Thermoresponsive Thylakoid‐Integrated Artificial Photosynthetic Hydrogel Films for Dynamic Regulation of Photo‐Biocatalytic Cofactor Regeneration

Abstract Efficient regeneration of biological energy carriers, such as cofactors of reduced nicotinamide adenine dinucleotide phosphate (NADPH) and adenosine triphosphate, is crucial for driving anabolic processes in artificial photosynthetic systems (APS). However, achieving spatiotemporally regulated cofactor regeneration under physiologically mild conditions remains a fundamental challenge. Here, a nature‐inspired, thermoresponsive photo‐biocatalytic photosynthetic hydrogel platform that enables dynamic regulation of photosynthetic cofactor regeneration through spatiotemporally tunable photo‐biocatalysis is presented. This intelligent system integrates native thylakoid (Tk) membranes as photo‐biocatalytic modules within poly(N‐isopropylacrylamide) (PNI) (Tk@PNI) hydrogel matrix, constructed via templated, ultraviolet‐initiated in situ polymerization process. The integrated thylakoids retain their intrinsic light‐harvesting and electron transport capabilities, enabling light‐driven NADPH regeneration via photoinduced proton‐coupled electron transfer (PCET). Simultaneously, the PNI network functions as a dynamic structural modulator, undergoing reversible volume‐phase transitions in response to temperature changes. This triggers reversible change in microenvironment and spatial constraint embedded thylakoid domains, allowing real‐time regulation of PCET kinetics and catalytic output. By coupling light‐driven energy conversion with thermoresponsive structural reorganization, this biohybrid platform achieves nonlinear, adaptive regulation of photosynthetic performance, sustaining efficient NADPH regeneration over 15–45 °C. This work provides a strategy to integrate stimuli‐responsive polymers with biological energy modules for programmable energy conversion and sustainable biochemical synthesis.