title: Light-powering Escherichia coli with proteorhodopsin.
abstract: Proteorhodopsin (PR) is a light-powered proton pump identified by community sequencing of ocean samples. Previous studies have established the ecological distribution and enzymatic activity of PR, but its role in powering cells and participation in ocean energy fluxes remains unclear. Here, we show that when cellular respiration is inhibited by depleting oxygen or by the respiratory poison azide, Escherichia coli cells expressing PR become light-powered. Illumination of these cells with light coinciding with PR's absorption spectrum creates a proton motive force (pmf) that turns the flagellar motor, yielding cells that swim when illuminated with green light. By measuring the pmf of individual illuminated cells, we quantify the coupling between light-driven and respiratory proton currents, estimate the Michaelis-Menten constant (Km) of PR (10(3) photons per second/nm2), and show that light-driven pumping by PR can fully replace respiration as a cellular energy source in some environmental conditions. Moreover, sunlight-illuminated PR+ cells are less sensitive to azide than PR- cells, consistent with PR+ cells possessing an alternative means of maintaining cellular pmf and, thus, viability. Proteorhodopsin allows Escherichia coli cells to withstand environmental respiration challenges by harvesting light energy.
authors.affiliation: Department of Physics, University of California, Berkeley, CA 94720, USA.
authors.forename: Jessica M
authors.initials: JM
authors.lastname: Walter
authors.forename: Derek
authors.initials: D
authors.lastname: Greenfield
authors.forename: Carlos
authors.initials: C
authors.lastname: Bustamante
authors.forename: Jan
authors.initials: J
authors.lastname: Liphardt
mesh_terms: D000693:Anaerobiosis; D001386:Azides / Q000494:pharmacology; D004734:Energy Metabolism; D004926:Escherichia coli / Q000378:metabolism* / Q000502:physiology; D005407:Flagella; D007700:Kinetics; D008027:Light; D009792:Oceans and Seas; D017494:Proton Pumps; D012243:Rhodopsin / Q000502:physiology*; D025604:Rhodopsins, Microbial
chemical_list: D001386:Azides; D017494:Proton Pumps; D025604:Rhodopsins, Microbial; C413548:proteorhodopsin; D012243:Rhodopsin