Designing Artificial Metalloenzymes by Tuning of the Environment beyond the Primary Coordination Sphere

Chemical Reviews, Год журнала: 2022, Номер 122(14), С. 11974 - 12045

Опубликована: Июль 11, 2022

Metalloenzymes catalyze a variety of reactions using limited number natural amino acids and metallocofactors. Therefore, the environment beyond primary coordination sphere must play an important role in both conferring tuning their phenomenal catalytic properties, enabling active sites with otherwise similar environments to perform diverse array biological functions. However, since interactions are numerous weak, it has been difficult pinpoint structural features responsible for activities native enzymes. Designing artificial metalloenzymes (ArMs) offers excellent basis elucidate roles these further develop practical catalysts. In this review, we highlight how secondary spheres ArMs influence metal binding catalysis, particular focus on use protein scaffolds as templates design by either rational aided computational modeling, directed evolution, or combination approaches. describing successes designing heme, nonheme Fe, Cu metalloenzymes, heteronuclear containing those other centers (including non-native ions metallocofactors), have summarized insights gained careful controls sphere, including hydrophobic hydrogen bonding interactions, allow generation respective systems approach, rival, and, few cases, exceed We also provided outlook remaining challenges field future directions that will deeper understanding coordintion be gained, turn guide broader more efficient ArMs.

Язык: Английский

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In Vivo Assembly of Artificial Metalloenzymes and Application in Whole‐Cell Biocatalysis**

Angewandte Chemie International Edition, Год журнала: 2021, Номер 60(11), С. 5913 - 5920

Опубликована: Янв. 15, 2021

Abstract We report the supramolecular assembly of artificial metalloenzymes (ArMs), based on Lactococcal multidrug resistance regulator (LmrR) and an exogeneous copper(II)–phenanthroline complex, in cytoplasm E. coli cells. A combination catalysis, cell‐fractionation, inhibitor experiments, supplemented with in‐cell solid‐state NMR spectroscopy, confirmed assembly. The ArM‐containing whole cells were active catalysis enantioselective Friedel–Crafts alkylation indoles Diels–Alder reaction azachalcone cyclopentadiene. Directed evolution resulted two different improved mutants for both reactions, LmrR_A92E_M8D LmrR_A92E_V15A, respectively. whole‐cell ArM system required no engineering microbial host, protein scaffold, or cofactor to achieve catalysis. consider this a key step towards integrating abiological biosynthesis generate hybrid metabolism.

Язык: Английский

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Using BpyAla to generate Copper Artificial Metalloenzymes: a catalytic and structural study

Catalysis Science & Technology, Год журнала: 2024, Номер unknown

Опубликована: Янв. 1, 2024

Artificial metalloenzymes (ArMs) have emerged as a promising avenue in the field of biocatalysis, offering new reactivity. However, their design remains challenging due to limited understanding protein dynamics and how introduced cofactors alter scaffold structure. Here we present structures catalytic activity novel copper ArMs capable (

Язык: Английский

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Unlocking Iminium Catalysis in Artificial Enzymes to Create a Friedel–Crafts Alkylase

ACS Catalysis, Год журнала: 2021, Номер 11(12), С. 6763 - 6770

Опубликована: Май 26, 2021

The construction and engineering of artificial enzymes consisting abiological catalytic moieties incorporated into protein scaffolds is a promising strategy to realize non-natural mechanisms in biocatalysis. Here, we show that incorporation the noncanonical amino acid para-aminophenylalanine (pAF) nonenzymatic scaffold LmrR creates proficient stereoselective enzyme (LmrR_pAF) for vinylogous Friedel–Crafts alkylation between α,β-unsaturated aldehydes indoles. pAF acts as residue, activating enal substrates toward conjugate addition via formation intermediate iminium ion species, while provides rate acceleration stereoinduction. Improved LmrR_pAF variants were identified by low-throughput directed evolution advised alanine-scanning obtain triple mutant provided higher yields enantioselectivities range aliphatic enals substituted Analysis Michaelis–Menten kinetics evolved mutants reveals different activities emerge evolutionary pathways diverge from one another specialize reactivity. Translating this iminium-based mechanism an enzymatic context will enable many more biocatalytic transformations inspired organocatalysis.

Язык: Английский

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Review on Recent Developments in Biocatalysts for Friedel–Crafts Reactions

ACS Catalysis, Год журнала: 2022, Номер 12(17), С. 10742 - 10763

Опубликована: Авг. 17, 2022

The Friedel–Crafts (F–C) reaction has been a fundamental pillar of both academic and industrial synthetic organic chemistry since its discovery in 1873. Its success is based on the versatility applicability F–C reactions for wide range substrates, there have an impressive number publications patents describing catalytic methods. asymmetric version was discovered about 100 years after seminal work by Friedel Crafts become major area research. While chemical methods with much-improved efficacies scopes discovered, still suffer from limitations. Biocatalysis potential to be best solution this challenge because excellent selectivity (enantio-, chemo-, regioselectivity) displayed enzymes. In last two decades, advancements molecular biology techniques, bioinformatics, high-throughput screening, directed evolution, process scale-up led biocatalysis becoming mature field. It therefore not surprising that researchers around globe developed several biocatalysts reactions. Herein, we review recent developments design use stereoselective strategies performing

Язык: Английский

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Biocatalytic Friedel‐Crafts Reactions

ChemCatChem, Год журнала: 2022, Номер 14(18)

Опубликована: Июль 11, 2022

Abstract Friedel‐Crafts alkylation and acylation reactions are important methodologies in synthetic industrial chemistry for the construction of aryl‐alkyl aryl‐acyl linkages that ubiquitous bioactive molecules. Nature also exploits these many biosynthetic processes. Much work has been done to expand application enzymes unnatural substrates through directed evolution. The promise such biocatalysts is their potential supersede inefficient toxic chemical approaches reactions, with mild operating conditions ‐ hallmark enzymes. Complementary created bio‐hybrid catalysts consisting anchored into biomolecular scaffolds, which display same desirable characteristics. In this Review, we summarise efforts, focussing on both mechanistic aspects considerations, concluding an overview frontiers field routes towards more efficient benign future humankind.

Язык: Английский

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Design of Artificial Enzymes: Insights into Protein Scaffolds

ChemBioChem, Год журнала: 2022, Номер 24(6)

Опубликована: Ноя. 24, 2022

Abstract The design of artificial enzymes has emerged as a promising tool for the generation potent biocatalysts able to promote new‐to‐nature reactions with improved catalytic performances, providing powerful platform wide‐ranging applications and better understanding protein functions structures. selection an appropriate scaffold plays key role in process. This review aims give general overview most common scaffolds that can be exploited enzymes. Several examples are discussed categorized according strategy used biocatalyst, namely functionalization natural enzymes, creation new site bearing wide hydrophobic pocket de novo design. is concluded by comparison these different methods our perspective on topic.

Язык: Английский

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Mechanistic and structural characterization of an iridium-containing cytochrome reveals kinetically relevant cofactor dynamics

Nature Catalysis, Год журнала: 2023, Номер 6(1), С. 39 - 51

Опубликована: Янв. 12, 2023

Язык: Английский

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Chemogenetic Evolution of Diversified Photoenzymes for Enantioselective [2 + 2] Cycloadditions in Whole Cells

Journal of the American Chemical Society, Год журнала: 2024, Номер 146(28), С. 19030 - 19041

Опубликована: Июль 8, 2024

Artificial photoenzymes with novel catalytic modes not found in nature are high demand; yet, they also present significant challenges the field of biocatalysis. In this study, a chemogenetic modification strategy is developed to facilitate rapid diversification photoenzymes. This integrates site-specific chemical conjugation various artificial photosensitizers into natural protein cavities and iterative mutagenesis cell lysates. Through rounds directed evolution, prominent visible-light-activatable photoenzyme variants were developed, featuring thioxanthone chromophore. They successfully enabled enantioselective [2 + 2] photocycloaddition 2-carboxamide indoles, class UV-sensitive substrates that traditionally challenging for known Furthermore, versatility demonstrated whole-cell photobiocatalysis, enabling efficient synthesis enantioenriched cyclobutane-fused indoline tetracycles. These findings significantly expand photophysical properties photoenzymes, critical factor enhancing their potential harnessing excited-state reactivity stereoselective transformations.

Язык: Английский

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Artificial enzymes with multiple active sites

Current Opinion in Green and Sustainable Chemistry, Год журнала: 2021, Номер 29, С. 100452 - 100452

Опубликована: Янв. 21, 2021

Artificial enzymes based on the modification of a protein structure for creation new active catalytic sites have experienced great boom in recent years. Multidisciplinary strategies genetic engineering, chemical or biological tools been successfully described to synthetize them. However, challenge has focused creating artificial with more than one site. This could represent direction application enzymatic sustainable chemistry. Actually, only few technologies designing two multiple sites. review article underlines these most significant advances.

Язык: Английский

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