{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,30]],"date-time":"2025-10-30T11:12:03Z","timestamp":1761822723569,"version":"build-2065373602"},"reference-count":28,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2004,2,19]],"date-time":"2004-02-19T00:00:00Z","timestamp":1077148800000},"content-version":"vor","delay-in-days":3885,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotech &amp;amp; Bioengineering"],"published-print":{"date-parts":[[1993,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Alcohol dehydrogenase (E. C. 1.1.1.1) from <jats:italic>Thermoanaerobium brockii<\/jats:italic> at 25\u00b0C and at 65\u00b0C is more active with secondary than primary alcohols. The enzyme utilizes NADP and NADPH as cosubstrates better than NAD and NADH. The maximum velocities (<jats:italic>V<\/jats:italic><jats:sub><jats:italic>m<\/jats:italic><\/jats:sub>) for secondary alcohols at 65\u00b0C are 10 to 100 times higher than those at 25\u00b0C, whereas the <jats:italic>K<\/jats:italic><jats:sub><jats:italic>m<\/jats:italic><\/jats:sub> values are more comparable.<\/jats:p><jats:p>At both 25\u00b0C and 65\u00b0C the substrate analogue 1,1,1,3,3,3\u2010hexafluoro\u20102\u2010propanol inhibited the oxidation of alcohol competitively with respect to cyclopentanol, and uncompetitively with respect to NADP. Dimethylsulfoxide inhibited the reduction of cyclopentanone competitively with respect to cyclopentanone, and uncompetitively with respect to NADPH. As a product inhibitor, NADP was competitive with respect to NADPH. These results demonstrate that the enzyme binds the nucleotide and then the alcohol or ketone to form a ternary complex which is converted to a product ternary complex that releases product and nucleotide in that order.<\/jats:p><jats:p>At 25\u00b0C, all aldehydes and ketones examined inhibited the enzyme at concentrations above their Michaelis constants. The substrate inhibition by cyclopentanone was incomplete, and it was uncompetitive with respect to NADPH. Furthermore, cyclopentanone as a product inhibitor showed intercept\u2010linear, slope\u2010parabolic inhibition with respect to cyclopentanol. These results indicate that cyclopentanone binds to the enzyme\u2013NADP complex at high concentrations. The resulting ternary complex slowly dissociates NADP and cyclopentanone.<\/jats:p><jats:p>At 65\u00b0C, all of the secondary alcohols, with the exception of cyclohexanol, show substrate activation at high concentration. Experiments in which NADP was the variable substrate and cyclopentanol as the constant\u2013variable substrate over a wide range of concentrations gave double reciprocal plots in which the intercepts showed substrate activation and the slopes showed substrate inhibition. These results indicate that the secondary alcohols bind to the enzyme\u2013NADPH complex at high concentrations and that the resulting ternary complex dissociates NADPH faster than the enzyme\u2013NADPH complex. \u00a9 1993 John Wiley &amp; Sons, Inc.<\/jats:p>","DOI":"10.1002\/bit.260420314","type":"journal-article","created":{"date-parts":[[2004,12,30]],"date-time":"2004-12-30T11:15:58Z","timestamp":1104405358000},"page":"367-375","source":"Crossref","is-referenced-by-count":10,"title":["Kinetic models for synthesis by a thermophilic alcohol dehydrogenase"],"prefix":"10.1002","volume":"42","author":[{"given":"Jonathan B.","family":"Ford","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"K. Janine","family":"Askins","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kenneth B.","family":"Taylor","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2004,2,19]]},"reference":[{"key":"e_1_2_1_2_2","doi-asserted-by":"crossref","first-page":"946","DOI":"10.1016\/S0021-9258(19)45698-6","article-title":"Isotope exchange studies on liver alcohol dehydrogenase with cyclohexanol and cyclohexanone as reactants","volume":"247","author":"Ainslie G. R.","year":"1972","journal-title":"J. Biol. Chem."},{"key":"e_1_2_1_3_2","doi-asserted-by":"publisher","DOI":"10.1139\/o90-135"},{"key":"e_1_2_1_4_2","first-page":"256","article-title":"Use of biological systems for the preparation of chiral molecules. 3","volume":"52","author":"Bolan A.","year":"1987","journal-title":"Application in pheromone synthesis. J. Org. 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Unusual stereoselectivity in the reduction of bicyclo(3.3.0)oct\u20102\u2010ene\u20108\u2010one byThermoanaerobium brockiialcohol dehydrogenase.J. Chem. Soc. Chem. 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