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Direct involvement of the CreA transcription factor in penicillin biosynthesis and expression of the pcbAB gene in Penicillium chrysogenum

  • Applied genetics and molecular biotechnology
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Abstract

The transcription factor CreA is the main regulator responsible for carbon repression in filamentous fungi. CreA is a wide domain regulator that binds to regulatory elements in the promoters of target genes to repress their transcription. Penicillin biosynthesis and the expression of penicillin biosynthetic genes are subject to carbon repression. However, evidence of the participation of CreA in this regulation is still lacking, and previous studies on the promoter of the pcbC gene of Aspergillus nidulans indicated the lack of involvement of CreA in its regulation. Here we present clear evidence of the participation of CreA in carbon repression of penicillin biosynthesis and expression of the pcbAB gene, encoding the first enzyme of the pathway, in Penicillium chrysogenum. Mutations in cis of some of the putative CreA binding sites present in the pcbAB gene promoter fused to a reporter gene caused an important increase in the measured enzyme activity in glucose-containing medium, whereas activity in the medium with lactose was not affected. An RNAi strategy was used to attenuate the expression of the creA gene. Transformants expressing a small interfering RNA for creA showed higher penicillin production, and this increase was more evident when glucose was used as carbon source. These results confirm that CreA plays an important role in the regulation of penicillin biosynthesis in P. chrysogenum and opens the possibility of its utilization to improve the industrial production of this antibiotic.

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Acknowledgments

This work was supported by a Grant of the European Union (EUROFUNGBASE, LSSG-CT-2005-018964). C. Cepeda-García received a PhD fellowship associated to a research project from the Ministry of Science and Technology of Spain (BIO2000-1726-C02-01). R. Domínguez-Santos was granted a fellowship from the Junta de Castilla y León (ORDEN EDU/1204/2010) cofinanced by the European Social Fund.

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The authors declare that they have no conflict of interest.

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Author notes

  1. Cristina Cepeda-García

    Present address: Centro Andaluz de Biología Molecular y Medicina Regenerativa (CABIMER), Parque Científico y Tecnológico Cartuja, Sevilla, Spain

Authors and Affiliations

  1. Área de Microbiología, Facultad de Ciencias Biológicas y Ambientales, Universidad de León, León, Spain

    Cristina Cepeda-García, Rebeca Domínguez-Santos, Francisco Fierro & Juan Francisco Martín

  2. Instituto de Biotecnología (INBIOTEC), Parque Científico de León, León, Spain

    Rebeca Domínguez-Santos, Carlos García-Estrada & Juan Francisco Martín

  3. Departamento de Microbiología, Facultad de Ciencias Básicas, Universidad de Pamplona, Pamplona, Colombia

    Ramón O. García-Rico & Angela Cajiao

  4. Departamento de Biotecnología, Universidad Autónoma Metropolitana-Unidad Iztapalapa, Avda. San Rafael Atlixco 186, Col. Vicentina, 09340, México, DF, Mexico

    Francisco Fierro

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  1. Cristina Cepeda-García
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Correspondence to Francisco Fierro.

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Cristina Cepeda-García, Rebeca Domínguez-Santos and Ramón O. García-Rico contributed equally to this work.

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Cepeda-García, C., Domínguez-Santos, R., García-Rico, R.O. et al. Direct involvement of the CreA transcription factor in penicillin biosynthesis and expression of the pcbAB gene in Penicillium chrysogenum . Appl Microbiol Biotechnol 98, 7113–7124 (2014). https://doi.org/10.1007/s00253-014-5760-1

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