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Biomimetic enzyme nanocomplexes and their use as antidotes and preventive measures for alcohol intoxication


Organisms have sophisticated subcellular compartments containing enzymes that function in tandem. These confined compartments ensure effective chemical transformation and transport of molecules, and the elimination of toxic metabolic wastes1,2. Creating functional enzyme complexes that are confined in a similar way remains challenging. Here we show that two or more enzymes with complementary functions can be assembled and encapsulated within a thin polymer shell to form enzyme nanocomplexes. These nanocomplexes exhibit improved catalytic efficiency and enhanced stability when compared with free enzymes. Furthermore, the co-localized enzymes display complementary functions, whereby toxic intermediates generated by one enzyme can be promptly eliminated by another enzyme. We show that nanocomplexes containing alcohol oxidase and catalase could reduce blood alcohol levels in intoxicated mice, offering an alternative antidote and prophylactic for alcohol intoxication.

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Figure 1: Synthesis of enzyme nanocomplexes.
Figure 2: Structure and enhanced activity and stability of enzyme nanocomplexes.
Figure 3: In vivo detoxifying capability of the catalase-containing enzyme nanocomplexes.
Figure 4: Efficacy of n(AOx–Cat) as a prophylactic and antidote for alcohol intoxication.


  1. Schoffelen, S. & van Hest, J. C. M. Multi-enzyme systems: bringing enzymes together in vitro. Soft Matter 8, 1736–1746 (2012).

    Article  CAS  Google Scholar 

  2. Conrado, R. J., Varner, J. D. & DeLisa, M. P. Engineering the spatial organization of metabolic enzymes: mimicking nature's synergy. Curr. Opin. Biotechnol. 19, 492–499 (2008).

    Article  CAS  Google Scholar 

  3. Yan, W., Aebersold, R. & Raines, E. W. Evolution of organelle-associated protein profiling. J. Proteom. 72, 4–11 (2009).

    Article  CAS  Google Scholar 

  4. Matsumoto, R. et al. A liposome-based energy conversion system for accelerating the multi-enzyme reactions. Phys. Chem. Chem. Phys. 12, 13904–13906 (2010).

    Article  CAS  Google Scholar 

  5. Van Dongen, S. F. M., Nallani, M., Cornelissen, J. J. L. M., Nolte, R. J. M. & van Hest, J. C. M. A three-enzyme cascade reaction through positional assembly of enzymes in a polymersome nanoreactor. Chem. Eur. J. 15, 1107–1114 (2009).

    Article  CAS  Google Scholar 

  6. Wilner, O. I. et al. Enzyme cascades activated on topologically programmed DNA scaffolds. Nature Nanotech. 4, 249–254 (2009).

    Article  CAS  Google Scholar 

  7. Niemeyer, C. M., Koehler, J. & Wuerdemann, C. DNA-directed assembly of bienzymic complexes from in vivo biotinylated NAD(P)H:FMN oxidoreductase and luciferase. ChemBioChem 3, 242–245 (2002).

    Article  CAS  Google Scholar 

  8. Fierobe, H-P. et al. Degradation of cellulose substrates by cellulosome chimeras. Substrate targeting versus proximity of enzyme components. J. Biol. Chem. 277, 49621–49630 (2002).

    Article  CAS  Google Scholar 

  9. Kristensen, C. et al. Metabolic engineering of dhurrin in transgenic Arabidopsis plants with marginal inadvertent effects on the metabolome and transcriptome. Proc. Natl Acad. Sci. USA 102, 1779–1784 (2005).

    Article  CAS  Google Scholar 

  10. Dueber, J. E. et al. Synthetic protein scaffolds provide modular control over metabolic flux. Nature Biotechnol. 27, 753–759 (2009).

    Article  CAS  Google Scholar 

  11. Wanders, R. J. A. & Waterham, H. R. Biochemistry of mammalian peroxisomes revisited. Annu. Rev. Biochem. 75, 295–332 (2006).

    Article  CAS  Google Scholar 

  12. Schrader, M. & Fahimi, H. D. Mammalian peroxisomes and reactive oxygen species. Histochem. Cell Biol. 122, 383–393 (2004).

    Article  CAS  Google Scholar 

  13. Sheikh, F. G., Pahan, K., Khan, M., Barbosa, E. & Singh, I. Abnormality in catalase import into peroxisomes leads to severe neurological disorder. Proc. Natl Acad. Sci. USA 95, 2961–2966 (1998).

    Article  CAS  Google Scholar 

  14. Scism, R. A. & Bachmann, B. O. Five-component cascade synthesis of nucleotide analogues in an engineered self-immobilized enzyme aggregate. ChemBioChem 11, 67–70 (2010).

    Article  CAS  Google Scholar 

  15. Bäumler, H. & Georgieva, R. Coupled enzyme reactions in multicompartment microparticles. Biomacromolecules 11, 1480–1487 (2010).

    Article  Google Scholar 

  16. Stempfer, G., Höll-Neugebauer, B., Kopetzki, E. & Rudolph, R. A fusion protein designed for noncovalent immobilization: stability, enzymatic activity, and use in an enzyme reactor. Nature Biotechnol. 14, 481–484 (1996).

    Article  CAS  Google Scholar 

  17. Yan, M. et al. A novel intracellular protein delivery platform based on single-protein nanocapsules. Nature Nanotech. 5, 48–53 (2009).

    Article  Google Scholar 

  18. Selvin, P. R. The renaissance of fluorescence resonance energy transfer. Nature Struct. Biol. 7, 730–734 (2000).

    Article  CAS  Google Scholar 

  19. Ellis, R. J. Macromolecular crowding: obvious but underappreciated. Trends Biochem. Sci. 26, 597–604 (2001).

    Article  CAS  Google Scholar 

  20. Sherman, M. R., Saifer, M. G. P. & Perez-Ruiz, F. PEG-uricase in the management of treatment-resistant gout and hyperuricemia. Adv. Drug Deliv. Rev. 60, 59–68 (2008).

    Article  CAS  Google Scholar 

  21. Kehrer, J. P. Free radicals as mediators of tissue injury and disease. Crit. Rev. Toxicol. 23, 21–48 (1993).

    Article  CAS  Google Scholar 

  22. Cochrane, J. Alcohol use in China. Alcohol Alcoholism 38, 537–542 (2003).

    Article  Google Scholar 

  23. Lee, K., Møller, L., Hardt, F., Haubek, A. & Jensen, E. Alcohol-induced brain damage and liver damage in young males. Lancet 2, 759–761 (1979).

    Article  CAS  Google Scholar 

  24. Jamaty, C. et al. Lipid emulsions in the treatment of acute poisoning: a systematic review of human and animal studies. Clin. Toxicol. 48, 1–27 (2010).

    Article  CAS  Google Scholar 

  25. Bertrand, N., Bouvet, C., Moreau, P. & Leroux, J-C. Transmembrane pH-gradient liposomes to treat cardiovascular drug intoxication. ACS Nano 4, 7552–7558 (2010).

    Article  CAS  Google Scholar 

  26. Shen, Y. et al. Dihydromyricetin as a novel anti-alcohol intoxication medication. J. Neurosci. 32, 390–401 (2012).

    Article  CAS  Google Scholar 

  27. Martins, S., Sarmento, B., Ferreira, D. C. & Souto, E. B. Lipid-based colloidal carriers for peptide and protein delivery—liposomes versus lipid nanoparticles. Int. J. Nanomed. 2, 595–607 (2007).

    CAS  Google Scholar 

  28. Averbakh, A. Z. et al. Flavin-dependent alcohol oxidase from the yeast Pichia pinus. Spatial localization of the coenzyme FAD in the protein structure: hot-tritium bombardment and ESR experiments. Biochem. J. 310, 601–604 (1995).

    Article  CAS  Google Scholar 

  29. Barry, R. E. & McGivan, J. D. Acetaldehyde alone may initiate hepatocellular damage in acute alcoholic liver disease. Gut 26, 1065–1069 (1985).

    Article  CAS  Google Scholar 

  30. Ji, C. & Kaplowitz, N. Betaine decreases hyperhomocysteinemia, endoplasmic reticulum stress, and liver injury in alcohol-fed mice. Gastroenterology 124, 1488–1499 (2003).

    Article  CAS  Google Scholar 

  31. Ji, C., Deng, Q. & Kaplowitz, N. Role of TNF-α in ethanol-induced hyperhomocysteinemia and murine alcoholic liver injury. Hepatology 40, 442–451 (2004).

    Article  CAS  Google Scholar 

  32. Ji, C. et al. Liver-specific loss of glucose-regulated protein 78 perturbs the unfolded protein response and exacerbates a spectrum of liver diseases in mice. Hepatology 54, 229–239 (2011).

    Article  CAS  Google Scholar 

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This work was partially supported by the Defense Threat Reducing Agency (DTRA), the National Institutes of Health (NIH, grants R01AA018846 and R01AA018612), the National Natural Science Foundation of China (NSFC, grants 81025018, 91127045 and 50830103) and the National Basic Research Program of China (973 Program, no. 2011CB932500).

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M.Y., L.S., W.C., O.Y., C.J. and Y.Lu conceived or designed the experiments. Y.Liu and J.D. performed the synthesis, characterization and data analysis. M.Lau, J.H., H.H. and J.L. performed the in vivo tests. S.L., W.W., X.Z. and H.W. performed the biodistribution and pharmacokinetic studies. Y.Liu, J.D., C.J. and Y.Lu co-wrote the paper. All authors discussed the results and commented on the manuscript.

Corresponding authors

Correspondence to Linqi Shi, Wei Chen, Cheng Ji or Yunfeng Lu.

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The authors declare no competing financial interests.

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Liu, Y., Du, J., Yan, M. et al. Biomimetic enzyme nanocomplexes and their use as antidotes and preventive measures for alcohol intoxication. Nature Nanotech 8, 187–192 (2013).

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