title: Identification and characterization of two families of F420 H2-dependent reductases from Mycobacteria that catalyse aflatoxin degradation.
abstract: Aflatoxins are polyaromatic mycotoxins that contaminate a range of food crops as a result of fungal growth and contribute to serious health problems in the developing world because of their toxicity and mutagenicity. Although relatively resistant to biotic degradation, aflatoxins can be metabolized by certain species of Actinomycetales. However, the enzymatic basis for their breakdown has not been reported until now. We have identified nine Mycobacterium smegmatis enzymes that utilize the deazaflavin cofactor F(420) H(2) to catalyse the reduction of the α,β-unsaturated ester moiety of aflatoxins, activating the molecules for spontaneous hydrolysis and detoxification. These enzymes belong to two previously uncharacterized F(420) H(2) dependent reductase (FDR-A and -B) families that are distantly related to the flavin mononucleotide (FMN) dependent pyridoxamine 5'-phosphate oxidases (PNPOxs). We have solved crystal structures of an enzyme from each FDR family and show that they, like the PNPOxs, adopt a split barrel protein fold, although the FDRs also possess an extended and highly charged F(420) H(2) binding groove. A general role for these enzymes in xenobiotic metabolism is discussed, including the observation that the nitro-reductase Rv3547 from Mycobacterium tuberculosis that is responsible for the activation of bicyclic nitroimidazole prodrugs belongs to the FDR-A family.
authors.affiliation: CSIRO Ecosystem Sciences, GPO Box 1700, Canberra, ACT 2601, Australia. m.taylor@csiro.au
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Taylor
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Jackson
authors.forename: David B
authors.initials: DB
authors.lastname: Tattersall
authors.forename: Nigel
authors.initials: N
authors.lastname: French
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Peat
authors.forename: Janet
authors.initials: J
authors.lastname: Newman
authors.forename: Lyndall J
authors.initials: LJ
authors.lastname: Briggs
authors.forename: Gauri V
authors.initials: GV
authors.lastname: Lapalikar
authors.forename: Peter M
authors.initials: PM
authors.lastname: Campbell
authors.forename: Colin
authors.initials: C
authors.lastname: Scott
authors.forename: Robyn J
authors.initials: RJ
authors.lastname: Russell
authors.forename: John G
authors.initials: JG
authors.lastname: Oakeshott
mesh_terms: D000348:Aflatoxins / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002384:Catalysis; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D005810:Multigene Family; D009161:Mycobacterium / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D010088:Oxidoreductases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D011485:Protein Binding; D012256:Riboflavin / Q000031:analogs & derivatives* / Q000378:metabolism
chemical_list: D000348:Aflatoxins; D001426:Bacterial Proteins; C015084:coenzyme F420; D010088:Oxidoreductases; D012256:Riboflavin
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