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Engineering a Metathesis-Catalyzing Artificial Metalloenzyme Based on HaloTag

Published: 12 May 2021
ACS Catalysis , Volume 11, pp 6343-6347; doi:10.1021/acscatal.1c01470

Abstract: Artificial metalloenzymes (ArMs) are created by embedding a synthetic metal catalyst into a protein scaffold. ArMs have the potential to merge the catalytic advantages of natural enzymes with the reaction scope of synthetic catalysts. The choice of the protein scaffold is of utmost importance to tune the activity of the ArM. Herein, we show the repurposing of HaloTag, a self-labeling protein widely used in chemical biology, to create an ArM scaffold for metathesis. This monomeric protein scaffold allows for covalent attachment of metathesis cofactors, and the resulting ArMs are capable of catalyzing ring-closing metathesis. Both chemical and genetic engineering were explored to determine the evolvability of the resulting ArM. Additionally, exploration of the substrate scope revealed a reaction with promising turnover numbers (>48) and conversion rates (>96%).
Keywords: artificial metalloenzymes / aqueous catalysis / metathesis / enzyme engineering / HaloTag

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