Strategies for the expression and characterization of artificial myoglobin-based carbene transferases.

Abstract:

:Myoglobin has recently emerged as a versatile metalloprotein scaffold for the design of efficient and selective biocatalysts for abiological carbene transfer reactions, including asymmetric cyclopropanation reactions. Over the past few years, our group has explored several strategies to modulate the carbene transfer reactivity of myoglobin-based catalysts, including the substitution of the native heme cofactor and conserved histidine axial ligand with non-native porphynoid ligands and alternative natural and unnatural amino acids as the metal-coordinating ligands, respectively. Herein, we report protocols for the generation and reconstitution in vitro and in vivo of myoglobin-based artificial carbene transferases incorporating non-native iron-porphynoid cofactors, also in combination with unnatural amino acids as the proximal ligand. These strategies are effective for imparting these myoglobin-based cyclopropanation biocatalysts with altered and improved function, including tolerance to aerobic conditions and improved reactivity toward electrondeficient olefins.

journal_name

Methods Enzymol

journal_title

Methods in enzymology

authors

Carminati DM,Moore EJ,Fasan R

doi

10.1016/bs.mie.2020.07.007

subject

Has Abstract

pub_date

2020-01-01 00:00:00

pages

35-61

eissn

0076-6879

issn

1557-7988

pii

S0076-6879(20)30282-2

journal_volume

644

pub_type

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