Abstract
Artificial enzymes, which are hybrids of proteins with abiological catalytic groups, have emerged as a powerful approach towards the creation of enzymes for new-to-nature reactions. Typically, only a single abiological catalytic moiety is incorporated. Here we introduce a design of an artificial enzyme that comprises two different abiological catalytic moieties and show that these can act synergistically to achieve high activity and enantioselectivity (up to >99% e.e.) in the catalysed Michael addition reaction. The design is based on the lactococcal multidrug resistance regulator as the protein scaffold and combines a genetically encoded unnatural p-aminophenylalanine residue (which activates an enal through iminium ion formation) and a supramolecularly bound Lewis acidic Cu(ii) complex (which activates the Michael donor by enolization and delivers it to one preferred prochiral face of the activated enal). This study demonstrates that synergistic combination of abiological catalytic groups is a robust way to achieve catalysis that is normally outside of the realm of artificial enzymes.
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All data that support the findings of this study are available in the Article and its Supplementary Information, or from the corresponding author on reasonable request.
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Acknowledgements
We thank R. B. Leveson-Gower for assistance in preparation of the figures and K. E. Splan for useful discussions. Support from the Netherlands Organisation for Scientific Research (NWO) (Vici grant no. 724.013.003) and the Ministry of Education, Culture and Science (Gravitation programme no. 024.001.035) is gratefully acknowledged.
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G.R. conceived and directed the project. Z.Z. performed the experimental work and analysed the data. The authors discussed the results and wrote the manuscript together.
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Supplementary Tables 1 and 2, Figs. 1–7, methods and references.
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Zhou, Z., Roelfes, G. Synergistic catalysis in an artificial enzyme by simultaneous action of two abiological catalytic sites. Nat Catal 3, 289–294 (2020). https://doi.org/10.1038/s41929-019-0420-6
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DOI: https://doi.org/10.1038/s41929-019-0420-6
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