Bioelectrocatalytic Cofactor Regeneration Coupled to CO2 Fixation in a Redox-Active Hydrogel for Stereoselective C-C Bond Formation.

The sustainable capture and conversion of carbon dioxide (CO2 ) is key to achieving a circular carbon economy. Bioelectrocatalysis, which aims at using renewable energies to power the highly specific, direct transformation of CO2 into value added products, holds promise to achieve this goal. However, the functional integration of CO2 -fixing enzymes onto electrode materials for the electrosynthesis of stereochemically complex molecules remains to be demonstrated. Here, we show the electricity-driven regio- and stereoselective incorporation of CO2 into crotonyl-CoA by an NADPH-dependent enzymatic reductive carboxylation. Co-immobilization of a ferredoxin NADP+ reductase and crotonyl-CoA carboxylase/reductase within a 2,2'-viologen-modified hydrogel enabled iterative NADPH recycling and stereoselective formation of (2S)-ethylmalonyl-CoA, a prospective intermediate towards multi-carbon products from CO2 , with 92±6 % faradaic efficiency and at a rate of 1.6±0.4 µmol cm-2 h-1 . This approach paves the way for realizing even more complex bioelectrocatalyic cascades in the future.

The electron as a probe to measure the thickness distributions of electroactive films.

Electron conducting films are ubiquitous in applications such as energy conversion, and their ability to fulfill their catalytic function can be greatly limited by inhomogeneities in their thickness or breaks within the film. Knowing the electroactive film thickness distribution would greatly facilitate optimization efforts, but techniques to measure this are lacking. Here, we present an electroanalytical method that provides the thickness distribution of the electrochemically accessible fraction of redox-active films in which the transfer of electrons is diffusional, i.e. by electron hopping. In this method, as the time scale of the experiment (the scan rate) is changed, the location of the diffusion layer boundary relative to the film roughness features is varied, allowing for the extraction of the film thickness distribution. In addition to being conveniently carried out in the solvated state, which is often the operational state of these conductive films, this approach is highly complementary to classical microscopy methods since it samples the entire modified electrode and is specific to the electroactive portions of the film. Therefore, this approach provides information on film morphology that is truly relevant for the catalytic processes being optimized, and thus can guide the optimization of catalyst integration in films towards macroscale cohesion and thickness homogeneity which are essential for optimal performances.

Suppressing hydrogen peroxide generation to achieve oxygen-insensitivity of a [NiFe] hydrogenase in redox active films.

Redox-active films were proposed as protective matrices for preventing oxidative deactivation of oxygen-sensitive catalysts such as hydrogenases for their use in fuel cells. However, the theoretical models predict quasi-infinite protection from oxygen and the aerobic half-life for hydrogenase-catalyzed hydrogen oxidation within redox films lasts only about a day. Here, we employ operando confocal microscopy to elucidate the deactivation processes. The hydrogen peroxide generated from incomplete reduction of oxygen induces the decomposition of the redox matrix rather than deactivation of the biocatalyst. We show that efficient dismutation of hydrogen peroxide by iodide extends the aerobic half-life of the catalytic film containing an oxygen-sensitive [NiFe] hydrogenase to over one week, approaching the experimental anaerobic half-life. Altogether, our data support the theory that redox films make the hydrogenases immune against the direct deactivation by oxygen and highlight the importance of suppressing hydrogen peroxide production in order to reach complete protection from oxidative stress.

Complete Protection of O2-Sensitive Catalysts in Thin Films.

Energy conversion schemes involving dihydrogen hold great potential for meeting sustainable energy needs, but widespread implementation cannot proceed without solutions that mitigate the cost of rare metal catalysts and the O2 instability of biological and bioinspired replacements. Recently, thick films (>100 µm) of redox polymers were shown to prevent O2 catalyst damage but also resulted in unnecessary catalyst load and mass transport limitations. Here we apply novel homogeneous thin films (down to 3 µm) that provide protection from O2 while achieving highly efficient catalyst utilization. Our empirical data are explained by modeling, demonstrating that resistance to O2 inactivation can be obtained for nonlimiting periods of time when the optimal thickness for catalyst utilization and current generation is achieved, even when using highly fragile catalysts such as the enzyme hydrogenase. We show that different protection mechanisms operate depending on the matrix dimensions and the intrinsic catalyst properties and can be integrated together synergistically to achieve stable H2 oxidation currents in the presence of O2, potentially enabling a plethora of practical applications for bioinspired catalysts under harsh oxidative conditions.

Preventing the coffee-ring effect and aggregate sedimentation by in situ gelation of monodisperse materials.

Drop-casting and inkjet printing are virtually the most versatile and cost-effective methods for depositing active materials on surfaces. However, drawbacks associated with the coffee-ring effect, as well as uncontrolled aggregation of the coating materials, have impeded the use of these methods for applications requiring high control of film properties. We now report on a simple method based on covalent cross-linking of monodisperse materials that enables the formation of thin films with homogeneous thicknesses and macroscale cohesion. The coffee-ring effect is impeded by triggering gelation of the coating materials via a thioacetate-disulfide transition which counterbalances the capillary forces induced by evaporation. Aggregates are prevented by monodisperse building blocks that ensure that the resulting gel resists sedimentation until complete droplet drying. This combined strategy yields an unprecedented level of homogeneity in the resulting film thickness in the 100 nm to 10 µm range. Moreover, macroscale cohesion is preserved as evidenced by the long-range charge transfer within the matrix. We highlight the impact of this method with bioelectrocatalysts for H2 and NADPH oxidation. Peak catalytic performances are reached at about 10-fold lower catalyst loading compared to conventional approaches owing to the high control on film cohesion and thickness homogeneity, thus setting new benchmarks in catalyst utilization.

Mechanism of protection of catalysts supported in redox hydrogel films.

The use of synthetic inorganic complexes as supported catalysts is a key route in energy production and in industrial synthesis. However, their intrinsic oxygen sensitivity is sometimes an issue. Some of us have recently demonstrated that hydrogenases, the fragile but very efficient biological catalysts of H2 oxidation, can be protected from O2 damage upon integration into a film of a specifically designed redox polymer. Catalytic oxidation of H2 produces electrons which reduce oxygen near the film/solution interface, thus providing a self-activated protection from oxygen [Plumeré et al., Nat Chem. 2014, 6, 822-827]. Here, we rationalize this protection mechanism by examining the time-dependent distribution of species in the hydrogenase/polymer film, using measured or estimated values of all relevant parameters and the numerical and analytical solutions of a realistic reaction-diffusion scheme. Our investigation sets the stage for optimizing the design of hydrogenase-polymer films, and for expanding this strategy to other fragile catalysts.