Enzymatic Nitrogen Insertion into Unactivated C–H Bonds

Journal of the American Chemical Society, Journal Year: 2022, Volume and Issue: 144(41), P. 19097 - 19105

Published: Oct. 4, 2022

Selective functionalization of aliphatic C–H bonds, ubiquitous in molecular structures, could allow ready access to diverse chemical products. While enzymatic oxygenation bonds is well established, the analogous nitrogen still unknown; nature reliant on preoxidized compounds for incorporation. Likewise, synthetic methods selective derivatization unbiased remain elusive. In this work, new-to-nature heme-containing nitrene transferases were used as starting points directed evolution enzymes selectively aminate and amidate unactivated C(sp3)–H sites. The desymmetrization methyl- ethylcyclohexane with divergent site selectivity offered demonstration. evolved these lineages are highly promiscuous show activity toward a wide array substrates, providing foundation further transferase function. Computational studies kinetic isotope effects (KIEs) consistent stepwise radical pathway involving an irreversible, enantiodetermining hydrogen atom transfer (HAT), followed by lower-barrier diastereoselectivity-determining rebound step. In-enzyme dynamics (MD) simulations reveal predominantly hydrophobic pocket favorable dispersion interactions substrate. By offering direct path from saturated precursors, present new biochemical logic accessing nitrogen-containing compounds.

Language: Английский

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Engineering Hydroxylase Activity, Selectivity, and Stability for a Scalable Concise Synthesis of a Key Intermediate to Belzutifan

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(13)

Published: Jan. 27, 2024

Biocatalytic oxidations are an emerging technology for selective C-H bond activation. While promising a range of oxidations, practical use enzymes catalyzing aerobic hydroxylation is presently limited by their substrate scope and stability under industrially relevant conditions. Here, we report the engineering application non-heme iron α-ketoglutarate-dependent dioxygenase direct stereo- regio-selective non-native fluoroindanone en route to oncology treatment belzutifan, replacing five-step chemical synthesis with enantioselective hydroxylation. Mechanistic studies indicated that formation desired product was enzyme overoxidation, these properties subsequently improved directed evolution, yielding biocatalyst capable >15,000 total turnovers. Highlighting industrial utility this biocatalyst, high-yielding, green, efficient oxidation demonstrated at kilogram scale belzutifan.

Language: Английский

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Engineering non-haem iron enzymes for enantioselective C(sp3)–F bond formation via radical fluorine transfer

Nature Synthesis, Journal Year: 2024, Volume and Issue: 3(8), P. 958 - 966

Published: March 28, 2024

Language: Английский

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Biocatalytic enantioselective C(sp3)–H fluorination enabled by directed evolution of non-haem iron enzymes

Nature Synthesis, Journal Year: 2024, Volume and Issue: 3(8), P. 967 - 975

Published: April 26, 2024

Language: Английский

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Overview of Amino Acid Modifications by Iron- and α-Ketoglutarate-Dependent Enzymes

ACS Catalysis, Journal Year: 2023, Volume and Issue: 13(7), P. 4853 - 4865

Published: March 27, 2023

Modified amino acids are important building blocks in both natural biomolecules and synthetic small-molecule drugs. Here, we review the roles of iron- α-ketoglutarate-dependent dioxygenases oxidative modification acids, focusing on C–H hydroxylation halogenation. Recent engineering approaches to alter substrate specificity, improve catalytic efficiency, install non-native activities also discussed, along with several applications complex molecule synthesis. We conclude a brief discussion recent discoveries unique cyclization by family members contemporary challenges expanding biocatalytic utility reaction scope enzyme family.

Language: Английский

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Non-Native Site-Selective Enzyme Catalysis

Chemical Reviews, Journal Year: 2023, Volume and Issue: 123(16), P. 10381 - 10431

Published: July 31, 2023

The ability to site-selectively modify equivalent functional groups in a molecule has the potential streamline syntheses and increase product yields by lowering step counts. Enzymes catalyze site-selective transformations throughout primary secondary metabolism, but leveraging this capability for non-native substrates reactions requires detailed understanding of limitations enzyme catalysis how these bounds can be extended protein engineering. In review, we discuss representative examples involving group manipulation C-H bond functionalization. We include illustrative native catalysis, our focus is on cases often using engineered enzymes. then use enzymes chemoenzymatic target-oriented synthesis conclude with survey tools techniques that could expand scope catalysis.

Language: Английский

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Mechanisms of C(sp3)–H and C=C selective oxidative heterofunctionalizations by non-heme Fe and Mn mimics of oxygenase enzymes

Coordination Chemistry Reviews, Journal Year: 2024, Volume and Issue: 508, P. 215793 - 215793

Published: March 18, 2024

Language: Английский

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Copper-dependent halogenase catalyses unactivated C−H bond functionalization

Nature, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 29, 2025

Language: Английский

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A New Age of Biocatalysis Enabled by Generic Activation Modes

JACS Au, Journal Year: 2024, Volume and Issue: 4(6), P. 2068 - 2080

Published: May 31, 2024

Biocatalysis is currently undergoing a profound transformation. The field moves from relying on nature's chemical logic to discipline that exploits generic activation modes, allowing for novel biocatalytic reactions and, in many instances, entirely new chemistry. Generic modes enable wide range of reaction types and played pivotal role advancing the fields organo- photocatalysis. This perspective aims summarize principal harnessed enzymes develop biocatalysts. Although extensively researched past, highlighted when applied within enzyme active sites, facilitate transformations have largely eluded efficient selective catalysis. advance attributed multiple tunable interactions substrate binding pocket precisely control competing pathways transition states. We will highlight cases synthetic methodologies achieved by engineered provide insights into potential future developments this rapidly evolving field.

Language: Английский

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Synthesis of non-canonical amino acids through dehydrogenative tailoring

Nature, Journal Year: 2024, Volume and Issue: 634(8033), P. 352 - 358

Published: Aug. 29, 2024

Language: Английский

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