Selective Biocatalytic N‐Methylation of Unsaturated Heterocycles DOI Creative Commons
Felipe Ospina, Kai H. Schülke, Jordi Soler Soler

et al.

Angewandte Chemie, Journal Year: 2022, Volume and Issue: 134(48)

Published: Oct. 7, 2022

Abstract Methods for regioselective N ‐methylation and ‐alkylation of unsaturated heterocycles with “off the shelf” reagents are highly sought‐after. This reaction could drastically simplify synthesis privileged bioactive molecules. Here we report engineered natural methyltransferases challenging ‐(m)ethylation heterocycles, including benzimidazoles, benzotriazoles, imidazoles indazoles. The reactions performed through a cyclic enzyme cascade that consists two using only iodoalkanes or methyl tosylate as simple reagents. method enables selective important molecules otherwise difficult to access, proceeds high regioselectivity (r.r. up >99 %), yield (up 99 on preparative scale, nearly equimolar concentrations starting materials.

Language: Английский

Non-Native Site-Selective Enzyme Catalysis DOI
Dibyendu Mondal, Harrison M. Snodgrass,

Christian A. Gomez

et al.

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

Citations

24
Chemical Synthesis of Modified RNA DOI Creative Commons

Laurin Flemmich,

Raphael Bereiter,

Ronald Micura

et al.

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(22)

Published: March 26, 2024

Ribonucleic acids (RNAs) play a vital role in living organisms. Many of their cellular functions depend critically on chemical modification. Methods to modify RNA controlled manner-both vitro and vivo-are thus essential evaluate understand biology at the molecular mechanistic levels. The diversity modifications, combined with size uniformity (made up only 4 nucleotides) makes its site-specific modification challenging task that needs be addressed by complementary approaches. One such approach is solid-phase synthesis. We discuss recent developments this field, starting new protection concepts ongoing effort overcome current limitations. continue selected modifications have posed significant challenges for incorporation into RNA. These include deazapurine bases required atomic mutagenesis elucidate aspects catalytic RNAs, containing xanthosine, N

Language: Английский

Citations

9
Engineering an O‐methyltransferase for the Regioselective Biosynthesis of Hesperetin Dihydrochalcone DOI Creative Commons
Andreas Kunzendorf, Bastian Zirpel, Lars Milke

et al.

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

Published: Aug. 21, 2023

Abstract Directed evolution of the O ‐methyltransferase ZgOMT from Zooshikella ganghwensis focusing on active site residues resulted in highly regioselective biocatalysts (regioisomeric ratios up to 99 : 1) for preparation taste hesperetin dihydrochalcone and related compounds. These newly constructed enzyme variants provide an attractive synthesis route para ‐methylation catechol scaffolds, which is challenging perform with high regioselectivity utilizing wild‐type ‐methyltransferases.

Language: Английский

Citations

17
Efficient Transferase Engineering for SAM Analog Synthesis from Iodoalkanes DOI Creative Commons
Kai H. Schülke,

Jana S. Fröse,

Alina Klein

et al.

ChemBioChem, Journal Year: 2024, Volume and Issue: 25(10)

Published: March 13, 2024

Abstract S ‐Adenosyl‐ l ‐methionine (SAM) is an important cosubstrate in various biochemical processes, including selective methyl transfer reactions. Simple methods for the (re)generation of SAM analogs could expand chemistry accessible with SAM‐dependent transferases and go beyond methylation Here we present efficient enzyme engineering strategy to synthesize different from “off‐the‐shelf” iodoalkanes through enzymatic alkylation ‐adenosyl‐ ‐homocysteine (SAH). This was achieved by mutating multiple hydrophobic structurally dynamic amino acids simultaneously. Combinatorial mutagenesis guided natural acid diversity generated a highly functional mutant library. approach increased speed as well scale providing panel optimized enzymes orders magnitude higher activities substrates just one round engineering. The exhibit catalytic efficiencies up 31 M −1 s , convert iodoalkanes, bearing cyclopropyl or aromatic moieties, catalyze ‐alkylation SAH very high stereoselectivities (>99 % de ). We further report throughput chromatographic screening system reliable rapid analog analysis. believe that described herein will advance field biocatalytic enabling regeneration reagents.

Language: Английский

Citations

8
Selective Biocatalytic N‐Methylation of Unsaturated Heterocycles DOI Creative Commons
Felipe Ospina, Kai H. Schülke, Jordi Soler Soler

et al.

Angewandte Chemie International Edition, Journal Year: 2022, Volume and Issue: 61(48)

Published: Oct. 7, 2022

Methods for regioselective N-methylation and -alkylation of unsaturated heterocycles with "off the shelf" reagents are highly sought-after. This reaction could drastically simplify synthesis privileged bioactive molecules. Here we report engineered natural methyltransferases challenging N-(m)ethylation heterocycles, including benzimidazoles, benzotriazoles, imidazoles indazoles. The reactions performed through a cyclic enzyme cascade that consists two using only iodoalkanes or methyl tosylate as simple reagents. method enables selective important molecules otherwise difficult to access, proceeds high regioselectivity (r.r. up >99 %), yield (up 99 on preparative scale, nearly equimolar concentrations starting materials.

Language: Английский

Citations

26
A biocatalytic platform for asymmetric alkylation of α-keto acids by mining and engineering of methyltransferases DOI Creative Commons
Shuyun Ju,

Kaylee P. Kuzelka,

R.T. Guo

et al.

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: Sept. 14, 2023

Abstract Catalytic asymmetric α-alkylation of carbonyl compounds represents a long-standing challenge in synthetic organic chemistry. Herein, we advance dual biocatalytic platform for the efficient alkylation α-keto acids. First, guided by our recently obtained crystal structures, develop SgvM VAV as general biocatalyst enantioselective methylation, ethylation, allylation and propargylation range acids with total turnover numbers (TTNs) up to 4,600. Second, mine family bacterial HMTs from Pseudomonas species sharing less than 50% sequence identities known evaluated their activities SAM regeneration. Our best performing HMT P. aeruginosa , Pa HMT, displays highest regeneration efficiencies (TTN 7,700) among characterized date. Together, synergistic use affords fully protocol methylation featuring record efficiency, providing solution notorious problem alkylation.

Language: Английский

Citations

13
ComparativeS-adenosyl-l-methionine analogue generation for selective biocatalytic Friedel-Crafts alkylation DOI
Arne Hoffmann, Kai H. Schülke, Stephan C. Hammer

et al.

Chemical Communications, Journal Year: 2023, Volume and Issue: 59(36), P. 5463 - 5466

Published: Jan. 1, 2023

Methyltransferases provide excellent specificity in late-stage alkylation of biomolecules. Their dependence on S-adenosyl-L-methionine (SAM) mandates efficient access to SAM analogues for biocatalytic applications. We directly compared halide methyltransferase (HMT) and methionine adenosyltransferase (MAT) explored their utility cascade reactions with NovO regioselective, Friedel-Crafts a coumarin. The HMT efficiently provided methylation, while the MAT also supplied high levels reactions.

Language: Английский

Citations

11
Engineering of Halide Methyltransferase BxHMT through Dynamic Cross‐Correlation Network Analysis DOI

Chun‐Yu Gao,

Guiying Yang,

Xu‐Wei Ding

et al.

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

Citations

4
Organophosphorus S-adenosyl-L-methionine mimetics: synthesis, stability, and substrate properties DOI Creative Commons
A. Yu. Rudenko, Sofia S. Mariasina,

Anastasiia K. Bolikhova

et al.

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

Citations

4
Enzymatic Reactions of S-Adenosyl-L-Methionine: Synthesis and Applications DOI
A. Yu. Rudenko, Sofia S. Mariasina,

Ratislav M. Ozhiganov

et al.

Biochemistry (Moscow), Journal Year: 2025, Volume and Issue: 90(S1), P. S105 - S134

Published: Feb. 1, 2025

Language: Английский

Citations

0