Protein expression in Yeast

Yeast expression systems like Pichia pastoris are preferred tools for high expression of properly folded secretory proteins and limited production of endogenous secretory proteins.

It is a cost-effective eukaryotic expression system that allows both secretion expression and intracellular expression. Post-translational modifications have been documented including glycosylation, phosphorylation, and acylation, that are key for biological activities. It is particularly suitable for the expression and preparation of eukaryotic functional proteins such as defensin, interleukin and cytokines. The often efficient secretion of properly folded proteins into the extracellular medium is often observed and allows production of high purity functional protein.

Service description

All services include multiple steps from gene to purified protein as described in the table below.

Leadtime is approximately 9-12 weeks.

Description	Resulting Material	Average turnaround (*)
Gene synthesis
- Codon optimisation - Gene synthesis - Cloning in standard pUC plasmid - Complete sequencing	Cloned, synthesized and verified gene	2 weeks
Subcloning for expression
- Plasmid prep - Subcloning - Clone selection - Complete sequencing	Plasmid vector with cloned target gene	2 weeks
Pilot expression
- Expression pilot and optimisation experiments - Choice of most suitable host - Adjust culture conditions - Determine yields and protein behavior	10-50 µg of semi-purified protein	2-3 weeks
Expression and purification
- Expression - Protein extraction - Protein purification - SDS-PAGE, WESTERN blot	Purified protein	2-3 weeks

(*) Average production time for average difficult project. Actual turn around time varies with individual project features.