A flavin-dependent halogenase from metagenomic analysis prefers bromination over chlorination

PLoS One. 2018 May 10;13(5):e0196797. doi: 10.1371/journal.pone.0196797. eCollection 2018.

Abstract

Flavin-dependent halogenases catalyse halogenation of aromatic compounds. In most cases, this reaction proceeds with high regioselectivity and requires only the presence of FADH2, oxygen, and halide salts. Since marine habitats contain high concentrations of halides, organisms populating the oceans might be valuable sources of yet undiscovered halogenases. A new Hidden-Markov-Model (HMM) based on the PFAM tryptophan halogenase model was used for the analysis of marine metagenomes. Eleven metagenomes were screened leading to the identification of 254 complete or partial putative flavin-dependent halogenase genes. One predicted halogenase gene (brvH) was selected, codon optimised for E. coli, and overexpressed. Substrate screening revealed that this enzyme represents an active flavin-dependent halogenase able to convert indole to 3-bromoindole. Remarkably, bromination prevails also in a large excess of chloride. The BrvH crystal structure is very similar to that of tryptophan halogenases but reveals a substrate binding site that is open to the solvent instead of being covered by a loop.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / metabolism
  • Binding Sites
  • Escherichia coli / metabolism
  • Flavin-Adenine Dinucleotide / analogs & derivatives
  • Flavin-Adenine Dinucleotide / metabolism
  • Flavins / metabolism*
  • Halogenation / physiology
  • Metagenomics / methods
  • Oceans and Seas
  • Oxidoreductases / metabolism*

Substances

  • Bacterial Proteins
  • Flavins
  • Flavin-Adenine Dinucleotide
  • 1,5-dihydro-FAD
  • Oxidoreductases
  • tryptophan halogenase

Grants and funding

The authors acknowledge support for the Article Processing Charge by the Deutsche Forschungsgemeinschaft and the Open Access Publication Fund of Bielefeld University. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.