Plant-Produced Viral Nanoparticles as a Functionalized Catalytic Support for Metabolic Engineering

Area of Science:

• Biotechnology
• Synthetic Biology
• Plant Science

Background:

• Substrate channeling enhances metabolic efficiency by localizing enzymes.
• Virus nanoparticles (VNPs) are promising scaffolds for supramolecular assembly.
• Plant-produced VNPs offer a sustainable platform for biotechnological applications.

Purpose of the Study:

• To develop functionalized VNPs as enzymatic hubs for plant metabolic engineering.
• To demonstrate the in vitro and in planta catalytic activity of enzyme-displaying VNPs.
• To establish a heterologous cannabinoid precursor biosynthetic pathway using VNP scaffolds.

Main Methods:

• Genetic fusion of E-coil peptides to VNP coat proteins (TBSV, PVX, HBc VLPs).
• K-coil peptide tagging of enzymes (lichenase, acyl-activating enzyme 1, olivetolic acid synthase, olivetolic acid cyclase).
• In vitro enzyme assays and in planta transient co-expression studies.

Main Results:

• Engineered E-coil VNPs successfully interacted with K-coil-tagged lichenase in vitro, demonstrating catalytic activity.
• Co-expression of K-coil-enzymes and E-coil-HBc VLPs established a functional cannabinoid precursor pathway in planta.
• VNP scaffolds significantly increased the yield of olivetolic acid glucoside compared to non-scaffolded systems.

Conclusions:

• Functionalized VNPs serve as effective enzymatic hubs for enhancing metabolic pathways.
• Plant-based VNP technology is a viable strategy for metabolic engineering applications.
• This approach holds potential for optimizing the production of valuable plant natural products.