title: Multi-genome analysis identifies functional and phylogenetic diversity of basidiomycete adenylate-forming reductases.
abstract: Among the invaluable benefits of basidiomycete genomics is the dramatically enhanced insight into the potential capacity to biosynthesize natural products. This study focuses on adenylate-forming reductases, which is a group of natural product biosynthesis enzymes that resembles non-ribosomal peptide synthetases, yet serves to modify one substrate, rather than to condense two or more building blocks. Phylogenetically, these reductases fall in four classes. The phylogeny of Heterobasidion annosum (Russulales) and Serpula lacrymans (Boletales) adenylate-forming reductases was investigated. We identified a previously unrecognized phylogenetic branch within class III adenylate-forming reductases. Three representatives were heterologously produced and their substrate preferences determined in vitro: NPS9 and NPS11 of S. lacrymans preferred l-threonine and benzoic acid, respectively, while NPS10 of H. annosum accepted phenylpyruvic acid best. We also investigated two class IV adenylate-forming reductases of Coprinopsis cinerea, which each were active with l-alanine, l-valine, and l-serine as substrates. Our results show that adenylate-forming reductases are functionally more diverse than previously recognized. As none of the natural products known from the species investigated in this study includes the identified substrates of their respective reductases, our findings may help further explore the diversity of these basidiomycete secondary metabolomes.
authors.affiliation: Friedrich-Schiller-Universität Jena, Department Pharmaceutical Microbiology at the Hans-Knöll-Institute, Winzerlaer Strasse 2, 07745 Jena, Germany.
authors.forename: Eileen
authors.initials: E
authors.lastname: Brandenburger
authors.affiliation: Friedrich-Schiller-Universität Jena, Department Pharmaceutical Microbiology at the Hans-Knöll-Institute, Winzerlaer Strasse 2, 07745 Jena, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Braga
authors.affiliation: ETH Zürich, Institute of Microbiology, Vladimir-Prelog-Weg 4, 8093 Zürich, Switzerland.
authors.forename: Anja
authors.initials: A
authors.lastname: Kombrink
authors.affiliation: Friedrich-Schiller-Universität Jena, Junior Research Group Synthetic Microbiology at the Hans-Knöll-Institute, Adolf-Reichwein-Strasse 23, 07745 Jena, Germany.
authors.forename: Gerald
authors.initials: G
authors.lastname: Lackner
authors.affiliation: Friedrich-Schiller-Universität Jena, Department Pharmaceutical Microbiology at the Hans-Knöll-Institute, Winzerlaer Strasse 2, 07745 Jena, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Gressler
authors.affiliation: ETH Zürich, Institute of Microbiology, Vladimir-Prelog-Weg 4, 8093 Zürich, Switzerland.
authors.forename: Markus
authors.initials: M
authors.lastname: Künzler
authors.affiliation: Friedrich-Schiller-Universität Jena, Department Pharmaceutical Microbiology at the Hans-Knöll-Institute, Winzerlaer Strasse 2, 07745 Jena, Germany. Electronic address: dirk.hoffmeister@leibniz-hki.de.
authors.forename: Dirk
authors.initials: D
authors.lastname: Hoffmeister
mesh_terms: D000363:Agaricales / Q000201:enzymology* / Q000235:genetics; D014644:Genetic Variation; D016681:Genome, Fungal; D010088:Oxidoreductases / Q000145:classification* / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D011994:Recombinant Proteins; D010088:Oxidoreductases
keywords: Coprinopsis; Genome; Heterobasidion; Nonribosomal peptide synthetase; Reductase; Serpula