Optimized Low-Voltage Electric Field Extraction Enhances the Recovery and Bioactivity of Cyclotides From Viola Ignobilis Publisher



Hashempour H ; Esmaeilpour Z ; Bagheroghli H ; Barani Pour S ; Sardroodi J J ; Ebadi M ; Esmaeili F ; Farhadpour M ; Didarian R ; Dogan Guzel F ; Lotfibakhshaiesh N ; Ghorbanpoor H ; Ebrahimi A ; Avci H
Source: Microchemical Journal Published:2026

Abstract

Cyclotides are ultra-stable plant peptides with strong therapeutic potential, but their low natural abundance and inefficient extraction limit pharmaceutical development. In this study, we introduce, for the first time, a low-voltage electric field (LV-EF) method for extracting bioactive Mobius cyclotides from Viola ignobilis. The key variables tested were voltage (20, 40, 60 V) and electrode gap (1.0–2.5 cm). Optimal conditions (20 V, 1.5 cm; field strength 13.33 V/cm) significantly improved cyclotide yield compared to conventional maceration and ultrasound-assisted extraction. Molecular dynamics simulations revealed that the electric field strengthens hydrogen-bonding between cyclotide Vigno 5 and the acetonitrile/water/formic acid solvent system, providing a mechanistic basis for enhanced extractability. Scanning electron microscopy confirmed greater cellular disruption under LV-EF. The optimized extract showed potent and selective cytotoxicity against PC-3 prostate cancer cells (IC₅₀ = 47.61 μg mL−1) via apoptosis, with minimal toxicity to normal L929 cells (IC₅₀ ≥ 300 μg mL−1). Practical implications include a scalable, low-cost, and green extraction platform for peptide-based therapeutics. Future work should focus on continuous-flow LV-EF systems and isolation of individual cyclotides for structure-activity studies. This establishes LV-EF as a superior technology for producing high-quality bioactive cyclotide extracts. © 2026 Elsevier B.V.