Light-Driven [FeFe] Hydrogenase Based H2 Production in E. coli: A Model Reaction for Exploring E. coli Based Semiartificial Photosynthetic Systems.

Biohybrid technologies like semiartificial photosynthesis are attracting increased attention, as they enable the combination of highly efficient synthetic light-harvesters with the self-healing and outstanding performance of biocatalysis. However, such systems are intrinsically complex, with multiple interacting components. Herein, we explore a whole-cell photocatalytic system for hydrogen (H2) gas production as a model system for semiartificial photosynthesis. The employed whole-cell photocatalytic system is based on Escherichia coli cells heterologously expressing a highly efficient, but oxygen-sensitive, [FeFe] hydrogenase. The system is driven by the organic photosensitizer eosin Y under broad-spectrum white light illumination. The direct involvement of the [FeFe] hydrogenase in the catalytic reaction is verified spectroscopically. We also observe that E. coli provides protection against O2 damage, underscoring the suitability of this host organism for oxygen-sensitive enzymes in the development of (photo) catalytic biohybrid systems. Moreover, the study shows how factorial experimental design combined with analysis of variance (ANOVA) can be employed to identify relevant variables, as well as their interconnectivity, on both overall catalytic performance and O2 tolerance.

Dinuclear 2,4-di(pyridin-2-yl)-pyrimidine based ruthenium photosensitizers for hydrogen photo-evolution under red light.

In this study, we report two dinuclear Ru(II) complexes C1 and C2 and compare them to their mononuclear analogues Ref1 and Ref2. The dinuclear species exhibit a much stronger absorption, longer excited-state lifetimes and higher luminescence quantum yields than the mononuclear complexes. In addition, C1 and C2 are easier to reduce. An estimation of the driving forces for the electron transfer processes relevant to photocatalytic hydrogen evolution suggests that C1 and Ref2 possess similar activity as photosensitizer (PS). Yet, the improved photophysical properties of C1 make it a more promising candidate for hydrogen evolution. In hydrogen evolution experiments, C1 indeed exhibits increased activity as PS, however, the catalytic system loses its activity after only a few hours. C2 is less active than the mononuclear complexes despite its superior photophysical properties. This observation is attributed to a lack of driving force for the electron transfer towards the catalyst. Further studies of the dinuclear complex C1 show that it is indeed the PS, which decomposes under the catalytic conditions, presumably due to the electron transfer towards the catalyst being the rate-limiting step.