Revising catalytic “acceleration” of enzymes on citrate-capped gold nanoparticles

Abstract In recent years, many papers have reported a catalytic “acceleration” of enzymes when immobilized on gold nanoparticles. Concordantly, gold nanoparticles are often considered as an inert and safe nanomaterial, and are widely used for various purposes, e.g., experiments with humans are being conducted in vivo. In this work we have carried out an in-detail study of catalytic properties of citrate-capped gold nanoparticles and gold nanoparticle-protein conjugates using three model proteins – enzymes glucose oxidase and catalase, and catalytically inactive protein bovine serum albumin. Catalytic properties were studied at different protein-nanoparticle ratios. UV-Vis, DLS, AFM and ζ potential measurements confirmed protein-nanoparticle conjugate formation. Catalytic activity measurements were conducted using oxygen electrode and the data were analyzed by modeling of reaction rate kinetics. The designed experiments demonstrated that gold nanoparticles form stable conjugates with all the investigated proteins, yet they do not increase catalytic activity of the investigated enzymes – in certain conditions gold nanoparticles mimic enzymatic reactions, which may be misattributed to accelerated enzymatic catalysis. Additionally, we present specific key points demonstrating why it may be difficult to differentiate between enzyme- and gold nanoparticle-catalyzed reactions, as well as suggest specific measurements enabling better differentiation. We do not claim that enzymes cannot be accelerated on nanoparticles in general, but rather emphasize, that experimental results demonstrating atypical catalytic performance of enzymes on nanoparticles should be interpreted with additional care, and a widely propagated view of “inert gold nanoparticles” should probably be reconsidered.