Isolation, Identification and Antibacterial Mechanism of the Main Antibacterial Component from Pickled and Dried Mustard (Brassica juncea Coss. var. foliosa Bailey)

Abstract

Our previous study showed that the ethyl acetate fraction (EAF) from an ethanolic extract of pickled and dried mustard (Brassica juncea Coss. var. foliosa Bailey) had significant antibacterial activity. Here, the EAF was further separated into seven sub-fractions by silica gel column chromatography. The antibacterial activities of the EAF and its sub-fractions against Staphylococcus aureus and Pseudomonas fluorescens were assessed using the agar diffusion method and double dilution method. Among the seven sub-fractions, the third sub-fraction (Fr 3) possessed the strongest antibacterial activity. The main component in Fr 3 was identified by GC-MS, UV-vis, FT-IR, HPLC, 1H NMR and 13C NMR techniques, and was found to be succinic acid. The content of succinic acid in Fr 3 was determined as 88.68% (w/w) by HPLC. Finally, the antibacterial mechanism of succinic acid against the tested strains was explored by determining the intracellular component leakage, measuring the cell particle size and observing the cell morphology. The results showed that succinic acid could damage the cell membrane structure and intracellular structure to increase the leakage of cell components and reduce the cell particle size. Our results suggest that succinic acid could be used in food industry to control bacterial contamination by S. aureus and P. fluorescens.