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Abstract Anthropogenic carbon dioxide (CO₂) release in the atmosphere from fossil fuel combustion has inspired scientists to study CO₂to biofuel conversion. Oxygenic phototrophs such as cyanobacteria have been used to produce biofuels using CO₂. However, oxygen generation during oxygenic photosynthesis adversely affects biofuel production efficiency. To producen-butanol (biofuel) from CO₂, here we introduce ann-butanol biosynthesis pathway into an anoxygenic (non-oxygen evolving) photoautotroph,Rhodopseudomonas palustrisTIE-1 (TIE-1). Using different carbon, nitrogen, and electron sources, we achieven-butanol production in wild-type TIE-1 and mutants lacking electron-consuming (nitrogen-fixing) or acetyl-CoA-consuming (polyhydroxybutyrate and glycogen synthesis) pathways. The mutant lacking the nitrogen-fixing pathway produce the highestn-butanol. Coupled with novel hybrid bioelectrochemical platforms, this mutant producesn-butanol using CO₂, solar panel-generated electricity, and light with high electrical energy conversion efficiency. Overall, this approach showcases TIE-1 as an attractive microbial chassis for carbon-neutraln-butanol bioproduction using sustainable, renewable, and abundant resources.