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

A New Age of Biocatalysis Enabled by Generic Activation Modes DOI Creative Commons
Shubhanshu Jain, Felipe Ospina, Stephan C. Hammer

et al.

JACS Au, Journal Year: 2024, Volume and Issue: 4(6), P. 2068 - 2080

Published: May 31, 2024

Biocatalysis is currently undergoing a profound transformation. The field moves from relying on nature's chemical logic to discipline that exploits generic activation modes, allowing for novel biocatalytic reactions and, in many instances, entirely new chemistry. Generic modes enable wide range of reaction types and played pivotal role advancing the fields organo- photocatalysis. This perspective aims summarize principal harnessed enzymes develop biocatalysts. Although extensively researched past, highlighted when applied within enzyme active sites, facilitate transformations have largely eluded efficient selective catalysis. advance attributed multiple tunable interactions substrate binding pocket precisely control competing pathways transition states. We will highlight cases synthetic methodologies achieved by engineered provide insights into potential future developments this rapidly evolving field.

Language: Английский

Citations

8
SETDB1 as a cancer target: Challenges and perspectives in drug design DOI
Haifa Hassanie, André Berndt Penteado, Larissa Costa de Almeida

et al.

RSC Medicinal Chemistry, Journal Year: 2024, Volume and Issue: 15(5), P. 1424 - 1451

Published: Jan. 1, 2024

The SETDB1 protein's structure and primary function are succinctly described in this review article. It also gives light on potential inhibitory mechanisms that, the future, can be employed as a cancer target.

Language: Английский

Citations

6
Recent developments in the enzymatic modifications of steroid scaffolds DOI Creative Commons
Huibin Wang, Ikuro Abe

Organic & Biomolecular Chemistry, Journal Year: 2024, Volume and Issue: 22(18), P. 3559 - 3583

Published: Jan. 1, 2024

This review highlights the recent advancements in enzymatic modifications of steroid scaffolds, emphasizing hydroxylation, ketoreduction, dehydrogenation, cascade reactions, and other modifications.

Language: Английский

Citations

6
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
Biomimetic S‐Adenosylmethionine Regeneration Starting from Multiple Byproducts Enables Biocatalytic Alkylation with Radical SAM Enzymes DOI Creative Commons

Lukas Gericke,

Dipali Mhaindarkar,

Lukas Karst

et al.

ChemBioChem, Journal Year: 2023, Volume and Issue: 24(9)

Published: March 21, 2023

Abstract S ‐Adenosylmethionine (SAM) is an enzyme cofactor involved in methylation, aminopropyl transfer, and radical reactions. This versatility renders SAM‐dependent enzymes of great interest biocatalysis. The usage SAM analogues adds to this diversity. However, high cost instability the impedes investigation these enzymes. While regeneration protocols from methyltransferase (MT) byproduct ‐adenosylhomocysteine are available, transferases not covered. Here, we report a set efficient one‐pot systems supply or regenerate for all three classes. systems’ flexibility showcased by transfer ethyl group with cobalamin‐dependent MT using ‐adenosylethionine as cofactor. shows potential (analogue) application diverse chemistry, well mechanistic studies analogues.

Language: Английский

Citations

13
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
Protein arginine methyltransferase 3: A crucial regulator in metabolic reprogramming and gene expression in cancers DOI Creative Commons
Shih‐Han Hsu, Wen‐Chun Hung

Cancer Letters, Journal Year: 2022, Volume and Issue: 554, P. 216008 - 216008

Published: Nov. 16, 2022

Post-translational modification (PTM) of proteins increases proteome diversity, which is critical for maintaining cellular homeostasis. The importance protein methylation in the regulation diverse biological processes has been highlighted past decades. Methylation arginine residue on catalyzed by members methyltransferase (PRMT) family. PRMTs play indispensable roles various pathways that regulate cancer development, progression, and drug response. In this review, we discuss role PRMT3, a member PRMT family, controlling oncogenic processes. Additionally, effects PRMT3 regulatory involved transcription, post-transcriptional control, ribosomal maturation, translation, synthesis, metabolic signaling are summarized. Moreover, recent progresses development inhibitors introduced. Overall, review highlights tumorigenesis discusses underlying mechanisms modulates metabolism gene expression. These results also provide molecular basis therapeutic modalities targeting PRMT3.

Language: Английский

Citations

19
Biocatalytic Fluoroalkylation Using Fluorinated S-Adenosyl-l-methionine Cofactors DOI
Wenping Ding,

Minqi Zhou,

Huayu Li

et al.

Organic Letters, Journal Year: 2023, Volume and Issue: 25(30), P. 5650 - 5655

Published: July 25, 2023

Modification of organic molecules with fluorine functionalities offers a critical approach to develop new pharmaceuticals. Here, we report multienzyme strategy for biocatalytic fluoroalkylation using S-adenosyl-l-methionine (SAM)-dependent methyltransferases (MTs) and fluorinated SAM cofactors prepared from ATP l-methionine analogues by an engineered human methionine adenosyltransferase hMAT2AI322A. This work introduces the first example 3,3-difluoroallylation. Importantly, this can be applied late-stage site-selective complex molecule vancomycin conversions up 99%.

Language: Английский

Citations

11
Promiscuous acyltransferases for ester and amide synthesis in aqueous solution DOI Creative Commons

Benjamin Baumert,

Hannes Meinert, Clemens Cziegler

et al.

Catalysis Today, Journal Year: 2024, Volume and Issue: 442, P. 114925 - 114925

Published: July 9, 2024

Hydrolases are well-known for hydrolyzing esters, amides, carbamates, peptides, or acid anhydrides in the presence of water. However, some them also capable catalyzing reverse reaction (condensation) under certain conditions aqueous systems. Hence, these enzymes called promiscuous hydrolases/acyltransferases. This review deals with their discovery, background information on mechanism action, and significant improvements by enzyme engineering to both enhance product formation decrease undesired hydrolysis targeted acyl products. Their applications biocatalysis exemplified synthesis a wide range esters amides systems, including preparative-scale processes combination hydrolases/acyltransferases other cascade reactions utilize alternative feedstocks from renewable resources, example. Complementary, use ATP-dependent amide synthesizing is covered. Together, represent practically useful alternatives well-established chemical reactions, operating solutions that appeal different industries.

Language: Английский

Citations

4
Molecular basis for azetidine-2-carboxylic acid biosynthesis DOI Creative Commons
Tim J. Klaubert,

Jonas Gellner,

Charles Bernard

et al.

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: Feb. 4, 2025

Language: Английский

Citations

0