Solar Water Splitting with a Hydrogenase Integrated in Photoelectrochemical Tandem Cells.

Hydrogenases (H2 ases) are benchmark electrocatalysts for H2 production, both in biology and (photo)catalysis in vitro. We report the tailoring of a p-type Si photocathode for optimal loading and wiring of H2 ase through the introduction of a hierarchical inverse opal (IO) TiO2 interlayer. This proton-reducing Si|IO-TiO2 |H2 ase photocathode is capable of driving overall water splitting in combination with a photoanode. We demonstrate unassisted (bias-free) water splitting by wiring Si|IO-TiO2 |H2 ase to a modified BiVO4 photoanode in a photoelectrochemical (PEC) cell during several hours of irradiation. Connecting the Si|IO-TiO2 |H2 ase to a photosystem II (PSII) photoanode provides proof of concept for an engineered Z-scheme that replaces the non-complementary, natural light absorber photosystem I with a complementary abiotic silicon photocathode.

Photoelectrocatalytic H2 evolution in water with molecular catalysts immobilised on p-Si via a stabilising mesoporous TiO2 interlayer.

The development of photoelectrodes capable of light-driven hydrogen evolution from water is an important approach for the storage of solar energy in the form of a chemical energy carrier. However, molecular catalyst-based photocathodes remain scarcely reported and typically suffer from low efficiencies and/or stabilities due to inadequate strategies for interfacing the molecular component with the light-harvesting material. In this study, we report the straightforward preparation of a p-silicon|mesoporous titania|molecular catalyst photocathode assembly that is active towards proton reduction in aqueous media with an onset potential of +0.4 V vs. RHE. The mesoporous TiO2 scaffold acts as an electron shuttle between the silicon and the catalyst, while also stabilising the silicon from passivation and enabling a high loading of molecular catalysts (>30 nmol (geometrical cm)-2). When a Ni bis(diphosphine)-based catalyst is anchored on the surface of the electrode, a high turnover number of ∼1 × 103 was obtained from photoelectrolysis under UV-filtered simulated solar irradiation at 1 Sun after 24 h at pH 4.5. Notwithstanding its aptitude for molecular catalyst immobilisation, the p-Si|TiO2 photoelectrode showed great versatility towards different catalysts and pH conditions, with photoelectrocatalytic H2 generation also being achieved with platinum and a hydrogenase as catalyst, highlighting the flexible platform it represents for many potential reductive catalysis transformations.

Photoelectrochemical Reduction of Carbon Dioxide to Methanol through a Highly Efficient Enzyme Cascade.

Natural photosynthesis is an effective route for the clean and sustainable conversion of CO2 into high-energy chemicals. Inspired by the natural process, a tandem photoelectrochemical (PEC) cell with an integrated enzyme-cascade (TPIEC) system was designed, which transfers photogenerated electrons to a multienzyme cascade for the biocatalyzed reduction of CO2 to methanol. A hematite photoanode and a bismuth ferrite photocathode were applied to fabricate the iron oxide based tandem PEC cell for visible-light-assisted regeneration of the nicotinamide cofactor (NADH). The cell utilized water as an electron donor and spontaneously regenerated NADH. To complete the TPIEC system, a superior three-dehydrogenase cascade system was employed in the cathodic part of the PEC cell. Under applied bias, the TPIEC system achieved a high methanol conversion output of 220 µm h-1 , 1280 µmol g-1 h-1 using readily available solar energy and water.

Sunlight-assisted, biocatalytic formate synthesis from CO2 and water using silicon-based photoelectrochemical cells.

We report on a silicon-based photoelectrochemical cell that integrates a formate dehydrogenase from Thiobacillus sp. (TsFDH) to convert CO2 to formate using water as an electron donor under visible light irradiation and an applied bias. Our current study suggests that the deliberate integration of biocatalysis to a light-harvesting platform could provide an opportunity to synthesize valuable chemicals with the use of earth-abundant materials and sustainable resources.

Cofactor-free light-driven whole-cell cytochrome P450 catalysis.

Cytochromes P450 can catalyze various regioselective and stereospecific oxidation reactions of non-functionalized hydrocarbons. Here, we have designed a novel light-driven platform for cofactor-free, whole-cell P450 photo-biocatalysis using eosin Y (EY) as a photosensitizer. EY can easily enter into the cytoplasm of Escherichia coli and bind specifically to the heme domain of P450. The catalytic turnover of P450 was mediated through the direct transfer of photoinduced electrons from the photosensitized EY to the P450 heme domain under visible light illumination. The photoactivation of the P450 catalytic cycle in the absence of cofactors and redox partners is successfully conducted using many bacterial P450s (variants of P450 BM3) and human P450s (CYPs 1A1, 1A2, 1B1, 2A6, 2E1, and 3A4) for the bioconversion of different substrates, including marketed drugs (simvastatin, lovastatin, and omeprazole) and a steroid (17ß-estradiol), to demonstrate the general applicability of the light-driven, cofactor-free system.

New platform for cytochrome p450 reaction combining in situ immobilization on biopolymer.

We describe an efficienct chemical conversion platform with in situ immobilization of P450-BM3 on poly(3-hydroxybutyrate) granules. Through fusion with phasin, P450-BM3 is easily immobilized on poly(3-hydroxybutyrate) granules in Escherichia coli. In our work, the immobilized P450 exhibited higher stability and catalytic activity compared to free P450 against changes of pH, temperature, and concentrations of urea and ions. Through quick recovery of immobilized enzyme, the P450-P(3HB) complex successfully catalyzed an O-dealkylation reaction several times with maintained activity. Using the robust P450-P(3HB) complex, we performed a P450-catalyzed reaction on a preparative reactor scale (100 mL) and high-level production (12.3 µM) of 7-hydroxycoumarine from 7-ethoxycoumarin could be achieved.

Silicon nanowire photocathodes for light-driven electroenzymatic synthesis.

The photoelectroenzymatic synthesis of chemical compounds employing platinum nanoparticle-decorated silicon nanowires (Pt-SiNWs) is presented. The Pt-SiNWs proved to be an efficient material for photoelectrochemical cofactor regeneration because the silicon nanowires absorbs a wide range of the solar spectrum while the platinum nanoparticle serve as an excellent catalyst for electron and proton transfer. By integrating the platform with redox enzymatic reaction, visible-light-driven electroenzymatic synthesis of L-glutamate was achieved. Compared to electrochemical and photochemical methods, this approach is free from side reactions caused by sacrificial electron donors and has the advantage of applying low potential to realize energy-efficient and sustainable synthesis of chemicals by a photoelectroenzymatic system.

Biocatalytic photosynthesis with water as an electron donor.

Efficient harvesting of unlimited solar energy and its conversion into valuable chemicals is one of the ultimate goals of scientists. With the ever-increasing concerns about sustainable growth and environmental issues, numerous efforts have been made to develop artificial photosynthetic process for the production of fuels and fine chemicals, thus mimicking natural photosynthesis. Despite the research progress made over the decades, the technology is still in its infancy because of the difficulties in kinetic coupling of whole photocatalytic cycles. Herein, we report a new type of artificial photosynthesis system that can avoid such problems by integrally coupling biocatalytic redox reactions with photocatalytic water splitting. We found that photocatalytic water splitting can be efficiently coupled with biocatalytic redox reactions by using tetracobalt polyoxometalate and Rh-based organometallic compound as hole and electron scavengers, respectively, for photoexcited [Ru(bpy)3](2+). Based on these results, we could successfully photosynthesize a model chiral compound (L-glutamate) using a model redox enzyme (glutamate dehydrogenase) upon in situ photoregeneration of cofactors.

Nanobiocatalytic assemblies for artificial photosynthesis.

Natural photosynthesis, a solar-to-chemical energy conversion process, occurs through a series of photo-induced electron transfer reactions in nanoscale architectures that contain light-harvesting complexes, protein-metal clusters, and many redox biocatalysts. Artificial photosynthesis in nanobiocatalytic assemblies aims to reconstruct man-made photosensitizers, electron mediators, electron donors, and redox enzymes for solar synthesis of valuable chemicals through visible light-driven cofactor regeneration. The key requirement in the design of biocatalyzed artificial photosynthetic process is an efficient and forward electron transfer between each photosynthetic component. This review describes basic principles in combining redox biocatalysis with photocatalysis, and highlights recent research outcomes in the development of nanobiocatalytic assemblies that can mimic natural photosystems I and II, respectively. Current issues in biocatalyzed artificial photosynthesis and future perspectives will be briefly discussed.

Near-infrared-light-driven artificial photosynthesis by nanobiocatalytic assemblies.

Artificial photosynthesis in nanobiocatalytic assemblies aims to reconstruct man-made photosensitizers, electron mediators, electron donors, and redox enzymes for solar synthesis of valuable chemicals through photochemical cofactor regeneration. Herein, we report, for the first time, on nanobiocatalytic artificial photosynthesis in near-infrared (NIR) light, which constitutes over 46% of the solar energy. For NIR-light-driven photoenzymatic synthesis, we synthesized silica-coated upconversion nanoparticles, Si-NaYF4:Yb,Er and Si-NaYF4:Yb,Tm, for efficient photon-conversion through Förster resonance energy transfer (FRET) with rose bengal (RB), a photosensitizer. We observed NIR-induced electron transfer by using linear sweep voltammetric analysis; this indicates that photoexcited electrons of RB/Si-NaYF4:Yb,Er are transferred to NAD+ through a Rh-based electron mediator. RB/Si-NaYF4:Yb,Er nanoparticles, which exhibit higher FRET efficiency due to more spectral overlap than RB/Si-NaYF4:Yb,Tm, perform much better in the photoenzymatic conversion.

Visible light-driven NADH regeneration sensitized by proflavine for biocatalysis.

Harvest time: Proflavine drives the reduction of NAD(+) in the presence of a Rh-based electron mediator. Photoregenerated NADH was enzymatically active for oxidation by NADH-dependent L-glutamate dehydrogenase for the synthesis of L-glutamate. This work suggests that proflavine has the potential to become an efficient light-harvesting component in biocatalytic photosynthesis driven by solar energy.

Photoenzymatic synthesis through sustainable NADH regeneration by SiO2-supported quantum dots.

Sustainable photochemical NADH regeneration and redox-enzymatic synthesis are accomplished by using CdS nanocrystals grown on the surface of SiO(2) beads. CdS nanocrystals grown on SiO(2) beads worked efficiently as a visible-light absorbing photocatalyst for in situ NADH regeneration with high catalytic activity and minimal loss of activity despite repeated uses.

Self-assembled, photoluminescent peptide hydrogel as a versatile platform for enzyme-based optical biosensors.

A self-assembled peptide hydrogel consisting of Fmoc-diphenylalanine has been employed as a biosensing platform through the encapsulation of enzyme bioreceptors (e.g., glucose oxidase or horseradish peroxidase) and fluorescent reporters (e.g., CdTe and CdSe quantum dots). Enzymes and quantum dots (QDs) were physically immobilized within the hydrogel matrix in situ in a single step by simply mixing aqueous solution containing QDs and enzymes with monomeric peptide (Fmoc-diphenylalanine) solution. By using atomic force microscopy and scanning transmission electron microscopy, we observed that the self-assembled peptide hydrogel had a three-dimensional network of nanofibers (with a diameter of approximately 70-90 nm) that physically hybridized with QDs and encapsulated enzyme bioreceptors with a minimal leakage. We successfully applied the peptide hydrogel to the detection of analytes such as glucose and toxic phenolic compounds by using a photoluminescence quenching of the hybridized QDs. The Michaelis-Menten constant (K(M)) of the photoluminescent peptide hydrogel was found to be 3.12 mM (GOx for glucose) and 0.82 mM (HRP for hydroquinone), respectively, which were much lower than those of conventional gel materials. These results suggest that the peptide hydrogel is an alternative optical biosensing platform with practical advantages such as simple fabrication via self-assembly, efficient diffusion of target analytes, and high encapsulation efficiencies for fluorescent reporters and bioreceptors.

Eosin Y-sensitized artificial photosynthesis by highly efficient visible-light-driven regeneration of nicotinamide cofactor.

Dye-sensitized photosynthesis: Eosin Y (EY), a dye photosensitizer, works efficiently as a molecular photoelectrode by catalyzing the visible-light-driven electron-transfer reaction for efficient regeneration of NADH through a photosensitizer-electron relay dyad. Injection of the photosensitized electron resulted in highly accelerated regeneration of NADH, which can be used by glutamate dehydrogenase for the photosynthesis of L-glutamate.