A novel, lactase-based selection and strain improvement strategy for recombinant protein expression in Kluyveromyces lactis.

ABSTRACT:

Background

The Crabtree-negative yeast species Kluyveromyces lactis has been established as an attractive microbial expression system for recombinant proteins at industrial scale. Its LAC genes allow for utilization of the inexpensive sugar lactose as a sole source of carbon and energy. Lactose efficiently induces the LAC4 promoter, which can be used to drive regulated expression of heterologous genes. So far, strain manipulation of K. lactis by homologous recombination was hampered by the high rate of non-homologous end-joining.

Results

Selection for growth on lactose was applied to target the insertion of heterologous genes downstream of the LAC4 promoter into the K. lactis genome and found to yield high numbers of positive transformants. Concurrent reconstitution of the β

SUBMITTER: Krijger JJ 

PROVIDER: S-EPMC3520740 | biostudies-literature | 2012 Aug

REPOSITORIES: biostudies-literature