{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T13:47:39Z","timestamp":1791208059650,"version":"4.1.0"},"reference-count":26,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2008,1,1]],"date-time":"2008-01-01T00:00:00Z","timestamp":1199145600000},"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":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2008,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The anoxic metabolism of cholesterol was studied in the denitrifying bacterium\n            <jats:italic>Sterolibacterium denitrificans<\/jats:italic>\n            , which was grown with cholesterol and nitrate. Cholest-4-en-3-one was identified before as the product of cholesterol dehydrogenase\/isomerase, the first enzyme of the pathway. The postulated second enzyme, cholest-4-en-3-one-\u0394\n            <jats:sup>1<\/jats:sup>\n            -dehydrogenase, was partially purified, and its N-terminal amino acid sequence and tryptic peptide sequences were determined. Based on this information, the corresponding gene was amplified and cloned and the His-tagged recombinant protein was overproduced, purified, and characterized. The recombinant enzyme catalyzes the expected \u0394\n            <jats:sup>1<\/jats:sup>\n            -desaturation (cholest-4-en-3-one to cholesta-1,4-dien-3-one) under anoxic conditions. It contains approximately one molecule of FAD per 62-kDa subunit and forms high molecular aggregates in the absence of detergents. The enzyme accepts various artificial electron acceptors, including dichlorophenol indophenol and methylene blue. It oxidizes not only cholest-4-en-3-one, but also progesterone (with highest catalytic efficiency, androst-4-en-3,17-dione, testosterone, 19-nortestosterone, and cholest-5-en-3-one. Two steroids, corticosterone and estrone, act as competitive inhibitors. The dehydrogenase resembles 3-ketosteroid-\u0394\n            <jats:sup>1<\/jats:sup>\n            -dehydrogenases from other organisms (highest amino acid sequence identity with that from\n            <jats:italic>Pseudoalteromonas haloplanktis<\/jats:italic>\n            ), with some interesting differences. Due to its catalytic properties, the enzyme may be useful in steroid transformations.\n          <\/jats:p>","DOI":"10.1128\/aem.01968-07","type":"journal-article","created":{"date-parts":[[2007,11,10]],"date-time":"2007-11-10T01:33:51Z","timestamp":1194658431000},"page":"107-113","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":49,"title":["Cholest-4-En-3-One-\u0394\n            <sup>1<\/sup>\n            -Dehydrogenase, a Flavoprotein Catalyzing the Second Step in Anoxic Cholesterol Metabolism"],"prefix":"10.1128","volume":"74","author":[{"given":"Yin-Ru","family":"Chiang","sequence":"first","affiliation":[{"name":"Mikrobiologie, Institut Biologie II, Albert-Ludwigs-Universita\u0308t Freiburg, Freiburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wael","family":"Ismail","sequence":"additional","affiliation":[{"name":"Mikrobiologie, Institut Biologie II, Albert-Ludwigs-Universita\u0308t Freiburg, Freiburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Se\u0301bastien","family":"Gallien","sequence":"additional","affiliation":[{"name":"Laboratoire de Spe\u0301ctrometrie de Masse Bio-Organique, CRNS, ECPM, Universite\u0301 Louis Pasteur, Strasbourg, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dimitri","family":"Heintz","sequence":"additional","affiliation":[{"name":"Laboratoire de Spe\u0301ctrometrie de Masse Bio-Organique, CRNS, ECPM, Universite\u0301 Louis Pasteur, Strasbourg, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alain","family":"Van Dorsselaer","sequence":"additional","affiliation":[{"name":"Laboratoire de Spe\u0301ctrometrie de Masse Bio-Organique, CRNS, ECPM, Universite\u0301 Louis Pasteur, Strasbourg, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Georg","family":"Fuchs","sequence":"additional","affiliation":[{"name":"Mikrobiologie, Institut Biologie II, Albert-Ludwigs-Universita\u0308t Freiburg, Freiburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","unstructured":"Current protocols in molecular biology 1987"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(76)90527-3"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M610963200"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1093\/oxfordjournals.jbchem.a124804"},{"key":"e_1_3_2_6_2","first-page":"103","year":"1999","unstructured":"Fersht, A. 1999. The basic equation of enzyme kinetics, p. 103-131. In M. R. Julet (ed.), Structure and mechanism in protein science. W. H. Freeman and Company, New York, NY.","journal-title":"Structure and mechanism in protein science"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.178.11.3322-3330.1996"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1007\/s002030050442"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.57.5.1386-1393.1991"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/0005-2760(86)90251-1"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/0167-4838(90)90010-D"},{"key":"e_1_3_2_12_2","first-page":"339","year":"1994","unstructured":"Kadode, M., H. Matsushita, K. Suzuki, and E. Itagaki. 1994. Essential arginine residue(s) of 3-ketosteroid-\u03941-dehydrogenase, p. 339-342. In K. Yagi (ed.), Flavins and flavoproteins. Walter de Gruyter & Co., Berlin, Germany.","journal-title":"Flavins and flavoproteins"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1002\/jobm.3620250713"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/227680a0"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0960-0760(00)00044-3"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.183.6.1899-1908.2001"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1093\/oxfordjournals.jbchem.a022195"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1002\/elps.1150110202"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1128\/aem.62.7.2405-2410.1996"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00163a023"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.173.22.7219-7227.1991"},{"key":"e_1_3_2_22_2","first-page":"31","year":"1991","unstructured":"Schreuder, H. A., J. M. Van der Laan, W.-G. J. Hol, and J. Drenth. 1991. The structure of p-hydroxybenzoate hydroxylase, p. 31-64. In F. M\u00fcller (ed.), Chemistry and biochemistry of flavoproteins,vol. II. CRC Press, Boca Raton, FL.","journal-title":"Chemistry and biochemistry of flavoproteins"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.82.4.1074"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1099\/ijs.0.02039-0"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1099\/00221287-148-10-3285"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.66.5.2029-2036.2000"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0605728104"}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.01968-07","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.01968-07","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,23]],"date-time":"2022-02-23T16:42:34Z","timestamp":1645634554000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AEM.01968-07"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2008,1]]},"references-count":26,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2008,1]]}},"alternative-id":["10.1128\/AEM.01968-07"],"URL":"https:\/\/doi.org\/10.1128\/aem.01968-07","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2008,1]]},"assertion":[{"value":"2007-08-28","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2007-10-31","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2008-01-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}