Biocatalytic Strategies for Nitration Reactions DOI Creative Commons
Xiling Wang, Matteo Aleotti, Mélanie Hall

et al.

JACS Au, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 16, 2024

Nitro compounds are key synthetic intermediates used as enabling tools in synthesis and found a large range of essential compounds, including pharmaceuticals, pesticides, various organic dyes. Despite recent methodological developments, the industrial preparation nitro still suffers from harsh reaction conditions, along with poor selectivity problematic environmental footprint. Although biological enzymatic methods exist, mild approaches for bionitration underexplored. Enzymes, their exquisite compatibility have potential to revolutionize way prepared. In this perspective, we systematically analyze currently available biological/enzymatic methods, oxidation an amine precursor or consisting direct oxidative nitration non-oxidative nitration. By examining both scope mechanism these reactions, aim present update on state-of-the-art while highlighting current challenges emerging field. The goal perspective is inspire innovation sustainable synthesis, providing chemists valuable guide.

Language: Английский

Semirationally Engineering an Efficient P450 Peroxygenase for Regio- and Enantioselective Hydroxylation of Steroids DOI
Shengxian Fan, Mingming Qin, Qian Wang

et al.

ACS Catalysis, Journal Year: 2025, Volume and Issue: unknown, P. 2977 - 2986

Published: Feb. 5, 2025

Language: Английский

Citations

2
Structure–Function Analysis of the Self-Sufficient CYP102 Family Provides New Insights into Their Biochemistry DOI Open Access
Tiara Padayachee, David C. Lamb, David R. Nelson

et al.

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(5), P. 2161 - 2161

Published: Feb. 28, 2025

Cytochromes P450 are a superfamily of heme-containing monooxygenases involved in variety oxidative metabolic reactions, primarily catalyzing the insertion an oxygen atom into C-H bond. CYP102 represents first example bacterial that can be classified as type II (eukaryotic-like) and functions catalytically self-sufficient enzyme. These unique features have made attractive system for studying structure function. However, overall picture specific amino acid residues crucial to functioning effect mutations on catalysis is yet reported. Such approach will aid protein engineering approaches used improve this To address research knowledge gap, we investigated 105 crystal structures study. We demonstrate active site highly dynamic flexible. Amino play critical roles substrate binding, orientation, anchoring were identified. Mutational studies highlighted acids provided possible bioengineering improvement strategies CYP102. Decoy molecules promising agent deceiving permitting non-native substrates site. Ru(II)-diimine photosensitizers zinc/cobalt (III) sepulchrate (Co(III)Sep) could alternative electron sources. The present study serves reference understanding structure-functional analysis family members precisely enzymes general. Significantly, work contributes effort develop improved enzyme, thereby advancing field potentially leading new industrial applications.

Language: Английский

Citations

0
Characterisation of the Self‐Sufficient Cytochrome P450 CYP116B234 From Rhodococcus globerulus and Its Suggested Native Role in 2‐Hydroxyphenylacetic Acid Metabolism DOI Creative Commons
Simran Kundral,

Heather Beamish,

Peter D. Giang

et al.

Microbial Biotechnology, Journal Year: 2025, Volume and Issue: 18(3)

Published: March 1, 2025

Cytochromes P450 (P450s) are exceptional biocatalysts that enable the selective oxidation of unactivated C-H bonds using molecular oxygen. Typically, auxiliary redox partner proteins deliver electrons from NAD(P)H to P450, enabling oxygen activation. However, associating native partners with P450s can be challenging, particularly when they genomically separated. Self-sufficient P450s, where heme domain is naturally fused partners, represent a simpler, single-protein system. Here, we present CYP116B234, novel self-sufficient Rhodococcus globerulus, comprising and phthalate-family oxygenase reductase (PFOR) domains. The gene encoding CYP116B234 was codon-optimised for heterologous expression in E. coli subsequently purified homogeneity. Spectroelectrochemical analysis electron paramagnetic resonance spectroscopy were performed determine potentials associated Fe-S FMN cofactors PFOR domain. binds efficiently oxidises substituted aromatic compound 2-hydroxyphenylacetic acid (2-HPA) homogentisic acid. Quantitative proteomics revealed R. globerulus grown on 2-HPA, suggesting role initiating 2-HPA degradation. This study presents new addition CYP116 family provides evidence their function.

Language: Английский

Citations

0
Catalytic Enantioselective Smiles Rearrangement Enabled by the Directed Evolution of P450 Radical Aryl Migratases DOI
Wen‐Zhen Fu,

Katherina Murcek,

Jasper Chen

et al.

Journal of the American Chemical Society, Journal Year: 2025, Volume and Issue: unknown

Published: April 1, 2025

Despite its synthetic potential, catalytic enantioselective Smiles rearrangement has remained elusive. Through the directed evolution of P450 radical aryl migratases (P450Smiles's), we describe first example rearrangement. A range racemic N-arylsulfonyl-α-chloroamides could be transformed by P450Smiles in an enantioconvergent manner, affording acyclic amide products possessing all-carbon quaternary stereocenter with excellent chemo- and enantioselectivity. Both electron-rich electron-deficient substituents were compatible migrating group, demonstrating this P450-catalyzed is insensitive to electronic properties group. Importantly, our evolved variants capable overriding innate cyclization activity N-alkyl amidyl intermediate, allowing chemoselective reductive formation products. Classical molecular dynamics (MD) simulations revealed unusual enzyme-controlled chemoselectivity stems from restricted conformation within enzyme active site, disfavoring pathway. This new-to-nature biocatalytic asymmetric showcases potential enzymatic enantioselectivity control over highly reactive intermediates eluding small-molecule catalysts.

Language: Английский

Citations

0
Selective Oxidation of Vitamin D3 Enhanced by Long‐Range Effects of a Substrate Channel Mutation in Cytochrome P450BM3 (CYP102A1) DOI Creative Commons
Wen-Yu Chen, John Lynch,

Claudia Bustamante

et al.

Chemistry - A European Journal, Journal Year: 2024, Volume and Issue: unknown

Published: Aug. 22, 2024

Abstract Vitamin D deficiency affects nearly half the population, with many requiring or opting for supplements vitamin 3 (VD ), precursor of (1α,25‐dihydroxyVD ). 25‐HydroxyVD , circulating form D, is a more effective supplement than VD but its synthesis complex. We report here engineering cytochrome P450 BM3 (CYP102A1) selective oxidation to 25‐hydroxyVD . Long‐range effects substrate‐channel mutation Glu435Ile promoted binding side chain close heme, enhancing activity that reached 6.62 g isolated from 1‐litre scale reaction (69.1 % yield; space‐time‐yield 331 mg/L/h).

Language: Английский

Citations

2
Indigo production identifies hotspots in cytochrome P450 BM3 for diversifying aromatic hydroxylation DOI
Douglas J. Fansher,

Jonathan N. Besna,

Joelle N. Pelletier

et al.

Faraday Discussions, Journal Year: 2024, Volume and Issue: 252, P. 29 - 51

Published: Jan. 1, 2024

Indigo (+) and indigo (−) single variants a combinatorial library, with mutations that enable the blue phenotype, were screened for their ability to hydroxylate panel of 12 aromatic compounds using 4-aminoantipyrine colorimetric assay.

Language: Английский

Citations

2
Design of a Genetically Programmable and Customizable Protein Scaffolding System for the Hierarchical Assembly of Robust, Functional Macroscale Materials DOI
R. Zhang,

Sun-Young Kang,

François Gaascht

et al.

ACS Synthetic Biology, Journal Year: 2024, Volume and Issue: 13(11), P. 3724 - 3745

Published: Oct. 31, 2024

Inspired by the properties of natural protein-based biomaterials, protein nanomaterials are increasingly designed with or engineered peptides building blocks. Few examples describe design functional materials for biotechnological applications that can be readily manufactured, amenable to functionalization, and exhibit robust assembly macroscale material formation. Here, we a protein-scaffolding system self-assembles into robust, suitable in vitro cell-free applications. By controlling coexpression Escherichia coli self-assembling scaffold blocks without modifications covalent attachment cross-linking cargo proteins, hybrid scaffolds spatially organized conjugation sites overproduced isolated. Cargo including enzymes, rapidly cross-linked onto formation materials. We show these used operation coimmobilized two-enzyme reaction recovered reused. believe this work will provide versatile platform scalable production customizable robustness required

Language: Английский

Citations

2
Concluding remarks: biocatalysis DOI Creative Commons
Uwe T. Bornscheuer

Faraday Discussions, Journal Year: 2024, Volume and Issue: 252, P. 507 - 515

Published: Jan. 1, 2024

Biocatalysis is a rapidly evolving field with increasing impact in organic synthesis, chemical manufacturing and medicine. The

Language: Английский

Citations

1
Chemoenzymatic Synthesis of the Cyclopiane Family of Diterpenoid Natural Products DOI Open Access
Tao Wang,

Zou Jian,

Kaibiao Wang

et al.

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 27, 2024

A three-stage chemoenzymatic synthesis of the cyclopiane family and related diterpenes is reported. Deoxyconidiogenol with a 6/5/5/5-fused tetracyclic skeleton was first produced by an engineered E. coli host harboring corresponding terpene cyclase PchDS. Ten were synthesized late-stage functionalization rings A, B D through direct redox operations, directed C-H activation, enzymatic hydroxylation, respectively. Skeletal diversification achieved taking advantage selective 1,2-alkyl migration cation generated chemically or enzymatically. Three cyclopiane-related skeletons, including spiro 5/5/5/5-tetracyclic spiroviolene, angular 5/6/5/5-fused ring system phomopsene, new linear amycolatene, either chemical skeletal transformation from skeleton, cyclases discovered genome mining.

Language: Английский

Citations

1
Design of a genetically programmable and customizable protein scaffolding system for the hierarchical assembly of robust, functional macroscale materials DOI Open Access
R. Zhang,

Sun-Young Kang,

François Gaascht

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: May 3, 2024

ABSTRACT Inspired by the properties of natural protein-based biomaterials, protein nanomaterials are increasingly designed with or engineered peptides, building blocks. Very few examples describe design functional materials for biotechnological applications that can be readily manufactured, amenable to functionalization, and exhibit robust assembly macroscale material formation. Here, we a protein-scaffolding system self-assembles into robust, suitable cell-free applications. By controlling co-expression in E. coli self-assembling scaffold blocks without modifications covalent attachment cross-linking cargo proteins, hybrid scaffolds spatially organized conjugation sites overproduced isolated. Cargo including enzymes, rapidly cross-linked onto formation materials. We show these used operation co-immobilized two-enzyme reaction recovered reused. believe this work will provide versatile platform scalable production customizable robustness required

Language: Английский

Citations

0