Research data supporting "Towards solar-powered growth of autotrophic Escherichia coli using photoelectrochemistry"

This deposit contains the figure files (TIF or PNG renderings of each panel as published) and the underlying source data (XLSX numerical tables for kinetic, electrochemical, and spectroscopic traces; mnova native files for raw 1H NMR spectra) for every figure in the main manuscript (MS) and Supporting Information (SI), plus the Table of Contents (TOC) graphic. Tables S1–S4 are summary tables embedded within the published Supporting Information PDF and are not deposited as separate files. Files are organised into two top-level folders, MS/ and SI/, each subdivided into one folder per figure (e.g. MS/Figure 3/, SI/Figure S22/), so that filenames map directly to the figure numbering used in the manuscript. Time-resolved bacterial OD600, formate, acetate, and electrochemical traces are reported as mean and standard deviation from three biological or technical replicates unless stated otherwise on the data sheet. Manuscript (Figures 1–5) Figure 1. Schematic of natural photosynthesis (1A) and the semi-biological platform pairing the BiVO4|TiCo photoanode with the OPV|IO-TiO2|FDH+CA photocathode (1B). Files: Figure 1.tif. Figure 2. Adaptive laboratory evolution of autotrophic E. coli on 60 mM formate over 27 serial transfers (1:10 dilution, 5% O2 / 10% CO2 / 85% N2). OD600 versus time for the parental (#1) and evolved (#27) strains. Files: Figure 2.tif, Figure 2.xlsx. Figure 3. (3A) Schematic of the IO-TiO2|FDH+CA cathode for bioelectrochemical formate synthesis and utilisation. (3B) Controlled potential electrolysis at −0.4 V vs. RHE over 10 h: current density and Faradaic yield of formate. (3C) OD600 of evolved E. coli and consumption of electrochemically generated formate over 6 days. Files: Figure 3.tif, Figure 3.xlsx. Figure 4. (4A) Schematic of the OPV-integrated photocathode for consecutive solar-driven biomass growth. (4B) Chopped-light photocurrent density and formate Faradaic yield at 0.6 V vs. RHE over 10 h. (4C) OD600 and formate consumption of evolved E. coli in the solar reactor effluent over 3 days. Files: Figure 4.tif, Figure 4.xlsx. Figure 5. (5A) Configuration of the semi-artificial leaf with OPV photobiocathode and BiVO4 photoanode. (5B) OD600 of E. coli and 13C-formate concentration under bias-free 1 sun illumination, tracking 13C turnover into biomass. Files: Figure 5.tif, Figure 5.xlsx. Supporting Information (Figures S1–S29) Figure S1. (S1A) Stationary-phase OD600 across 27 ALE transfers. (S1B) Growth and formate consumption kinetics of #1 versus #27. (S1C) Plate-reader growth profiles of #1, #27, and the pitA-reverted control #27-pitAWT in M9 + 40 mM formate at 37 °C. Files: Figure S1.png, Figure S1 A-B.xlsx, Figure S1 C.xlsx. Figure S2. Scanning electron microscopy of IO-TiO2 on Ti foil at 5 kV: (S2A) surface, view field 2 μm; (S2B) cross-section, view field 64 μm. Files: Figure S2.png, Figure S2A.tif, Figure S2B.TIF. Figure S3. Preliminary growth of E. coli on a carbon-felt|TiO2|FDH cathode catholyte. (S3A) OD600 over 3 days; (S3B) formate concentration; (S3C–D) 1H NMR spectra of Day 0 and Day 3 supernatants in the glycerol region. Files: Figure S3.png, Figure S3.xlsx. Figure S4. Switch from PEC-generated formate to glycerol metabolism. (S4A) Formate concentration over 4 days; (S4B) OD600 over 2 days; (S4C–E) 1H NMR spectra of the post-reaction electrolyte showing glycerol depletion. Files: Figure S4.png, Figure S4A-B.xlsx, Figure S4C-D.mnova, Figure S4C.png, Figure S4D.png, Figure S4E.mnova, Figure S4E.png. Figure S5. Electrochemistry with FDH purified through a 50 kDa MWCO filter on IO-TiO2. (S5A) PFV before CPE; (S5B) 10 h CPE at −0.4 V vs. RHE; (S5C) PFV of pristine FDH for comparison; (S5D) PFV after CPE; (S5E) post-CPE 1H NMR with pure-glycerol reference. Files: Figure S5.png, Figure S5A-D.xlsx, Figure S5E_Pure glycerol.mnova, Figure S5E_Reaction 10h.mnova. Figure S6. Electrochemistry with FDH purified through a 30 kDa MWCO filter on IO-TiO2. (S6A) PFV before CPE; (S6B) 7 h CPE at −0.4 V vs. RHE; (S6C) PFV after CPE; (S6D) post-CPE 1H NMR with pure-glycerol reference. Files: Figure S6.png, Figure S6A-C.xlsx, Figure S6D_pure glycerol.mnova, Figure S6D_Reaction 10h.mnova. Figure S7. 1H NMR of the post-CPE electrolyte after the optimised three-cycle 30 kDa wash protocol, with commercial glycerol reference. Files: Figure S7.png, Figure S7_pure glycerol.mnova, Figure S7_Reaction 10h.mnova. Figure S8. Reference 1H NMR spectra. (S8A–B) Post-CPE electrolyte with purified and pristine FDH; (S8C) commercial TRIS-HCl; (S8D) glycerol; (S8E) DTT. Files: Figure S8.png, Figure S8.mnova. Figure S9. Zoomed-in 1H NMR of TRIS-HCl, showing two minor resonances not visible in the broad-range spectrum of S8C. Files: Figure S9.png, Figure S9.mnova. Figure S10. Photographs of the H-cell setup: (S10A) electrochemistry; (S10B) PEC front view; (S10C) PEC side view. Files: Figure S10.png, Figure S10A.png, Figure S10B.png, Figure S10C.png. Figure S11. Linear-sweep PFV scans of purified FDH on IO-TiO2 before (S11A) and after (S11B) 10 h CPE at −0.4 V vs. RHE. Files: Figure S11.png, Figure S11.xlsx. Figure S12. Nyquist plot from electrochemical impedance spectroscopy of bare IO-TiO2 versus IO-TiO2|FDH+CA at open-circuit potential. Files: Figure S12.png, Figure S12.xlsx. Figure S13. Triplicate 10 h CPE traces at −0.4 V vs. RHE for three independent IO-TiO2|FDH+CA cathodes (500 pmol FDH + 100 pmol CA). Files: Figure S13.png, Figure S13.xlsx. Figure S14. ΔOD600 of evolved E. coli batch cultures across initial formate concentrations of 14, 23, 31, and 51 mM, defining the working range for the integrated system. Files: Figure S14.png, Figure S14.xlsx. Figure S15. Growth profile of evolved E. coli in the post-electrolysis electrolyte without supplementary trace elements, showing the trace-element requirement. Files: Figure S15.png, Figure S15.xlsx. Figure S16. OD600, formate consumption, and acetate production of evolved E. coli at varying initial headspace O2 (1–10%) over 6 days, with the metabolic shift to fermentation under hypoxia. Files: Figure S16.png, Figure S16.xlsx. Figure S17. PFV of OPV|IO-TiO2|FDH+CA photocathode under chopped, continuous, and dark illumination, before (S17A) and after (S17B) 10 h CPE at 0.6 V vs. RHE. Files: Figure S17.png, Figure S17.xlsx. Figure S18. External quantum efficiency spectra of the OPV|IO-TiO2|FDH+CA photocathode at 0 V and 0.6 V vs. RHE. Files: Figure S18.png, Figure S18.xlsx. Figure S19. Triplicate 10 h CPE scans of OPV|IO-TiO2|FDH+CA photocathodes at 0.6 V vs. RHE under chopped 1 sun illumination (50 min on, 10 min off). Files: Figure S19.png, Figure S19.xlsx. Figure S20. Cyclic voltammetry of the BiVO4|TiCo photoanode under simulated 1 sun (AM1.5G, >400 nm UV filter) at 37 °C with stirring. Files: Figure S20.png, Figure S20.xlsx. Figure S21. PFV scans of the integrated semi-artificial leaf in two-electrode mode under chopped, continuous, and dark illumination, showing onset voltage near −0.6 V. Files: Figure S21.png, Figure S21.xlsx. Figure S22. Triplicate bias-free CPE traces and O2 evolution profiles of the semi-artificial leaf in E. coli-containing electrolyte over 20 h. Files: Figure S22.png, Figure S22.xlsx. Figure S23. OD600 and formate concentration in the integrated leaf system with (S23A) and without (S23B) trace element supplementation; arrows mark the headspace exchange to 5% O2 / 10% CO2 / 85% N2. Files: Figure S23.png, Figure S23.xlsx. Figure S24. Final OD600 of E. coli after 3 days in the semi-artificial leaf system under simulated solar illumination versus a paired dark control (n = 3, p = 0.023). Files: Figure S24.png, Figure S24.xlsx. Figure S25. SEM images of the IO-TiO2 electrode after the 3-day biomass production experiment at view fields of 10 μm (S25A) and 2.5 μm (S25B), 5 kV. Files: Figure S25.png, Figure S25A.TIF, Figure S25B.TIF. Figure S26. BiVO4|TiCo photoanode after 3-day incubation with E. coli. (S26A–B) SEM at 4 and 2 μm view fields; (S26C–D) backscattered electron images for compositional contrast; (S26E) energy-dispersive X-ray spectrum showing salt deposition from the bacterial medium. Files: Figure S26.png, Figure S26A.TIF, Figure S26B.TIF, Figure S26C-D.TIF, Figure S26E.png. Figure S27. 1H NMR spectra of the semi-artificial leaf electrolyte supplied with 13CO2, showing the time-dependent emergence and consumption of the 13C-formate doublet (peak integrations exported as XLSX). Files: Figure S27.png, Figure S27.xlsx. Figure S28. Schematic of the proposed metabolic pathway for acetate synthesis under hypoxic conditions. Files: Figure S28.png. Figure S29. 13C-isotope tracing under 0–5% O2. (S29A, S29C) Representative 1H NMR spectra at day 2 and day 13 of incubation. (S29B, S29D) Quantification of OD600 with 12C/13C-formate and 12C/13C-acetate at the matching time points. Files: Figure S29.png, Figure S29.xlsx. Table of Contents graphic TOC. Graphical abstract for the published article (3.25 in. × 1.75 in., 300 dpi). Files: TOC.tif. Keywords semi-artificial photosynthesis; biohybrid photoelectrochemistry; CO2 reduction; formate; tungsten formate dehydrogenase; carbonic anhydrase; inverse opal TiO2; organic photovoltaic; PCE10:EH-IDTBR; BiVO4 photoanode; autotrophic Escherichia coli; adaptive laboratory evolution; pitA; reductive glycine pathway; Calvin–Benson–Bassham cycle; 13C isotope tracing; semi-artificial leaf; solar fuels; formate bioeconomy. Software / usage instructions XLSX files: Microsoft Excel; or LibreOffice Calc (free, open-source, www.libreoffice.org). TIF and PNG files: any standard image viewer; ImageJ / Fiji (free, open-source, imagej.net) for quantitative image analysis of SEM micrographs. mnova files: MestReNova by Mestrelab Research (proprietary, mestrelab.com); a free MestReNova Reader is available from the same site for read-only inspection of spectra. Where the integrated peak areas underpin a figure panel (e.g. Figures S27, S29), the processed values are also provided in the corresponding XLSX file so that the data can be re-used without the proprietary software.