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Functional analysis of Kluyveromyces lactis carboxylic acids permeases: heterologous expression of KlJEN1 and KlJEN2 genes

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Abstract

The present work describes a detailed physiological and molecular characterization of the mechanisms of transport of carboxylic acids in Kluyveromyces lactis. This yeast species presents two homologue genes to JEN1 of Saccharomyces cerevisiae: KlJEN1 encodes a monocarboxylate permease and KlJEN2 encodes a dicarboxylic acid permease. In the strain K. lactis GG1888, expression of these genes does not require an inducer and activity for both transport systems was observed in glucose-grown cells. To confirm their key role for carboxylic acids transport in K. lactis, null mutants were analyzed. Heterologous expression in S. cerevisiae has been performed and chimeric fusions with GFP showed their proper localization in the plasma membrane. S. cerevisiae jen1Δ cells transformed with KlJEN1 recovered the capacity to use lactic acid, as well as to transport labeled lactic acid by a mediated mechanism. When KlJEN2 was heterologously expressed, S. cerevisiae transformants gained the ability to transport labeled succinic and malic acids by a mediated mechanism, exhibiting, however, a poor growth in malic acid containing media. The results confirmed the role of KlJen1p and KlJen2p as mono and dicarboxylic acids permeases, respectively, not subjected to glucose repression, being fully functional in S. cerevisiae.

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Acknowledgments

This study was supported by the Portuguese grant POCTI/BIO/38106/2001 (Eixo 2, Medida 2.3, QCAIII—FEDER).

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Authors and Affiliations

  1. Instituto Superior de Ciências da Saúde-Norte (ISCSN), Rua Central da Gandra 1317 4585-116 Gandra, Paredes, Portugal

    Odília Queirós

  2. Centro de Biologia, Departamento de Biologia, Universidade do Minho, Campus de Gualtar, 4710-057, Braga, Portugal

    Leonor Pereira, Sandra Paiva & Margarida Casal

  3. Instituto de Biologia Molecular e Celular (IBMC), Universidade do Porto, R. Campo Alegre 823, 4150-180, Porto, Portugal

    Odília Queirós, Leonor Pereira & Pedro Moradas-Ferreira

  4. Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto, Largo Professor Abel Salazar 2, 4099-003, Porto, Portugal

    Pedro Moradas-Ferreira

Authors
  1. Odília Queirós
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  2. Leonor Pereira
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  3. Sandra Paiva
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  4. Pedro Moradas-Ferreira
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  5. Margarida Casal
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Corresponding author

Correspondence to Margarida Casal.

Additional information

Communicated by K. Breunig.

O. Queirós and L. Pereira contributed equally to this work.

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Queirós, O., Pereira, L., Paiva, S. et al. Functional analysis of Kluyveromyces lactis carboxylic acids permeases: heterologous expression of KlJEN1 and KlJEN2 genes. Curr Genet 51, 161–169 (2007). https://doi.org/10.1007/s00294-006-0107-9

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  • DOI: https://doi.org/10.1007/s00294-006-0107-9

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