Abstract
We report a genetic approach for the overproduction of a Rhizopus oryzae lipase (ROL) expressed in Pichia pastoris under the alcohol oxidase 1 promoter, Paoxi- It is based on the development of a stable P. pastoris strain containing multiple copies of the heterologous gene, ROL. This strain was obtained after transforming P.pastoris with the expression vector pPICZαA-ROL, and subsequent selection of strains containing multiple vector insertion events in the presence of higher Zeocin concentrations. The final ROL multicopy strains showed higher lipolytic activity and were isogenic with regard to the single copy ROL strain genetic background (i.e., His+, Mut+), being therefore suitable for high density cultures using a defined synthetic medium.
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Serrano, A., Cereghino, G.L., Ferrer, P., Cregg, J.M., Valero, F. (2001). Overexpression of a Rhizopus Oryzae Lipase in Pichia Pastoris Strains Containing Multiple Copies of the Target Gene. In: Merten, OW., et al. Recombinant Protein Production with Prokaryotic and Eukaryotic Cells. A Comparative View on Host Physiology. Springer, Dordrecht. https://doi.org/10.1007/978-94-015-9749-4_18
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DOI: https://doi.org/10.1007/978-94-015-9749-4_18
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