Development and Characterization of the Peroxisome for Metabolic Engineering

Engineered metabolic pathways often suffer from undesirable interactions with the production host’s native cellular processes. Evolution has solved the problem of metabolic crosstalk by segregating distinct cellular functions into membrane-bound organelles.  By improving the modularity and efficiency of protein import into the yeast peroxisome, determining the natural chemical permeability of the peroxisomal membrane, and characterizing both the rate and capacity of protein import, we have transformed the peroxisome into a useful option for compartmentalization and sequestration of proteins and pathways.  With this new, characterized system, we demonstrate successful compartmentalization of a model pathway and inducible, rapid sequestration of essential proteins.  These advances will serve as the basis for turning the peroxisome into a well-characterized synthetic organelle for use in limiting crosstalk in pathways and protein sequestration.