Genomic Determinants Encode the Reactivity and Regioselectivity of Flavin-Dependent Halogenases in Bacterial Genomes and Metagenomes

Jeon, Jehyun; Lee, Jaehee; Jung, Se Min; Shin, Jae Hong; Song, Woon Ju; Rho, Mina

doi:10.1128/msystems.00053-21

Cited by 4 publications

(2 citation statements)

References 80 publications

(114 reference statements)

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“…These profiling efforts have determined previously unknown data on their binding sites, halide specificity, regioselectivity and substrate specificity. Novel and unlikely substrate scopes were unveiled, especially related to large, three-dimensionally complex compounds, a task which would previously require several rounds of directed evolution to accomplish ( Fisher et al, 2019 ; Jeon et al, 2021 ). Additional evidence in favor of sequence-function correlation is an in vitro prototyping workflow for screening a library of O -methyltransferase enzymes ( Haslinger et al, 2021 ).…”

Section: Discussion and Perspectivesmentioning

confidence: 99%

Late-stage diversification of bacterial natural products through biocatalysis

Lazic,

Filipovic,

Pantelic

et al. 2024

Front. Bioeng. Biotechnol.

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Bacterial natural products (BNPs) are very important sources of leads for drug development and chemical novelty. The possibility to perform late-stage diversification of BNPs using biocatalysis is an attractive alternative route other than total chemical synthesis or metal complexation reactions. Although biocatalysis is gaining popularity as a green chemistry methodology, a vast majority of orphan sequenced genomic data related to metabolic pathways for BNP biosynthesis and its tailoring enzymes are underexplored. In this review, we report a systematic overview of biotransformations of 21 molecules, which include derivatization by halogenation, esterification, reduction, oxidation, alkylation and nitration reactions, as well as degradation products as their sub-derivatives. These BNPs were grouped based on their biological activities into antibacterial (5), antifungal (5), anticancer (5), immunosuppressive (2) and quorum sensing modulating (4) compounds. This study summarized 73 derivatives and 16 degradation sub-derivatives originating from 12 BNPs. The highest number of biocatalytic reactions was observed for drugs that are already in clinical use: 28 reactions for the antibacterial drug vancomycin, followed by 18 reactions reported for the immunosuppressive drug rapamycin. The most common biocatalysts include oxidoreductases, transferases, lipases, isomerases and haloperoxidases. This review highlights biocatalytic routes for the late-stage diversification reactions of BNPs, which potentially help to recognize the structural optimizations of bioactive scaffolds for the generation of new biomolecules, eventually leading to drug development.

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Section: Discussion and Perspectivesmentioning

confidence: 99%

Late-stage diversification of bacterial natural products through biocatalysis

Lazic,

Filipovic,

Pantelic

et al. 2024

Front. Bioeng. Biotechnol.

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“…At the C-terminus of the flavin binding loop is a Glu (Glu46 in AbeH, Glu49 in BorH), the Ca of which acts as a pivot point about which the Glu side chain and peptide backbone exchange positions when the flavin binding loop moves from open (Glu side chain points towards vacant isoalloxazine binding site) to closed (Glu side chain flips out of binding site to make room for isoalloxazine).Residues immediately C-terminal to the flavin binding loop (Phe 49/Ser50 in AbeH/PyrH, Val52/Pro53 in BorH/Thal) have been implicated in the inhibition of Trp binding, as they move into the Trp binding site when the flavin binding loop closes and move out when the loop opens, due to backbone shifts caused by the flipping of the adjacent Glu46/Glu49. This region is variously described as the Trp gate(37) and the Trp 1 region(38). Closing of the Trp gate can be seen when comparing the crystal structures of AbeH and PyrH .…”

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confidence: 99%

Crystallographic and thermodynamic evidence of negative cooperativity of flavin and tryptophan binding in the flavin-dependent halogenases AbeH and BorH

Ashaduzzaman,

Lingkon,

De Silva

et al. 2023

Preprint

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The flavin-dependent halogenase AbeH produces 5-chlorotryptophan in the biosynthetic pathway of the chlorinated bisindole alkaloid BE-54017. We report that in vitro, AbeH (assisted by the flavin reductase AbeF) can chlorinate and brominate tryptophan as well as other indole derivatives and substrates with phenyl and quinoline groups. We solved the X-ray crystal structures of AbeH alone and complexed with FAD, as well as crystal structures of the tryptophan- 6-halogenase BorH alone, in complex with 6-chlorotryptophan, and in complex with FAD and tryptophan. Partitioning of FAD and tryptophan into different chains of BorH and failure to incorporate tryptophan into AbeH/FAD crystals suggested that flavin and tryptophan binding are negatively coupled in both proteins. ITC and fluorescence quenching experiments confirmed the ability of both AbeH and BorH to form binary complexes with FAD or tryptophan and the inability of tryptophan to bind to AbeH/FAD or BorH/FAD complexes. FAD could not bind to BorH/tryptophan complexes, but FAD appears to displace tryptophan from AbeH/tryptophan complexes in an endothermic entropically-driven process.

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Trendbericht Biochemie 2022: Späte Funktionalisierung mit Biokatalysatoren aus Naturstoffsynthesen

Haslinger¹,

Schmidt²

2022

Nachr Chem

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Genomic Determinants Encode the Reactivity and Regioselectivity of Flavin-Dependent Halogenases in Bacterial Genomes and Metagenomes

Abstract: Halogenases are playing an important role as tailoring enzymes in biosynthetic pathways. Flavin-dependent tryptophan halogenases (Trp-FDHs) are among the enzymes that have broad substrate scope and high selectivity.

Cited by 4 publications

References 80 publications

Late-stage diversification of bacterial natural products through biocatalysis

Late-stage diversification of bacterial natural products through biocatalysis

Crystallographic and thermodynamic evidence of negative cooperativity of flavin and tryptophan binding in the flavin-dependent halogenases AbeH and BorH

Trendbericht Biochemie 2022: Späte Funktionalisierung mit Biokatalysatoren aus Naturstoffsynthesen

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