Biochemical Studies of a Cyanobacterial Halogenase Support the Involvement of a Dimetal Cofactor DOI Creative Commons
M. Wang, Nathaniel R. Glasser, Mrutyunjay A. Nair

et al.

Biochemistry, Journal Year: 2025, Volume and Issue: unknown

Published: April 29, 2025

Halogenation is a prominent transformation in natural product biosynthesis, with over 5000 halogenated products known to date. Biosynthetic pathways accomplish the synthetic challenge of selective halogenation, especially at unactivated sp3 carbon centers, using halogenase enzymes. The CylC, discovered as part cylindrocyclophane (cyl) biosynthetic pathway, performs highly chlorination reaction on an center and proposed use dimetal cofactor. Putative halogenases are widely distributed across cyanobacterial pathways. However, rigorous vitro biochemical structural characterization these enzymes has been challenging. Here, we report additional bioinformatic analyses putative newly identified CylC homologue. Site-directed mutagenesis identifies conserved metal-binding residues, Mössbauer spectroscopy provides direct evidence for presence diiron cofactor halogenases. These insights suggest mechanistic parallels between mononuclear nonheme iron halogenases, potential guide further engineering this unique subfamily metalloenzymes.

Language: Английский

Fungal RiPPs Side Chain Macrocyclization Catalyzed by Copper-Dependent DUF3328 Enzyme DOI
Chen‐Yu Chiang, M. Ohashi, Yi Tang

et al.

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

Published: March 3, 2025

Fungal enzymes containing domain of unknown function (DUF) 3328 have been implicated in the oxidative maturation ribosomally synthesized and post-translationally modified peptides (RiPPs). We report here functional characterization one such enzyme, AprY, involved biosynthesis RiPP asperipin-2a. Biochemical reconstitution AprY showed enzyme catalyzes two consecutive C-O cross-linking reactions between tyrosines beta-carbons residues core hexapeptide. The side-chain macrocyclization activities are copper- oxygen-dependent do not require a leader peptide sequence.

Language: Английский

Citations

0
Biochemical Studies of a Cyanobacterial Halogenase Support the Involvement of a Dimetal Cofactor DOI Creative Commons
M. Wang, Nathaniel R. Glasser, Mrutyunjay A. Nair

et al.

Biochemistry, Journal Year: 2025, Volume and Issue: unknown

Published: April 29, 2025

Halogenation is a prominent transformation in natural product biosynthesis, with over 5000 halogenated products known to date. Biosynthetic pathways accomplish the synthetic challenge of selective halogenation, especially at unactivated sp3 carbon centers, using halogenase enzymes. The CylC, discovered as part cylindrocyclophane (cyl) biosynthetic pathway, performs highly chlorination reaction on an center and proposed use dimetal cofactor. Putative halogenases are widely distributed across cyanobacterial pathways. However, rigorous vitro biochemical structural characterization these enzymes has been challenging. Here, we report additional bioinformatic analyses putative newly identified CylC homologue. Site-directed mutagenesis identifies conserved metal-binding residues, Mössbauer spectroscopy provides direct evidence for presence diiron cofactor halogenases. These insights suggest mechanistic parallels between mononuclear nonheme iron halogenases, potential guide further engineering this unique subfamily metalloenzymes.

Language: Английский

Citations

0