Virus particles are probably the most precisely defined nanometre-sized objects that can be formed by protein self-assembly. Although their natural function is the storage and transport of genetic material, they have more recently been applied as scaffolds for mineralization and as containers for the encapsulation of inorganic compounds1,2. The reproductive power of viruses has been used to develop versatile analytical methods, such as phage display, for the selection and identification of (bio)active compounds3. To date, the combined use of self-assembly and reproduction has not been used for the construction of catalytic systems. Here we describe a self-assembled system based on a plant virus that has its coat protein genetically modified to provide it with a lipase enzyme. Using single-object and bulk catalytic studies, we prove that the virus-anchored lipase molecules are catalytically active. This anchored biocatalyst, unlike man-made supported catalysts, has the capability to reproduce itself in vivo, generating many independent catalytically active copies.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1.

    et al. Virus-based toolkit for the directed synthesis of magnetic and semiconducting nanowires. Science 303, 213–217 (2004).

  2. 2.

    & Host-guest encapsulation of materials by assembled virus protein cages. Nature 393, 152–155 (1998).

  3. 3.

    et al. Virus-enabled synthesis and assembly of nanowires for lithium ion battery electrodes. Science 312, 885–888 (2006).

  4. 4.

    & Renaissance of immobilized catalysts. New types of polymer-supported catalysts, ‘microencapsulated catalysts’, which enable environmentally benign and powerful high-throughput organic synthesis. Chem. Commun. 449–460 (2003).

  5. 5.

    Immobilizing enzymes: How to create more suitable biocatalysts. Angew. Chem. Int. Edn 42, 3336–3337 (2003).

  6. 6.

    , , & Further characterization of the krebs tricarboxylic-acid cycle metabolon. J. Biol. Chem. 262, 1786–1790 (1987).

  7. 7.

    Respiratory chain supercomplexes of mitochondria and bacteria. Biochim. Biophys. Acta-Bioenerg. 1555, 154–159 (2002).

  8. 8.

    et al. Directed evolution of novel polymerase activities: Mutation of a DNA polymerase into an efficient RNA polymerase. Proc. Natl Acad. Sci. USA 99, 6597–6602 (2002).

  9. 9.

    & Yeast cell-surface display—applications of molecular display. Appl. Microbiol. Biotechnol. 64, 28–40 (2004).

  10. 10.

    Autodisplay: efficient bacterial surface display of recombinant proteins. Appl. Microbiol. Biotechnol. 69, 607–614 (2006).

  11. 11.

    , & Surface features of potato virus X from fiber diffraction. Virology 300, 291–295 (2002).

  12. 12.

    & In Particle Structure in the Plant Viruses vol. 4 (ed. Milne, R. C.) 51–83 (Plenum Press, New York, 1988).

  13. 13.

    , & One biocatalyst—Many applications: The use of Candida antarctica B-lipase in organic synthesis. Biocatal. Biotransform. 16, 181–204 (1998).

  14. 14.

    , , , & Displacement of the equilibrium in lipase catalyzed transesterification in ethyl octanoate by continous evaporation of ethanol. Biotechnol. Lett. 14, 263–268 (1992).

  15. 15.

    , , , & Thioethyl-octanoate, vinyl-octanoate, ethyl-octanoate esters and octanoic-acid as acyl donors in lipase-catalyzed acyl transfer-reactions. Biocatalysis 9, 105–114 (1994)

  16. 16.

    et al. In situ spatial organization of potato virus A coat protein subunits as assessed by tritium bombardment. J. Virol. 75, 9696–9702 (2001)

  17. 17.

    et al. Analysis of the aphthovirus 2A/2B polyprotein ‘cleavage’ mechanism indicates not a proteolytic reaction, but a novel translational effect: a putative ribosomal ‘skip’. J. Gen. Virol. 82, 1013–1025 (2001)

  18. 18.

    et al. Assembly and movement of a plant virus carrying a green fluorescent protein overcoat. Proc. Natl Acad. Sci. USA 93, 6286–6290 (1996)

  19. 19.

    , & Protein Methods (Wiley-Liss, New York, 1996).

  20. 20.

    et al. Do enzymes sleep and work? Chem. Commun. 935–940 (2006).

  21. 21.

    et al. Single-enzyme kinetics of CALB-catalyzed hydrolysis. Angew. Chem. Int. Edn 44, 560–564 (2005).

  22. 22.

    et al. Stretched exponential decay and correlations in the catalytic activity of fluctuating single lipase molecules. Proc. Natl Acad. Sci. USA 102, 2368–2372 (2005).

  23. 23.

    et al. Use of modified plum pox virus coat protein genes developed to limit heteroencapsidation-associated risks in transgenic plants. J. Gen. Virol. 79, 1509–1517 (1998)

  24. 24.

    et al. Aphid transmission of a non-aphid-transmissible strain of zucchini yellow mosaic potyvirus from transgenic plants expressing the capsid protein of plum pox potyvirus. Mol. Plant-Microbe Interact. 6, 403–406 (1993).

Download references


The authors acknowledge the financial support provided through the European Community's Human Potential Programme under contract HPRN-CT-2001-00188 (SMASHYBIO). R.N. acknowledges financial support from the Royal Netherlands Academy of Sciences. We thank S. Chapman (Scottish Crop Research Institute, Invergowrie, Dundee, UK) for providing us with the pTXS.GFP-CP vector. We are indebted to H. Rogniaux for mass spectroscopy analysis.

Author information


  1. Interactions Plante Virus, UMR GDPP, IBVM, INRA, BP 81, F-33883, Villenave d'Ornon, France

    • Noëlle Carette
    •  & Thierry Michon
  2. Institute for Molecules and Materials, Radboud University Nijmegen, PO Box 9010, 6500 GL, Nijmegen, The Netherlands

    • Hans Engelkamp
    • , Eric Akpa
    • , Peter C. M. Christianen
    • , Jan C. Maan
    • , Alan E. Rowan
    • , Roeland J. M. Nolte
    •  & Jan C. M. Van Hest
  3. DSM Research, PO Box 18, 6160 MD, Geleen, The Netherlands

    • Sebastien J. Pierre
    •  & Jens C. Thies
  4. University of Durham, Department of Chemistry, South Road, Durham DH1 3LE, UK

    • Neil R. Cameron
  5. Technische Universität München, Chemie Department, Lehrstuhl für Makromolekulare Stoffe, Lichtenbergstrasse 4, 85747 Garching, Germany

    • Ralf Weberskirch


  1. Search for Noëlle Carette in:

  2. Search for Hans Engelkamp in:

  3. Search for Eric Akpa in:

  4. Search for Sebastien J. Pierre in:

  5. Search for Neil R. Cameron in:

  6. Search for Peter C. M. Christianen in:

  7. Search for Jan C. Maan in:

  8. Search for Jens C. Thies in:

  9. Search for Ralf Weberskirch in:

  10. Search for Alan E. Rowan in:

  11. Search for Roeland J. M. Nolte in:

  12. Search for Thierry Michon in:

  13. Search for Jan C. M. Van Hest in:


T.M., J.H., N.R.C., J.T., and R.W. conceived and designed the experiments. N.C., H.E., E.A. and S.P. performed the experiments. N.C., H.E., P.C., R.N., T.M., and J.K. analysed the data. P.C., J.M., and A.R., contributed analysis tools. N.C., H.E., R.M., T.M., and J.K., co-wrote the paper. All authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Thierry Michon or Jan C. M. Van Hest.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary figures S1—S3 and Table S1

About this article

Publication history






Further reading