Identification of Fluorinases from Streptomyces sp MA37, Norcardia brasiliensis, and Actinoplanes sp N902-109 by Genome Mining

Authors

  • Dr. Hai Deng,

    Corresponding author
    1. UK Marine Biodiscovery Centre, Department of Chemistry, University of Aberdeen, Meston Walk, Aberdeen AB24 3UE (UK)
    • Hai Deng, UK Marine Biodiscovery Centre, Department of Chemistry, University of Aberdeen, Meston Walk, Aberdeen AB24 3UE (UK)

      David O'Hagan, School of Chemistry and Biomedical Sciences Research Centre, University of St Andrews, North Haugh, St Andrews KY169ST (UK)

    Search for more papers by this author
    • These authors contributed equally to this work.

  • Dr. Long Ma,

    1. School of Chemistry and Biomedical Sciences Research Centre, University of St Andrews, North Haugh, St Andrews KY169ST (UK)
    Search for more papers by this author
    • These authors contributed equally to this work.

  • Nouchali Bandaranayaka,

    1. School of Chemistry and Biomedical Sciences Research Centre, University of St Andrews, North Haugh, St Andrews KY169ST (UK)
    Search for more papers by this author
    • These authors contributed equally to this work.

  • Zhiwei Qin,

    1. UK Marine Biodiscovery Centre, Department of Chemistry, University of Aberdeen, Meston Walk, Aberdeen AB24 3UE (UK)
    Search for more papers by this author
  • Greg Mann,

    1. School of Chemistry and Biomedical Sciences Research Centre, University of St Andrews, North Haugh, St Andrews KY169ST (UK)
    Search for more papers by this author
  • Dr. Kwaku Kyeremeh,

    1. Department of Chemistry, FGO Torto Building, University of Ghana, P.O. Box LG56, Legon-Accra (Ghana)
    Search for more papers by this author
  • Dr. Yi Yu,

    1. Key Laboratory of Combinatory Biosynthesis and Drug Discovery (Ministry of Education), School of Pharmaceutical Sciences, Wuhan University, 185 East Lake Road, Wuhan 430071 (P. R. China)
    Search for more papers by this author
  • Dr. Thomas Shepherd,

    1. The James Hutton Institute, Invergowrie, Dundee, DD2 5DA (UK)
    Search for more papers by this author
  • Prof. James H. Naismith,

    1. School of Chemistry and Biomedical Sciences Research Centre, University of St Andrews, North Haugh, St Andrews KY169ST (UK)
    Search for more papers by this author
  • Prof. David O'Hagan

    Corresponding author
    1. School of Chemistry and Biomedical Sciences Research Centre, University of St Andrews, North Haugh, St Andrews KY169ST (UK)
    • Hai Deng, UK Marine Biodiscovery Centre, Department of Chemistry, University of Aberdeen, Meston Walk, Aberdeen AB24 3UE (UK)

      David O'Hagan, School of Chemistry and Biomedical Sciences Research Centre, University of St Andrews, North Haugh, St Andrews KY169ST (UK)

    Search for more papers by this author

Abstract

The fluorinase is an enzyme that catalyses the combination of S-adenosyl-L-methionine (SAM) and a fluoride ion to generate 5′-fluorodeoxy adenosine (FDA) and L-methionine through a nucleophilic substitution reaction with a fluoride ion as the nucleophile. It is the only native fluorination enzyme that has been characterised. The fluorinase was isolated in 2002 from Streptomyces cattleya, and, to date, this has been the only source of the fluorinase enzyme. Herein, we report three new fluorinase isolates that have been identified by genome mining. The novel fluorinases from Streptomyces sp. MA37, Nocardia brasiliensis, and an Actinoplanes sp. have high homology (80–87 % identity) to the original S. cattleya enzyme. They all possess a characteristic 21-residue loop. The three newly identified genes were overexpressed in E. coli and shown to be fluorination enzymes. An X-ray crystallographic study of the Streptomyces sp. MA37 enzyme demonstrated that it is almost identical in structure to the original fluorinase. Culturing of the Streptomyces sp. MA37 strain demonstrated that it not only also elaborates the fluorometabolites, fluoroacetate and 4-fluorothreonine, similar to S. cattleya, but this strain also produces a range of unidentified fluorometabolites. These are the first new fluorinases to be reported since the first isolate, over a decade ago, and their identification extends the range of fluorination genes available for fluorination biotechnology.

Ancillary