Fig. 6: Construction and evaluation of energy adapter in E. coli.
a Constructing energy adapter by assembling the PserA promoter-based energy responder and the regulatory protein (UbiV and NadD)-based protein capacitor with a low degree of charging. Abbreviation: RBSL, low strength ribosome binding site; EGFP, enhanced green fluorescent protein; mKate, a basic red fluorescent protein; TEV-site, TEV recognition site; TVMV-site, TVMV recognition site; A-IDP, artificial intrinsically disordered protein; PserA, promoter serA; RBS, ribosome binding site; TEV, tobacco etch virus protease; ATPS, ATP sulfurylase; IPTG, isopropyl-β-D-thiogalactopyranoside. b, c Laser confocal microscopy images and pixel intensity analysis to verify the effectiveness of an energy adapter that senses and regulates the intracellular ATP levels in standby and running modes. d Constructing energy adapter by assembling the PfepA promoter-based energy responder and the regulatory protein (UbiV and NadD)-based protein capacitor with a low degree of charging. Abbreviation: PfepA, promoter fepA; TVMV, tobacco vein mottling virus protease; NuoE, NADH dehydrogenase complex subunit E. e, f Laser confocal microscopy images and pixel intensity analysis to verify the effectiveness of energy adapter that senses and regulates the intracellular NADH levels in standby and running modes. The white scale bar at the bottom right of the microscope image represented 1.0 μm (b, e). The average pixel intensity data within the white dashed circle was collected by software Image J in microscope image (b, e). The white dashed circles represented the outlines of the cytoplasmic region and droplet region in the microscope image, respectively (b, e). Values were shown as mean ± s.d. from three biological replicates (c, f). All experiments were repeated independently three times with similar results (b, e). *p < 0.05, **p < 0.01, ***p < 0.001, n.s., no significance, as determined by two-sided t test. Source data are provided as a Source Data file.
