title: The npgA/ cfwA gene encodes a putative 4'-phosphopantetheinyl transferase which is essential for penicillin biosynthesis in Aspergillus nidulans.
abstract: Non-ribosomal peptide synthetases, polyketides and fatty acid synthetases have a modular organisation of multi-enzymatic activities. In all of them, the acyl or peptidyl carrier proteins have 4'-phosphopantetheine (P-pant) as an essential prosthetic group. This is added by 4'-phosphopantetheinyl transferases (PPTases) that derive the P-pant group from coenzyme A. While many PPTases of varying specificity have now been isolated from a number of bacteria, a filamentous fungal PPTase has yet to be characterised. Through database searching of the Aspergillus fumigatus genome sequence against Sfp from Bacillus subtilis, we identified a unique sequence which appears to encode a PPTase, as deduced from conserved residues considered important in PPTases. The PPTase candidate was used to search the NCBI data base and an unexpected homologue in A. nidulans was identified as npgA. Mutations in this gene (cfwA/ npgA) were identified previously as leading to defects in growth and pigmentation. To test whether the temperature-sensitive cfwA2 mutation impairs penicillin biosynthesis, which is dependent on the delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase, bioassays with B. calidolactis were set up at permissive and non-permissive temperatures. The cfwA2 mutant did not produce penicillin at the non-permissive temperature. Since no other PPTase homologue has been detected in the A. fumigatus genome to date, the data suggest that a single enzyme may be able to transfer the cofactor to a broad range of enzymes with acyl or peptidyl carrier protein domains.
authors.affiliation: Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield, S10 2TN, UK. keszenman-pereyra@sheffield.ac.uk
authors.forename: David
authors.initials: D
authors.lastname: Keszenman-Pereyra
authors.forename: Stephen
authors.initials: S
authors.lastname: Lawrence
authors.forename: Mohammed-E
authors.initials: ME
authors.lastname: Twfieg
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Price
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Turner
mesh_terms: D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000378:metabolism*; D001483:Base Sequence; D001681:Biological Assay; D017931:DNA Primers; D008969:Molecular Sequence Data; D010406:Penicillins / Q000096:biosynthesis*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017855:Transferases (Other Substituted Phosphate Groups) / Q000235:genetics* / Q000378:metabolism
chemical_list: D017931:DNA Primers; D010406:Penicillins; D017855:Transferases (Other Substituted Phosphate Groups); C021364:holo-(acyl-carrier-protein) synthase