title: Understanding How Nanoparticle Attachment Enhances Phosphotriesterase Kinetic Efficiency.
abstract: As a specific example of the enhancement of enzymatic activity that can be induced by nanoparticles, we investigate the hydrolysis of the organophosphate paraoxon by phosphotriesterase (PTE) when the latter is displayed on semiconductor quantum dots (QDs). PTE conjugation to QDs underwent extensive characterization including structural simulations, electrophoretic mobility shift assays, and dynamic light scattering to confirm orientational and ratiometric control over enzyme display which appears to be necessary for enhancement. PTE hydrolytic activity was then examined when attached to ca. 4 and 9 nm diameter QDs in comparison to controls of freely diffusing enzyme alone. The results confirm that the activity of the QD conjugates significantly exceeded that of freely diffusing PTE in both initial rate (∼4-fold) and enzymatic efficiency (∼2-fold). To probe kinetic acceleration, various modified assays including those with increased temperature, presence of a competitive inhibitor, and increased viscosity were undertaken to measure the activation energy and dissociation rates. Cumulatively, the data indicate that the higher activity is due to an acceleration in enzyme-product dissociation that is presumably driven by the markedly different microenvironment of the PTE-QD bioconjugate's hydration layer. This report highlights how a specific change in an enzymatic mechanism can be both identified and directly linked to its enhanced activity when displayed on a nanoparticle. Moreover, the generality of the mechanism suggests that it could well be responsible for other examples of nanoparticle-enhanced catalysis.
authors.affiliation: American Society for Engineering Education , Washington, DC 20036, United States.
authors.forename: Joyce C
authors.initials: JC
authors.lastname: Breger
authors.forename: Mario G
authors.initials: MG
authors.lastname: Ancona
authors.forename: Scott A
authors.initials: SA
authors.lastname: Walper
authors.affiliation: Sotera Defense Solutions, Inc. 7230 Lee DeForest Drive, Columbia, Maryland 21046, United States.
authors.forename: Eunkeu
authors.initials: E
authors.lastname: Oh
authors.affiliation: Sotera Defense Solutions, Inc. 7230 Lee DeForest Drive, Columbia, Maryland 21046, United States.
authors.forename: Kimihiro
authors.initials: K
authors.lastname: Susumu
authors.forename: Michael H
authors.initials: MH
authors.lastname: Stewart
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Deschamps
authors.forename: Igor L
authors.initials: IL
authors.lastname: Medintz
mesh_terms: D055162:Biocatalysis; D019187:Cadmium Compounds / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D008958:Models, Molecular; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D009578:Nitrobenzenes / Q000737:chemistry; D010261:Paraoxon / Q000737:chemistry*; D010316:Particle Size; D044345:Phosphoric Triester Hydrolases / Q000737:chemistry*; D045663:Quantum Dots / Q000737:chemistry* / Q000648:ultrastructure; D018036:Selenium Compounds / Q000737:chemistry; D013816:Thermodynamics
chemical_list: D019187:Cadmium Compounds; D004800:Enzymes, Immobilized; D009578:Nitrobenzenes; D018036:Selenium Compounds; C058667:cadmium selenide; D044345:Phosphoric Triester Hydrolases; C013154:4-nitrophenetol; D010261:Paraoxon
keywords: Km; Michaelis−Menten; bionanotechnology; enzyme; kcat; kinetics; nanoparticle; nerve agent; paraoxon; phosphotriesterase; quantum dot; rate