Molecular characterization of a fungal gene paralogue of the penicillin penDE gene of Penicillium chrysogenum
[EN] Background. Penicillium chrysogenum converts isopenicillin N (IPN) into hydrophobic penicillins by means of the peroxisomal IPN acyltransferase (IAT), which is encoded by the penDE gene. In silico analysis of the P. chrysogenum genome revealed the presence of a gene, Pc13g09140, initially descr...
| Autores: | , , , , , |
|---|---|
| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2009 |
| País: | España |
| Institución: | Universidad de León |
| Repositorio: | BULERIA. Repositorio Institucional de la Universidad de León |
| OAI Identifier: | oai:buleria.unileon.es:10612/24277 |
| Acceso en línea: | https://bmcmicrobiol.biomedcentral.com/articles/10.1186/1471-2180-9-104 https://hdl.handle.net/10612/24277 |
| Access Level: | acceso abierto |
| Palabra clave: | Toxicología Fungal gene penDE gene Penicillium chrysogenum |
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Molecular characterization of a fungal gene paralogue of the penicillin penDE gene of Penicillium chrysogenumGarcía Estrada, CarlosVaca, InmaculadaUllán, Ricardo Vicentevan den Berg, Marco ABovenberg, Roel ALMartín Martín, Juan FranciscoToxicologíaFungal genepenDE genePenicillium chrysogenum[EN] Background. Penicillium chrysogenum converts isopenicillin N (IPN) into hydrophobic penicillins by means of the peroxisomal IPN acyltransferase (IAT), which is encoded by the penDE gene. In silico analysis of the P. chrysogenum genome revealed the presence of a gene, Pc13g09140, initially described as paralogue of the IAT-encoding penDE gene. We have termed this gene ial because it encodes a protein with high similarity to IAT (IAL for IAT-Like). We have conducted an investigation to characterize the ial gene and to determine the role of the IAL protein in the penicillin biosynthetic pathway. Results The IAL contains motifs characteristic of the IAT such as the processing site, but lacks the peroxisomal targeting sequence ARL. Null ial mutants and overexpressing strains indicated that IAL lacks acyltransferase (penicillin biosynthetic) and amidohydrolase (6-APA forming) activities in vivo. When the canonical ARL motif (leading to peroxisomal targeting) was added to the C-terminus of the IAL protein (IALARL) by site-directed mutagenesis, no penicillin biosynthetic activity was detected. Since the IAT is only active after an accurate self-processing of the preprotein into α and β subunits, self-processing of the IAL was tested in Escherichia coli. Overexpression experiments and SDS-PAGE analysis revealed that IAL is also self-processed in two subunits, but despite the correct processing, the enzyme remained inactive in vitro.Conclusion: No activity related to the penicillin biosynthesis was detected for the IAL. Sequence comparison among the P. chrysogenum IAL, the A. nidulans IAL homologue and the IAT, revealed that the lack of enzyme activity seems to be due to an alteration of the essential Ser309 in the thioesterase active site. Homologues of the ial gene have been found in many other ascomycetes, including non-penicillin producers. Our data suggest that like in A. nidulans, the ial and penDE genes might have been formed from a single ancestral gene that became duplicated during evolution, although a separate evolutive origin for the ial and penDE genes, is also discussed.SIBioMed CentralToxicologiaFacultad de Veterinaria2009info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttps://bmcmicrobiol.biomedcentral.com/articles/10.1186/1471-2180-9-104https://hdl.handle.net/10612/24277reponame:BULERIA. Repositorio Institucional de la Universidad de Leóninstname:Universidad de LeónIngléshttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:buleria.unileon.es:10612/242772026-06-24T12:43:27Z |
| dc.title.none.fl_str_mv |
Molecular characterization of a fungal gene paralogue of the penicillin penDE gene of Penicillium chrysogenum |
| title |
Molecular characterization of a fungal gene paralogue of the penicillin penDE gene of Penicillium chrysogenum |
| spellingShingle |
Molecular characterization of a fungal gene paralogue of the penicillin penDE gene of Penicillium chrysogenum García Estrada, Carlos Toxicología Fungal gene penDE gene Penicillium chrysogenum |
| title_short |
Molecular characterization of a fungal gene paralogue of the penicillin penDE gene of Penicillium chrysogenum |
| title_full |
Molecular characterization of a fungal gene paralogue of the penicillin penDE gene of Penicillium chrysogenum |
| title_fullStr |
Molecular characterization of a fungal gene paralogue of the penicillin penDE gene of Penicillium chrysogenum |
| title_full_unstemmed |
Molecular characterization of a fungal gene paralogue of the penicillin penDE gene of Penicillium chrysogenum |
| title_sort |
Molecular characterization of a fungal gene paralogue of the penicillin penDE gene of Penicillium chrysogenum |
| dc.creator.none.fl_str_mv |
García Estrada, Carlos Vaca, Inmaculada Ullán, Ricardo Vicente van den Berg, Marco A Bovenberg, Roel AL Martín Martín, Juan Francisco |
| author |
García Estrada, Carlos |
| author_facet |
García Estrada, Carlos Vaca, Inmaculada Ullán, Ricardo Vicente van den Berg, Marco A Bovenberg, Roel AL Martín Martín, Juan Francisco |
| author_role |
author |
| author2 |
Vaca, Inmaculada Ullán, Ricardo Vicente van den Berg, Marco A Bovenberg, Roel AL Martín Martín, Juan Francisco |
| author2_role |
author author author author author |
| dc.contributor.none.fl_str_mv |
Toxicologia Facultad de Veterinaria |
| dc.subject.none.fl_str_mv |
Toxicología Fungal gene penDE gene Penicillium chrysogenum |
| topic |
Toxicología Fungal gene penDE gene Penicillium chrysogenum |
| description |
[EN] Background. Penicillium chrysogenum converts isopenicillin N (IPN) into hydrophobic penicillins by means of the peroxisomal IPN acyltransferase (IAT), which is encoded by the penDE gene. In silico analysis of the P. chrysogenum genome revealed the presence of a gene, Pc13g09140, initially described as paralogue of the IAT-encoding penDE gene. We have termed this gene ial because it encodes a protein with high similarity to IAT (IAL for IAT-Like). We have conducted an investigation to characterize the ial gene and to determine the role of the IAL protein in the penicillin biosynthetic pathway. Results The IAL contains motifs characteristic of the IAT such as the processing site, but lacks the peroxisomal targeting sequence ARL. Null ial mutants and overexpressing strains indicated that IAL lacks acyltransferase (penicillin biosynthetic) and amidohydrolase (6-APA forming) activities in vivo. When the canonical ARL motif (leading to peroxisomal targeting) was added to the C-terminus of the IAL protein (IALARL) by site-directed mutagenesis, no penicillin biosynthetic activity was detected. Since the IAT is only active after an accurate self-processing of the preprotein into α and β subunits, self-processing of the IAL was tested in Escherichia coli. Overexpression experiments and SDS-PAGE analysis revealed that IAL is also self-processed in two subunits, but despite the correct processing, the enzyme remained inactive in vitro.Conclusion: No activity related to the penicillin biosynthesis was detected for the IAL. Sequence comparison among the P. chrysogenum IAL, the A. nidulans IAL homologue and the IAT, revealed that the lack of enzyme activity seems to be due to an alteration of the essential Ser309 in the thioesterase active site. Homologues of the ial gene have been found in many other ascomycetes, including non-penicillin producers. Our data suggest that like in A. nidulans, the ial and penDE genes might have been formed from a single ancestral gene that became duplicated during evolution, although a separate evolutive origin for the ial and penDE genes, is also discussed. |
| publishDate |
2009 |
| dc.date.none.fl_str_mv |
2009 |
| dc.type.none.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
| format |
article |
| status_str |
publishedVersion |
| dc.identifier.none.fl_str_mv |
https://bmcmicrobiol.biomedcentral.com/articles/10.1186/1471-2180-9-104 https://hdl.handle.net/10612/24277 |
| url |
https://bmcmicrobiol.biomedcentral.com/articles/10.1186/1471-2180-9-104 https://hdl.handle.net/10612/24277 |
| dc.language.none.fl_str_mv |
Inglés |
| language_invalid_str_mv |
Inglés |
| dc.rights.none.fl_str_mv |
http://creativecommons.org/licenses/by/4.0/ info:eu-repo/semantics/openAccess |
| rights_invalid_str_mv |
http://creativecommons.org/licenses/by/4.0/ |
| eu_rights_str_mv |
openAccess |
| dc.publisher.none.fl_str_mv |
BioMed Central |
| publisher.none.fl_str_mv |
BioMed Central |
| dc.source.none.fl_str_mv |
reponame:BULERIA. Repositorio Institucional de la Universidad de León instname:Universidad de León |
| instname_str |
Universidad de León |
| reponame_str |
BULERIA. Repositorio Institucional de la Universidad de León |
| collection |
BULERIA. Repositorio Institucional de la Universidad de León |
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15,812429 |