A set of aspartyl protease-deficient strains for improved expression of heterologous proteins in Kluyveromyces lactis.

ABSTRACT: Secretion of recombinant proteins is a common strategy for heterologous protein expression using the yeast Kluyveromyces lactis. However, a common problem is degradation of a target recombinant protein by secretory pathway aspartyl proteases. In this study, we identified five putative pfam00026 aspartyl proteases encoded by the K. lactis genome. A set of selectable marker-free protease deletion mutants was constructed in the prototrophic K. lactis GG799 industrial expression strain background using a PCR-based dominant marker recycling method based on the Aspergillus nidulans acetamidase gene (amdS). Each mutant was assessed for its secretion of protease activity, its health and growth characteristics, and its ability to efficiently produce heterologous proteins. In particular, despite hav

SUBMITTER: Ganatra MB 

PROVIDER: S-EPMC3041862 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature