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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Engineering of Halide Methyltransferase BxHMT through Dynamic Cross‐Correlation Network Analysis

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(25)

Published: April 16, 2024

Halide methyltransferases (HMTs) provide an effective way to regenerate S-adenosyl methionine (SAM) from homocysteine and reactive electrophiles, such as methyl iodide (MeI) toluene sulfonate (MeOTs). As compared with MeI, the cost-effective unnatural substrate MeOTs can be accessed directly cheap abundant alcohols, but shows only limited reactivity in SAM production. In this study, we developed a dynamic cross-correlation network analysis (DCCNA) strategy for quickly identifying hot spots influencing catalytic efficiency of enzyme, applied it evolution HMT Paraburkholderia xenovorans. Finally, optimal mutant, M4 (V55T/C125S/L127T/L129P), exhibited remarkable improvement, specific activity 4.08 U/mg towards MeOTs, representing 82-fold increase wild-type (WT) enzyme. Notably, also demonstrated positive impact on ability other donors. The structural mechanism behind enhanced enzyme was uncovered by molecular dynamics simulations. Our work not contributes promising biocatalyst regeneration SAM, offers efficient engineering.

Language: Английский

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Organophosphorus S-adenosyl-L-methionine mimetics: synthesis, stability, and substrate properties

Frontiers in Chemistry, Journal Year: 2024, Volume and Issue: 12

Published: Aug. 1, 2024

S-Adenosyl-l-methionine (SAM)-mediated methylation of biomolecules controls their function and regulates numerous vital intracellular processes. Analogs SAM with a reporter group in place the S-methyl are widely used to study these However, many analogs chemically unstable that largely limits practical application. We have developed new compound, SAM-PH , which contains an H-phosphinic (-P(O)(H)OH) instead carboxylic group. SAM-P H is significantly more stable than SAM, retains functional activity catechol-O-methyltransferase methyltransferase WBSCR27 reactions. The last associated Williams-Beuren syndrome. Rac-SAM-P was synthesized chemically, while (R,S)-SAM-P its were prepared enzymatically either from methionine (Met-PH) or analog S-adenosyl-l-homocysteine (SAH-P ) using adenosyltransferase 2A halide methyltransferases, respectively. SAH-P undergoes glycoside bond cleavage presence methylthioadenosine nucleosidase like natural SAH. Thus, promising tools for investigating methyltransferases incorporating groups into substrates.

Language: Английский

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Sustainable chemo-enzymatic NADP(H) synthesis from biomass-derived xylose, polyphosphate, and nicotinamide

Green Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

Chemo-enzymatic NAD(P)H synthetic strategy from biomass-derived crude xylose and nicotinamide.

Language: Английский

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Enzymatic Synthesis of l‐Methionine Analogues and Application in a Methyltransferase Catalysed Alkylation Cascade**

Chemistry - A European Journal, Journal Year: 2023, Volume and Issue: 29(46)

Published: May 26, 2023

Chemical modification of small molecules is a key step for the development pharmaceuticals. S-adenosyl-l-methionine (SAM) analogues are used by methyltransferases (MTs) to transfer alkyl, allyl and benzyl moieties chemo-, stereo- regioselectively onto nucleophilic substrates, enabling an enzymatic way specific derivatisation wide range molecules. l-Methionine required synthesis SAM analogues. Most these not commercially available. In nature, O-acetyl-l-homoserine sulfhydrolases (OAHS) catalyse l-methionine from or l-homocysteine, methyl mercaptan. Here, we investigated substrate scope ScOAHS Saccharomyces cerevisiae production l-homocysteine organic thiols. The promiscuous enzyme was synthesise nine different with modifications on thioether residue up conversion 75 %. combined established MT dependent three-enzyme alkylation cascade, allowing in total seven two substrates. For ethylation, nearly doubled new four-enzyme indicating beneficial effect situ ScOAHS.

Language: Английский

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Expanding the Substrate Scope of N‐ and O‐Methyltransferases from Plants for Chemoselective Alkylation**

ChemCatChem, Journal Year: 2023, Volume and Issue: 15(22)

Published: Aug. 17, 2023

Abstract Methylation reactions are of significant interest when generating pharmaceutically active molecules and building blocks for other applications. Synthetic methylating reagents often toxic unselective due to their high reactivity. S ‐Adenosyl‐ l ‐methionine (SAM)‐dependent methyltransferases (MTs) present a chemoselective environmentally friendly alternative. The anthranilate N ‐MT from Ruta graveolens ( Rg ANMT) is involved in acridone alkaloid biosynthesis, anthranilate. Although it known methylate substrates only at the N‐ position, closest relatives with respect amino acid sequence similarities over 60 % O ‐MTs catalysing methylation reaction caffeate derivatives containing hydroxyl groups (CaOMTs). In this study, we investigated substrate range ANMT CaOMT Prunus persica Pp CaOMT) using compounds both, an amino‐ group (aminophenols) as possible methyl acceptors. For both enzymes, was highly chemoselective. Furthermore, cofactor situ enabled transfer alkyl chains onto aminophenols, leading enlarged pool products. Selected MT were performed preparative biocatalytic scale vitro vivo experiments resulting yields up 62 %.

Language: Английский

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Methyltransferases from RiPP pathways: shaping the landscape of natural product chemistry

Beilstein Journal of Organic Chemistry, Journal Year: 2024, Volume and Issue: 20, P. 1652 - 1670

Published: July 18, 2024

This review article aims to highlight the role of methyltransferases within context ribosomally synthesised and post-translationally modified peptide (RiPP) natural products. Methyltransferases play a pivotal in biosynthesis diverse products with unique chemical structures bioactivities. They are highly chemo-, regio-, stereoselective allowing methylation at various positions. The different possible acceptor regions peptides described this article. Furthermore, we will discuss potential application these as powerful biocatalytic tools synthesis other bioactive compounds. By providing an overview options available, is intended emphasise RiPP their impact on field product chemistry.

Language: Английский

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Non‐canonical nucleosides: Biomimetic triphosphorylation, incorporation into mRNA and effects on translation and structure

FEBS Journal, Journal Year: 2023, Volume and Issue: 290(20), P. 4899 - 4920

Published: June 17, 2023

Recent advances in mRNA therapeutics demand efficient toolkits for the incorporation of nucleoside analogues into suitable downstream applications. Herein, we report application a versatile enzyme cascade triphosphorylation broad range analogues, including unprotected nucleobases containing chemically labile moieties. Our biomimetic system was preparation triphosphates adenosine, cytidine, guanosine, uridine and non-canonical core structures, as determined by capillary electrophoresis coupled to mass spectrometry. This enabled us establish an workflow transcribing purifying functional these combined with spectrometric verification analogue incorporation. methodology allows analyses how that are commercially unavailable affect properties: The translational fidelity produced demonstrated incorporated adenosine impact recoding. For SARS-CoV-2 frameshifting site, pseudoknot structure using circular dichroism spectroscopy allowed insight pharmacologically active 7-deazaadenosine destabilises RNA secondary structure, consistent observed changes recoding efficiency.

Language: Английский

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Back to the Future of Metabolism—Advances in the Discovery and Characterization of Unknown Biocatalytic Functions and Pathways

Life, Journal Year: 2024, Volume and Issue: 14(3), P. 364 - 364

Published: March 10, 2024

The architecture, organization, and functioning of biocatalytic reaction networks, which are coded in the cell-specific genome work together small space biological cells, a fascinating feature life evolved over more than 3 billion years. Knowledge about diversity functions metabolic pathways sustaining on our planet is highly important, especially as currently occurring loss biodiversity considered planetary boundary that at high risk, knowledge current organisms should be gained before they become extinct. In addition to well-known enzymatic reactions involved biochemical pathways, enzyme universe offers numerous opportunities for discovering novel pathways. Maintaining thousands molecules properly within may exposed various kinds external hazards, environmental stress, side reactions, or non-enzymatic chemical key keeping cellular healthy. This review aims outline advances assigning protein sequences discovery

Language: Английский

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Engineering of Halide Methyltransferase BxHMT through Dynamic Cross‐Correlation Network Analysis

Angewandte Chemie, Journal Year: 2024, Volume and Issue: 136(25)

Published: April 16, 2024

Abstract Halide methyltransferases (HMTs) provide an effective way to regenerate S ‐adenosyl methionine (SAM) from homocysteine and reactive electrophiles, such as methyl iodide (MeI) toluene sulfonate (MeOTs). As compared with MeI, the cost‐effective unnatural substrate MeOTs can be accessed directly cheap abundant alcohols, but shows only limited reactivity in SAM production. In this study, we developed a dynamic cross‐correlation network analysis (DCCNA) strategy for quickly identifying hot spots influencing catalytic efficiency of enzyme, applied it evolution HMT Paraburkholderia xenovorans . Finally, optimal mutant, M4 (V55T/C125S/L127T/L129P), exhibited remarkable improvement, specific activity 4.08 U/mg towards MeOTs, representing 82‐fold increase wild‐type (WT) enzyme. Notably, also demonstrated positive impact on ability other donors. The structural mechanism behind enhanced enzyme was uncovered by molecular dynamics simulations. Our work not contributes promising biocatalyst regeneration SAM, offers efficient engineering.

Language: Английский

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