Rewiring Escherichia coli to transform formate into methyl groups DOI Creative Commons
Michael K. F. Mohr, Ari Satanowski, Steffen N. Lindner

et al.

Microbial Cell Factories, Journal Year: 2025, Volume and Issue: 24(1)

Published: March 7, 2025

Biotechnological applications are steadily growing and have become an important tool to reinvent the synthesis of chemicals pharmaceuticals for lower dependence on fossil resources. In order sustain this progression, new feedstocks biotechnological hosts be explored. One-carbon (C1-)compounds, including formate, derived from CO2 or organic waste accessible in large quantities with renewable energy, making them promising candidates. Previous studies showed that introducing formate assimilation machinery Methylorubrum extorquens into Escherichia coli allows through C1-tetrahydrofolate (C1-H4F) metabolism. Applying route assimilation, we here investigated utilisation value-added building blocks E. using S-adenosylmethionine (SAM)-dependent methyltransferases (MT). We first used a two-vector system link SAM-dependent methylation three different MTs BL21. By feeding isotopically labelled methylated products 51-81% 13C-labelling could obtained without substantial changes conversion rates. Focussing improvement product formation one MT, analysed engineered C1-auxotrophic strain C1S. Screening concentrations allowed doubling rate comparison not formate-supplemented BL21 share more than 70% formate-derived methyl groups. Within study transformation groups is demonstrated coli. Our findings support can improve availability usable C1-compounds and, as result, increase whole-cell Using starting point, introduction additional auxiliary enzymes ideas make energy-efficient discussed future applications.

Language: Английский

Methyltransferases: Functions and Applications DOI Creative Commons
Eman M. M. Abdelraheem,

Benjamin Thair,

Romina Fernández Varela

et al.

ChemBioChem, Journal Year: 2022, Volume and Issue: 23(18)

Published: June 13, 2022

In this review the current state-of-the-art of S-adenosylmethionine (SAM)-dependent methyltransferases and SAM are evaluated. Their structural classification diversity is introduced key mechanistic aspects presented which then detailed further. Then, catalytic as a target for drugs, approaches to utilise cofactor in synthesis with different supply regeneration The use analogues also described. Finally O-, N-, C- S-MTs, their synthetic applications potential compound diversification given.

Language: Английский

Citations

95
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
Methyl transfer in psilocybin biosynthesis DOI Creative Commons
Jesse Hudspeth, Kai Rogge,

Sebastian Dörner

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: March 28, 2024

Abstract Psilocybin, the natural hallucinogen produced by Psilocybe (“magic”) mushrooms, holds great promise for treatment of depression and several other mental health conditions. The final step in psilocybin biosynthetic pathway, dimethylation tryptophan-derived intermediate norbaeocystin, is catalysed PsiM. Here we present atomic resolution (0.9 Å) crystal structures PsiM trapped at various stages its reaction cycle, providing detailed insight into SAM-dependent methylation mechanism. Structural phylogenetic analyses suggest that derives from epitranscriptomic N 6 -methyladenosine writers METTL16 family, which further supported observation bound substrates physicochemically mimic RNA. Inherent limitations ancestral monomethyltransferase scaffold hamper efficiency assembly leave incapable catalysing trimethylation to aeruginascin. results our study will support bioengineering efforts aiming create novel variants with improved therapeutic properties.

Language: Английский

Citations

11
The effect of exercise and physical activity on skeletal muscle epigenetics and metabolic adaptations DOI
Gregg S. Mallett

European Journal of Applied Physiology, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 8, 2025

Language: Английский

Citations

1
Analytical strategies for quantifying methylated selenium species in staple crops: methods, and emerging techniques DOI Creative Commons
Angstone Thembachako Mlangeni,

Fatema Jagot,

Sydney Namaumbo

et al.

CHINESE JOURNAL OF ANALYTICAL CHEMISTRY (CHINESE VERSION), Journal Year: 2025, Volume and Issue: unknown, P. 100511 - 100511

Published: Feb. 1, 2025

Language: Английский

Citations

1
Synthetic Reagents for Enzyme‐Catalyzed Methylation DOI Open Access

Xiaojin Wen,

Florian Leisinger,

Viviane Leopold

et al.

Angewandte Chemie International Edition, Journal Year: 2022, Volume and Issue: 61(41)

Published: Aug. 22, 2022

Late-stage methylation is a key technology in the development of pharmaceutical compounds. Methyltransferase biocatalysis may provide powerful options to insert methyl groups into complex molecules with high regio- and chemoselectivity. The challenge large-scale application methyltransferases their dependence on S-adenosylmethionine (SAM) as stoichiometric, thus exceedingly expensive co-substrate. As solution this problem, we others have explored use halides reagents for situ regeneration SAM. However, need handle volatile electrophiles, such iodide (MeI), also hamper applications at scale. more practical solution, now developed an enzyme-catalyzed process SAM toluene sulfonate. Herein, describe enzymes from thiopurine methyltransferase family that accept sulfate- sulfonate-based donors convert S-adenosylhomocysteine efficiencies rival MeI-based reactions.

Language: Английский

Citations

36
A SAM analogue-utilizing ribozyme for site-specific RNA alkylation in living cells DOI Creative Commons
Takumi Okuda, Ann‐Kathrin Lenz, Florian Seitz

et al.

Nature Chemistry, Journal Year: 2023, Volume and Issue: 15(11), P. 1523 - 1531

Published: Sept. 4, 2023

Post-transcriptional RNA modification methods are in high demand for site-specific labelling and analysis of functions. In vitro-selected ribozymes attractive tools research have the potential to overcome some limitations chemoenzymatic approaches with repurposed methyltransferases. Here we report an alkyltransferase ribozyme that uses a synthetic, stabilized S-adenosylmethionine (SAM) analogue catalyses transfer propargyl group specific adenosine target RNA. Almost quantitative conversion was achieved within 1 h under wide range reaction conditions vitro, including physiological magnesium ion concentrations. A genetically encoded version SAM analogue-utilizing (SAMURI) expressed HEK293T cells, intracellular propargylation confirmed by fluorescent labelling. SAMURI is general tool installation smallest tag azide-alkyne click chemistry, which can be further functionalized fluorophores, affinity tags or other functional probes.

Language: Английский

Citations

22
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
Enzymatic Fluoroethylation by a Fluoroethyl Selenium Analogue of S-Adenosylmethionine DOI

Nanhai Yu,

Huimin Zhao, Wenrui Wang

et al.

ACS Catalysis, Journal Year: 2024, Volume and Issue: 14(8), P. 6211 - 6216

Published: April 9, 2024

Fluorine is a unique element with important roles in medicinal chemistry, agrochemistry, and materials chemistry. The fluoroethyl group an fluoroalkyl functional unit that widely used clinical drugs, 19F probes 18F PET diagnostic drugs. Chemo- regioselective fluoroethylation difficult chemical synthesis. To date, no enzymatic reaction for selective has been reported. Based on the widespread natural methyl donor S-adenosine-l-methionine (SAM), we designed synthesized SAM analogue (FEt-SAM). A stability study revealed FEt-SAM was very labile under physiological conditions gave fluorine-elimination product vinyl-SAM. We circumvented this problem by replacing S Se to give Se-adenosyl-l-selenomethionine (FEt-SeAM). By using halide methyltransferase (HMT) its mutant situ production of FEt-SeAM, created cascade reactions HMT methyltransferases fluoroethylated several O-, N-, S-, C-nucleophiles. For did not recognize FEt-SeAM well, such as DnrK NovO, simple mutagenesis conserved hydrophobic residues (Leu Ile) binding pocket smaller amino acids significantly increased activities. Therefore, have provided useful tool late-stage products This method could also be enzymatically prepare NMR tests.

Language: Английский

Citations

8