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


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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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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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.

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Correspondence to Michael H Hecht.

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The authors declare no competing financial interests.

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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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