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Engineering a Metathesis-Catalyzing Artificial Metalloenzyme Based on HaloTag.

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%).

SUBMITTER: Fischer S

PROVIDER: S-EPMC8154321 | biostudies-literature | 2021 May

REPOSITORIES: biostudies-literature

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Engineering a Metathesis-Catalyzing Artificial Metalloenzyme Based on HaloTag.

Fischer Sandro S Ward Thomas R TR Liang Alexandria D AD

ACS catalysis 20210512 10

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 scaffol ...[more]

PMID: 34055452

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