyzed by CuNiRs. Cryoreduction‐EPR is broadly applicable for the study of electron transfer in other redox enzymes and paves the way to explore transient states in multiple redox‐center containing proteins (homo and hetero metal ions).

Study of Electron Transfer Reactions Associated with Benzyl Viologen-β-Cyclodextrin Complexation in Buffer Solution of pH 7: Equilibrium and Kinetic Aspects

In this study, complex formation of benzyl viologen dication (BzV2+) with β-cyclodextrin (β-CD), in its different oxidation states, had been studied in buffer solution of pH 7, through cyclic voltammetry. In buffer solution of pH 7, extensive deposition of benzyl viologen neutral (BzVº) has been found. Further to that first redox process of BzV2+ was found reversible in buffer solution of pH 7. Benzyl viologen mono-cation (BzV+•) was found to interact with β-CD reversibly according to EC mechanism. Equilibrium constant of BzV+•-nβ-CD complex in buffer solution of pH 7, has been calculated as 5.49 M-n. The value of n was found to be 0.2. The bimolecular rate constant for the complex formation of BzV+• with β-CD was found to be 6.44 M-ns− in buffer solution of pH 7. This study is based upon an electrochemical approach to obtain the bonding equilibrium constant, the rate constants of formation and the association numbers of the complex formed between reduced forms of viologen and β-CD. This study explore the electron transfer reaction mechanism of benzyl viologen with β-CD to extend their applications at neutral pH.

Oxidation-State Constrained Density Functional Theory for the Study of Electron-Transfer Reactions.

We propose a new constrained density functional theory (CDFT) approach which directly controls the oxidation state of the target atoms. In this new approach called oxidation-state constrained density functional theory (OS-CDFT), the eigenvalues of the occupation matrix obtained from projecting the Kohn-Sham wave functions onto the valence orbitals are constrained to obtain the desired oxidation states. This approach is particularly useful to study electron transfer problems in transition metal-containing systems due to the multivalent nature of the transition metal ions. The calculation of the forces on the ions and of the coupling constant was implemented under the OS-CDFT scheme to allow efficient and accurate study of electron transfer reactions. We demonstrated the application of this method in the study of different electron transfer reactions including the aqueous ferrous-ferric self-exchange reaction, polaron hopping in the TiO2 anatase and bismuth vanadate, and photoexcited electron transfer in the sapphire.

Projection-Based Wavefunction-in-DFT Embedding.

Complex chemical systems present challenges to electronic structure theory stemming from large system sizes, subtle interactions, coupled dynamical time scales, and electronically nonadiabatic effects. New methods are needed to perform reliable, rigorous, and affordable electronic structure calculations for simulating the properties and dynamics of such systems. This Account reviews projection-based quantum embedding for electronic structure, which provides a formally exact method for density functional theory (DFT) embedding. The method also provides a rigorous and accurate approach for describing a small part of a chemical system at the level of a correlated wavefunction (WF) method while the remainder of the system is described at the level of DFT. A key advantage of projection-based embedding is that it can be formulated in terms of an extremely simple level-shift projection operator, which eliminates the need for any optimized effective potential calculation or kinetic energy functional approximation while simultaneously ensuring that no extra programming is needed to perform WF-in-DFT embedding with an arbitrary WF method. The current work presents the theoretical underpinnings of projection-based embedding, describes use of the method for combining wavefunction and density functional theories, and discusses technical refinements that have improved the applicability and robustness of the method. Applications of projection-based WF-in-DFT embedding are also reviewed, with particular focus on recent work on transition-metal catalysis, enzyme reactivity, and battery electrolyte decomposition. In particular, we review the application of projection-based embedding for the prediction of electrochemical potentials and reaction pathways in a Co-centered hydrogen evolution catalyst. Projection-based WF-in-DFT calculations are shown to provide quantitative accuracy while greatly reducing the computational cost compared with a reference coupled cluster calculation on the full system. Additionally, projection-based WF-in-DFT embedding is used to study the mechanism of citrate synthase; it is shown that projection-based WF-in-DFT largely eliminates the sensitivity of the potential energy landscape to the employed DFT exchange-correlation functional. Finally, we demonstrate the use of projection-based WF-in-DFT to study electron transfer reactions associated with battery electrolyte decomposition. Projection-based WF-in-DFT embedding is used to calculate the oxidation potentials of neat ethylene carbonate (EC), neat dimethyl carbonate (DMC), and 1:1 mixtures of EC and DMC in order to overcome qualitative inaccuracies in the electron densities and ionization energies obtained from conventional DFT methods. By further embedding the WF-in-DFT description in a molecular mechanics point-charge environment, this work enables an explicit description of the solvent and ensemble averaging of the solvent configurations. Looking forward, we anticipate continued refinement of the projection-based embedding methodology as well as its increasingly widespread application in diverse areas of chemistry, biology, and materials science.

Electrochemical STM Tip-Enhanced Raman Spectroscopy Study of Electron Transfer Reactions of Covalently Tethered Chromophores on Au(111)

The ability to study electron transfer reactions at the solid–liquid interface with nanometer resolution has the potential to critically improve our understanding of electrocatalytic processes. However, few techniques are capable of studying electrode surfaces in situ at the nanoscale. We study the redox reactions of Nile Blue (NB) covalently tethered to an Au(111) electrode using in situ tip-enhanced Raman spectroscopy (TERS) and show that TERS amplitude decreases reversibly as NB is reduced. The potential dependent TERS intensity allows us to associate an electrochemical wave with the loss of electronic resonance of NB and another with the peak of fluorescence of tethered NB, which we tentatively attribute to the disassembly of on-surface NB aggregates. The study of the electrochemical activity of immobile adsorbates at the solid–liquid interface with TERS is an essential step toward the realization of in situ spectroscopic mapping at the nanoscale.

Introducing the mean field approximation to CDFT/MMpol method: Statistically converged equilibrium and nonequilibrium free energy calculation for electron transfer reactions in condensed phases

A new theoretical method to study electron transfer reactions in condensed phases is proposed by introducing the mean-field approximation into the constrained density functional theory/molecular mechanical method with a polarizable force field (CDFT/MMpol). The method enables us to efficiently calculate the statistically converged equilibrium and nonequilibrium free energies for diabatic states in an electron transfer reaction by virtue of the mean field approximation that drastically reduces the number of CDFT calculations. We apply the method to the system of a formanilide-anthraquinone dyad in dimethylsulfoxide, in which charge recombination and cis-trans isomerization reactions can take place, previously studied by the CDFT/MMpol method. Quantitative agreement of the driving force and the reorganization energy between our results and those from the CDFT/MMpol calculation and the experimental estimates supports the utility of our method. The calculated nonequilibrium free energy is analyzed by its decomposition into several contributions such as those from the averaged solute-solvent electrostatic interactions and the explicit solvent electronic polarization. The former contribution is qualitatively well described by a model composed of a coarse-grained dyad in a solution in the linear response regime. The latter contribution reduces the reorganization energy by more than 10 kcal/mol.

The study of electron transfer reactions in a dendrimeric assembly: proper utilization of dendrimer fluorescence.

The PAMAM dendrimer with a cage like structure acts as an excellent electron donor in the presence of an electron deficient molecule. Electron transfer (ET) causes significant quenching of dendrimer fluorescence. Trapping of quencher molecules within the dendrimer cage helps ET to take place through an expeditious route. Utilization of intrinsic fluorescence and sensing applications of dendrimers have been established here.

Quantum Calculations of Electron Tunneling in Respiratory Complex III.

The most detailed and comprehensive to date study of electron transfer reactions in the respiratory complex III of aerobic cells, also known as bc1 complex, is reported. In the framework of the tunneling current theory, electron tunneling rates and atomistic tunneling pathways between different redox centers were investigated for all electron transfer reactions comprising different stages of the proton-motive Q-cycle. The calculations reveal that complex III is a smart nanomachine, which under certain conditions undergoes conformational changes gating electron transfer, or channeling electrons to specific pathways. One-electron tunneling approximation was adopted in the tunneling calculations, which were performed using hybrid Broken-Symmetry (BS) unrestricted DFT/ZINDO levels of theory. The tunneling orbitals were determined using an exact biorthogonalization scheme that uniquely separates pairs of tunneling orbitals with small overlaps out of the remaining Franck-Condon orbitals with significant overlap. Electron transfer rates in different redox pairs show exponential distance dependence, in agreement with the reported experimental data; some reactions involve coupled proton transfer. Proper treatment of a concerted two-electron bifurcated tunneling reaction at the Q(o) site is given.

Characterization of the free energy dependence of an interprotein electron transfer reaction by variation of pH and site-directed mutagenesis

The interprotein electron transfer (ET) reactions of the cupredoxin amicyanin, which mediates ET from the tryptophan tryptophylquinone (TTQ) cofactor of methylamine dehydrogenase to cytochrome c-551i have been extensively studied. However, it was not possible to perform certain key experiments in that native system. This study examines the ET reaction from reduced amicyanin to an alternative electron acceptor, the diheme protein MauG. It was possible to vary the ΔG° for this ET reaction by simply changing pH to determine the dependence of kET on ΔG°. A P94A mutation of amicyanin significantly altered its oxidation–reduction midpoint potential value. It was not possible to study the ET from reduced P94A amicyanin to cytochrome c-551i in the native system because that reaction was kinetically coupled. However, the reaction from reduced P94A amicyanin to MauG was a true ET reaction and it was possible to determine values of reorganization energy (λ) and electronic coupling for the reactions of this variant as well as native amicyanin. Comparison of the λ values associated with the ET reactions between amicyanin and the TTQ of methylamine dehydrogenase, the diheme center of MauG and the single heme of cytochrome c-551i, provides insight into the factors that dictate the λ values for the respective reactions. These results demonstrate how study of ET reactions with alternative redox partner proteins can complement and enhance our understanding of the reactions with the natural redox partners, and further our understanding of mechanisms of protein ET reactions.

Thermoluminescence as a complementary technique for the toxicological evaluation of chemicals in photosynthetic organisms

Thermoluminescence is a simple technique very useful for studying electron transfer reactions on photosystem II (standard thermoluminescence) or the level of lipid peroxidation in membranes (high temperature thermoluminescence) in photosynthetic organisms. Both techniques were used to investigate the effects produced on Chlorella vulgaris cells by six compounds: the chemical intermediates bromobenzene and diethanolamine, the antioxidant propyl gallate, the semiconductor indium nitrate, the pesticide sodium monofluoroacetate and the antimalarial drug chloroquine. Electron transfer activity of the photosystem II significantly decreased after the exposure of Chlorella cells to all the six chemicals used. Lipid peroxidation was slightly decreased by the antioxidant propyl gallate, not changed by indium nitrate and very potently stimulated by diethanolamine, chloroquine, sodium monofluoroacetate and bromobenzene. For five of the chemicals studied (not bromobenzene) there is a very good correlation between the cytotoxic effects in Chlorella cells measured by the algal growth inhibition test, and the inhibition of photosystem II activity. The results suggest that one very important effect of these chemicals in Chlorella cells is the inhibition of photosynthetic metabolism by the blocking of photosystem II functionality. In the case of sodium monofluoroacetate, diethanolamine and chloroquine this inhibition seems to be related with the induction of high level of lipid peroxidation in cells that may alter the stability of photosystem II. The results obtained by both techniques supply information that can be used as a supplement to the growth inhibition test and allows a more complete assessment of the effects of a chemical in photosynthetic organisms of aquatic ecosystems.

Generation of Counter Ion Radical (Br2•–) and Its Reactions in Water-in-Oil (CTAB or CPB)/n-Butanol/Cyclohexane/Water) Microemulsion

Herein we report the generation of counterion radicals and their reactions in quaternary water-in-oil microemulsion. Hydrated electrons in the microemulsion CTAB/H(2)O/n-butanol/cyclohexane have a remarkably short half-life (∼1 μs) and lower yield as compared to that in the pure water system. Electrons are solvated in two regions: one is the water core and other the interface; however, the electrons in the water core have a shorter half-life than those in the interface. The decay of the solvated electrons in the interface is found to be water content dependent and it has been interpreted in terms of increased interfacial fluidity with the increase in water content of the microemulsion. Interestingly another species, dibromide radical anion (Br(2)(•-)) in CTAB and CPB microemulsions have been observed after the electron beam irradiation. Assuming that the extinction coefficient of the radicals is the same as that in the aqueous solution, the yields of the radicals per 100 eV are 0.29 and 0.48 for the Br(2)(•-) radical in CTAB and CPB containing microemulsions (W(0) = 40), respectively, under N(2)O saturated conditions. Further, we intended to study electron transfer reactions, which occur at and through the interface. The reaction of the Br(2)(•-) radical anion with ABTS [2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)] has been studied to generate the ABTS radical in the water core, and further, its reaction has been investigated with the water-insoluble molecule vitamin E (tocopherol) and water-soluble vitamin C (ascorbic acid). In the present study, we were able to show that, even for molecules which are completely insoluble in water, ABTS scavenging assay is possible by pulse radiolysis technique. Furthermore, these results show that it is possible to follow the reaction of the hydrated inorganic radical with solutes dissolved in the organic phase in a microemulsion without use of a phase transfer catalyst.

An STM Study of the pH Dependent Redox Activity of a Two-Dimensional Hydrogen Bonding Porphyrin Network at an Electrochemical Interface

Studying electron transfer reactions of porphyrin molecules is important for a wide range of applications including biology, molecular devices, artificial photosynthesis, information storage, and fuel cells. It is known that porphyrins adsorbed in a self-assembled monolayer at an electrochemical interface may lose their electrochemical activity. However, the mechanism of the suppressed electrochemical activity is not clear. In this article, the electrochemical behavior of the two-dimensional network structures of 5,10,15,20-tetrakis(4-carboxylphenyl)-21H,23H-porphyrin (TCPP) molecules, formed via intermolecular hydrogen bonding on Au(111), was investigated by electrochemical scanning tunneling microscopy (EC-STM). Three types of domains, including a square network with molecules trapped inside, square packing, and hexagonal close-packing structures have been observed under various pH conditions. The difference in STM contrast between oxidized and reduced TCPP allows the slow electrochemical reduction of adsorbed TCPP to be visualized by STM. For the first time, the pH dependent reduction of porphyrins was imaged by EC-STM, revealing the mechanism of porphyrin slow reduction at electrochemical interfaces. TCPP reduction can be accelerated either by tuning the working electrode potential to a more negative value or by lowering the H(+) concentration. A redox reaction model was proposed based on the pH dependent reduction of TCPP to elucidate the fundamental aspects of porphyrin redox reactions.

Self-assembled multilayers of polyethylenimine, DNA and gold nanoparticles. A study of electron transfer reaction

The present manuscript describes studies of the electron transfer kinetics at gold electrodes modified by electrostatic self-assemblies of polyethylenimine (PEI), DNA and gold nanoparticles (NP) by Scanning Electrochemical Microscopy (SECM). Two redox mediators of similar structure, ferrocenemethanol (FcOH), and ferrocenecarboxylic acid (FcCOOH) were used to evaluate the effect of the electrode modification on the electron transfer process. For both redox probes, the observed electrochemical behavior was dependent of the charge of the external layer of the self-assembled structure. The corresponding apparent heterogeneous rate constant, k0, was determined. The effect of NP adsorption was also evaluated. Independently of the mediator used, an increase of the k0 was observed when NPs were incorporated, and the surfaces presented a conductive behavior similar to the bare gold electrode. SECM images using FcOH as redox mediator were also recorded. Variations in the normalized currents permitted to evaluate differences of the surface electroactivity due to the polymers and/or nanoparticles adsorption.

Block-Localized Density Functional Theory (BLDFT), Diabatic Coupling, and Their Use in Valence Bond Theory for Representing Reactive Potential Energy Surfaces.

A multistate density functional theory in the framework of the valence bond model is described. The method is based on a block-localized density functional theory (BLDFT) for the construction of valence-bond-like diabatic electronic states and is suitable for the study of electron transfer reactions and for the representation of reactive potential energy surfaces. The method is equivalent to a valence bond theory with the treatment of the localized configurations by using density functional theory (VBDFT). In VBDFT, the electron densities and energies of the valence bond states are determined by BLDFT. A functional estimate of the off-diagonal matrix elements of the VB Hamiltonian is proposed, making use of the overlap integral between Kohn-Sham determinants and the exchange-correlation functional for the ground state substituted with the transition (exchange) density. In addition, we describe an approximate approach, in which the off-diagonal matrix element is computed by wave function theory using block-localized Kohn-Sham orbitals. The key feature is that the electron density of the adiabatic ground state is not directly computed nor used to obtain the ground-state energy; the energy is determined by diagonalization of the multistate valence bond Hamiltonian. This represents a departure from the standard single-determinant Kohn-Sham density functional theory. The multistate VBDFT method is illustrated by the bond dissociation of H2+ and a set of three nucleophilic substitution reactions in the DBH24 database. In the dissociation of H2+, the VBDFT method yields the correct asymptotic behavior as the two protons stretch to infinity, whereas approximate functionals fail badly. For the S(N)2 nucleophilic substitution reactions, the hybrid functional B3LYP severely underestimates the barrier heights, while the approximate two-state VBDFT method overcomes the self-interaction error, and overestimates the barrier heights. Inclusion of the ionic state in a three-state model, VBDFT(3), significantly improves the computed barrier heights, which are found to be in accord with accurate results. The BLDFT method is a versatile theory that can be used to analyze conventional DFT results to gain insight into chemical bonding properties, and it is illustrated by examining the intricate energy contributions to the ion-dipole complex stabilization.