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Engineering a Metathesis-Catalyzing Artificial Metalloenzyme Based on HaloTag

Fischer, Sandro; Ward, Thomas R; Liang, Alexandria Deliz (2021). Engineering a Metathesis-Catalyzing Artificial Metalloenzyme Based on HaloTag. ACS Catalysis, 11(10):6343-6347.

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%).

Additional indexing

Item Type:Journal Article, not_refereed, original work
Communities & Collections:07 Faculty of Science > Department of Chemistry
Dewey Decimal Classification:540 Chemistry
Scopus Subject Areas:Physical Sciences > Catalysis
Physical Sciences > General Chemistry
Uncontrolled Keywords:Catalysis, General Chemistry
Language:English
Date:21 May 2021
Deposited On:09 Jan 2023 08:03
Last Modified:26 Jun 2025 01:56
Publisher:American Chemical Society (ACS)
ISSN:2155-5435
OA Status:Green
Free access at:PubMed ID. An embargo period may apply.
Publisher DOI:https://doi.org/10.1021/acscatal.1c01470
PubMed ID:34055452
Project Information:
  • Funder: SNSF
  • Grant ID: 182046
  • Project Title: Directed Evolution of Artificial Metalloproteins and Metalloenzymes
  • Funder: H2020
  • Grant ID: 694424
  • Project Title: Directed Evolution of Artificial Metalloenzymes for In Vivo Applications
  • Funder: Marie Skłodowska-Curie
  • Grant ID: H2020-MSCA-IF-2017
  • Project Title:
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  • Licence: Creative Commons: Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
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