{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T10:25:24Z","timestamp":1761560724209,"version":"build-2065373602"},"reference-count":29,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2008,9,5]],"date-time":"2008-09-05T00:00:00Z","timestamp":1220572800000},"content-version":"vor","delay-in-days":2804,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotechnology Progress"],"published-print":{"date-parts":[[2001,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>We report on the performance of cross\u2010linked enzyme microcrystals (CLECs) of subtilisin Carlsberg in supercritical fluids (SC\u2010fluids). The catalytic activity of CLECs in SC\u2010ethane was found to be 2\u2010 to 10\u2010fold greater than in hexane under the same conditions, using CLECs dried by propanol washing. Air\u2010dried CLECs and lyophilized powders showed much lower activities, reflecting the same hydration hysteresis effects as in organic solvents. Reaction rates were much lower in SC\u2010CO<jats:sub>2<\/jats:sub>, especially at higher water activity, probably as a result of acid\u2010base effects of carbonic acid on the enzyme.<\/jats:p>","DOI":"10.1021\/bp000148m","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T00:56:31Z","timestamp":1027644991000},"page":"355-358","source":"Crossref","is-referenced-by-count":17,"title":["Supercritical Fluids Are Superior Media for Catalysis by Cross\u2010Linked Enzyme Microcrystals of Subtilisin Carlsberg"],"prefix":"10.1002","volume":"17","author":[{"given":"Nuno","family":"Fontes","sequence":"first","affiliation":[]},{"given":"M. Concei\u00e7\u00e3o","family":"Almeida","sequence":"additional","affiliation":[]},{"given":"Silvia","family":"Garcia","sequence":"additional","affiliation":[]},{"given":"C\u00e9lia","family":"Peres","sequence":"additional","affiliation":[]},{"given":"Johann","family":"Partridge","sequence":"additional","affiliation":[]},{"given":"Peter J.","family":"Halling","sequence":"additional","affiliation":[]},{"given":"Susana","family":"Barreiros","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2008,9,5]]},"reference":[{"key":"e_1_2_5_2_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja00044a064"},{"key":"e_1_2_5_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.94.9.4250"},{"key":"e_1_2_5_4_2","doi-asserted-by":"publisher","DOI":"10.1021\/jo9618334"},{"key":"e_1_2_5_5_2","unstructured":"SC\u2010fluids are substances above their critical point that exhibit a tunable solvation ability and improved mass transfer relative to conventional liquid solvents."},{"key":"e_1_2_5_6_2","doi-asserted-by":"publisher","DOI":"10.1021\/cr970040u"},{"key":"e_1_2_5_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0141-0229(97)00241-X"},{"key":"e_1_2_5_8_2","doi-asserted-by":"publisher","DOI":"10.1021\/ie970933c"},{"key":"e_1_2_5_9_2","doi-asserted-by":"publisher","DOI":"10.3109\/07388559509150531"},{"key":"e_1_2_5_10_2","unstructured":"The assumption is that the activity coefficient of water in the SCF\u2010fluid mixture is constant. For evidence supporting this assumption in the systems considered see refs 5b and 5c."},{"key":"e_1_2_5_11_2","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19970720)55:2<367::AID-BIT14>3.0.CO;2-E"},{"key":"e_1_2_5_12_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja961383x"},{"key":"e_1_2_5_13_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.22.12918"},{"key":"e_1_2_5_14_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.89.3.1100"},{"key":"e_1_2_5_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-4838(98)00086-7"},{"key":"e_1_2_5_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0141-0229(96)00204-9"},{"volume-title":"Biochemical Engineering Fundamentals","year":"1986","author":"Bailey J. E.","key":"e_1_2_5_17_2"},{"key":"e_1_2_5_17_3","unstructured":"\u03a6 is a dimensionless parameter defined as \u03a6 = [vo(DesSo)](Vp\/Ap)2 wherevois the observed initial rate Desis the effective diffusion coefficient of APEE Sois the bulk APEE concentration andVpandApare the volume and external surface area of the catalyst particles respectively.Desis a function of the diffusion coefficient of APEE in free solution (Do) of particle porosity and tortuosity and the radii of the particle pore and the APEE molecule. Microscopic observation of the CLECs revealed great heterogeneity in particle shapes and sizes. Typical needlelike particles were present but the dominant particles were rod\u2010shaped with an average diameter of 2 \u03bcm and an average length of 3 \u03bcm. Kasche et al. (20) calculatedDesof a substrate in CLECs of a different enzyme and obtained a value about 10% lower thanDo. We thus assumed thatDeswould vary betweenDoand 0.l \u00d7 Do.Doof solutes in organic solvents including hexane at 40 \u00b0C are typically of the order of 10\u22125cm2s\u22121(21) about 1 order of magnitude lower than in SC\u2010fluids (22)."},{"key":"e_1_2_5_18_2","doi-asserted-by":"publisher","DOI":"10.1002\/bit.260391114"},{"key":"e_1_2_5_19_2","doi-asserted-by":"crossref","unstructured":"Partridge J.;Halling P. J.;Moore B. D.Practical route to high activity enzyme preparations for synthesis in organic media.Chem. Commun.1998 841\u2013842.","DOI":"10.1039\/a800408k"},{"key":"e_1_2_5_20_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.24.10969"},{"key":"e_1_2_5_20_3","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19970220)53:4<351::AID-BIT1>3.0.CO;2-M"},{"key":"e_1_2_5_21_2","unstructured":"The degree of carbamate formation increases with decreasingawand increasing pH. See ref 6."},{"key":"e_1_2_5_22_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja961596m"},{"key":"e_1_2_5_23_2","unstructured":"The ionization state of the enzyme may be controlled in the nonaqueous phase by"},{"key":"e_1_2_5_23_3","doi-asserted-by":"publisher","DOI":"10.1016\/0167-4838(94)90203-8"},{"key":"e_1_2_5_23_4","doi-asserted-by":"publisher","DOI":"10.1021\/ja972635c"},{"key":"e_1_2_5_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-7799(99)01322-0"},{"key":"e_1_2_5_25_2","doi-asserted-by":"publisher","DOI":"10.1002\/aic.690010222"},{"key":"e_1_2_5_26_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-662-07380-3"}],"container-title":["Biotechnology Progress"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1021%2Fbp000148m","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/aiche.onlinelibrary.wiley.com\/doi\/pdf\/10.1021\/bp000148m","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:31:47Z","timestamp":1760034707000},"score":1,"resource":{"primary":{"URL":"https:\/\/aiche.onlinelibrary.wiley.com\/doi\/10.1021\/bp000148m"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2001,1]]},"references-count":29,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2001,1]]}},"alternative-id":["10.1021\/bp000148m"],"URL":"https:\/\/doi.org\/10.1021\/bp000148m","archive":["Portico"],"relation":{},"ISSN":["8756-7938","1520-6033"],"issn-type":[{"type":"print","value":"8756-7938"},{"type":"electronic","value":"1520-6033"}],"subject":[],"published":{"date-parts":[[2001,1]]}}}