Discovery, Characterization and Synthetic Application of a Promiscuous Nonheme Iron Biocatalyst with Dual Hydroxylase/Desaturase Activity DOI
Hao Xu,

Jidong Zhao,

Hans Renata

et al.

Angewandte Chemie, Journal Year: 2024, Volume and Issue: 136(48)

Published: Aug. 29, 2024

Abstract Alpha‐ketoglutarate‐dependent dioxygenases (αKGDs) have recently emerged as useful biocatalysts for C−H oxidation and functionalization. In this work, we characterized a new αKGD from aculene biosynthesis, AneA, which displays broad promiscuity toward number of substrates with different ring systems. Unexpectedly, AneA was found to be capable both desaturation hydroxylation require an amino ester motif on its substrate productive catalysis. Insights gathered the functional characterization substrate‐activity profiling enabled development chemoenzymatic strategy several complex sesquiterpenoids.

Language: Английский

tert‐Butyl as a Functional Group: Non‐Directed Catalytic Hydroxylation of Sterically Congested Primary C−H Bonds DOI Creative Commons
Siu‐Chung Chan, Andrea Palone, Massimo Bietti

et al.

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(28)

Published: May 1, 2024

Abstract The tert ‐butyl group is a common aliphatic motif extensively employed to implement steric congestion and conformational rigidity in organic organometallic molecules. Because of the combination high bond dissociation energy (~100 kcal mol −1 ) limited accessibility, absence directing groups, neither radical nor approaches are effective for chemical modification C−H bonds. Herein we overcome these limits by employing highly electrophilic manganese catalyst, [Mn( CF3 bpeb)(OTf) 2 ], that operates strong hydrogen donor solvent nonafluoro‐ alcohol (NFTBA) catalytically activates peroxide generate powerful manganese‐oxo species effectively oxidizes Leveraging on interplay steric, electronic, medium torsional effects, site‐selective product chemoselective hydroxylation accomplished with broad reaction scope, delivering primary alcohols as largely dominant products preparative yields. Late‐stage at sites demonstrated 6 densely functionalized molecules pharmaceutical interest. This work uncovers novel disconnection approach, harnessing potential functional strategic synthetic planning complex molecular architectures.

Language: Английский

Citations

2
Rational Design of the Spatial Effect in a Fe(II)/α‐Ketoglutarate‐Dependent Dioxygenase Reverses the Regioselectivity of C(sp3)−H Bond Hydroxylation in Aliphatic Amino Acids DOI

Chenni Zheng,

Wanqing Wei, Jian Wen

et al.

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(32)

Published: May 25, 2024

Abstract The hydroxylation of remote C( sp 3 )−H bonds in aliphatic amino acids yields crucial precursors for the synthesis high‐value compounds. However, accurate regulation regioselectivity continues to be a common challenge chemosynthesis and biosynthesis. In this study, Fe(II)/α‐ketoglutarate‐dependent dioxygenase from Bacillus subtilis (BlAH) was mined found catalyze at γ δ sites acids. Crystal structure analysis, molecular dynamics simulations, quantum chemical calculations revealed that regulated by spatial effect BlAH. Based on these results, BlAH reconstructed stabilize transition state site acids, thereby successfully reversing site. For example, L‐Homoleucine ( 5 ) reversed (1 : 12) (>99 1). present study not only expands toolbox biocatalysts regioselective functionalization bonds, but also provides theoretical guidance precision‐driven modification similarly complex molecules.

Language: Английский

Citations

2
Immunoglobulin E Epitope Mapping and Structure–Allergenicity Relationship Analysis of Crab Allergen Scy p 9 DOI

Xinrong He,

Yang Yang, Yexin Chen

et al.

Journal of Agricultural and Food Chemistry, Journal Year: 2023, Volume and Issue: unknown

Published: Nov. 6, 2023

Filamin C is an allergen of Scylla paramamosain (Scy p 9), and six IgE linear epitopes the allergenic predominant region had previously been validated. However, epitope structure-allergenicity relationship Scy 9 are unclear. In this study, a hydrophobic bond was found to be important factor conformation maintaining. The critical amino acids in predicted conformational were mutated, IgE-binding capacity surface hydrophobicity four mutants (E216A, T270A, Y699A, V704A) reduced compared 9. Ten verified with synthetic peptides, among which L-AA187-205 strongest capacity. addition, mapped protruding tertiary structure, conducive binding exhibited high conservation filamin genes. Overall, these data provided basis for mapping

Language: Английский

Citations

4
Algal Kainoid Synthases Exhibit Substrate-Dependent Hydroxylation and Cyclization Activities DOI Creative Commons
Austin Hopiavuori, Shaun M. K. McKinnie

ACS Chemical Biology, Journal Year: 2023, Volume and Issue: 18(12), P. 2457 - 2463

Published: Dec. 4, 2023

FeII/α-ketoglutarate-dependent dioxygenases (Fe/αKG) make up a large enzyme family that functionalize C–H bonds on diverse organic substrates. Although Fe/αKG homologues catalyze an array of chemically useful reactions, hydroxylation typically predominates. Microalgal DabC uniquely forms novel C–C bond to construct the bioactive pyrrolidine ring in domoic acid biosynthesis; however, we have identified this kainoid synthase exclusively performs stereospecific reaction its cis substrate regioisomer. Mechanistic and kinetic analyses with native alternative substrates 20-fold rate increase radical cyclization over β-hydroxylation no observable 1,5-hydrogen atom transfer. Moreover, dual activity was conserved among macroalgal RadC1 KabC provided insight into recognition reactivity trends. Investigation substrate-dependent chemistry improves our understanding synthases their biocatalytic application.

Language: Английский

Citations

4
Expression, purification, and biochemical characterization of micro- and macroalgal kainoid synthases DOI

Austin R. Hopiavuori,

Radcliff T Huffman,

Shaun M. K. McKinnie

et al.

Methods in enzymology on CD-ROM/Methods in enzymology, Journal Year: 2024, Volume and Issue: unknown, P. 233 - 258

Published: Jan. 1, 2024

Language: Английский

Citations

1
Protein3D: Enabling analysis and extraction of metal‐containing sites from the Protein Data Bank with molSimplify DOI Creative Commons

Freya Edholm,

Aditya Nandy, Clorice R. Reinhardt

et al.

Journal of Computational Chemistry, Journal Year: 2023, Volume and Issue: 45(6), P. 352 - 361

Published: Oct. 24, 2023

Metalloenzymes catalyze a wide range of chemical transformations, with the active site residues playing key role in modulating reactivity and selectivity. Unlike smaller synthetic catalysts, metalloenzyme is embedded larger protein, which makes interrogation electronic properties geometric features quantum mechanical calculations challenging. Here we implement ability to fetch crystallographic structures from Protein Data Bank analyze metal binding sites program molSimplify. We show usefulness newly created protein3D class extract local environment around non-heme iron enzymes containing two histidine motif prepare 372 for calculations. Our implementation serves expand systems molSimplify can be used will enable high-throughput study metal-containing proteins.

Language: Английский

Citations

3
One-pot chemoenzymatic syntheses of non-canonical amino acids DOI Creative Commons
Tsung-Han Chao, Xiangyu Wu, Hans Renata

et al.

Journal of Industrial Microbiology & Biotechnology, Journal Year: 2024, Volume and Issue: 51

Published: Jan. 1, 2024

Abstract Despite their prevalent use in drug discovery and protein biochemistry, non-canonical amino acids are still challenging to synthesize through purely chemical means. In recent years, biocatalysis has emerged as a transformative paradigm for small-molecule synthesis. One strategy further empower is it combination with modern reactions take advantage of the strengths each method enable access structural motifs that were previously unattainable using alone. this Mini-Review, we highlight several case studies feature synergistic enzymatic transformations one pot novel acids. One-Sentence Summary This Mini-Review highlights

Language: Английский

Citations

0
Engineering Hydroxylase Activity, Selectivity, and Stability for a Scalable Concise Synthesis of a Key Intermediate to Belzutifan DOI
Wai Ling Cheung‐Lee, Joshua N. Kolev, John A. McIntosh

et al.

Angewandte Chemie, Journal Year: 2024, Volume and Issue: 136(13)

Published: Jan. 27, 2024

Abstract 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

Citations

0
Affinity-peptide-mediated multi-enzyme self-assembly system enhances dioxygenase catalyzing C–H hydroxylation via in situ α-ketoglutarate generation and H2O2 elimination DOI
Huan Liu, Lunjie Wu, Jie Gu

et al.

Molecular Catalysis, Journal Year: 2024, Volume and Issue: 555, P. 113873 - 113873

Published: Jan. 29, 2024

Language: Английский

Citations

0
tert‐Butyl as a Functional Group: Non‐Directed Catalytic Hydroxylation of Sterically Congested Primary C−H Bonds DOI Creative Commons
Siu‐Chung Chan, Andrea Palone, Massimo Bietti

et al.

Angewandte Chemie, Journal Year: 2024, Volume and Issue: 136(28)

Published: May 1, 2024

Abstract The tert ‐butyl group is a common aliphatic motif extensively employed to implement steric congestion and conformational rigidity in organic organometallic molecules. Because of the combination high bond dissociation energy (~100 kcal mol −1 ) limited accessibility, absence directing groups, neither radical nor approaches are effective for chemical modification C−H bonds. Herein we overcome these limits by employing highly electrophilic manganese catalyst, [Mn( CF3 bpeb)(OTf) 2 ], that operates strong hydrogen donor solvent nonafluoro‐ alcohol (NFTBA) catalytically activates peroxide generate powerful manganese‐oxo species effectively oxidizes Leveraging on interplay steric, electronic, medium torsional effects, site‐selective product chemoselective hydroxylation accomplished with broad reaction scope, delivering primary alcohols as largely dominant products preparative yields. Late‐stage at sites demonstrated 6 densely functionalized molecules pharmaceutical interest. This work uncovers novel disconnection approach, harnessing potential functional strategic synthetic planning complex molecular architectures.

Language: Английский

Citations

0