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Rachiplusia nu larva as a biofactory to achieve high level expression of horseradish peroxidase

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Abstract

A process based on orally-infected Rachiplusia nu larvae as biological factories for expression and one-step purification of horseradish peroxidase isozyme C (HRP-C) is described. The process allows obtaining high levels of pure HRP-C by membrane chromatography purification. The introduction of the partial polyhedrin homology sequence element in the target gene increased HRP-C expression level by 2.8-fold whereas it increased 1.8-fold when the larvae were reared at 27°C instead of at 24°C, summing up a 4.6-fold overall increase in the expression level. Additionally, HRP-C purification by membrane chromatography at a high flow rate greatly increase D the productivity without affecting the resolution. The Vmax and Km values of the recombinant HRP-C were similar to those of the HRP from Armoracia rusticana roots.

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Acknowledgments

This work was supported by grants from the Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina (PIP 00052) and the Universidad de Buenos Aires (B 043).

FJW, OC and MVM are career researchers from the Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina.

We thank Ms. M. Fogar and Ms. M. Simonella from INTA Sáenz Peña for provision of the larvae.

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Authors and Affiliations

  1. Cátedra de Microbiología Industrial y Biotecnología, Facultad de Farmacia y Bioquímica (UBA), Junín 956 (1113), Ciudad Autónoma de Buenos Aires, Argentina

    Lucía Virginia Romero, Alexandra Marisa Targovnik, Federico Javier Wolman, Osvaldo Cascone & María Victoria Miranda

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  1. Lucía Virginia Romero
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  2. Alexandra Marisa Targovnik
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  3. Federico Javier Wolman
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  4. Osvaldo Cascone
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  5. María Victoria Miranda
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Correspondence to Osvaldo Cascone.

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Romero, L.V., Targovnik, A.M., Wolman, F.J. et al. Rachiplusia nu larva as a biofactory to achieve high level expression of horseradish peroxidase. Biotechnol Lett 33, 947–956 (2011). https://doi.org/10.1007/s10529-011-0540-9

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  • DOI: https://doi.org/10.1007/s10529-011-0540-9

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