Identification and characterisation of antimicrobial compound produced by probiotic Alkalihalobacillus clausii 088AE.

Abstract

In the present study, Alkalihalobacillus clausii 088AE was evaluated for the production of antimicrobial compounds, their characterisation, and identification. The results showed that, 48-h-old A. clausii supernatant is able to inhibit the growth of indicator bacteria Micrococcus luteus MTCC 106T (7.0 ± 0.51 mm). The cultivation of 088AE on solid media along with XAD16N beads for 5 days, and isopropanol extraction of antimicrobial compound thereof showed enhanced antimicrobial activity (19.67 ± 0.58 mm) against indicator strain. Further purification with a reversed-phase C18 cartridge yielded a powder with 389.12 ± 10.08 μg of protein per mg. The UV spectra of the sample revealed the characteristic aromatic amino acid peaks at 262 and 276 nm. The results of triple-quadrupole mass spectrometry showed a peak of 1055 m/z, confirming the existence of 10542+, i.e., 2108 Da, suggestive of class I lantibiotic clausin. Furthermore, in silico analysis confirmed the presence of genes responsible for the production of lanthipeptide clausin. The clausin is stable in the presence of proteases (trypsin and pepsin), high temperature (100 °C), pH up to 11 and showed antimicrobial activity against Listeria monocytogenes ATCC 19115, M. luteus, and Staphylococcus aureus ATCC 6538P. Therefore, it could be useful to control gut infections caused by Gram-positive pathogens.