Fig. 5: Construction of E. coli photosynthesis system with an energy adapter.
a Relative abundance of mass isotopomers for intracellular formate, DHA, MET, and pyruvate in strain TC-F1 with blue light illumination. Abbreviation: DHA, dihydroxyacetone; MET, methylglyoxal. b Schematic diagram of E. coli photosynthesis system with an energy adapter to dynamically match light reaction (NPM*) and dark reaction (ADRP). Abbreviation: ATP/ADP, adenosine triphosphate/diphosphate; NADH/NAD+, nicotinamide adenine dinucleotide reduced/oxidized; PserA, promoter serA; PfepA, promoter fepA; TEV, tobacco etch virus protease; TVMV, tobacco vein mottling virus protease; A-IDP, artificial intrinsically disordered protein; RBS, ribosome binding site; TEV-site, TEV recognition site; TVMV-site, TVMV recognition site; UQ, ubiquinone; NMN, nicotinamide mononucleotide; e−, electron. c, d The effectiveness of nine candidate promoters in sensing ATP levels (c) and NADH levels (d) was evaluated by detecting EGFP (IPTG induction for 4 h) and mKate (IPTG induction for 2 h) fluorescence intensity, respectively. IPTG could induce the expression of ATPS and NuoE to consume ATP and NADH, thereby decreasing the intracellular ATP and NADH levels. Gradient concentrations of IPTG were added to generate different intracellular ATP and NADH levels. Abbreviation: T7, T7 promoter; IPTG, isopropyl-β-D-thiogalactopyranoside; ATPS, ATP sulfurylase; NuoE, NADH dehydrogenase complex subunit E; PATP, promoter that senses intracellular ATP levels; PNADH, promoter that senses intracellular NADH levels; EGFP, enhanced green fluorescent protein; mKate, a basic red fluorescent protein. e The effectiveness of the protein capacitor was evaluated by laser confocal microscopy images and pixel intensity analysis. 1 and 0 signals represented the aggregation and release of protein, respectively. (1, 1), (0, 1), (1, 0), and (0, 0) represented the four states of protein capacitor. The white scale bar at the bottom right of the microscope image represented 2.5 μm. The pixel intensity data was collected by laser confocal microscopy software LAS AF Lite along the track of a white solid arrow in the microscope image. Cells pointed by white triangular arrows had significant differences in microscope images. Abbreviation: RBSH, high strength ribosome binding site. Values were shown as mean ± s.d. from three biological replicates (a, c, d). All experiments were repeated independently three times with similar results (e). Source data are provided as a Source Data file.
