{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T16:08:22Z","timestamp":1791043702001,"version":"4.1.0"},"reference-count":25,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2009,4,13]],"date-time":"2009-04-13T00:00:00Z","timestamp":1239580800000},"content-version":"vor","delay-in-days":2569,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2002,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The hyperthermophilic bacterium <jats:italic>Thermotoga maritima<\/jats:italic> encodes a gene sharing sequence similarities with several known genes for alkaline phosphatase (AP). The putative gene was isolated and the corresponding protein expressed in <jats:italic>Escherichia coli,<\/jats:italic> with and without a predicted signal sequence. The recombinant protein showed phosphatase activity toward the substrate <jats:italic>p<\/jats:italic>\u2010nitrophenyl\u2010phosphate with a k<jats:sub>cat<\/jats:sub> of 16 s<jats:sup>\u22121<\/jats:sup> and a K<jats:sub>m<\/jats:sub> of 175 \u03bcM at a pH optimum of 8.0 when assayed at 25\u00b0C. <jats:italic>T. maritima<\/jats:italic> phosphatase activity increased at high temperatures, reaching a maximum k<jats:sub>cat<\/jats:sub> of 100 s<jats:sup>\u22121<\/jats:sup>, with a K<jats:sub>m<\/jats:sub> of 93 \u03bcM at 65\u00b0C. Activity was stable at 65\u00b0C for &gt;24 h and at 90\u00b0C for 5 h. Phosphatase activity was dependent on divalent metal ions, specifically Co(II) and Mg(II). Circular dichroism spectra showed that the enzyme gains secondary structure on addition of these metals. Zinc, the most common divalent metal ion required for activity in known APs, was shown to inhibit the <jats:italic>T. maritima<\/jats:italic> phosphatase enzyme at concentrations above 0.3 moles Zn: 1 mole monomer. All activity was abolished in the presence of 0.1 mM EDTA. The <jats:italic>T. maritima<\/jats:italic> AP primary sequence is 28% identical when compared with <jats:italic>E. coli<\/jats:italic> AP. Based on a structural model, the active sites are superimposable except for two residues near the <jats:italic>E. coli<\/jats:italic> AP Mg binding site, D153 and K328 (<jats:italic>E. coli<\/jats:italic> numbering) corresponding to histidine and tryptophan in <jats:italic>T. maritima<\/jats:italic> AP, respectively. Sucrose\u2010density gradient sedimentation experiments showed that the protein exists in several quaternary forms predominated by an octamer.<\/jats:p>","DOI":"10.1110\/ps.4260102","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T23:59:58Z","timestamp":1027727998000},"page":"903-911","source":"Crossref","is-referenced-by-count":50,"title":["Alkaline phosphatase from the hyperthermophilic bacterium <i>T. maritima<\/i> requires cobalt for activity"],"prefix":"10.1002","volume":"11","author":[{"given":"Cheryl L.","family":"Wojciechowski","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James P.","family":"Cardia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evan R.","family":"Kantrowitz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2009,4,13]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-4838(01)00247-3"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/0006-3002(60)91282-8"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0968-0004(99)01427-9"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.1150181505"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(99)01448-9"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00409880"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.172.2.735-740.1990"},{"key":"e_1_2_7_9_1","first-page":"1077","article-title":"Bacillus subtilis alkaline phosphatases III and IV","volume":"266","author":"Hulett M.F.","year":"1991","journal-title":"J. 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