Low pH-Based Method to Increase the Yield of Plant-Derived Nanoparticles from Fresh Ginger Rhizomes

Abstract

Plant-derived nanoparticles (PDNPs) are naturally occurring exosome-like nanovesicles derived from dietary plants containing key plant bioactives. Ginger-derived PDNPs have a therapeutic effect on alcohol-induced liver injury, inflammatory bowel disease, and colon cancer. PDNPs are conventionally purified by differential ultracentrifugation, a technique not amenable for scale up. We have recently developed a polyethylene glycol (PEG) 6000-based method for cost-effective purification of ginger PDNPs, with comparable efficiency to differential ultracentrifugation (Sci. Rep. 2020, 10 (1), 4456.). Herein, we report a 4–5-fold higher ginger PDNP recovery when PEG precipitation was carried out in low pH conditions (pH 4 and 5). Low pH-derived ginger PDNPs were smaller in size without an overt change in zeta potential. The spontaneous intracellular entry and protection against oxidative stress in A431 cells were similar between ginger PDNPs purified under low, neutral, and alkaline pH. Low-pH purified ginger PDNPs had higher levels of total polyphenolic content compared to PDNPs purified under neutral and alkaline pH. Recently, ginger PDNP-derived microRNAs have been shown to exhibit cross-kingdom regulation by targeting human, gut microbiome, and viral transcripts. Using qRT-PCR, we also verified the presence of miRNAs that were predicted to target SARS-CoV-2 in ginger PDNPs purified under low pH. Thus, we have developed a method to purify ginger PDNPs in high yields by using low-pH conditions without affecting the major bioactive contents of PDNPs.

Characterization of the MicroRNA Profile of Ginger Exosome-like Nanoparticles and Their Anti-Inflammatory Effects in Intestinal Caco-2 Cells

“…37 Others extracted GELNs using PEG6000 based on the principle of polymer precipitation. 38 However, there is still a lack of available extraction methods at present. In this work, we attempted to extract GELNs by differential centrifugation combined with the commercial kit, and the feasibility of this method was confirmed via characterization of GELNs by TEM and dynamic light scattering (DLS).…”

“…Although some scholars purified the exosome-like nanoparticles from sunflower seeds via selective extraction of apoplastic fluid by vacuum infiltration procedure . Others extracted GELNs using PEG6000 based on the principle of polymer precipitation . However, there is still a lack of available extraction methods at present.…”

Isolation and characterization of ginseng-derived exosome-like nanoparticles with sucrose cushioning followed by ultracentrifugation

“…In comparison with a previous isolation method of plant-derived exosome-like nanoparticles from carrot, apple, lemon, grape, grapefruits, tea leaves, cabbage, ginger, and garlic [32][33][34][35][36][37][38], we established the double fractionation of GDEs by sucrose-based isolation to purify GDEs from vesicle aggregates or contaminated proteins for reproducible high-quality exosome-like nanoparticles. To collect homogeneously sized GDEs with high yield, we first isolated non-contaminated intact EVs from ginseng juice through sucrose cushion of different densities, which can prevent the degradation of EVs during repeated highforce centrifugations.…”

The Emerging Role of Plant-Derived Exosomes-Like Nanoparticles in Immune Regulation and Periodontitis Treatment

“…PEG methods are related to pH. Suresh et al reported a higher yield of PELNs when PEG precipitation was carried out in pH 4 and 5 (58). PELNs can be internalized in plant cells, mammalian cells, fungi, and bacteria (28,(45)(46)(47).…”