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General features to enhance enzymatic activity of poly(ethylene terephthalate) hydrolysis


Poly(ethylene terephthalate) (PET) is the most abundant polyester plastic and a major contributor to plastic pollution. IsPETase, from the PET-assimilating bacterium Ideonella sakaiensis, is a unique PET-hydrolytic enzyme that shares high sequence identity to canonical cutinases, but shows substrate preference towards PET and exhibits higher PET-hydrolytic activity at ambient temperature. Structural analyses suggest that IsPETase harbours a substrate-binding residue, W185, with a wobbling conformation and a highly flexible W185-locating β6-β7 loop. Here, we show that these features result from the presence of S214 and I218 in IsPETase, whose equivalents are strictly His and Phe, respectively, in all other homologous enzymes. We found that mutating His/Phe residues to Ser/Ile could enhance the PET-hydrolytic activity of several IsPETase-like enzymes. In conclusion, the Ser/Ile mutations should provide an important strategy to improve the activity of potential PET-hydrolytic enzymes with properties that may be useful for various applications.

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Fig. 1: IsPETase-unique structural features.
Fig. 2: The PET-hydrolytic activity of wild-type and mutated type IIb PL enzymes.
Fig. 3: Time-course of PET-hydrolytic activity of enzymes produced in E. coli and P. pastoris.
Fig. 4: The PET-hydrolytic activity of wild-type and mutated type IIa PL enzymes and type I cutinases.

Data availability

Source data are provided with this paper. The atomic coordinates and structure factors of the reported structures have been deposited in the Protein Data Bank under accession codes 7CY0 for IsPETase S214H and 7CWQ for BurPL. The web links of previously reported protein structures that were utilized for analyses and comparisons in this study are provided in the figure legends. Data and other findings of this study are available from the corresponding author on reasonable request.


  1. 1.

    Geyer, B., Lorenz, G. & Kandelbauer, A. Recycling of poly(ethylene terephthalate)—a review focusing on chemical methods. Express Polym. Lett. 10, 559–586 (2016).

    CAS  Article  Google Scholar 

  2. 2.

    Borrelle, S. B. et al. Predicted growth in plastic waste exceeds efforts to mitigate plastic pollution. Science 369, 1515–1518 (2020).

    CAS  Article  Google Scholar 

  3. 3.

    Lau, W. W. Y. et al. Evaluating scenarios toward zero plastic pollution. Science 369, 1455–1461 (2020).

    CAS  Article  Google Scholar 

  4. 4.

    Chen, C.-C., Dai, L., Ma, L. & Guo, R.-T. Enzymatic degradation of plant biomass and synthetic polymers. Nat. Rev. Chem. 4, 114–126 (2020).

    Article  Google Scholar 

  5. 5.

    Wei, R. & Zimmermann, W. Microbial enzymes for the recycling of recalcitrant petroleum-based plastics: how far are we? Microb. Biotechnol. 10, 1308–1322 (2017).

    CAS  Article  Google Scholar 

  6. 6.

    George, N. & Kurian, T. Recent developments in the chemical recycling of postconsumer poly(ethylene terephthalate) waste. Ind. Eng. Chem. Res. 53, 14185–14198 (2014).

    CAS  Article  Google Scholar 

  7. 7.

    Kawai, F., Kawabata, T. & Oda, M. Current knowledge on enzymatic PET degradation and its possible application to waste stream management and other fields. Appl. Microbiol. Biotechnol. 103, 4253–4268 (2019).

    CAS  Article  Google Scholar 

  8. 8.

    Yoshida, S. et al. A bacterium that degrades and assimilates poly(ethylene terephthalate). Science 351, 1196–1199 (2016).

    CAS  Article  Google Scholar 

  9. 9.

    Taniguchi, I. et al. Biodegradation of PET: current status and application aspects. ACS Catal. 9, 4089–4105 (2019).

    CAS  Article  Google Scholar 

  10. 10.

    Han, X. et al. Structural insight into catalytic mechanism of PET hydrolase. Nat. Commun. 8, 2106 (2017).

    Article  Google Scholar 

  11. 11.

    Austin, H. P. et al. Characterization and engineering of a plastic-degrading aromatic polyesterase. Proc. Natl Acad. Sci. USA 115, E4350–E4357 (2018).

    CAS  Article  Google Scholar 

  12. 12.

    Chen, C. C., Han, X., Ko, T. P., Liu, W. & Guo, R. T. Structural studies reveal the molecular mechanism of PETase. FEBS J. 285, 3717–3723 (2018).

    CAS  Article  Google Scholar 

  13. 13.

    Fecker, T. et al. Active site flexibility as a hallmark for efficient PET degradation by I. sakaiensis PETase. Biophys. J. 114, 1302–1312 (2018).

    CAS  Article  Google Scholar 

  14. 14.

    Joo, S. et al. Structural insight into molecular mechanism of poly(ethylene terephthalate) degradation. Nat. Commun. 9, 382 (2018).

    Article  Google Scholar 

  15. 15.

    Liu, B. et al. Protein crystallography and site-direct mutagenesis analysis of the poly(ethylene terephthalate) hydrolase PETase from Ideonella sakaiensis. ChemBioChem 19, 1471–1475 (2018).

    CAS  Article  Google Scholar 

  16. 16.

    Bollinger, A. et al. A novel polyester hydrolase from the marine bacterium Pseudomonas aestusnigri—structural and functional insights. Front. Microbiol. 11, 114 (2020).

    Article  Google Scholar 

  17. 17.

    Shental-Bechor, D. & Levy, Y. Effect of glycosylation on protein folding: a close look at thermodynamic stabilization. Proc. Natl Acad. Sci. USA 105, 8256–8261 (2008).

    CAS  Article  Google Scholar 

  18. 18.

    Xu, Z., Cen, Y.-K., Zou, S.-P., Xue, Y.-P. & Zheng, Y.-G. Recent advances in the improvement of enzyme thermostability by structure modification. Crit. Rev. Biotechnol. 40, 83–98 (2020).

    CAS  Article  Google Scholar 

  19. 19.

    Meilleur, C., Hupé, J.-F., Juteau, P. & Shareck, F. Isolation and characterization of a new alkali-thermostable lipase cloned from a metagenomic library. J. Ind. Microbiol. Biotechnol. 36, 853–861 (2009).

    CAS  Article  Google Scholar 

  20. 20.

    Danso, D. et al. New insights into the function and global distribution of polyethylene terephthalate (PET)-degrading bacteria and enzymes in marine and terrestrial metagenomes. Appl. Environ. Microbiol. 84, e02773-17 (2018).

    Article  Google Scholar 

  21. 21.

    Sulaiman, S. et al. Isolation of a novel cutinase homolog with polyethylene terephthalate-degrading activity from leaf-branch compost by using a metagenomic approach. Appl. Environ. Microbiol. 78, 1556–1562 (2012).

    CAS  Article  Google Scholar 

  22. 22.

    Müller, R.-J., Schrader, H., Profe, J., Dresler, K. & Deckwer, W.-D. Enzymatic degradation of poly(ethylene terephthalate): rapid hydrolyse using a hydrolase from T. fusca. Macromol. Rapid Commun. 26, 1400–1405 (2005).

    Article  Google Scholar 

  23. 23.

    Hiraga, K., Taniguchi, I., Yoshida, S., Kimura, Y. & Oda, K. Biodegradation of waste PET: a sustainable solution for dealing with plastic pollution. EMBO Rep. 20, e49365 (2019).

    CAS  PubMed  PubMed Central  Google Scholar 

  24. 24.

    Son, H. F. et al. Rational protein engineering of thermo-stable PETase from Ideonella sakaiensis for highly efficient PET degradation. ACS Catal. 9, 3519–3526 (2019).

    CAS  Article  Google Scholar 

  25. 25.

    Tournier, V. et al. An engineered PET depolymerase to break down and recycle plastic bottles. Nature 580, 216–219 (2020).

    CAS  Article  Google Scholar 

  26. 26.

    Ru, J., Huo, Y. & Yang, Y. Microbial degradation and valorization of plastic wastes. Front. Microbiol. 11, 442 (2020).

    Article  Google Scholar 

  27. 27.

    Otwinowski, Z. & Minor, W. Processing of X-ray diffraction data collected in oscillation mode. Meth. Enzymol. 276, 307–326 (1997).

    CAS  Article  Google Scholar 

  28. 28.

    McCoy, A. J. et al. Phaser crystallographic software. J. Appl. Crystallogr. 40, 658–674 (2007).

    CAS  Article  Google Scholar 

  29. 29.

    Murshudov, G. N., Vagin, A. A. & Dodson, E. J. Refinement of macromolecular structures by the maximum-likelihood method. Acta Crystallogr. D Biol. Crystallogr. 53, 240–255 (1997).

    CAS  Article  Google Scholar 

  30. 30.

    Emsley, P. & Cowtan, K. Coot: model-building tools for molecular graphics. Acta Crystallogr. D Biol. Crystallogr. 60, 2126–2132 (2004).

    Article  Google Scholar 

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This work was supported by the National Key Research and Development Program of China (2019YFA0706900, R.-T.G.), the National Natural Science Foundation of China (31870790, R.-T.G.; 31971205, C.-C.C.; 31800662, X.H.; 32000899, L.Z.) and the Natural Science Foundation Innovative Group Project of Hubei Province (2020CFA011, R.-T.G.). We thank X-ray crystallography facility of State Key Laboratory of Biocatalysis and Enzyme Engineering and National Synchrotron Radiation Research Center (NSRRC) for access to beamlines TPS-05A and TLS-15A1, which contributed to the synchrotron data collection.

Author information




R.-T.G., C.-C.C. and L.M. conceived the experiments. X.H., X.L., P.J., D.N., S.L., Y.Q., H.H. and W.Z. performed the experiments. C.-C.C., L.D., W.L., J.M., Y.Y. and L.Z. analysed the data and discussed the results. C.-C.C., R.-T.G. and J.-W.H. wrote the manuscript. R.-T.G., L.D. and J.-W.H. supervised the project.

Corresponding authors

Correspondence to Jian-Wen Huang or Longhai Dai or Rey-Ting Guo.

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The authors declare no competing interests.

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Peer review information Nature Catalysis thanks Kohei Oda, César A. Ramírez-Sarmiento and Mitch H. Weiland for their contribution to the peer review of this work.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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Chen, CC., Han, X., Li, X. et al. General features to enhance enzymatic activity of poly(ethylene terephthalate) hydrolysis. Nat Catal 4, 425–430 (2021).

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