In summary, we have investigated the kinetics of the significantly enhanced activity of an AuNP-bound enzyme. The immobilization of lipase onto the nanoparticles’ surfaces, without any surface modification, provided colloidal stability that allowed us to determine the coverage of the particles with the enzyme and assay the enzymatic activity. The efficiency of enhanced catalytic activity with colloidal stability has been optimized by regularizing the ratios of AuNPs to enzyme. The lipase capping the AuNPs catalyzed the hydrolysis of pNPP through the same kinetic model as that of the free enzyme, with the product’s release being the rate-limiting step, but with increased catalytic activity, as evidenced by lower values of the activation energy and KM. We ascribe this behavior to the enhanced substrate selectivity of the AuNP-bound, thereby decreasing the activation energy through enhancing the rate constants leading to the formation of the ES complex. Our findings suggest that the ability of AuNPs to manipulate catalytic activity might become an important nanobiotechnological tool for optimizing clinical studies and improving drug delivery.
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