Reconstitution of the Final Steps in the Biosynthesis of Valanimycin Reveals the Origin of Its Characteristic Azoxy Moiety DOI
Ziyang Zheng, Jin Xiong,

Junling Bu

et al.

Angewandte Chemie, Journal Year: 2023, Volume and Issue: 136(1)

Published: Nov. 15, 2023

Abstract Valanimycin is an azoxy‐containing natural product isolated from the fermentation broth of Streptomyces viridifaciens MG456‐hF10. While biosynthesis valanimycin has been partially characterized, how azoxy group constructed remains obscure. Herein, membrane protein VlmO and putative hydrazine synthetase ForJ formycin biosynthetic pathway are demonstrated to catalyze N−N bond formation converting O ‐( l ‐seryl)‐isobutyl hydroxylamine into N ‐(isobutylamino)‐ ‐serine. Subsequent installation shown be catalyzed by non‐heme diiron enzyme VlmB in a reaction which single VlmO/ForJ oxidized four electrons yield group. The catalytic cycle appears begin with resting μ‐oxo diferric complex VlmB, as supported Mössbauer spectroscopy. This study also identifies d ‐serine alternative substrate for leading two regioisomers. reactions kinase VlmJ lyase VlmK during final steps established well. was thus fully reconstituted vitro using enzymes VlmO/ForJ, VlmK. Importantly, VlmB‐catalyzed represents first example enzyme‐catalyzed expected proceed atypical mechanism.

Language: Английский

Structural Insights into the N–N Bond-Formation Mechanism of the Heme-Dependent Piperazate Synthase KtzT DOI

Yunyun Yang,

Yanling Li, Licheng Yao

et al.

ACS Catalysis, Journal Year: 2025, Volume and Issue: 15(2), P. 1265 - 1273

Published: Jan. 7, 2025

N–N bond formation plays a critical role in the synthesis of organic compounds and has broad applications producing dyes, pharmaceuticals, functional materials. However, is challenging due to nucleophilicity nitrogen. Here, we determined crystal structures heme-dependent enzyme, KtzT, which catalyzes cyclization l-N5-hydroxyornithine (l-N5-OH-Orn) yield l-piperazate (l-piz) by linking two intramolecular nitrogen atoms. The complex structure KtzTC197A with l-N5-OH-Orn reveals substrate-interaction network, validated through mutagenesis experiments. Notably, N5 atom substrate directly coordinates heme iron, precluding oxygen binding. This supports prior knowledge that KtzT an oxygen-independent reaction. Intriguingly, exhibits distinct conformations our crystals. Based on distance between atoms product accommodation pose KtzTC197A/l-piz structure, conformation 2 likely productive pose, while more extended 1 may be transient state facilitating entry into catalytic tunnel. A potential pathway also proposed. These findings offer structural insights for developing bio- metal-catalyzed methods formation.

Language: Английский

Citations

2
Enzymatic synthesis of azide by a promiscuous N-nitrosylase DOI
Antonio Del Rio Flores, Rui Zhai, David W. Kastner

et al.

Nature Chemistry, Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 27, 2024

Language: Английский

Citations

7
Identification of the Azaserine Biosynthetic Gene Cluster Implicates Hydrazine as an Intermediate to the Diazo Moiety DOI
Zi-Wang Wei, Haruka Niikura, Menghua Wang

et al.

Organic Letters, Journal Year: 2023, Volume and Issue: 25(22), P. 4061 - 4065

Published: May 26, 2023

Azaserine (1) is a natural product and nonproteinogenic amino acid containing diazo group. Here we report the biosynthetic gene cluster for 1 from Glycomyces harbinensis. We then use isotopic feeding, deletion, biochemical experiments to support pathway whereby hydrazinoacetic (2) peptidyl carrier protein-loaded serine (3) are intermediates on route final 1.

Language: Английский

Citations

11
Identification and Analysis of the Biosynthetic Gene Cluster for the Hydrazide-Containing Aryl Polyene Spinamycin DOI
Seiji Kawai,

Akito Yamada,

Danyao Du

et al.

ACS Chemical Biology, Journal Year: 2023, Volume and Issue: 18(8), P. 1821 - 1828

Published: July 27, 2023

Natural products containing nitrogen-nitrogen (N-N) bonds have attracted much attention because of their bioactivities and chemical features. Several recent studies revealed the nitrous acid-dependent N-N bond-forming machinery. However, catalytic mechanisms hydrazide synthesis using acid remain unknown. Herein, we focused on spinamycin, a hydrazide-containing aryl polyene produced by Streptomyces albospinus JCM3399. In S. genome, discovered putative spinamycin biosynthetic gene (spi) cluster genes that encode type II polyketide synthase for secondary metabolism-specific biosynthesis pathway. A inactivation experiment showed this was responsible biosynthesis. feeding stable isotope-labeled sodium nitrite analysis acid-synthesizing enzymes in vitro strongly indicated one nitrogen atoms group derived from acid. substrate specificity SpiA3, which is loading starter onto synthase, bond formation occurs after loading. AMP-dependent ligase SpiA7 catalyzes diazotization an amino benzene ring without hydroxy group, resulting highly reactive diazo intermediate, may be key step formation. Therefore, propose overall pathway spinamycin. This study expands our knowledge microbial metabolism.

Language: Английский

Citations

11
C(sp3)–H Carboxylation via Carbene/Photoredox Cooperative Catalysis DOI

Cullen R. Schull,

Jing Cao,

Sophia R. Mitton-Fry

et al.

ACS Catalysis, Journal Year: 2025, Volume and Issue: 15(2), P. 1287 - 1293

Published: Jan. 7, 2025

C(sp3)–H bond functionalization is a powerful strategy for the synthesis of organic compounds due their abundance in simple starting materials. Photoredox catalysis has led to diverse array enabling activation strategies; however, general platform direct carboxylic acid derivatives remains elusive. Disclosed herein development cooperative NHC/photoredox-catalyzed esterification transformation. This method enables access benzylic, α-heteroatom, and formal β-esterification products moderate high yields under mild reaction conditions.

Language: Английский

Citations

0
Structure and mechanism of haem-dependent nitrogen–nitrogen bond formation in piperazate synthase DOI
M.A. Higgins,

Xinjie Shi,

Jordi Soler Soler

et al.

Nature Catalysis, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 13, 2025

Language: Английский

Citations

0
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 DOI Creative Commons
Yusuke Shikai, Seiji Kawai, Yohei Katsuyama

et al.

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

Citations

10
Bacterial Hydrazine Biosynthetic Pathways Featuring Cupin/Methionyl tRNA Synthetase‐like Enzymes DOI
Kenichi Matsuda, Toshiyuki Wakimoto

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

Citations

3
Phylogeny‐guided Characterization of Bacterial Hydrazine Biosynthesis Mediated by Cupin/methionyl tRNA Synthetase‐like Enzymes DOI
Kenichi Matsuda, Yuto Nakahara, Atina Rizkiya Choirunnisa

et al.

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

Citations

2
Reconstitution of the Final Steps in the Biosynthesis of Valanimycin Reveals the Origin of Its Characteristic Azoxy Moiety DOI
Ziyang Zheng, Jin Xiong,

Junling Bu

et al.

Angewandte Chemie International Edition, Journal Year: 2023, Volume and Issue: 63(1)

Published: Nov. 15, 2023

Valanimycin is an azoxy-containing natural product isolated from the fermentation broth of Streptomyces viridifaciens MG456-hF10. While biosynthesis valanimycin has been partially characterized, how azoxy group constructed remains obscure. Herein, membrane protein VlmO and putative hydrazine synthetase ForJ formycin biosynthetic pathway are demonstrated to catalyze N-N bond formation converting O-(l-seryl)-isobutyl hydroxylamine into N-(isobutylamino)-l-serine. Subsequent installation shown be catalyzed by non-heme diiron enzyme VlmB in a reaction which single VlmO/ForJ oxidized four electrons yield group. The catalytic cycle appears begin with resting μ-oxo diferric complex VlmB, as supported Mössbauer spectroscopy. This study also identifies N-(isobutylamino)-d-serine alternative substrate for leading two regioisomers. reactions kinase VlmJ lyase VlmK during final steps established well. was thus fully reconstituted vitro using enzymes VlmO/ForJ, VlmK. Importantly, VlmB-catalyzed represents first example enzyme-catalyzed expected proceed atypical mechanism.

Language: Английский

Citations

6