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Continuous Production of a Novel Lysozyme via Secretion from the Yeast, Pichia pastoris

Bio/Technology volume 7, pages 160–164 (1989)Cite this article

Abstract

We have cloned a cDNA for bovine lysozyme c2, a novel lysozyme characteristic of the cow stomach and expressed its protein product by secretion using its native signal sequence in the methylotrophic yeast, Pichia pastoris. A semi–continuous fermentation process used for production resulted in extremely high cell densities and product concentrations. During fed–batch fermentation, bovine lysozyme concentrations exceeded 550 mg/liter, and cell densities exceeded 120 g dry cells/liter. In continuous culture, bovine lysozyme and cell concentrations remained constant at approximately 350 mg/liter and 100 g dry cells/liter, respectively. The recombinant product is a single biologically active species that constitutes >80% of the protein in media from expressing cells; the signal sequence of the pre–lysozyme is accurately processed.

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Author notes

  1. M. E. Digan: Corresponding author.

Authors and Affiliations

  1. The Salk Institute Biotechnology/Industrial Associates, Inc., La Jolla, California, 92037

    M. E. Digan, S. V. Lair, R. A. Brierley, R. S. Siegel, M. E. Williams, S. B. Ellis, P. A. Kellaris, S. A. Provow, W. S. Craig, G. Veliçelebi, M. M. Harpold & G. P. Thill

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  1. M. E. Digan
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  2. S. V. Lair
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  3. R. A. Brierley
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  4. R. S. Siegel
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  5. M. E. Williams
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  6. S. B. Ellis
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  7. P. A. Kellaris
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  9. W. S. Craig
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  11. M. M. Harpold
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  12. G. P. Thill
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Digan, M., Lair, S., Brierley, R. et al. Continuous Production of a Novel Lysozyme via Secretion from the Yeast, Pichia pastoris. Nat Biotechnol 7, 160–164 (1989). https://doi.org/10.1038/nbt0289-160

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