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The genome sequences of two lignin\u2010degrading bacteria, <jats:italic>Ochrobactrum<\/jats:italic> sp., and <jats:italic>Paenibacillus<\/jats:italic> sp., contain no B\u2010type DyP peroxidases implicated in lignin degradation in other bacteria, but contain putative multicopper oxidase genes. Multi\u2010copper oxidase CueO from <jats:italic>Ochrobactrum<\/jats:italic> sp. was expressed and reconstituted as a recombinant laccase\u2010like enzyme, and kinetically characterized. <jats:italic>Ochrobactrum<\/jats:italic> CueO shows activity for oxidation of \u03b2\u2010aryl ether and biphenyl lignin dimer model compounds, generating oxidized dimeric products, and shows activity for oxidation of Ca\u2010lignosulfonate, generating vanillic acid as a low molecular weight product. The crystal structure of <jats:italic>Ochrobactrum<\/jats:italic> CueO (OcCueO) has been determined at 1.1\u00a0\u00c5 resolution (<jats:styled-content style=\"fixed-case\">PDB<\/jats:styled-content>: <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.rcsb.org\/pdb\/search\/structidSearch.do?structureId=6EVG\">6EVG<\/jats:ext-link>), showing a four\u2010coordinate mononuclear type I copper center with ligands His495, His434 and Cys490 with Met500 as an axial ligand, similar to that of <jats:italic>Escherichia\u00a0coli<\/jats:italic> CueO and bacterial azurin proteins, whereas fungal laccase enzymes contain a three\u2010coordinate type I copper metal center. A trinuclear type 2\/3 copper cluster was modeled into the active site, showing similar structure to <jats:italic>E.\u00a0coli<\/jats:italic> CueO and fungal laccases, and three solvent channels leading to the active site. Site\u2010directed mutagenesis was carried out on amino acid residues found in the solvent channels, indicating the importance for residues Asp102, Gly103, Arg221, Arg223, and Asp462 for catalytic activity. The work identifies a new bacterial multicopper enzyme with activity for lignin oxidation, and implicates a role for bacterial laccase\u2010like multicopper oxidases in some lignin\u2010degrading bacteria.<\/jats:p><\/jats:sec><jats:sec><jats:title>Database<\/jats:title><jats:p>Structural data are available in the PDB under the accession number <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.rcsb.org\/pdb\/search\/structidSearch.do?structureId=6EVG\">6EVG<\/jats:ext-link>.<\/jats:p><\/jats:sec>","DOI":"10.1111\/febs.14437","type":"journal-article","created":{"date-parts":[[2018,3,25]],"date-time":"2018-03-25T18:21:29Z","timestamp":1522002089000},"page":"1684-1700","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":75,"title":["Structural and functional characterisation of multi\u2010copper oxidase CueO from lignin\u2010degrading bacterium <i>Ochrobactrum<\/i> sp. reveal its activity towards lignin model compounds and lignosulfonate"],"prefix":"10.1111","volume":"285","author":[{"given":"Rommel Santiago","family":"Granja\u2010Travez","sequence":"first","affiliation":[{"name":"Department of Chemistry University of Warwick Coventry UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachael C.","family":"Wilkinson","sequence":"additional","affiliation":[{"name":"School of Life Sciences University of Warwick Coventry UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriela Felix","family":"Persinoti","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio Nacional de Ci\u00eancia e Tecnologia do Bioetanol (CTBE) Centro Nacional de Pesquisa em Energia e Materiais (CNPEM) Campinas S\u00e3o Paulo Brasil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabio M.","family":"Squina","sequence":"additional","affiliation":[{"name":"Programa de Processos Tecnol\u00f3gicos e Ambientais Universidade de Sorocaba Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vilmos","family":"F\u00fcl\u00f6p","sequence":"additional","affiliation":[{"name":"School of Life Sciences University of Warwick Coventry UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy D. 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