{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T20:49:37Z","timestamp":1789505377190,"version":"build-2803163510"},"reference-count":45,"publisher":"Portland Press Ltd.","issue":"2","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2009,1,15]]},"abstract":"<jats:p>Enzymatic activation of PAA (phenylacetic acid) to phenylacetyl-CoA is an important step in the biosynthesis of the \u03b2-lactam antibiotic penicillin G by the fungus Penicillium chrysogenum. CoA esters of PAA and POA (phenoxyacetic acid) act as acyl donors in the exchange of the aminoadipyl side chain of isopenicillin N to produce penicillin G or penicillin V. The phl gene, encoding a PCL (phenylacetate\u2013CoA ligase), was cloned in Escherichia coli as a maltose-binding protein fusion and the biochemical properties of the enzyme were characterized. The recombinant fusion protein converted PAA into phenylacetyl-CoA in an ATP- and magnesium-dependent reaction. PCL could also activate POA, but the catalytic efficiency of the enzyme was rather low with kcat\/Km values of 0.23\u00b10.06 and 7.8\u00b11.2\u00a0mM\u22121\u00b7s\u22121 for PAA and POA respectively. Surprisingly, PCL was very efficient in catalysing the conversion of trans-cinnamic acids to the corresponding CoA thioesters [kcat\/Km=(3.1\u00b10.4)\u00d7102\u00a0mM\u22121\u00b7s\u22121 for trans-cinnamic acid]. Of all the substrates screened, medium-chain fatty acids, which also occur as the side chains of the natural penicillins F, DF, H and K, were the best substrates for PCL. The high preference for fatty acids could be explained by a homology model of PCL that was constructed on the basis of sequence similarity with the Japanese firefly luciferase. The results suggest that PCL has evolved from a fatty-acid-activating ancestral enzyme that may have been involved in the \u03b2-oxidation of fatty acids.<\/jats:p>","DOI":"10.1042\/bj20081257","type":"journal-article","created":{"date-parts":[[2008,10,8]],"date-time":"2008-10-08T10:53:25Z","timestamp":1223463205000},"page":"467-476","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":51,"title":["Characterization of a phenylacetate\u2013CoA ligase from <i>Penicillium chrysogenum<\/i>"],"prefix":"10.1042","volume":"417","author":[{"given":"Martijn\u00a0J.","family":"Koetsier","sequence":"first","affiliation":[{"name":"Biochemical Laboratory, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, Groningen NL-9747 AG, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter\u00a0A.","family":"Jekel","sequence":"additional","affiliation":[{"name":"Biochemical Laboratory, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, Groningen NL-9747 AG, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marco\u00a0A.","family":"van\u00a0den\u00a0Berg","sequence":"additional","affiliation":[{"name":"DSM Anti-Infectives, P.O. Box 425, Delft NL-2600 AK, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roel\u00a0A.\u00a0L.","family":"Bovenberg","sequence":"additional","affiliation":[{"name":"DSM Anti-Infectives, P.O. Box 425, Delft NL-2600 AK, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dick\u00a0B.","family":"Janssen","sequence":"additional","affiliation":[{"name":"Biochemical Laboratory, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, Groningen NL-9747 AG, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","published-online":{"date-parts":[[2008,12,23]]},"reference":[{"key":"2021112214154267200_B1","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1111\/j.1432-1033.1993.tb18038.x","article-title":"The isopenicillin-N acyltransferase of Penicillium chrysogenum has isopenicillin-N amidohydrolase, 6-aminopenicillanic acid acyltransferase and penicillin amidase activities, all of which are encoded by the single penDE gene","volume":"215","author":"Alvarez","year":"1993","journal-title":"Eur. J. Biochem."},{"key":"2021112214154267200_B2","doi-asserted-by":"crossref","first-page":"684","DOI":"10.7164\/antibiotics.43.684","article-title":"Biosynthesis of benzylpenicillin (G), phenoxymethylpenicillin (V) and octanoylpenicillin (K) from glutathione S-derivatives","volume":"43","author":"Ferrero","year":"1990","journal-title":"J. Antibiot."},{"key":"2021112214154267200_B3","doi-asserted-by":"crossref","first-page":"5474","DOI":"10.1016\/S0021-9258(18)42790-1","article-title":"Isolation and characterization of the acetyl-CoA synthetase from Penicillium chrysogenum: involvement of this enzyme in the biosynthesis of penicillins","volume":"267","author":"Mart\u00ednez-Blanco","year":"1992","journal-title":"J. Biol. 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U.S.A."},{"key":"2021112214154267200_B23","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1021\/cr00002a004","article-title":"A survey of Hammett substituent constants and resonance and field parameters","volume":"91","author":"Hansch","year":"1991","journal-title":"Chem. Rev."},{"key":"2021112214154267200_B24","doi-asserted-by":"crossref","first-page":"1900","DOI":"10.1039\/B301513K","article-title":"Acylation is rate-limiting in glucosylasparaginase-catalyzed hydrolysis of N4-(4\u2032-substituted phenyl)-L-asparagines","volume":"1","author":"Du","year":"2003","journal-title":"Org. Biomol. Chem."},{"key":"2021112214154267200_B25","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1128\/br.20.3.133-147.1956","article-title":"The metabolism of the Penicillia in relation to penicillin biosynthesis","volume":"20","author":"Arnstein","year":"1956","journal-title":"Bacteriol. 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Bacteriol."},{"key":"2021112214154267200_B28","doi-asserted-by":"crossref","first-page":"2866","DOI":"10.1021\/bi0271603","article-title":"The 1.75\u00a0\u00c5 crystal structure of acetyl-CoA synthetase bound to adenosine-5\u2032-propylphosphate and coenzyme A","volume":"42","author":"Gulick","year":"2003","journal-title":"Biochemistry"},{"key":"2021112214154267200_B29","doi-asserted-by":"crossref","first-page":"1425","DOI":"10.1021\/bi035911a","article-title":"Crystal structure of yeast acetyl-coenzyme A synthetase in complex with AMP","volume":"43","author":"Jogl","year":"2004","journal-title":"Biochemistry"},{"key":"2021112214154267200_B30","doi-asserted-by":"crossref","first-page":"8670","DOI":"10.1021\/bi049384m","article-title":"Crystal structure of 4-chlorobenzoate:CoA ligase\/synthetase in the unliganded and aryl substrate-bound states","volume":"43","author":"Gulick","year":"2004","journal-title":"Biochemistry"},{"key":"2021112214154267200_B31","doi-asserted-by":"crossref","first-page":"31717","DOI":"10.1074\/jbc.M400100200","article-title":"Structural basis of the substrate-specific two-step catalysis of long chain fatty acyl-CoA synthetase dimer","volume":"279","author":"Hisanaga","year":"2004","journal-title":"J. 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Acta"},{"key":"2021112214154267200_B41","doi-asserted-by":"crossref","first-page":"1838","DOI":"10.1007\/s00018-003-3076-x","article-title":"Fatty acid metabolism in Saccharomyces cerevisiae","volume":"60","author":"Van\u00a0Roermund","year":"2003","journal-title":"Cell. Mol. Life Sci."},{"key":"2021112214154267200_B42","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1128\/am.5.6.386-391.1957","article-title":"Some factors influencing the production of certain biosynthetic penicillins","volume":"5","author":"MacMorine","year":"1957","journal-title":"Appl. Microbiol."},{"key":"2021112214154267200_B43","doi-asserted-by":"crossref","first-page":"2052","DOI":"10.1021\/ja01161a049","article-title":"Precursors for aliphatic penicillins","volume":"72","author":"Thorn","year":"1950","journal-title":"J. Am. Chem. Soc."},{"key":"2021112214154267200_B44","doi-asserted-by":"crossref","first-page":"16358","DOI":"10.1016\/S0021-9258(17)46231-4","article-title":"The cluster of penicillin biosynthetic genes: identification and characterization of the pcbAB gene encoding the \u03b1-aminoadipyl-cysteinyl-valine synthetase and linkage to the pcbC and penDE genes","volume":"265","author":"Diez","year":"1990","journal-title":"J. Biol. Chem."},{"key":"2021112214154267200_B45","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1111\/j.1365-2583.2006.00646.x","article-title":"Cloning and characterization of the homologous genes of firefly luciferase in the mealworm beetle, Tenebrio molitor","volume":"15","author":"Oba","year":"2006","journal-title":"Insect Mol. Biol."}],"container-title":["Biochemical Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/417\/2\/467\/655961\/bj4170467.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/portlandpress.com\/biochemj\/article-pdf\/417\/2\/467\/655961\/bj4170467.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,11,22]],"date-time":"2021-11-22T17:04:08Z","timestamp":1637600648000},"score":1,"resource":{"primary":{"URL":"https:\/\/portlandpress.com\/biochemj\/article\/417\/2\/467\/44790\/Characterization-of-a-phenylacetate-CoA-ligase"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2008,12,23]]},"references-count":45,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2009,1,15]]}},"URL":"https:\/\/doi.org\/10.1042\/bj20081257","relation":{},"ISSN":["0264-6021","1470-8728"],"issn-type":[{"value":"0264-6021","type":"print"},{"value":"1470-8728","type":"electronic"}],"subject":[],"published":{"date-parts":[[2008,12,23]]}}}