{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T10:17:00Z","timestamp":1789208220802,"version":"build-2803163510"},"reference-count":31,"publisher":"American Society for Microbiology","issue":"13","license":[{"start":{"date-parts":[[2008,7,1]],"date-time":"2008-07-01T00:00:00Z","timestamp":1214870400000},"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":[[2008,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The gene encoding a novel alcohol dehydrogenase (ADH) that belongs to the short-chain dehydrogenase\/reductase (SDR) superfamily was identified in the extremely thermophilic, halotolerant gram-negative eubacterium\n            <jats:italic>Thermus thermophilus<\/jats:italic>\n            HB27. The\n            <jats:italic>T. thermophilus<\/jats:italic>\n            ADH gene (\n            <jats:italic>adh<\/jats:italic>\n            <jats:sub>Tt<\/jats:sub>\n            ) was heterologously overexpressed in\n            <jats:italic>Escherichia coli<\/jats:italic>\n            , and the protein (ADH\n            <jats:sub>Tt<\/jats:sub>\n            ) was purified to homogeneity and characterized. ADH\n            <jats:sub>Tt<\/jats:sub>\n            is a tetrameric enzyme consisting of identical 26,961-Da subunits composed of 256 amino acids. The enzyme has remarkable thermophilicity and thermal stability, displaying activity at temperatures up to \u223c73\u00b0C and a 30-min half-inactivation temperature of \u223c90\u00b0C, as well as good tolerance to common organic solvents. ADH\n            <jats:sub>Tt<\/jats:sub>\n            has a strict requirement for NAD(H) as the coenzyme, a preference for reduction of aromatic ketones and \u03b1-keto esters, and poor activity on aromatic alcohols and aldehydes. This thermophilic enzyme catalyzes the following reactions with Prelog specificity: the reduction of acetophenone, 2,2,2-trifluoroacetophenone, \u03b1-tetralone, and \u03b1-methyl and \u03b1-ethyl benzoylformates to (\n            <jats:italic>S<\/jats:italic>\n            )-(\u2212)-1-phenylethanol (&gt;99% enantiomeric excess [ee]), (\n            <jats:italic>R<\/jats:italic>\n            )-\u03b1-(trifluoromethyl)benzyl alcohol (93% ee), (\n            <jats:italic>S<\/jats:italic>\n            )-\u03b1-tetralol (&gt;99% ee), methyl (\n            <jats:italic>R<\/jats:italic>\n            )-(\u2212)-mandelate (92% ee), and ethyl (\n            <jats:italic>R<\/jats:italic>\n            )-(\u2212)-mandelate (95% ee), respectively, by way of an efficient in situ NADH-recycling system involving 2-propanol and a second thermophilic ADH. This study further supports the critical role of the D37 residue in discriminating NAD(H) from NADP(H) in members of the SDR superfamily.\n          <\/jats:p>","DOI":"10.1128\/aem.00217-08","type":"journal-article","created":{"date-parts":[[2008,5,4]],"date-time":"2008-05-04T01:46:23Z","timestamp":1209865583000},"page":"3949-3958","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":62,"title":["Purification and Characterization of a Novel Recombinant Highly Enantioselective Short-Chain NAD(H)-Dependent Alcohol Dehydrogenase from\n            <i>Thermus thermophilus<\/i>"],"prefix":"10.1128","volume":"74","author":[{"given":"Angela","family":"Pennacchio","sequence":"first","affiliation":[{"name":"Istituto di Biochimica delle Proteine, Consiglio Nazionale delle Ricerche, Via P. Castellino 111, I-80131 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Biagio","family":"Pucci","sequence":"additional","affiliation":[{"name":"Istituto di Biochimica delle Proteine, Consiglio Nazionale delle Ricerche, Via P. Castellino 111, I-80131 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"Secundo","sequence":"additional","affiliation":[{"name":"Istituto di Chimica e del Riconoscimento Molecolare, Consiglio Nazionale delle Ricerche, Via M. Bianco 9, I-20131 Milano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"La Cara","sequence":"additional","affiliation":[{"name":"Istituto di Biochimica delle Proteine, Consiglio Nazionale delle Ricerche, Via P. Castellino 111, I-80131 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mose\u0300","family":"Rossi","sequence":"additional","affiliation":[{"name":"Istituto di Biochimica delle Proteine, Consiglio Nazionale delle Ricerche, Via P. Castellino 111, I-80131 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlo A.","family":"Raia","sequence":"additional","affiliation":[{"name":"Istituto di Biochimica delle Proteine, Consiglio Nazionale delle Ricerche, Via P. Castellino 111, I-80131 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi049736p"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M202160200"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1093\/protein\/11.10.925"},{"key":"e_1_3_2_5_2","first-page":"277","volume":"15","year":"1998","unstructured":"Fontana, A., V. De Filippis, P. Polverino de Laureto, E. Scaramella, and M. Zambonin. 1998. Rigidity of thermophilic enzymes, p. 277-294. In A. Ballestreros, F. J. Plou, J. L. Iborra, and P. J. Halling (ed.), Stability and stabilization in biocatalysis,vol. 15. Elsevier Sciences, Amsterdam, The Netherlands.","journal-title":"Stability and stabilization in biocatalysis"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0957-4166(00)80234-7"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20041539"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/1357-2725(95)00138-7"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(03)00857-X"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt956"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi051596b"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-7799(98)01207-4"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1271\/bbb.70.418"},{"key":"e_1_3_2_14_2","first-page":"248","volume":"10","year":"1976","unstructured":"Jones, J. B., and J. F. Beck. 1976. Applications of biochemical systems in organic chemistry, p. 248-401. In J. B. Jones, C. J. Sih, and D. Perlman (ed.), Techniques of chemistry series, part I,vol. 10. John Wiley & Sons, New York, NY.","journal-title":"Techniques of chemistry series, part I"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1432-1033.2002.03130.x"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja00261a026"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0040-4039(00)89018-8"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1038\/20981"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.1998.1750"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1038\/227680a0"},{"key":"e_1_3_2_21_2","unstructured":"GraFit version 5.0.11. 1992"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0403337101"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.72.1.233-238.2006"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(03)00081-0"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0040-4020(01)00689-5"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1351\/pac196409010119"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.177.17.4947-4962.1995"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(01)31056-X"},{"key":"e_1_3_2_29_2","unstructured":"Molecular cloning: a laboratory manual 1989"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2005.04.029"},{"key":"e_1_3_2_31_2","doi-asserted-by":"publisher","DOI":"10.1021\/jo034808o"},{"key":"e_1_3_2_32_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi062110"}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.00217-08","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.00217-08","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,23]],"date-time":"2022-02-23T16:35:13Z","timestamp":1645634113000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AEM.00217-08"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2008,7]]},"references-count":31,"journal-issue":{"issue":"13","published-print":{"date-parts":[[2008,7]]}},"alternative-id":["10.1128\/AEM.00217-08"],"URL":"https:\/\/doi.org\/10.1128\/aem.00217-08","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2008,7]]},"assertion":[{"value":"2008-01-24","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2008-04-27","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2008-07-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}