Purification and characterization of a bacteriocin-like compound (Lichenin) produced anaerobically by Bacillus licheniformis isolated from water buffalo

Abstract

Aims: To characterize a bacteriocin‐like factor from Bacillus licheniformis 26 L‐10/3RA isolated from buffalo rumen. Methods and Results: The culture supernatant exhibited the antibacterial activity against a number of indicator organisms in a cut‐well agar assay under anaerobic conditions. The inhibitory component was purified by following ammonium sulphate precipitation, gel filtration and ion exchange chromatography and confirmed to be a single peptide. A single band on tricine‐sodium dodecyl sulphate‐polyacrylamide gel electrophoresis confirmed that the peptide was purified to homogeneity and having an estimated molecular mass of approximately 1400 dalton. Complete amino acid sequence of the peptide yielded 12 amino acids from the N‐terminal end (ISLEICXIFHDN). No homology with previously reported bacteriocins was observed and has been designated as Lichenin. Lichenin was found to be hydrophobic, sensitive to atmospheric oxygen, retained biological activity even after boiling for 10 min and was active over a pH range of 4·0–9·0. Conclusions: The Lichenin represents the first anaerobiosis specific expression of bacteriocin‐like compound isolated from Bacillus licheniformis 26 L‐10/3RA of buffalo rumen origin. Significance and Impact of the Study: Lichenin could be a potential condidate for manipulating the rumen function at molecular level intended for improving the productivity of the ruminant.

Bovicin HC5, a bacteriocin from Streptococcus bovis HC5

“…HPLC indicated that the acidic NaCl extracts had some inactive peptides, but the active peptide could be purified and concentrated by rechromatography. The active peptide could not be silver stained, and other workers have noted a similar phenomenon (Carolissen-Mackay et al, 1997 ;Pattnaik et al, 2001 ;Villani et al, 1995). However, it reacted with Coomassie stain after the gel had been fixed with glutaraldehyde (Fig.…”

Purification and partial characterization of an antimicrobial peptide produced by Bacillus sp. strain P45, a bacterium from the Amazon basin fish Piaractus mesopotamicus

“…(Table 1). The inhibitory spectrum of this substance is rather similar to that of some BLS, such as lichenin (Pattnaik et al, 2001), entomocin 9 (Cherif et al, 2003) and BLS P40 .…”

“…Most bacteriocins produced by Bacillus have a higher molecular weight, like thuricin 439A of 2,919.9 Da, thuricin 439B of 2,803.8 Da (Ahern et al, 2003), and subtilosin A of 3,398.9 Da (Stein, 2005). However, the molecular mass of this peptide was close to a lipopeptide antibiotic of 1,500 Da produced by B. subtilis EN 63-1 (Bechard et al, 1998) and lichenin produced by Bacillus licheniformis of 1,400 Da (Pattnaik et al, 2001). The cluster of peaks differing by 14 Da suggests the presence of a molecule containing a fatty acid chain of variable length (CH 2 ϭ14 Da).…”

Antagonistic Bacillus cereus TC-1 Isolated from Solar Salt Work in Southern India

“…In vitro antagonistic activity of Bacillus cereus TC-1 against the five pathogenic Vibrio sp was tabulated in Table 2 Bacillus is an interesting genus to investigate for antimicrobial activity since Bacillus species production a diverse array of antimicrobial peptides representing several different basic chemical structures 23 , with a distinct diversity in their inhibitory activities against a variety of micro organisms 24 25 . Pseudoalteromonas sp.…”