{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T08:02:25Z","timestamp":1777536145254,"version":"3.51.4"},"reference-count":28,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2005,2,1]],"date-time":"2005-02-01T00:00:00Z","timestamp":1107216000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2005,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The bifunctional wax ester synthase\/acyl coenzyme A (acyl-CoA):diacylglycerol acyltransferase (WS\/DGAT) from\n            <jats:italic>Acinetobacter<\/jats:italic>\n            sp. strain ADP1 (formerly\n            <jats:italic>Acinetobacter calcoaceticus<\/jats:italic>\n            ADP1) mediating the biosyntheses of wax esters and triacylglycerols was used for the in vivo and in vitro biosynthesis of thio wax esters and dithio wax esters. For in vitro biosynthesis, 5\u2032His\n            <jats:sub>6<\/jats:sub>\n            WS\/DGAT comprising an N-terminal His\n            <jats:sub>6<\/jats:sub>\n            tag was purified from the soluble protein fraction of\n            <jats:italic>Escherichia coli<\/jats:italic>\n            Rosetta(DE3)pLysS (pET23a::\n            <jats:italic>\n              5\u2032His\n              <jats:sub>6<\/jats:sub>\n            <\/jats:italic>\n            <jats:italic>atf<\/jats:italic>\n            ). By employing SP-Sepharose high-pressure and Ni-nitrilotriacetic acid fast-protein liquid chromatographies, a 19-fold enrichment with a final specific activity of 165.2 nmol mg of protein\n            <jats:sup>\u22121<\/jats:sup>\n            min\n            <jats:sup>\u22121<\/jats:sup>\n            was achieved by using 1-hexadecanol and palmitoyl-CoA as substrates. Incubation of purified 5\u2032His\n            <jats:sub>6<\/jats:sub>\n            WS\/DGAT with 1-hexadecanethiol and palmitoyl-CoA as substrates resulted in the formation of palmitic acid hexadecyl thio ester (10.4% relative specific activity of a 1-hexadecanol control). Utilization of 1,8-octanedithiol and palmitoyl-CoA as substrates led to the formation of 1-\n            <jats:italic>S<\/jats:italic>\n            -monopalmitoyloctanedithiol and minor amounts of 1,8-\n            <jats:italic>S<\/jats:italic>\n            -dipalmitoyloctanedithiol (59.3% relative specific activity of a 1-hexadecanol control). The latter dithio wax ester was efficiently produced when 1-\n            <jats:italic>S<\/jats:italic>\n            -monopalmitoyloctanedithiol and palmitoyl-CoA were used as substrates (13.4% specific activity relative to that of a 1-hexadecanol control). For the in vivo biosynthesis of thio wax esters, the knockout mutant\n            <jats:italic>Acinetobacter<\/jats:italic>\n            sp. strain ADP1\n            <jats:italic>acr1<\/jats:italic>\n            \u03a9Km, which is unable to produce fatty alcohols, was used. Cultivation of\n            <jats:italic>Acinetobacter<\/jats:italic>\n            sp. strain ADP1\n            <jats:italic>acr1<\/jats:italic>\n            \u03a9Km in the presence of gluconate, 1-hexadecanethiol, and oleic acid in nitrogen-limited mineral salts medium resulted in the accumulation of unusual thio wax esters that accounted for around 1.19% (wt\/wt) of the cellular dry weight and consisted mainly of oleic acid hexadecyl thioester as revealed by gas chromatography-mass spectrometry.\n          <\/jats:p>","DOI":"10.1128\/aem.71.2.790-796.2005","type":"journal-article","created":{"date-parts":[[2005,2,3]],"date-time":"2005-02-03T18:24:28Z","timestamp":1107455068000},"page":"790-796","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":47,"title":["Thio Wax Ester Biosynthesis Utilizing the Unspecific Bifunctional Wax Ester Synthase\/Acyl Coenzyme A:Diacylglycerol Acyltransferase of\n            <i>Acinetobacter<\/i>\n            sp. Strain ADP1"],"prefix":"10.1128","volume":"71","author":[{"given":"Stefan","family":"Uthoff","sequence":"first","affiliation":[{"name":"Institut fu\u0308r Molekulare Mikrobiologie und Biotechnologie, Westfa\u0308lische Wilhelms-Universita\u0308t Mu\u0308nster"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tim","family":"Sto\u0308veken","sequence":"additional","affiliation":[{"name":"Institut fu\u0308r Molekulare Mikrobiologie und Biotechnologie, Westfa\u0308lische Wilhelms-Universita\u0308t Mu\u0308nster"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nikolaus","family":"Weber","sequence":"additional","affiliation":[{"name":"Institut fu\u0308r Lipidforschung, Bundesforschungsanstalt fu\u0308r Erna\u0308hrung und Lebensmittel, Mu\u0308nster, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Klaus","family":"Vosmann","sequence":"additional","affiliation":[{"name":"Institut fu\u0308r Lipidforschung, Bundesforschungsanstalt fu\u0308r Erna\u0308hrung und Lebensmittel, Mu\u0308nster, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Erika","family":"Klein","sequence":"additional","affiliation":[{"name":"Institut fu\u0308r Lipidforschung, Bundesforschungsanstalt fu\u0308r Erna\u0308hrung und Lebensmittel, Mu\u0308nster, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rainer","family":"Kalscheuer","sequence":"additional","affiliation":[{"name":"Institut fu\u0308r Molekulare Mikrobiologie und Biotechnologie, Westfa\u0308lische Wilhelms-Universita\u0308t Mu\u0308nster"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexander","family":"Steinbu\u0308chel","sequence":"additional","affiliation":[{"name":"Institut fu\u0308r Molekulare Mikrobiologie und Biotechnologie, Westfa\u0308lische Wilhelms-Universita\u0308t Mu\u0308nster"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-002-1135-0"},{"key":"e_1_3_1_3_2","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1002\/lipi.19970990704","volume":"99","year":"1997","unstructured":"Alvarez, H. 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