CIC bioGUNE
Published December 14, 2020 | Version v1
Journal article Open

Efficient Lewis acid catalysis of an abiological reaction in a de novo protein scaffold

Description

New enzyme catalysts are usually engineered by repurposing the active sites of natural proteins. Here we show that design and directed evolution can be used to transform a non-natural, functionally naïve zinc-binding protein into a highly active catalyst for an abiological hetero-Diels–Alder reaction. The artificial metalloenzyme achieves >104 turnovers per active site, exerts absolute control over reaction pathway and product stereochemistry, and displays a chemical proficiency (1/KTS = 2.9 × 1010 M-1) that exceeds all previously characterized Diels–Alderases. These properties capitalize on effective Lewis acid catalysis, a chemical strategy for accelerating Diels–Alder reactions common in the laboratory but so far unknown in nature. Extension of this approach to other metal ions and other de novo scaffolds may propel the design field in exciting new directions.
 

Notes

We thank M. Solar and N. Trapp from the Small Molecule Crystallography Center at ETH Zürich for structure elucidation. We also thank B. Blattmann from the Protein Crystallization Center at the University of Zürich, the teams from the Mass Spectrometry service and the NMR facility at the Laboratory for Organic Chemistry at ETH Zürich and the staff at the Swiss Light Source at the Paul Scherrer Institute for technical support. This work was supported by the Swiss National Science Foundation (SNSF) and ETH Zürich. G.J.-O. acknowledges grants from the Spanish Ministry of Science and Innovation (MCI) co-financed with FEDER funds (RTI2018-099592-B-C22 and RYC-2013-14706) and the Mizutani Foundation for Glycoscience (200077).

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