Via overexpression of selected chaperones, ARTES provides the tool to engineer and optimise secretion of recombinant proteins in yeast.
In the course of this study, selected chaperones such as Calnexin or SEB, but also other “helper” proteins like the processing enzyme KEX2 can be co-integrated into a genome of the production cell line in order to increase the intracellular level of these endogeneous host proteins. Using this approach, ARTES has been able to improve the extracellular level of all recombinant target proteins tested so far approximately two- to threefold. These improvements of productivities could be verified in high cell density fermentations.
Initially established for the Hansenula technology platform, the Chaperone Study can also be applied to other yeast production cell lines. During the course of this study, suitable chaperone expression vectors will be transformed and integrated into the genome of the production host. Positive transformants will initially be selected via chaperone specific PCR, followed by quantitative analysis of the level of product increase.