Artificial Self-assembling Nanocompartment for Organizing Metabolic Pathways in Yeast

Area of Science:

• Synthetic biology
• Cellular compartmentalization
• Metabolic engineering

Background:

• Metabolic pathways are often compartmentalized within cells to prevent unwanted interactions and optimize enzyme activity.
• Cellular compartments are valuable tools in synthetic biology for studying enzyme function and for metabolic engineering applications.

Purpose of the Study:

• To develop and validate Murine polyomavirus virus-like particles (MPyV VLPs) as novel intracellular compartments for budding yeast (Saccharomyces cerevisiae).
• To demonstrate the utility of MPyV VLPs for metabolic engineering by encapsulating a key enzyme in d-glucaric acid biosynthesis.

Main Methods:

• Utilized VP1 protein for self-assembly of the VLP shell and VP2C for cargo protein encapsulation.
• Engineered VP1 variants for enhanced cargo capture and subcellular localization.
• Encapsulated myo-inositol oxygenase (MIOX), an unstable enzyme, within MPyV VLPs in yeast.

Main Results:

• Encapsulated destabilized green fluorescent protein (GFP) was protected from degradation.
• Engineered VP1 showed improved cargo capture and localization.
• Yeast strains with encapsulated MIOX produced approximately 20% more d-glucaric acid compared to controls, with improved growth and reduced MIOX accumulation.

Conclusions:

• MPyV VLPs serve as effective artificial biocatalytic compartments in yeast, participating in metabolic pathways.
• The MPyV VLP system represents a promising new tool for yeast synthetic biology and metabolic engineering.