{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,18]],"date-time":"2025-10-18T10:03:29Z","timestamp":1760781809677},"reference-count":0,"publisher":"Portland Press Ltd.","issue":"2","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[1997,12,1]]},"abstract":"<jats:p>Rat liver peroxisomes contain two multifunctional enzymes: (1) perMFE-1 [2-enoyl-CoA hydratase 1\/\u03943,\u03942-enoyl-CoA isomerase\/(S)-3-hydroxyacyl-CoA dehydrogenase] and (2) perMFE-2 [2-enoyl-CoA hydratase 2\/(R)-3-hydroxyacyl-CoA dehydrogenase]. To investigate the role of the hydratase activity of perMFE-2 in \u03b2-oxidation, a truncated version of perMFE-2 was expressed in Escherichia coli as a recombinant protein. The protein catalyses the hydration of straight-chain (2E)-enoyl-CoAs to (3R)-hydroxyacyl-CoAs, but it is devoid of hydratase 1 [(2E)-enoyl-CoA to (3S)-hydroxyacyl-CoA] and (3R)-hydroxyacyl-CoA dehydrogenase activities. The purified enzyme (46 kDa hydratase 2) can be stored as an active enzyme for at least half a year. The recombinant enzyme hydrates (24E)-3\u03b1,7\u03b1,12\u03b1-trihydroxy- 5\u03b2-cholest-24-enoyl-CoA to (24R,25R)-3\u03b1,7\u03b1,12\u03b1,24-tetrahydroxy-5\u03b2-cholestanoyl-CoA, which has previously been characterized as a physiological intermediate in bile acid synthesis. The stereochemistry of the products indicates that the hydration reaction catalysed by the enzyme proceeds via a syn mechanism. A monofunctional 2-enoyl-CoA hydratase 2 has not been observed as a wild-type protein. The recombinant 46 kDa hydratase 2 described here survives in a purified form under storage, thus being the first protein of this type amenable to application as a tool in metabolic studies.<\/jats:p>","DOI":"10.1042\/bj3280377","type":"journal-article","created":{"date-parts":[[2015,8,10]],"date-time":"2015-08-10T21:57:14Z","timestamp":1439243834000},"page":"377-382","update-policy":"http:\/\/dx.doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":34,"title":["Recombinant 2-enoyl-CoA hydratase derived from rat peroxisomal multifunctional enzyme 2: role of the hydratase reaction in bile acid synthesis"],"prefix":"10.1042","volume":"328","author":[{"given":"Yong-Mei","family":"QIN","sequence":"first","affiliation":[{"name":"Biocenter Oulu, University of Oulu, Linnanmaa, FIN-90570 Oulu, Finland"},{"name":"Department of Biochemistry, University of Oulu, Linnanmaa, FIN-90570 Oulu, Finland"}]},{"given":"M. Antti","family":"HAAPALAINEN","sequence":"additional","affiliation":[{"name":"Biocenter Oulu, University of Oulu, Linnanmaa, FIN-90570 Oulu, Finland"},{"name":"Department of Biochemistry, University of Oulu, Linnanmaa, FIN-90570 Oulu, Finland"}]},{"given":"Demara","family":"CONRY","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Southwest Missouri State University, Springfield, MO 65804, U.S.A."}]},{"given":"A. Dean","family":"CUEBAS","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Southwest Missouri State University, Springfield, MO 65804, U.S.A."}]},{"given":"J. Kalervo","family":"HILTUNEN","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Oulu, Linnanmaa, FIN-90570 Oulu, Finland"},{"name":"Biocenter Oulu, University of Oulu, Linnanmaa, FIN-90570 Oulu, Finland"}]},{"given":"K. Dmitry","family":"NOVIKOV","sequence":"additional","affiliation":[{"name":"Biocenter Oulu, University of Oulu, Linnanmaa, FIN-90570 Oulu, Finland"},{"name":"Department of Biochemistry, University of Oulu, Linnanmaa, FIN-90570 Oulu, Finland"}]}],"member":"288","container-title":["Biochemical Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/328\/2\/377\/629922\/bj3280377.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/328\/2\/377\/629922\/bj3280377.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,24]],"date-time":"2021-11-24T00:38:32Z","timestamp":1637714312000},"score":1,"resource":{"primary":{"URL":"https:\/\/portlandpress.com\/biochemj\/article\/328\/2\/377\/34280\/Recombinant-2-enoyl-CoA-hydratase-derived-from-rat"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1997,12,1]]},"references-count":0,"journal-issue":{"issue":"2","published-print":{"date-parts":[[1997,12,1]]}},"URL":"https:\/\/doi.org\/10.1042\/bj3280377","relation":{},"ISSN":["0264-6021","1470-8728"],"issn-type":[{"value":"0264-6021","type":"print"},{"value":"1470-8728","type":"electronic"}],"subject":[],"published":{"date-parts":[[1997,12,1]]}}}