title: Insights into the Biosynthesis of Dehydroalanines in Goadsporin.
abstract: Dehydroalanines in goadsporin are proposed to be formed by GodF and GodG, which show slight homology to the N-terminal glutamylation and C-terminal elimination domains, respectively, of LanB, a class I lanthipeptide dehydratase. Although similar, separated-type LanBs are conserved among thiopeptides and indispensable for their biosynthesis and biological activities, these enzymes had not yet been characterized. Here, we identified goadsporin B, which has unmodified Ser4 and Ser14, from both godF and godG disruptants. The godG disruptant also produced goadsporin C, a glutamylated-Ser4 variant of goadsporin B. These results suggested that dehydroalanines are formed by glutamylation and glutamate elimination. NMR analysis revealed for the first time that the glutamyl group was attached to a serine via an ester bond, by the catalysis of LanB-type enzymes. Our findings provide insights into the function of separated-type LanBs involved in the biosynthesis of goadsporin and thiopeptides.
authors.affiliation: Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
authors.forename: Taro
authors.initials: T
authors.lastname: Ozaki
authors.affiliation: Department of Biotechnology, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama, 939-0398, Japan.
authors.forename: Yukari
authors.initials: Y
authors.lastname: Kurokawa
authors.affiliation: Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
authors.forename: Shohei
authors.initials: S
authors.lastname: Hayashi
authors.affiliation: Department of Biotechnology, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama, 939-0398, Japan.
authors.forename: Naoya
authors.initials: N
authors.lastname: Oku
authors.affiliation: Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
authors.forename: Shumpei
authors.initials: S
authors.lastname: Asamizu
authors.affiliation: Department of Biotechnology, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama, 939-0398, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Igarashi
authors.affiliation: Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan. aonaka@mail.ecc.u-tokyo.ac.jp.|Department of Biotechnology, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama, 939-0398, Japan. aonaka@mail.ecc.u-tokyo.ac.jp.
authors.forename: Hiroyasu
authors.identifier: http://orcid.org/0000-0003-4536-1881
authors.initials: H
authors.lastname: Onaka
mesh_terms: D039903:Actinobacteria / Q000378:metabolism; D000409:Alanine / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006836:Hydro-Lyases / Q000737:chemistry / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins; D005810:Multigene Family; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D021241:Spectrometry, Mass, Electrospray Ionization; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; C449027:goadsporin; C015102:dehydroalanine; D006836:Hydro-Lyases; D000409:Alanine
keywords: RiPPs; biosynthesis; dehydroalanine; glutamylation; goadsporin; natural products