Post-translational enzyme modification by the phosphopantetheinyl transferase is required for lysine and penicillin biosynthesis but not for roquefortine or fatty acid formation in Penicillium chrysogenum.
NRPSs (non-ribosomal peptide synthetases) and PKSs (polyketide synthases) require post-translational phosphopantetheinylation to become active. This reaction is catalysed by a PPTase (4'-phosphopantetheinyl transferase). The ppt gene of Penicillium chrysogenum, encoding a protein that shares 50% similarity with the stand-alone large PPTases, has been cloned. This gene is present as a single copy in the genome of the wild-type and high-penicillin-producing strains (containing multiple copies of the penicillin gene cluster). Amplification of the ppt gene produced increases in isopenicillin N and benzylpenicillin biosynthesis. A PPTase-defective mutant (Wis54-PPT(-)) was obtained. It required lysine and lacked pigment and penicillin production, but it still synthesized normal levels of roquefortine. The biosynthesis of roquefortine does not appear to involve PPTase-mediated modification of the synthesizing enzymes. The PPT(-) mutant did not require fatty acids, which indicates that activation of the fatty acid synthase is performed by a different PPTase. Complementation of Wis54-PPT(-) with the ppt gene restored lysine biosynthesis, pigmentation and penicillin production, which demonstrates the wide range of processes controlled by this gene.
García-Estrada
Carlos
C
Instituto de Biotecnología de León. Parque Científico de León, Av. Real, 1, 24006, León, Spain.
Ullán
Ricardo V
RV
Velasco-Conde
Tania
T
Godio
Ramiro P
RP
Teijeira
Fernando
F
Vaca
Inmaculada
I
Feltrer
Raúl
R
Kosalková
Katarina
K
Mauriz
Elba
E
Martín
Juan F
JF
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015152:Blotting, Northern; D015139:Blotting, Southern; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D005227:Fatty Acids / Q000096:biosynthesis; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D016681:Genome, Fungal / Q000235:genetics; D006576:Heterocyclic Compounds, 4 or More Rings / Q000378:metabolism; D007211:Indoles / Q000378:metabolism; D008239:Lysine / Q000096:biosynthesis*; D010406:Penicillins / Q000096:biosynthesis*; D010408:Penicillium chrysogenum / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D010879:Piperazines / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D017855:Transferases (Other Substituted Phosphate Groups) / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D005227:Fatty Acids; D005656:Fungal Proteins; D006576:Heterocyclic Compounds, 4 or More Rings; D007211:Indoles; D010406:Penicillins; D010879:Piperazines; C082662:phosphopantetheinyl transferase; C012536:roquefortine; D017855:Transferases (Other Substituted Phosphate Groups); D008239:Lysine