{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T01:04:40Z","timestamp":1790989480432,"version":"4.1.0"},"reference-count":24,"publisher":"National Academy of Sciences","issue":"17","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[1999,8,17]]},"abstract":"<jats:p>\n                    The catalytic activity of four lyophilized oxidative\n enzymes\u2014horseradish peroxidase, soybean peroxidase,\n                    <jats:italic>Caldariomyces fumago<\/jats:italic>\n                    chloroperoxidase, and mushroom\n polyphenol oxidase\u2014is much lower when directly suspended in organic\n solvents containing little water than when they are introduced into the\n same largely nonaqueous media by first dissolving them in water and\n then diluting with anhydrous solvents. The lower the water content of\n the medium, the greater this discrepancy becomes. The mechanism of this\n phenomenon was found to arise from reversible denaturation of the\n oxidases on lyophilization: because of its conformational rigidity, the\n denatured enzyme exhibits very limited activity when directly suspended\n in largely nonaqueous media but renatures and thus yields much higher\n activity if first redissolved in water. Two independent means were\n discovered for dramatically minimizing the lyophilization-induced\n inactivation, both involving the addition of certain types of\n excipients to the aqueous enzyme solution before lyophilization. The\n first group of excipients consists of phenolic and aniline substrates\n as well as other hydrophobic compounds; these presumably bind to the\n hydrophobic pocket of the enzyme active site, thereby preventing its\n collapse during dehydration. The second group consists of general\n lyoprotectants such as polyols and polyethylen glycol that apparently\n preserve the overall enzyme structure during dehydration. The\n activation effects of such excipients can reach into the tens and\n hundreds of fold. Moreover, the activations afforded by the two\n excipient groups are additive, resulting in up to a complete protection\n against lyophilization-induced inactivation when representatives of the\n two are present together.\n                  <\/jats:p>","DOI":"10.1073\/pnas.96.17.9475","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T10:38:21Z","timestamp":1027679901000},"page":"9475-9478","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":87,"title":["Striking activation of oxidative enzymes suspended in nonaqueous media"],"prefix":"10.1073","volume":"96","author":[{"given":"Lizhong","family":"Dai","sequence":"first","affiliation":[{"name":"Department of Chemistry, Massachusetts Institute of Technology,\r Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander M.","family":"Klibanov","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Massachusetts Institute of Technology,\r Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[1999,8,17]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-94-011-0611-5"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-7799(97)01013-5"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.24.10969"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0040-4020(97)00477-8"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1271\/bbb.63.73"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-091302-2.50033-5"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(82)90114-2"},{"key":"e_1_3_3_8_2","first-page":"6378","volume":"33","author":"Poli S","year":"1994","unstructured":"S Poli, M Gullotti, C Selvaggini, T Beringhelli Biochemistry 33, 6378\u20136386 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