Exquisite Complex Reaction Cascade in the Natural 1,2,4-Triazine Assembly

Journal of the American Chemical Society, Journal Year: 2025, Volume and Issue: unknown

Published: March 29, 2025

1,2,4-Triazine ring is a scaffold widely found in biologically active compounds, but how nature makes it remains enigmatic. In this study, we unveil the complex enzymatic and nonenzymatic cascade reactions that assemble 1,2,4-triazine moiety structures of natural products pseudoiodinine toxoflavin. Through biochemical studies, isotope labeling, application substrate analogues, propose plausible pathway for assembly from common precursor riboflavin biosynthesis. This process involves four two-electron oxidation steps, C-N bond formation, decarboxylation, N-N forming step catalyzed by metal-dependent WD40-repeat (WDR) protein. study thus not only provides first biocatalytic route also identifies previously unrecognized catalytic role large WDR protein family.

Language: Английский

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Metabolomics and Genomics Enable the Discovery of a New Class of Nonribosomal Peptidic Metallophores from a Marine Micromonospora

Journal of the American Chemical Society, Journal Year: 2022, Volume and Issue: 145(1), P. 58 - 69

Published: Dec. 19, 2022

Although microbial genomes harbor an abundance of biosynthetic gene clusters, there remain substantial technological gaps that impair the direct correlation newly discovered clusters and their corresponding secondary metabolite products. As example one approach designed to minimize or bridge such gaps, we employed hierarchical clustering analysis principal component (

Language: Английский

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In vitro characterization of nonribosomal peptide synthetase-dependent O-(2-hydrazineylideneacetyl)serine synthesis indicates a stepwise oxidation strategy to generate the α-diazo ester moiety of azaserine

Chemical Science, Journal Year: 2023, Volume and Issue: 14(33), P. 8766 - 8776

Published: Jan. 1, 2023

Azaserine, a natural product containing diazo group, exhibits anticancer activity. In this study, we investigated the biosynthetic pathway to azaserine. The putative azaserine gene (azs) cluster, which contains 21 genes, including those responsible for hydrazinoacetic acid (HAA) synthesis, was discovered using bioinformatics analysis of Streptomyces fragilis genome. Azaserine produced by heterologous expression azs cluster in albus. vitro enzyme assays recombinant Azs proteins revealed as follows. AzsSPTF and carrier protein (CP) AzsQ are used synthesize 2-hydrazineylideneacetyl (HDA) moiety attached from HAA. AzsD transfers HDA C-terminal CP domain AzsN. heterocyclization (Cy) nonribosomal peptide synthetase AzsO synthesizes O-(2-hydrazineylideneacetyl)serine (HDA-Ser) its l-serine moiety-attached thioesterase AzsB hydrolyzes it yield HDA-Ser, appears be converted oxidation. Bioinformatics Cy showed that has conserved DxxxxD motif; however, two amino residues (Thr Asp) important substituted Asn. Site-directed mutagenesis Asp motif (D193 D198) Asn (N414 N447) indicated these four ester bond synthesis. These results azasrine is synthesized stepwise oxidation HAA provided another strategy biosynthesize group.

Language: Английский

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Identification of the p-coumaric acid biosynthetic gene cluster in Kutzneria albida: insights into the diazotization-dependent deamination pathway

Beilstein Journal of Organic Chemistry, Journal Year: 2024, Volume and Issue: 20, P. 1 - 11

Published: Jan. 2, 2024

Recently, we identified the biosynthetic gene cluster of avenalumic acid (

Language: Английский

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Bacterial Hydrazine Biosynthetic Pathways Featuring Cupin/Methionyl tRNA Synthetase‐like Enzymes

ChemBioChem, Journal Year: 2024, Volume and Issue: 25(9)

Published: March 9, 2024

Nitrogen-Nitrogen (N-N) bond-containing functional groups in natural products and synthetic drugs play significant roles exerting biological activities. The mechanisms of N-N bond formation organic molecules have garnered increasing attention over the decades. Recent advances illuminated various enzymatic nonenzymatic strategies, our understanding construction is rapidly expanding. A group didomain proteins with zinc-binding cupin/methionyl-tRNA synthetase (MetRS)-like domains, also known as hydrazine synthetases, generates amino acid-based hydrazines, which serve key biosynthetic precursors diverse functionalities such hydrazone, diazo, triazene, pyrazole, pyridazinone groups. In this review, we summarize current knowledge on pathways employing unique bond-forming machinery.

Language: Английский

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1,6-Conjugate addition of in situ generated aryldiazenes to p-quinone methides

Organic & Biomolecular Chemistry, Journal Year: 2024, Volume and Issue: 22(27), P. 5636 - 5645

Published: Jan. 1, 2024

Herein we report a transition-metal free, base-mediated 1,6-conjugate addition of aryldiazenes to para -quinone methides ( p -QMs).

Language: Английский

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Conserved Enzymatic Cascade for Bacterial Azoxy Biosynthesis

Journal of the American Chemical Society, Journal Year: 2023, Volume and Issue: 145(49), P. 27131 - 27139

Published: Nov. 29, 2023

Azoxy compounds exhibit a wide array of biological activities and possess distinctive chemical properties. Although there has been considerable interest in the biosynthetic mechanisms azoxy metabolites, enzymatic basis responsible for bond formation remained largely enigmatic. In this study, we unveil enzyme cascade that constructs valanimycin biosynthesis. Our research demonstrates pair metalloenzymes, comprising membrane-bound hydrazine synthase nonheme diiron synthase, collaborate to convert an unstable pathway intermediate product through hydrazine-azo-azoxy pathway. Additionally, by characterizing homologues from other metabolite pathways, propose two-enzyme could represent conserved strategy bacteria. These findings provide significant mechanistic insights into N–N should facilitate targeted isolation bioactive genome mining.

Language: Английский

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Biosynthesis of Fungal Natural Products Involving Two Separate Pathway Crosstalk

Journal of Fungi, Journal Year: 2022, Volume and Issue: 8(3), P. 320 - 320

Published: March 21, 2022

Fungal natural products (NPs) usually possess complicated structures, exhibit satisfactory bioactivities, and are an outstanding source of drug leads, such as the cholesterol-lowering lovastatin immunosuppressive mycophenolic acid. The fungal NPs biosynthetic genes always arranged within one single gene cluster (BGC). However, a rare but fascinating phenomenon that crosstalk between two separate BGCs is indispensable to some dimeric biosynthesis has attracted increasing attention. hybridization not only increases structural complexity chemical diversity NPs, also expands scope bioactivities. More importantly, underlying mechanism for this process poorly understood needs further exploration, especially determination each building block construction identification enzyme(s) catalyzing precursors coupling processes Diels–Alder cycloaddition Michael addition. In review, we summarized produced by functional discrete BGCs, highlighted their processes, which might shed new light on genome mining with unprecedented frameworks, provide valuable insights into investigation mysterious mechanisms constructed collaboration BGCs.

Language: Английский

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Phylogeny‐guided Characterization of Bacterial Hydrazine Biosynthesis Mediated by Cupin/methionyl tRNA Synthetase‐like Enzymes

ChemBioChem, Journal Year: 2024, Volume and Issue: 25(7)

Published: Feb. 26, 2024

Cupin/methionyl-tRNA synthetase (MetRS)-like didomain enzymes catalyze nitrogen-nitrogen (N-N) bond formation between Nω-hydroxylamines and amino acids to generate hydrazines, key biosynthetic intermediates of various natural products containing N-N bonds. While the combination these two building blocks leads creation diverse hydrazine products, full extent their structural diversity remains largely unknown. To explore this, we herein conducted phylogeny-guided genome-mining related pathways consisting enzymes: flavin-dependent Nω-hydroxylating monooxygenases (NMOs) that produce Nω-hydroxylamine precursors cupin/MetRS-like couple with via A phylogenetic analysis identified unexplored sequence spaces enzyme families. The biochemical characterization NMOs demonstrated capabilities Nω-hydroxylamines, including those previously not known as Furthermore, five new novel combinations blocks, one non-amino acid blocks: 1,3-diaminopropane putrescine. This study substantially expanded variety forming mediated by enzymes.

Language: Английский

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Production of Phenyldiazene Derivatives Using the Biosynthetic Pathway of an Aromatic Diazo Group‐Containing Natural Product from an Actinomycete

ChemBioChem, Journal Year: 2024, Volume and Issue: 26(1)

Published: Oct. 18, 2024

Abstract The diazo group is an important functional in organic synthesis because it confers high reactivity to the compounds and has been applied various chemical reactions, such as Sandmeyer reaction, Wolff rearrangement, cyclopropanation, C−N bond formation with active methylene compounds. Previously, we revealed that 3‐diazoavenalumic acid (3‐DAA), which potentially produced by several actinomycete species contains aromatic group, a biosynthetic intermediate of avenalumic acid. In this study, aimed construct production system for phenyldiazene derivatives adding culture 3‐DAA‐producing recombinant actinomycete. First, acetoacetanilide its derivatives, have are raw materials arylide yellow dyes, were individually added When their metabolites analyzed, each expected compound phenyldiazenyl moiety was detected extract. Moreover, established one‐pot vitro enzymatic same using highly reactive diazotase, CmaA6. These results showed natural products attractive tool expanding structural diversity both vivo .

Language: Английский

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