Caractérisation des polycétones synthases intervenant dans la biosynthèse d’ochratoxine A, d’acide pénicillique, d’asperlactone et d’isoasperlactone chez aspergillus westerdijkiae

作者: Nafees Bacha

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摘要: Aspergillus westerdijkiaem qui est recemment demembre d'A. ochraceus un producteur principal de plusieurs composes type polycetone d'importance economique. Ces incluent l’ochratoxin A, mellein, l'acide penicillique, asperlactone et l’isoasperlactone quelques intermediaires comme 6- methylsalicylique l’acide orsellinique. La biosynthese ces metabolites catalysee par groupe d'enzymes connues la synthases (PKSs). Ce travail a ete vise pour cloner caracterise fonctionnellement les differentes genes des PKS i.e. aoks1, aolc35-12 aomsas, polyketide synthases-non ribosomal peptide synthase (PKS-NRPS) aolc35-6, chez A. westerdijkiae. ont inactives l'insertion du gene d’hygromycine B phosphotransferase d’Escherichia coli dans le genome westerdijkiae, obtenir mutantes aoΔks1, aoΔlc35-12, aoΔmsas aoΔlc35-6. Les mutants aoΔks1 aoΔlc35-12 trouves deficients d’ochratoxin mais produisaient encore mellein. A notre connaissance, c’est premiere fois que nous avons impliquees d’OTA, sachant etait propose litterature intermediaire, joue cune role biosynthesis l'OTA. Ensuite mutant n'a pas seulement perdu capacite produire isoasperlactone asperlactone, aussi il ne produit l’intermediaire acide 6-methylsalicylique. Base sur experiences caracterisation genetique complementation chimiques, shema hypothetique d’asperlactone lequel 6-methylsalicylique, diepoxide aspyrone jouent d’intermediaires. techniques knock-out reverse transcription PCR (RT-PCR) montre seulle PKS-NRPS « aolc35-6 » identifie westerdijkiae codant intermediaire inconnu(s) pourrait inciter l'expression aomsas implique d'acide orsellinique penicillique. ABSTRACT : which is recently dismembered from ochraceusm the producer of several economically important metabolites. These include ochratoxin penicillic acid, and some intermediates like orsellinic acid 6-methylsalicylic acid. The these catalyzed by group enzymes known as This work was aimed to clone functionally characterized various synthasesnon in were inactivated insertion Escherichia hygromycin obtain aoΔlc35-6 mutants. found deficient but are still producing To our knowledge, we for first time involved OTA biosynthesis, with information about mellein proposed literature be an intermediate OTA. Further not only lost capacity produce also nature product Based on genetic characterization chemical experiments, have hypothetical pathway mentioning that diepoxid asperlactone. Gene knockout technique shown so far identified encoding certain unknown intermediate(s) induces expression

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