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      A Marine Viral Halogenase that Iodinates Diverse Substrates

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          Abstract

          Oceanic cyanobacteria are the most abundant oxygen-generating phototrophs on our planet, and therefore, important to life. These organisms are infected by viruses called cyanophages, recently shown to encode metabolic genes that modulate host photosynthesis, phosphorus cycling and nucleotide metabolism. Herein, we report the characterisation of a wild type flavin-dependent viral halogenase (VirX1) from a cyanophage. Notably, halogenases have been previously associated with secondary metabolism, tailoring natural products. Exploration of this viral halogenase reveals it capable of regioselective halogenation of a diverse range of substrates, with a preference for forming aryl iodide species; this has potential implications for the metabolism of the infected host. Until recently, a flavin-dependent halogenase (FDH) capable of iodination in vitro had not been reported. VirX1 is interesting from a biocatalytic perspective showing strikingly broad substrate flexibility, and a clear preference for iodination, as illustrated by kinetic analysis. These factors together render it an attractive tool for synthesis.

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          Most cited references24

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          Biosynthesis of polybrominated aromatic organic compounds by marine bacteria

          Polybrominated diphenyl ethers (PBDEs) and polybrominated bipyrroles are natural products that bioaccumulate in the marine food chain. PBDEs have attracted widespread attention due to their persistence in the environment and potential toxicity to humans. However, the natural origins of PBDE biosynthesis are not known. Here we report marine bacteria as producers of PBDEs and establish a genetic and molecular foundation for their production that unifies paradigms for the elaboration of bromophenols and bromopyrroles abundant in marine biota. We provide biochemical evidence of marine brominase enzymes revealing decarboxylative-halogenation enzymology previously unknown among halogenating enzymes. Biosynthetic motifs discovered in our study were used to mine sequence databases to discover unrealized marine bacterial producers of organobromine compounds.
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            Enzymatic Halogenation and Dehalogenation Reactions: Pervasive and Mechanistically Diverse.

            Naturally produced halogenated compounds are ubiquitous across all domains of life where they perform a multitude of biological functions and adopt a diversity of chemical structures. Accordingly, a diverse collection of enzyme catalysts to install and remove halogens from organic scaffolds has evolved in nature. Accounting for the different chemical properties of the four halogen atoms (fluorine, chlorine, bromine, and iodine) and the diversity and chemical reactivity of their organic substrates, enzymes performing biosynthetic and degradative halogenation chemistry utilize numerous mechanistic strategies involving oxidation, reduction, and substitution. Biosynthetic halogenation reactions range from simple aromatic substitutions to stereoselective C-H functionalizations on remote carbon centers and can initiate the formation of simple to complex ring structures. Dehalogenating enzymes, on the other hand, are best known for removing halogen atoms from man-made organohalogens, yet also function naturally, albeit rarely, in metabolic pathways. This review details the scope and mechanism of nature's halogenation and dehalogenation enzymatic strategies, highlights gaps in our understanding, and posits where new advances in the field might arise in the near future.
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              Tryptophan 7-halogenase (PrnA) structure suggests a mechanism for regioselective chlorination.

              Chlorinated natural products include vancomycin and cryptophycin A. Their biosynthesis involves regioselective chlorination by flavin-dependent halogenases. We report the structural characterization of tryptophan 7-halogenase (PrnA), which regioselectively chlorinates tryptophan. Tryptophan and flavin adenine dinucleotide (FAD) are separated by a 10 angstrom-long tunnel and bound by distinct enzyme modules. The FAD module is conserved in halogenases and is related to flavin-dependent monooxygenases. On the basis of biochemical studies, crystal structures, and by analogy with monooxygenases, we predict that FADH2 reacts with O2 to make peroxyflavin, which is decomposed by Cl-. The resulting HOCl is guided through the tunnel to tryptophan, where it is activated to participate in electrophilic aromatic substitution.
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                Author and article information

                Journal
                101499734
                Nat Chem
                Nat Chem
                Nature chemistry
                1755-4330
                1755-4349
                10 September 2019
                14 October 2019
                December 2019
                14 April 2020
                : 11
                : 12
                : 1091-1097
                Affiliations
                [1 ] School of Chemistry, University of St Andrews, North Haugh, St Andrews, Fife, KY16 9ST, UK Biomedical Sciences Research Complex, University of St Andrews, North Haugh, St Andrews, Fife, KY16 9ST, UK
                [2 ]Department of Chemistry, University of York, Heslington, York YO10 5DD, UK
                [3 ]Syngenta, Jealott’s Hill International Research Centre, Bracknell, Berkshire, RG42 6EY, UK
                [4 ]QEDDI, Staff House Road, The University of Queensland, Brisbane, QLD 4072, Australia
                [5 ]Division of Structural Biology, Wellcome Trust Centre of Human Genomics, Roosevelt Drive, Oxford, OX3 7BN; Research Complex at Harwell, Rutherford Laboratory, Didcot, OX11 0FA; The Rosalind Franklin Institute, Didcot OX11 0FA
                Author notes
                Correspondence and requests for materials should be addressed to R.J.M.G. RJMG@ 123456St-Andrews.ac.uk
                Article
                EMS84344
                10.1038/s41557-019-0349-z
                6875430
                31611633
                f74123d2-3d5c-49c7-9a7a-4d86dae43fc9

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