RRC ID 89089
Author Shi J, Zhao Z, Yang J, Cheng Z, Li H, Liu Y, Zhao G, Wu M, Du YL.
Title Genome mining and characterization of a heme-dependent enzyme catalyzing intermolecular Nitrogen-Nitrogen bond formation in hydrazinosuccinic acid biosynthesis.
Journal Synth Syst Biotechnol
Abstract Nitrogen-nitrogen (N-N) bond-forming enzymes are rare but play vital roles in both primary and secondary metabolism. Guided by a nitric oxide synthase (NOS)-based genome mining strategy, we report the discovery and characterization of a new heme-dependent enzyme system that catalyzes intermolecular N-N bond formation. Using both in vivo and in vitro reconstitution approaches, we demonstrated that a protein complex, comprising a heme enzyme and a 2[4Fe-4S] ferredoxin partner, mediates the coupling of the α-amine group of l-aspartate with inorganic nitrogen oxide species, such as nitrite or nitric oxide, to generate hydrazinosuccinic acid, a key biosynthetic precursor in several natural product pathways. Structural modeling and site-directed mutagenesis suggest a plausible catalytic mechanism involving the formation of a reactive nitrogen intermediate, potentially a heme-bound nitrene species. These findings reveal a new family of N-N bond-forming biocatalysts that leverage inorganic nitrogen sources, offering valuable tools for genome mining and the synthetic biology.
Volume 10(4)
Pages 1421-1427
Published 2025-12-1
DOI 10.1016/j.synbio.2025.08.006
PII S2405-805X(25)00117-6
PMID 40918557
PMC PMC12408219
Resource
General Microbes JCM35029