Adsorption and Separation of the Biomolecular Motors of F_OF₁‐ATPase‐Embedded Chromatophores Using Titanium Dioxide Microsphere

The biomolecular motor F_OF₁-ATPase-embedded chromatophore, a biomolecular motor loaded into a lipid bilayer of chromatophores derived from biocells demonstrates significant potential for applications in various biomedical fields, such as targeted drug delivery within tumor microenvironments, biological tissue penetration, and biosensor detection. However, conventional purification strategies relying on gradient/ultracentrifugation remain hampered by prohibitive costs, technical complexity, and scalability constraints, critically limiting their biomedical translation. Here, we present a paradigm-shifting approach utilizing titanium dioxide (TiO₂) microspheres for efficient chromatophore isolation via Lewis acid-base interactions. Through constructing chromatophore-TiO₂ complexes, we systematically investigated adsorption mechanisms using isotherm modeling and FTIR spectroscopy, revealing that 7.11%-8.84% of interfacial interactions originated from physisorption. This novel strategy achieved 93.3% ± 3.21% separation efficiency and 90.7% ± 5.77% recovery rates-surpassing conventional centrifugation by 2.1-fold in operational efficiency while maintaining chromatophore integrity. Crucially, the preserved bio functionality of FoF₁-ATPase post-separation was validated through sustained proton gradient-driven ATP (adenosine triphosphate) synthesis. Our findings establish TiO₂-based adsorption as a robust alternative for biomotor purification and elucidate fundamental principles governing nanobiointerfaces between inorganic matrices and membrane-embedded molecular machines. This work provides a universal platform adaptable for diverse biofilm-encapsulated agents, bridging critical gaps between laboratory-scale development and clinical-scale production of advanced bionanodevices.