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 properties1,2,3,4. 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.
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This work was supported by the Council for Chemical Sciences of the Netherlands Organization for Scientific Research (NWO-CW) through a TOP grant to R.J.M.N. and Veni & Vidi Innovative Research Grants to J.J.L.M.C., and by the Royal Netherlands' Academy for Arts and Sciences (R.J.M.N.). The protein Dronpa was a generous gift from J. Hofkens, University of Leuven (Belgium).
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Comellas-Aragonès, M., Engelkamp, H., Claessen, V. et al. A virus-based single-enzyme nanoreactor. Nature Nanotech 2, 635–639 (2007) doi:10.1038/nnano.2007.299
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