Inducible Amplification of Gene Copy Number and Heterologous Protein Production in the Yeast Kluyveromyces lactis

Abstract

Heterologous protein production can be doubled by increasing the copy number of the corresponding heterologous gene. We constructed a host-vector system in the yeast Kluyveromyces lactis that was able to induce copy number amplification of pKD1 plasmid-based vectors upon expression of an integrated copy of the plasmid recombinase gene. We increased the production and secretion of two heterologous proteins, glucoamylase from the yeast Arxula adeninivorans and mammalian interleukin-1β, following gene dosage amplification when the heterologous genes were carried by pKD1-based vectors. The choice of the promoters for expression of the integrated recombinase gene and of the episomal heterologous genes are critical for the mitotic stability of the host-vector system.

Optimization of recombinant fungal laccase production with strains of the yeastKluyveromyces lactis from the pyruvate decarboxylase promoter

“…Protein production is also affected by the dosage of the heterologous gene and its expression. An inducible system for gene dosage increase has been described for vectors based on the stable and multicopy plasmid pKD1 (Morlino et al, 1999). In terms of regulation of the expression of the heterologous gene, commonly used inducible promoters are the PHO5 promoter of S. cerevisiae and the LAC4 promoter of K. lactis.…”

Heterologous protein production in : physiological effects and fermentative strategies

“…transformed with a multicopy expression vector [22]. K. lactis has been reported to produce a higher level of human IL-1b with the use of a multicopy expression system [23]. The exploitation of plasmids based on the sequences of the 2-lm-like endogenous Z. bailii plasmid, currently under development in our laboratory, could improve the production from Z. bailii hosts [12,24].…”

The apyrase KlYnd1p ofKluyveromyces lactisaffects glycosylation, secretion, and cell wall properties

“…The plasmids used in this study were pYG132, carrying a cassette for the expression of secreted rHSA driven by the native preprosignal sequence under the control of the ADH4 promoter (Saliola et al, 1999), and the the K. lactis-Escherichia coli shuttle vector pGM-GAM, containing the GAA gene from the yeast Arxula adeninivorans (Morlino et al, 1999). Yeast transformations were carried out by electroporation (Sambrook et al, 2001).…”