title: Engineering of Penicillium chrysogenum for fermentative production of a novel carbamoylated cephem antibiotic precursor.
abstract: Penicillium chrysogenum was successfully engineered to produce a novel carbamoylated cephalosporin that can be used as a synthon for semi-synthetic cephalosporins. To this end, genes for Acremonium chrysogenum expandase/hydroxylase and Streptomyces clavuligerus carbamoyltransferase were expressed in a penicillinG high-producing strain of P.chrysogenum. Growth of the engineered strain in the presence of adipic acid resulted in production of adipoyl-7-amino-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid (ad7-ACCCA) and of several adipoylated pathway intermediates. A combinatorial chemostat-based transcriptome study, in which the ad7-ACCCA-producing strain and a strain lacking key genes in beta-lactam synthesis were grown in the presence and absence of adipic acid, enabled the dissection of transcriptional responses to adipic acid per se and to ad7-ACCCA production. Transcriptome analysis revealed that adipate catabolism in P.chrysogenum occurs via beta-oxidation and enabled the identification of putative genes for enzymes involved in mitochondrial and peroxisomal beta-oxidation pathways. Several of the genes that showed a specifically altered transcript level in ad7-ACCCA-producing cultures were previously implicated in oxidative stress responses.
authors.affiliation: Department of Biotechnology, Delft University of Technology, The Netherlands.
authors.forename: Diana M
authors.initials: DM
authors.lastname: Harris
authors.forename: Ilja
authors.initials: I
authors.lastname: Westerlaken
authors.forename: Dick
authors.initials: D
authors.lastname: Schipper
authors.forename: Zita A
authors.initials: ZA
authors.lastname: van der Krogt
authors.forename: Andreas K
authors.initials: AK
authors.lastname: Gombert
authors.forename: John
authors.initials: J
authors.lastname: Sutherland
authors.forename: Leonie M
authors.initials: LM
authors.lastname: Raamsdonk
authors.forename: Marco A
authors.initials: MA
authors.lastname: van den Berg
authors.forename: Roel A L
authors.initials: RA
authors.lastname: Bovenberg
authors.forename: Jack T
authors.initials: JT
authors.lastname: Pronk
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Daran
mesh_terms: D000164:Acremonium / Q000502:physiology*; D002511:Cephalosporins / Q000096:biosynthesis*; D024861:Genetic Enhancement / Q000379:methods*; D010408:Penicillium chrysogenum / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis; D013302:Streptomyces / Q000502:physiology*
chemical_list: D002511:Cephalosporins; D011994:Recombinant Proteins