{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,30]],"date-time":"2026-10-30T18:46:45Z","timestamp":1793386005990,"version":"4.5.0"},"reference-count":34,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2012,4,15]],"date-time":"2012-04-15T00:00:00Z","timestamp":1334448000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Bacteriol"],"published-print":{"date-parts":[[2012,4,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The molecular basis for the ability of bacteria to live on caffeine as a sole carbon and nitrogen source is unknown.\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Pseudomonas putida<\/jats:named-content>\n            CBB5, which grows on several purine alkaloids, metabolizes caffeine and related methylxanthines via sequential\n            <jats:italic>N<\/jats:italic>\n            -demethylation to xanthine. Metabolism of caffeine by CBB5 was previously attributed to one broad-specificity methylxanthine\n            <jats:italic>N<\/jats:italic>\n            -demethylase composed of two subunits, NdmA and NdmB. Here, we report that NdmA and NdmB are actually two independent Rieske nonheme iron monooxygenases with\n            <jats:italic>N<\/jats:italic>\n            <jats:sub>1<\/jats:sub>\n            - and\n            <jats:italic>N<\/jats:italic>\n            <jats:sub>3<\/jats:sub>\n            -specific\n            <jats:italic>N<\/jats:italic>\n            -demethylation activity, respectively. Activity for both enzymes is dependent on electron transfer from NADH via a redox-center-dense Rieske reductase, NdmD. NdmD itself is a novel protein with one Rieske [2Fe-2S] cluster, one plant-type [2Fe-2S] cluster, and one flavin mononucleotide (FMN) per enzyme. All\n            <jats:italic>ndm<\/jats:italic>\n            genes are located in a 13.2-kb genomic DNA fragment which also contained a formaldehyde dehydrogenase.\n            <jats:italic>ndmA<\/jats:italic>\n            ,\n            <jats:italic>ndmB<\/jats:italic>\n            , and\n            <jats:italic>ndmD<\/jats:italic>\n            were cloned as His\n            <jats:sub>6<\/jats:sub>\n            fusion genes, expressed in\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Escherichia coli<\/jats:named-content>\n            , and purified using a Ni-NTA column. NdmA-His\n            <jats:sub>6<\/jats:sub>\n            plus His\n            <jats:sub>6<\/jats:sub>\n            -NdmD catalyzed\n            <jats:italic>N<\/jats:italic>\n            <jats:sub>1<\/jats:sub>\n            -demethylation of caffeine, theophylline, paraxanthine, and 1-methylxanthine to theobromine, 3-methylxanthine, 7-methylxanthine, and xanthine, respectively. NdmB-His\n            <jats:sub>6<\/jats:sub>\n            plus His\n            <jats:sub>6<\/jats:sub>\n            -NdmD catalyzed\n            <jats:italic>N<\/jats:italic>\n            <jats:sub>3<\/jats:sub>\n            -demethylation of theobromine, 3-methylxanthine, caffeine, and theophylline to 7-methylxanthine, xanthine, paraxanthine, and 1-methylxanthine, respectively. One formaldehyde was produced from each methyl group removed. Activity of an\n            <jats:italic>N<\/jats:italic>\n            <jats:sub>7<\/jats:sub>\n            -specific\n            <jats:italic>N<\/jats:italic>\n            -demethylase, NdmC, has been confirmed biochemically. This is the first report of bacterial\n            <jats:italic>N<\/jats:italic>\n            -demethylase genes that enable bacteria to live on caffeine. These genes represent a new class of Rieske oxygenases and have the potential to produce biofuels, animal feed, and pharmaceuticals from coffee and tea waste.\n          <\/jats:p>","DOI":"10.1128\/jb.06637-11","type":"journal-article","created":{"date-parts":[[2012,2,11]],"date-time":"2012-02-11T05:30:54Z","timestamp":1328938254000},"page":"2041-2049","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":105,"title":["Novel, Highly Specific\n            <i>N<\/i>\n            -Demethylases Enable Bacteria To Live on Caffeine and Related Purine Alkaloids"],"prefix":"10.1128","volume":"194","author":[{"given":"Ryan M.","family":"Summers","sequence":"first","affiliation":[{"name":"Department of Chemical and Biochemical Engineering, University of Iowa, Iowa City, Iowa, USA"},{"name":"The Center for Biocatalysis and Bioprocessing, University of Iowa, Coralville, Iowa, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tai Man","family":"Louie","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biochemical Engineering, University of Iowa, Iowa City, Iowa, USA"},{"name":"The Center for Biocatalysis and Bioprocessing, University of Iowa, Coralville, Iowa, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chi-Li","family":"Yu","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biochemical Engineering, University of Iowa, Iowa City, Iowa, USA"},{"name":"The Center for Biocatalysis and Bioprocessing, University of Iowa, Coralville, Iowa, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lokesh","family":"Gakhar","sequence":"additional","affiliation":[{"name":"Protein Crystallography Facility, University of Iowa, Iowa City, Iowa, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kailin C.","family":"Louie","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biochemical Engineering, University of Iowa, Iowa City, Iowa, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mani","family":"Subramanian","sequence":"additional","affiliation":[{"name":"Department of Chemical and Biochemical Engineering, University of Iowa, Iowa City, Iowa, USA"},{"name":"The Center for Biocatalysis and Bioprocessing, University of Iowa, Coralville, Iowa, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1038\/nature01363","article-title":"Human and bacterial oxidative demethylases repair alkylation damage in both RNA and DNA","volume":"421","author":"Aas PA","year":"2003","unstructured":"AasPA . 2003. 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