A virus-based single-enzyme nanoreactor.
Most enzyme studies are carried out in bulk aqueous solution, at the so-called ensemble level, but more recently studies have appeared in which enzyme activity is measured at the level of a single molecule, revealing previously unseen properties. To this end, enzymes have been chemically or physically anchored to a surface, which is often disadvantageous because it may lead to denaturation. In a natural environment, enzymes are present in a confined reaction space, which inspired us to develop a generic method to carry out single-enzyme experiments in the restricted spatial environment of a virus capsid. We report here the incorporation of individual horseradish peroxidase enzymes in the inner cavity of a virus, and describe single-molecule studies on their enzymatic behaviour. These show that the virus capsid is permeable for substrate and product and that this permeability can be altered by changing pH.
Comellas-Aragonès
Marta
M
Institute for Molecules and Materials, Radboud University Nijmegen, Toernooiveld 1, 6525 ED Nijmegen, The Netherlands.
Engelkamp
Hans
H
Claessen
Victor I
VI
Sommerdijk
Nico A J M
NA
Rowan
Alan E
AE
Christianen
Peter C M
PC
Maan
Jan C
JC
Verduin
Benedictus J M
BJ
Cornelissen
Jeroen J L M
JJ
Nolte
Roeland J M
RJ
D032701:Biomimetics / Q000379:methods*; D019149:Bioreactors; D002213:Capsid / Q000737:chemistry* / Q000648:ultrastructure; D004800:Enzymes, Immobilized / Q000737:chemistry*; D006735:Horseradish Peroxidase / Q000737:chemistry*; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure; D036103:Nanotechnology / Q000379:methods*; D010539:Permeability
D004800:Enzymes, Immobilized; D006735:Horseradish Peroxidase