A de novo enzyme catalyzes a life-sustaining reaction in Escherichia coli

Abstract

Producing novel enzymes that are catalytically active in vitro and biologically functional in vivo is a key goal of synthetic biology. Here we describe Syn-F4, the first de novo protein that meets both criteria. Purified Syn-F4 hydrolyzes the siderophore ferric enterobactin, and expression of Syn-F4 allows an inviable strain of Escherichia coli to grow in iron-limited medium. These findings demonstrate that entirely new sequences can provide life-sustaining enzymatic functions in living organisms.

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Figure 1: Syn-F4 catalyzes the hydrolysis of ferric enterobactin in vivo and in vitro.
Figure 2: Mutagenesis of Syn-F4 reveals conserved residues.

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Acknowledgements

We thank J. Eng for assistance with LC–MS and E. Nolan (MIT) for providing D-enterobactin. This work was funded by NSF Grant MCB-1409402, and by the Department of Defense (DoD) through the National Defense Science & Engineering Graduate Fellowship (NDSEG) Program to A.E.D.

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Contributions

A.E.D., G.S.M., and M.H.H. initiated the project idea. All authors contributed to the direction of the project. A.E.D. generated reagents and performed experiments. K.M.D. created the chromosomal Δfes::syn-F4 E. coli strain. G.S.M. generated protein models. All authors contributed to data analysis and discussion of results. A.E.D. and M.H.H. drafted the manuscript, with input from all the authors.

Corresponding author

Correspondence to Michael H Hecht.

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The authors declare no competing financial interests.

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Supplementary Figures 1–3 and Supplementary Notes 1–2 (PDF 1876 kb)

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Donnelly, A., Murphy, G., Digianantonio, K. et al. A de novo enzyme catalyzes a life-sustaining reaction in Escherichia coli. Nat Chem Biol 14, 253–255 (2018). https://doi.org/10.1038/nchembio.2550

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