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Engineering of NADPH Supply Boosts Photosynthesis-Driven Biotransformations.

ABSTRACT: Light-driven biocatalysis in recombinant cyanobacteria provides highly atom-efficient cofactor regeneration via photosynthesis, thereby remediating constraints associated with sacrificial cosubstrates. However, despite the remarkable specific activities of photobiocatalysts, self-shading at moderate-high cell densities limits efficient space-time-yields of heterologous enzymatic reactions. Moreover, efficient integration of an artificial electron sink into the tightly regulated network of cyanobacterial electron pathways can be highly challenging. Here, we used C=C bond reduction of 2-methylmaleimide by the NADPH-dependent ene-reductase YqjM as a model reaction for light-dependent biotransformations. Time-resolved NADPH fluorescence spectroscopy allowed direct monitoring of in-cell YqjM activity and revealed differences in NADPH steady-state levels and oxidation kinetics between different genetic constructs. This effect correlates with specific activities of whole-cells, which demonstrated conversions of >99%. Further channelling of electrons toward heterologous YqjM by inactivation of the flavodiiron proteins (Flv1/Flv3) led to a 2-fold improvement in specific activity at moderate cell densities, thereby elucidating the possibility of accelerating light-driven biotransformations by the removal of natural competing electron sinks. In the best case, an initial product formation rate of 18.3 mmol h-1 L-1 was reached, allowing the complete conversion of a 60 mM substrate solution within 4 h.

SUBMITTER: Assil-Companioni L

PROVIDER: S-EPMC7574619 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Engineering of NADPH Supply Boosts Photosynthesis-Driven Biotransformations.

Assil-Companioni Leen L Büchsenschütz Hanna C HC Solymosi Dániel D Dyczmons-Nowaczyk Nina G NG Bauer Kristin K F KKF Wallner Silvia S Macheroux Peter P Allahverdiyeva Yagut Y Nowaczyk Marc M MM Kourist Robert R

ACS catalysis 20200904 20

Light-driven biocatalysis in recombinant cyanobacteria provides highly atom-efficient cofactor regeneration <i>via</i> photosynthesis, thereby remediating constraints associated with sacrificial cosubstrates. However, despite the remarkable specific activities of photobiocatalysts, self-shading at moderate-high cell densities limits efficient space-time-yields of heterologous enzymatic reactions. Moreover, efficient integration of an artificial electron sink into the tightly regulated network of ...[more]

PMID: 33101760

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Project description:Drought during senescence has become more common in Mediterranean climates in recent years. Chenopodium quinoa Willd has been identified as tolerant to poor soil conditions and drought. Previous observations have found that sufficient nitrogen (N) supply mitigates yield losses under terminal drought conditions. However, there is no understanding of the mechanisms behind this effect. We hypothesized that N up-regulates both photosynthetic and photoprotective elements during drought-induced senescence, alleviating the negative impact of drought on yield. The role of N supply and terminal drought on photoprotection was tested using three Chilean quinoa genotypes from different climatic zones: Faro, UdeC9, and BO78. Plants were grown under high nitrogen (HN) or low nitrogen (LN) conditions and subjected to terminal drought at the onset of senescence. Photosynthetic and photochemical and non-photochemical processes were evaluated at both the onset of drought and after 15 days of drought conditions. N supplementation modified most of the physiological parameters related to photochemical dissipation of energy, photosynthesis, and yield in quinoa. In contrast, water restriction did not affect photosynthesis in quinoa, and its effect on yield was dependent on the genotype. A significant interaction N × G was observed in photosynthesis, relative water content, protein content, Fv/Fm, and chlorophylls. In general, Faro was able to maintain higher levels of these attributes under LN conditions than UdeC9 and BO78. In addition, the interacting effects of N × W regulated the level of most pigments in quinoa as well as the photoprotective induction of non-photochemical quenching (NPQ) during senescence. During terminal drought at LN conditions, Faro presented a larger NPQ induction under drought conditions than UdeC9 and BO78, which was supported by a larger zeaxanthin content and de-epoxidation state of the xanthophyll pool. Interestingly, BO78 did not induce NPQ in response to drought-induced senescence but instead enhanced the content of betacyanins. This response needs to be researched in future works. Finally, we observed that LN supply reduced the correlationship between the de-epoxidation state of the xanthophyll cycle and NPQ. This could be an indication that N supply not only compromised the capacity for photosynthetic performance in quinoa plants, but also affected the plasticity of thermal dissipation, restricting further changes during drought-induced senescence.

Dataset Information

Engineering of NADPH Supply Boosts Photosynthesis-Driven Biotransformations.

Publications

Engineering of NADPH Supply Boosts Photosynthesis-Driven Biotransformations.

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