title: Elucidation of the biosynthesis of the methane catalyst coenzyme F430.
abstract: Methane biogenesis in methanogens is mediated by methyl-coenzyme M reductase, an enzyme that is also responsible for the utilization of methane through anaerobic methane oxidation. The enzyme uses an ancillary factor called coenzyme F430, a nickel-containing modified tetrapyrrole that promotes catalysis through a methyl radical/Ni(ii)-thiolate intermediate. However, it is unclear how coenzyme F430is synthesized from the common primogenitor uroporphyrinogen iii, incorporating 11 steric centres into the macrocycle, although the pathway must involve chelation, amidation, macrocyclic ring reduction, lactamization and carbocyclic ring formation. Here we identify the proteins that catalyse the biosynthesis of coenzyme F430from sirohydrochlorin, termed CfbA-CfbE, and demonstrate their activity. The research completes our understanding of how the repertoire of tetrapyrrole-based pigments are constructed, permitting the development of recombinant systems to use these metalloprosthetic groups more widely.
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent CT2 7NJ, UK.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Moore
authors.affiliation: Institute of Biochemistry, Leipzig University, Brüderstrasse 34, 04103 Leipzig, Germany.
authors.forename: Sven T
authors.initials: ST
authors.lastname: Sowa
authors.affiliation: Institute of Biochemistry, Leipzig University, Brüderstrasse 34, 04103 Leipzig, Germany.
authors.forename: Christopher
authors.initials: C
authors.lastname: Schuchardt
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent CT2 7NJ, UK.
authors.forename: Evelyne
authors.initials: E
authors.lastname: Deery
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent CT2 7NJ, UK.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Lawrence
authors.affiliation: Institute of Biochemistry, Leipzig University, Brüderstrasse 34, 04103 Leipzig, Germany.
authors.forename: José Vazquez
authors.initials: JV
authors.lastname: Ramos
authors.affiliation: Institute of Analytical Chemistry, Leipzig University, Linnéstrasse 3, 04103 Leipzig, Germany.
authors.forename: Susan
authors.initials: S
authors.lastname: Billig
authors.affiliation: Institute of Analytical Chemistry, Leipzig University, Linnéstrasse 3, 04103 Leipzig, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Birkemeyer
authors.affiliation: Department of Chemistry and Department of Biosciences, Durham University, Durham DH1 3LE, UK.
authors.forename: Peter T
authors.initials: PT
authors.lastname: Chivers
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent CT2 7NJ, UK.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Howard
authors.affiliation: Manchester Institute of Biotechnology, School of Chemistry, University of Manchester, 131 Princess Street, Manchester M1 7DN, UK.
authors.forename: Stephen E J
authors.initials: SE
authors.lastname: Rigby
authors.affiliation: Institute of Biochemistry, Leipzig University, Brüderstrasse 34, 04103 Leipzig, Germany.
authors.forename: Gunhild
authors.initials: G
authors.lastname: Layer
authors.affiliation: School of Biosciences, University of Kent, Giles Lane, Canterbury, Kent CT2 7NJ, UK.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Warren
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism; D055162:Biocatalysis; D053898:Biosynthetic Pathways / Q000235:genetics; D003067:Coenzymes / Q000096:biosynthesis* / Q000737:chemistry; D008190:Lyases / Q000235:genetics / Q000378:metabolism; D008665:Metalloporphyrins / Q000737:chemistry / Q000378:metabolism*; D008697:Methane / Q000031:analogs & derivatives / Q000096:biosynthesis* / Q000378:metabolism; D017021:Methanosarcina barkeri / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D005810:Multigene Family; D009532:Nickel / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D045725:Tetrapyrroles / Q000096:biosynthesis* / Q000737:chemistry; D014578:Uroporphyrins / Q000737:chemistry / Q000378:metabolism
chemical_list: D003067:Coenzymes; D008665:Metalloporphyrins; D045725:Tetrapyrroles; D014578:Uroporphyrins; C051224:methyl radical; C014188:sirohydrochlorin; C023540:factor F430; D009532:Nickel; D010088:Oxidoreductases; C021572:methyl coenzyme M reductase; D000581:Amidohydrolases; D008190:Lyases; D008697:Methane