7 research outputs found
Organization of the roquefortine/meleagrin biosynthetic gene cluster and transcriptomic analysis.
<p>(A) Roquefortine/meleagrin biosynthetic gene cluster and their orthologs in phylogenetically relative species. Homologous proteins are indicated with the same color. (B) Microarray analysis of the roquefortine biosynthetic genes in <i>P. chrysogenum</i> DS54555 using shake flask culture conditions in the absence (−) or presence (+) phenylacetic acid (PAA). (C) Correlation between the expression level of <i>roqA</i> and the concentration of the product HTD (<b>1)</b> present in the growth media. The concentration of <b>1</b> was determined by HPLC-UV-MS.</p
Total ion chromatogram for culture broth of <i>P. chrysogenum</i>.
<p>Total ion chromatogram (TIC, black) and normalized extracted ion chromatograms (EIC, colored) of secondary metabolites from the roquefortine/meleagrin pathway in the culture broth of <i>P. chrysogenum</i> DS54555. HTD (<b>1</b>, 8.8 min), DHTD (<b>2</b>, 9.4 min), glandicoline A (<b>5</b>, 16.8 min), roquefortine D (<b>3</b>, 18.3 min), glandicoline B (<b>6</b>, 18.4 min), meleagrin (<b>7</b>, 19.6 min), roquefortine C (<b>4</b>, 21.4 min).</p
Change in production of roquefortine/meleagrin metabolites after precursor addition compared to production in cultures without addition.
<p>Colored cells show mean levels that are significant (P<0.05) different.</p
Southern blot analysis for deletion of the genes in the roquefortine/meleagrin pathway.
<p>Southern blot hybridization was performed with total DNA extracted from <i>P. chrysogenum</i> DS54555 strains with a deletion of the following genes: <i>roqA</i> (A), <i>roqR</i> (B), <i>roqD</i> (C), <i>roqM</i> (D), <i>roqO</i> (E), <i>roqN</i> (F) and <i>roqT</i> (G). The DNA was digested with the restriction enzymes as indicated in the schemes.</p
Internal standard corrected concentrations (RR = response ratio) of secondary metabolites from roquefortine/meleagrin pathway.
<p>The metabolite concentrations in the culture broth of the Δ<i>roqR</i> (A), Δ<i>roqD</i> (B), Δ<i>roqM</i> (C), Δ<i>roqO</i> (D), Δ<i>roqN</i> (E) and Δ<i>roqT</i> (F) strains was compared to the host strain <i>P. chrysogenum</i> DS54555.</p
Födoval av torsk (Gadus morrhua L.) i Skagerrak och Kattegatt under februari 1981 /
<div><p>Profiling and structural elucidation of secondary metabolites produced by the filamentous fungus <i>Penicillium chrysogenum</i> and derived deletion strains were used to identify the various metabolites and enzymatic steps belonging to the roquefortine/meleagrin pathway. Major abundant metabolites of this pathway were identified as histidyltryptophanyldiketopiperazine (HTD), dehydrohistidyltryptophanyldi-ketopiperazine (DHTD), roquefortine D, roquefortine C, glandicoline A, glandicoline B and meleagrin. Specific genes could be assigned to each enzymatic reaction step. The nonribosomal peptide synthetase RoqA accepts L-histidine and L-tryptophan as substrates leading to the production of the diketopiperazine HTD. DHTD, previously suggested to be a degradation product of roquefortine C, was found to be derived from HTD involving the cytochrome P450 oxidoreductase RoqR. The dimethylallyltryptophan synthetase RoqD prenylates both HTD and DHTD yielding directly the products roquefortine D and roquefortine C without the synthesis of a previously suggested intermediate and the involvement of RoqM. This leads to a branch in the otherwise linear pathway. Roquefortine C is subsequently converted into glandicoline B with glandicoline A as intermediates, involving two monooxygenases (RoqM and RoqO) which were mixed up in an earlier attempt to elucidate the biosynthetic pathway. Eventually, meleagrin is produced from glandicoline B involving a methyltransferase (RoqN). It is concluded that roquefortine C and meleagrin are derived from a branched biosynthetic pathway.</p></div