Direct Observation of Assembly and Function of Trigger‐Responsive Lipase Biohybrid Materials

Protein‐polymer biohybrids mark a cutting edge of advanced materials for high performance under diverse conditions by combining the stability and adaptability of polymers with the remarkable functionality of proteins. The potential of stimuli‐responsive polymers for precise, trigger‐responsive control over protein‐polymer biohybrid assembly and function has yet to be fully attained. Here, pH‐ and temperature‐responsive lipase‐polymer biohybrid materials at both the supramolecular and single‐molecule levels are thoroughly characterized. Besides conventional ensemble measurements, the stimuli‐triggered reversibility of the biohybrid material self‐assembly is extensively characterized, and the enzymatic activity at the fundamental level of individual nanoparticles is recorded. The findings demonstrate that the stimuli‐responsive materials not only exhibit a twofold increased activity as compared to native lipases but also maintain trigger‐responsive control over activity by temperature. Direct single‐particle recordings show a correlation of the biohybrid size to its activity, revealing individual enzymes to display higher activity in smaller nanoparticles. The biohybrids exhibit impressive long‐term stability, retaining in some cases almost 95% of the activity after 1‐year of storage at 4 °C as compared to native lipases retaining merely ≈40–60% under the same conditions. The improved stability and activity accompanied by triggered responsiveness highlight significant potential for tightly controlled biocatalysis and sustainable industrial applications.