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 Enzymoljournal_title
Methods in enzymologyauthors
Carminati DM,Moore EJ,Fasan Rdoi
10.1016/bs.mie.2020.07.007subject
Has Abstractpub_date
2020-01-01 00:00:00pages
35-61eissn
0076-6879issn
1557-7988pii
S0076-6879(20)30282-2journal_volume
644pub_type
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