Bioassay-guided Fractionation and Biological Activities of Antimycin A and 4-Hydroxybenzoic Acid Isolated from Nocardiopsis sp. Strain LC-9

Abstract

Nocardiopsis sp. strain LC-9 was isolated from freshwater sediments and explored for its varied bioactive traits. Initially, ethyl acetate extract of strain LC-9 at varied concentrations showed pronounced antibacterial activities. After column chromatography, fraction F2 and F3 of the extract were identified as prominent fractions in terms of antimicrobial activities with low minimum inhibitory concentration values. Antioxidant activities of fraction F2 and F3 revealed remarkable scavenging of free radicals with low IC₅₀ values (DPPH - 417.86 ± 0.24 μg/mL, ABTS - 431.6 ± 0.90 μg/mL, and FRAP - 404.36 ± 0.18 μg/mL). Fractions F2 and F3 were further characterized by UV spectrum, Fourier transform infrared spectroscopy, nuclear magnetic resonance, and liquid chromatography-mass spectrometry, and were identified as Antimycin A and 4-hydroxybenzoic acid. The compounds were further tested for anticancer activity against MCF-7 cells. The MTT assay showed reduced viability of MCF-7 cells with an increase in concentration of compounds. The IC₅₀ values for Antimycin A and 4-hydroxybenzoic acid were 9.6 ± 0.7 μg/mL and 20.8 ± 0.4 μg/mL, respectively. Staining techniques confirmed the apoptosis mechanism. Finally, molecular docking (against targeted proteins of bacteria, fungus, and cancer cells) and molecular dynamics confirmed the pharmaceutical efficacy of the purified compounds.