Biomanufacturing of Tomato-Derived Nanovesicles

Area of Science:

• Biotechnology and Biomaterials Science
• Plant Biology and Biochemistry

Background:

• Micro- and nano-sized vesicles (MVs and NVs) from edible plants are promising green, sustainable, and biocompatible materials for advanced delivery vectors.
• Isolating pure vesicles from complex plant matrices presents a significant challenge, balancing yield and purity.

Purpose of the Study:

• To develop and optimize methods for bulk production and purification of MVs and NVs from tomato (Solanum lycopersicum L.) fruit.
• To characterize the physical, morphological, and biocargo profiles of isolated tomato MVs and NVs.

Main Methods:

• Differential ultracentrifugation (dUC) was employed for bulk isolation of MVs and NVs from tomato fruit.
• Purification was achieved using sucrose density gradient ultracentrifugation (gUC) or size-exclusion chromatography (SEC).
• Proteomics and in silico analyses were performed on SEC-purified vesicles.

Main Results:

• MVs and NVs exhibited similar protein and phospholipid cargo, with phosphatidic acid being the most abundant phospholipid.
• SEC effectively removed co-purifying matrix components, yielding a high number of particles (2.6 × 10¹⁵) and protein (6.9 ± 1.5 mg) per kilogram of tomato.
• dUC/gUC separated vesicles based on buoyant density, and proteomics identified abundant lipoxygenase (LOX), ATPases, and heat shock proteins (HSPs) in SEC-purified vesicles.

Conclusions:

• Combined dUC and SEC offers an efficient method for high-yield purification of tomato MVs and NVs.
• The characterized vesicles possess cargo profiles suggesting potential for various biotechnological applications, including drug delivery.