Reduced function of a phenylacetate-oxidizing cytochrome p450 caused strong genetic improvement in early phylogeny of penicillin-producing strains.
The single-copy pahA gene from Penicillium chrysogenum encodes a phenylacetate 2-hydroxylase that catalyzes the first step of phenylacetate catabolism, an oxidative route that decreases the precursor availability for penicillin G biosynthesis. PahA protein is homologous to cytochrome P450 monooxygenases involved in the detoxification of xenobiotic compounds, with 84% identity to the Aspergillus nidulans homologue PhacA. Expression level of pahA displays an inverse correlation with the penicillin productivity of the strain and is subject to induction by phenylacetic acid. Gene expression studies have revealed a reduced oxidative activity of the protein encoded by pahA genes from penicillin-overproducing strains of P. chrysogenum compared to the activity conferred by phacA of A. nidulans. Sequencing and expression of wild-type pahA from P. chrysogenum NRRL 1951 revealed that an L181F mutation was responsible for the reduced function in present industrial strains. The mutation has been tracked down to Wisconsin 49-133, a mutant obtained at the Department of Botany of the University of Wisconsin in 1949, at the beginning of the development of the Wisconsin family of strains.
Rodríguez-Sáiz
M
M
Laboratorio de Biotecnología, Antibióticos S. A., 24009 León, Spain.
Barredo
J L
JL
Moreno
M A
MA
Fernández-Cañón
J M
JM
Peñalva
M A
MA
Díez
B
B
D001233:Aspergillus nidulans / Q000201:enzymology / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism*; D004352:Drug Resistance, Microbial; D005656:Fungal Proteins; D015966:Gene Expression Regulation, Fungal; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism; D010406:Penicillins / Q000096:biosynthesis*; D010408:Penicillium chrysogenum / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D010648:Phenylacetates / Q000378:metabolism* / Q000494:pharmacology; D010802:Phylogeny; D014158:Transcription, Genetic
D005656:Fungal Proteins; D010406:Penicillins; D010648:Phenylacetates; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D010105:Oxygenases; C436302:phenylacetate 2-hydroxylase; C025136:phenylacetic acid