Towards high atom economy in whole-cell redox biocatalysis: up-scaling light-driven cyanobacterial ene-reductions in a flat panel photobioreactor.

Abstract

Light-driven biotransformations in recombinant cyanobacteria benefit from the atom-efficient regeneration of reaction equivalents like NADPH from water and light by oxygenic photosynthesis. The self-shading of photosynthetic cells throughout the reaction volume, along with the need for extended light paths, limits adequate light supply and significantly restricts the potential for upscaling. Here, we present a flat panel photobioreactor (1 cm optical path length) as a scalable system to provide efficient illumination at high cell densities. The genes of five ene-reductases from different classes were expressed in Synechocystis sp. PCC 6803. The strains were characterised in the light-driven reduction of a set of prochiral substrates. With specific activities up to 150 U g_CDW ^-1 under standard conditions in small-scale reactions, the recombinant strains harbouring the ene-reductases TsOYE C25G I67T and OYE3 showed the highest specific activities observed so far in photobiotransformations and were selected for the up-scale in the flat panel photobioreactor in 120 mL-scale. The strain producing OYE3 exhibited a specific activity as high as 56.1 U g_CDW ^-1. The corresponding volumetric productivity of 1 g L^-1 h^-1 compares favourably to other photosynthesis-driven processes. This setup facilitated the conversion of 50 mM over approximately 8 hours to an isolated yield of 87%. The atom economy of 88% compares favourably to the use of the sacrificial co-substrates glucose and formic acid with 49% and 78%, respectively. Determination of the complete E-Factor of 203 including water reveals that the volumetric yield and water required for cultivation are crucial for the sustainability. In summary, our results point out key factors for the sustainability of light-driven whole-cell biotransformations, and provide a solid basis for future optimisation and up-scale campaigns of photosynthesis-driven bioproduction.