Identification, Purification and Characterization of Laterosporulin, a Novel Bacteriocin Produced by Brevibacillus sp. Strain GI-9

Abstract

BackgroundBacteriocins are antimicrobial peptides that are produced by bacteria as a defense mechanism in complex environments. Identification and characterization of novel bacteriocins in novel strains of bacteria is one of the important fields in bacteriology.Methodology/FindingsThe strain GI-9 was identified as Brevibacillus sp. by 16 S rRNA gene sequence analysis. The bacteriocin produced by strain GI-9, namely, laterosporulin was purified from supernatant of the culture grown under optimal conditions using hydrophobic interaction chromatography and reverse-phase HPLC. The bacteriocin was active against a wide range of Gram-positive and Gram-negative bacteria. MALDI-TOF experiments determined the precise molecular mass of the peptide to be of 5.6 kDa and N-terminal sequencing of the thermo-stable peptide revealed low similarity with existing antimicrobial peptides. The putative open reading frame (ORF) encoding laterosporulin and its surrounding genomic region was fished out from the draft genome sequence of GI-9. Sequence analysis of the putative bacteriocin gene did not show significant similarity to any reported bacteriocin producing genes in database.ConclusionsWe have identified a bacteriocin producing strain GI-9, belonging to the genus Brevibacillus sp. Biochemical and genomic characterization of laterosporulin suggests it as a novel bacteriocin with broad spectrum antibacterial activity.

Characterization of the Antimicrobial Peptide Penisin, a Class Ia Novel Lantibiotic from Paenibacillus sp. Strain A3

“…The antimicrobial substance was purified from CFB from 1,000 ml of nutrient broth in which strain A3 was grown for 48 h on a rotary shaker at 30°C (180 rpm). The crude extract was obtained by incubating CFB and 2% preactivated Diaion HP20 resin (Sigma, USA) as mentioned earlier (21). This crude extract was purified by gel filtration chromatography using a manually packed Sephadex G-50 column (50 mM NaCl with a flow rate of 1.0 ml/min), and fractions showing antimicrobial activity were collected and concentrated.…”

“…The membrane was finally rinsed multiple times in Milli-Q water. The excised peptide band was subjected to N-terminal sequencing as mentioned earlier (21). The partial sequence obtained was used to find the resembling sequences available in the database (http://aps .unmc.edu/AP/main.php).…”

Laterosporulin10: a novel defensin like Class IId bacteriocin from Brevibacillus sp. strain SKDU10 with inhibitory activity against microbial pathogens

“…There are about 200 bacteriocins reported with diverse amino acid sequences that are available at different bacteriocin databases, including www.bactibase.pfba-lab-tun.org. Though AMPs are predominantly reported from the strains belonging to the genera Lactobacillus and Bacillus (Klaenhammer, 1993;Teixeira et al, 2013;Zhao et al, 2012), members of other genera such as Paenibacillus and Brevibacillus were also found to produce bacteriocins (Baindara et al, 2015;Singh et al, 2012). Accordingly, members of the genus Brevibacillus, specifically, strains of species B. laterosporus are known for their beneficial functions (Ruiu et al, 2014) and antimicrobial substance production.…”

Interstrain Interactions between Bacteria Isolated from Vacuum-Packaged Refrigerated Beef

“…Baindara et al (41) characterized two antimicrobial peptides produced by a B. subtilis strain, which showed antagonistic properties against Gram-positive bacteria, including S. aureus and Listeria monocytogenes. Other Bacillus species have been reported to produce bacteriocins and biosurfactants (42,43); the bacteriocins inhibited the growth of a large range of Gram-positive and Gramnegative bacteria. Bacillus subtilis E0g strongly inhibited most Gram-positive targets, including C. maltaromaticum D8c, B. thermosphacta A0b, Bacillus sp.…”