{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T00:12:57Z","timestamp":1789776777445,"version":"4.0.1"},"reference-count":26,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2017,1,1]],"date-time":"2017-01-01T00:00:00Z","timestamp":1483228800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"US Department of Energy Office of Biological and Environmental Research","award":["DE-SC0006630"],"award-info":[{"award-number":["DE-SC0006630"]}]},{"DOI":"10.13039\/100001024","name":"Roy J. Carver Charitable Trust","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100001024","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2017,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Gene expression in methanotrophs has been shown to be affected by the availability of a variety of metals, most notably copper-regulating expression of alternative forms of methane monooxygenase. A copper-binding compound, or chalkophore, called methanobactin plays a key role in copper uptake in methanotrophs. Methanobactin is a ribosomally synthesized and posttranslationally modified peptide (RiPP) with two heterocyclic rings with an associated thioamide for each ring, formed from X-Cys dipeptide sequences that bind copper. The gene coding for the precursor polypeptide of methanobactin,<jats:italic>mbnA<\/jats:italic>, is part of a gene cluster, but the role of other genes in methanobactin biosynthesis is unclear. To begin to elucidate the function of these genes, we constructed an unmarked deletion of<jats:italic>mbnABCMN<\/jats:italic>in<jats:named-content content-type=\"genus-species\">Methylosinus trichosporium<\/jats:named-content>OB3b and then homologously expressed<jats:italic>mbnABCM<\/jats:italic>using a broad-host-range cloning vector to determine the function of<jats:italic>mbnN<\/jats:italic>, annotated as coding for an aminotransferase. Methanobactin produced by this strain was found to be substantially different from wild-type methanobactin in that the C-terminal methionine was missing and only one of the two oxazolone rings was formed. Rather, in place of the N-terminal 3-methylbutanoyl-oxazolone-thioamide group, a leucine and a thioamide-containing glycine (Gly-\u03a8) were found, indicating that MbnN is used for deamination of the N-terminal leucine of methanobactin and that this posttranslational modification is critical for closure of the N-terminal oxazolone ring in<jats:named-content content-type=\"genus-species\">M. trichosporium<\/jats:named-content>OB3b. These studies provide new insights into methanobactin biosynthesis and also provide a platform for understanding the function of other genes in the methanobactin gene cluster.<\/jats:p><jats:p><jats:bold>IMPORTANCE<\/jats:bold>Methanotrophs, microbes that play a critical role in the carbon cycle, are influenced by copper, with gene expression and enzyme activity changing as copper levels change. Methanotrophs produce a copper-binding compound, or chalkophore, called methanobactin for copper uptake, and methanobactin plays a key role in controlling methanotrophic activity. Methanobactin has also been shown to be effective in the treatment of Wilson disease, an autosomal recessive disorder where the human body cannot correctly assimilate copper. It is important to characterize the methanobactin biosynthesis pathway to understand how methanotrophs respond to their environment as well as to optimize the use of methanobactin for the treatment of copper-related diseases such as Wilson disease. Here we show that<jats:italic>mbnN<\/jats:italic>, encoding an aminotransferase, is involved in the deamination of the N-terminal leucine and necessary for the formation of one but not both of the heterocyclic rings in methanobactin that are responsible for copper binding.<\/jats:p>","DOI":"10.1128\/aem.02619-16","type":"journal-article","created":{"date-parts":[[2016,10,22]],"date-time":"2016-10-22T03:41:21Z","timestamp":1477107681000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":23,"title":["An Aminotransferase Is Responsible for the Deamination of the N-Terminal Leucine and Required for Formation of Oxazolone Ring A in Methanobactin of Methylosinus trichosporium OB3b"],"prefix":"10.1128","volume":"83","author":[{"given":"Wenyu","family":"Gu","sequence":"first","affiliation":[{"name":"Department of Civil and Environmental Engineering, University of Michigan, Ann Arbor, Michigan, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bipin S.","family":"Baral","sequence":"additional","affiliation":[{"name":"Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alan A.","family":"DiSpirito","sequence":"additional","affiliation":[{"name":"Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeremy D.","family":"Semrau","sequence":"additional","affiliation":[{"name":"Department of Civil and Environmental Engineering, University of Michigan, Ann Arbor, Michigan, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jinorgbio.2006.08.017"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi051815t"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1098322"},{"key":"e_1_3_3_5_2","doi-asserted-by":"crossref","first-page":"3077","DOI":"10.1111\/1462-2920.12150","article-title":"Methanobactin and MmoD work in concert to act as the \u2018copper-switch\u2019 in methanotrophs","volume":"15","author":"Semrau JD","year":"2013","unstructured":"Semrau JD JagadevanS DiSpiritoAA KhalifaA ScanlanJ BergmanBH FreemeierBC BaralBS BandowNS VorobevA HaftDH VuilleumierS MurrellJC. 2013. 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