Living Spatially Organized Hybrid Condensates for Efficient Light-Driven CO2-to-Chemical Conversion

Abstract Light-driven CO2-to-chemical conversion presents a sustainable route for biomanufacturing, yet the performance of photosynthetic biohybrid systems remains fundamentally constrained by disorganized biotic–abiotic interfaces, which result in inefficient photoelectron transfer, low carbon production, and substantial cellular toxicity. Here, we report a spatially organized intracellular biohybrid system that enables the light-powered conversion of CO2 to 2,3-butanediol inEscherichia coli. Inspired by the algal pyrenoid, we constructed a TiO2/RuBisCO hybrid condensate (TRHC) by integrating protein liquid–liquid phase separation (LLPS) with in situ biomimetic mineralization. This spatially defined condensate physically clusters light-harvesting and carbon-fixing modules, thereby facilitating direct photoelectron transfer and contributing to improved long-term cellular robustness under continuous illumination. The TRHC achieved an apparent quantum yield of 12.4% and reduced the light-induced cellular mortality by 64%. By providing a robust photoelectron supply for CO2 assimilation, the integrated system attains a 2,3-butanediol yield of 1.38 mol/mol from the consumed glucose. Moreover, under illuminated fed-batch fermentation, the platform maintains sustained biosynthetic activity over 5 days, producing 62.11 g/L of 2,3-butanediol. This LLPS-guided strategy offers a generalizable framework for engineering efficient biotic-abiotic interfaces in solar-driven biomanufacturing.