Exploiting Self-Processing 2A Peptides for Multigene Expression in Pichia pastoris

Metabolic engineering in Komagataella phaffii (reclassified from Pichia pastoris) often requires the implementation and expression of more than one (heterologous) gene. Instead of placing each gene under the separate control of a promoter, we present in this chapter a strategy for polycistronic pathway expression based on self-processing 2A peptides. This approach bears several advantages for multienzyme pathway expression such as compact pathway design, fast and simple pathway assembly, the generation of stable production strains, as well as the possibility for fine-tuning the expression of pathway genes.