In Vitro and In Silico Screening and Characterization of Antimicrobial Napin Bioactive Protein in Brassica juncea and Moringa oleifera

Abstract

The study aimed to investigate the antibacterial activity of Mustard (Brassica juncea) and Moringa (Moringa oleifera) leaf extracts and coagulant protein for their potential application in water treatment. Bacterial cell aggregation and growth kinetics studies were employed for thirteen bacterial strains with different concentrations of leaf extracts and coagulant protein. Moringa oleifera leaf extract (MOS) and coagulant protein showed cell aggregation against ten bacterial strains, whereas leaf extract alone showed growth inhibition of five bacterial strains for up to 6 h and five bacterial strains for up to 3 h. Brassica juncea leaf extract (BJS) showed growth inhibition for up to 6 h, and three bacterial strains showed inhibition for up to 3 h. The highest inhibition concentration with 2.5 mg/mL was 19 mm, and furthermore, the minimum inhibitory concentration (MIC) (0.5 mg/mL) and MBC (1.5 mg/mL) were determined to have a higher antibacterial effect for <3 KDa peptides. Based on LCMS analysis, napin was identified in both MOS and BJS; furthermore, the mode of action of napin peptide was determined on lipoprotein X complex (LpxC) and four-chained structured binding protein of bacterial type II topoisomerase (4PLB). The docking analysis has exhibited moderate to potent inhibition with a range of dock score −912.9 Kcal/mol. Thus, it possesses antibacterial-coagulant potential bioactive peptides present in the Moringa oleifera purified protein (MOP) and Brassica juncea purified protein (BJP) that could act as an effective antimicrobial agent to replace currently available antibiotics. The result implies that MOP and Brassica juncea purified coagulant (BJP) proteins may perform a wide degree of antibacterial functions against different pathogens.

Characterization of a Novel Antimicrobial Peptide Isolated from Moringa oleifera Seed Protein Hydrolysates and Its Membrane Damaging Effects on Staphylococcus aureus

“…oleifera flour. − In a previous study, the coagulant proteins, the compound aglycone of deoxy-Niazimicine, and the phenolic compounds from M. oleifera leaves and flour showed antibacterial activities against thirteen pathogenic microorganisms . Dasgupta et al (a) found that the cationic protein from M.…”

Novel Antimicrobial Peptides from Saline Environments Active against E. faecalis and S. aureus: Identification, Characterisation and Potential Usage

“…Synthetic peptides have been proposed to be the ideal inhibitors of enzyme activity either alone or in combination with small-molecule drugs [40]. Chandrashekar S. et al conducted a study, where the protein NAP1 inhibited a S. aureus gyrase-AM8191 complex [41]. Molecular modelling study revealed that peptide P1 and P3 interacted with Topoisomerase IV of E. faecalis and formed hydrogen bonds with Ans650 as a reference antibiotic used in the treatment-levofloxacin.…”

Moringa oleifera: Recent Insights for Its Biochemical and Medicinal Applications

“…Peptides of hypotensive efect were isolated from MO as angiotensin-converting enzyme (ACE), Gly-Leu-Phe-Phe (GLFF), and renin peptides Leu-Gly-Phe-Phe (LGF) [51]. Many peptides of antimicrobial agents have been purifed from MO proteins [215]. Another peptide (<7.5 kDa) has been isolated from MO and possessed an efcient antimicrobial activity [216].…”