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

• Published in: Plants (Basel) Vol. 13; no. 4; p. 503
• Main Authors: Sator, Christian, Lico, Chiara, Pannucci, Elisa, Marchetti, Luca, Baschieri, Selene, Warzecha, Heribert, Santi, Luca
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.02.2024; MDPI
• Subjects: Agricultural biotechnology; Antigens; Biosynthesis; Bushy stunt; cannabinoids; Catalysis; Catalyst supports; Chemical properties; Coils; Enzymes; Genetic engineering; Hepatitis B; Metabolic engineering; Metabolism; Metabolites; Methods; multi-enzymatic assemblies; Nanoparticles; Natural products; olivetolic acid; Peptides; Physiological aspects; plant metabolic engineering; plant virus nanoparticles; Plant viruses; plant-produced virus-like particles; Potatoes; Proteins; Scaffolds; Self-assembly; Signal transduction; Substrates; Tomatoes; Virus diseases of plants; Virus-like particles; Viruses; Yeast; Germany; cannabinoids; multi-enzymatic assemblies; olivetolic acid; plant-produced virus-like particles; plant virus nanoparticles; plant metabolic engineering
• ISSN: 2223-7747; 2223-7747
• DOI: 10.3390/plants13040503

Substrate channeling could be very useful for plant metabolic engineering; hence, we propose that functionalized supramolecular self-assembly scaffolds can act as enzymatic hubs able to perform reactions in close contiguity. Virus nanoparticles (VNPs) offer an opportunity in this context, and we present a functionalization strategy to display different enzymes on the outer surface of three different VNPs produced in plants. Tomato bushy stunt virus (TBSV) and Potato virus X (PVX) plant viruses were functionalized by the genetic fusion of the E-coil peptide coding sequence to their respective coat proteins genes, while the enzyme lichenase was tagged with the K-coil peptide. Immobilized E-coil VNPs were able to interact with the plant-produced functionalized lichenase, and catalysis was demonstrated by employing a lichenase assay. To prove this concept , the Hepatitis B core (HBc) virus-like particles (VLPs) were similarly functionalized by genetic fusion with the E-coil sequence, while acyl-activating enzyme 1, olivetolic acid synthase, and olivetolic acid cyclase enzymes were tagged with the K-coil. The transient co-expression of the K-coil-enzymes together with E-coil-VLPs allowed the establishment of the heterologous cannabinoid precursor biosynthetic pathway. Noteworthy, a significantly higher yield of olivetolic acid glucoside was achieved when the scaffold E-coil-VLPs were employed.