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Effect of pH on the stability of Pleurotus eryngii versatile peroxidase during heterologous production in Emericella nidulans

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Abstract

Complementary DNA (cDNA) encoding the new versatile peroxidase from the ligninolytic basidiomycete Pleurotus eryngii has been expressed in the ascomycete Emericella nidulans. In recombinant E. nidulans cultures, the pH reached values as high as 8.3, correlating with a sharp decrease in peroxidase activity. Peroxidase was rapidly inactivated at alkaline pH, but was comparatively stable at acidic pH. The peroxidase inactivation in alkaline buffer could be reversed by adding Ca2+ and lowering the pH. However, reactivation did not result after incubating the enzyme in non-buffered E. nidulans cultures that reached pH 7.5. To optimize recombinant peroxidase production, the effect of controlling the pH in E. nidulans bioreactor cultures was studied. An extended growth period, and a significant increase in the recombinant peroxidase level (5.3-fold higher activity than in the bioreactor without pH control) was obtained when the pH was maintained at 6.8, showing that culture pH is an important parameter for recombinant peroxidase production.

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Acknowledgements

The authors thank M.A. Peñalva (CIB, CSIC, Madrid) for providing the E. nidulans strain and the plasmid palcA. This work was partially supported by the EU contract “Fungal metalloenzymes oxidizing aromatic compounds of industrial interest” (QLK3-99-590), the Comunidad de Madrid, and the Spanish Commission of Science and Technology (BIO98-610 and BIO99-908).

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Authors and Affiliations

  1. Department of Chemical Engineering, Institute of Technology, 15706, Santiago de Compostela, Spain

    T. A. Lú-Chau, G. Feijoo & J. M. Lema

  2. Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, 28040, Madrid, Spain

    F. J. Ruiz-Dueñas, S. Camarero, M. J. Martínez & A. T. Martínez

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  1. T. A. Lú-Chau
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  2. F. J. Ruiz-Dueñas
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  3. S. Camarero
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  4. G. Feijoo
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  5. M. J. Martínez
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  6. J. M. Lema
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  7. A. T. Martínez
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Correspondence to T. A. Lú-Chau.

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Lú-Chau, T.A., Ruiz-Dueñas, F.J., Camarero, S. et al. Effect of pH on the stability of Pleurotus eryngii versatile peroxidase during heterologous production in Emericella nidulans. Bioprocess Biosyst Eng 26, 287–293 (2004). https://doi.org/10.1007/s00449-004-0365-1

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  • DOI: https://doi.org/10.1007/s00449-004-0365-1

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