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Secretion of Kluyveromyces lactis Cu/Zn SOD: strategies for enhanced production

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Abstract

The Kluyveromyces lactis Cu/Zn SOD gene (SOD1) was fused with the toxin K1 signal sequence to obtain extracellular production of superoxide dismutase. Kluyveromyces marxianus L3 and K. lactis MW98-8C strains were transformed and compared as hosts for the secretion. The effects of the media composition were evaluated: In K. lactis, the highest volumetric activity was obtained in YKK synthetic medium in the presence of Cu2+/Zn2+ cofactors (9.6 kU l−1). In K. marxianus, active SOD was produced only in YPD medium supplemented with Cu2+ and Zn2+ (8.8 kU l−1). In order to improve the production of secreted active SOD in K. lactis, the SOD1 copper carrier (CCS1) was overexpressed and targeted to the secretory apparatus. A positive effect was observed only when K. lactis was grown in a medium without Cu2+/Zn2+ supplement. The best performing culture conditions for K. lactis and K. marxianus recombinant strains were successfully applied to two laboratory-scale fed-batch processes, and volumetric SOD activities increased up to 19.4 and 24.1 kU l−1, respectively.

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Acknowledgments

This work was partially supported by the Istituto Pasteur Cenci Bolognetti and by the AST 2008.

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Author notes

  1. Stefano Raimondi

    Present address: Department of Chemistry, University of Modena and Reggio Emilia, Via Campi 183, 41125, Modena, Italy

Authors and Affiliations

  1. Department of Developmental and Cell Biology, University LA SAPIENZA, P.le Aldo Moro 5, 00185, Rome, Italy

    Stefano Raimondi, Daniela Uccelletti & Claudio Palleschi

  2. Department of Chemistry, University of Modena and Reggio Emilia, Via Campi 183, 41125, Modena, Italy

    Alberto Amaretti, Alan Leonardi & Maddalena Rossi

Authors
  1. Stefano Raimondi
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  2. Daniela Uccelletti
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  5. Claudio Palleschi
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  6. Maddalena Rossi
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Correspondence to Maddalena Rossi.

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Raimondi, S., Uccelletti, D., Amaretti, A. et al. Secretion of Kluyveromyces lactis Cu/Zn SOD: strategies for enhanced production. Appl Microbiol Biotechnol 86, 871–878 (2010). https://doi.org/10.1007/s00253-009-2353-5

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  • DOI: https://doi.org/10.1007/s00253-009-2353-5

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