Fig. 1.
Isolation and identification of Brevibacillus laterosporus. (A) Gram staining of five strains of lateral spore-forming bacteria; (B) Antibacterial activity of the sterile supernatants of the culture fluids of five suspected strains; (C) 1% gel electrophoresis of the 16S rRNA of the isolates; M – DNAmarker; N – negative control; (D) Phylogenetic tree constructed based on partial sequences of the 16S rRNA gene. Red circles mark the positions of strains XJ-24-1, XJ-24-2, XJ-24-3, XJ-24-4 and XJ-24-5 isolated in this experiment
Fig. 2.
Determination of antibacterial activity of the Brevibacillus laterosporus XJ-24-3 isolate. (A) Bacteriostatic activity of bacteria-free supernatant and B. laterosporus XJ-24-3 against methicillin-resistant Staphylococcus aureus (MRSA), S. aureus, Listeria monocytogenes, Bacillus cereus, E. coli and Klebsiella pneumoniae; (B) The observation of MRSA treated by cultural supernatant of B. laterosporus XJ-24-3 using a scanning electron microscope; b1 – untreated MRSA; b2 and b3 – MRSA treated by the supernatant of B. laterosporus XJ-24-3 for 1 h and 2 h, respectively
Fig. 3.
Genome map of the Brevibacillus laterosporus XJ-24-3 isolate. Kbp – kilobase pairs; rRNA – ribosomal RNA; tRNA – transfer RNA; sRNA – small RNA; CRISPR – clustered regularly interspaced short palindromic repeats; GC – guanine and cytosine; C G – Cluster of rthologous Groups
Fig. 4A and B.
Functional annotation of the genome of the Brevibacillus laterosporus XJ-24-3 isolate. (A) GO annotation; (B) KEGG analysis annotation
Fig. 4C, D and E.
Statistical analysis of secondary metabolic gene clusters in the genome of the Brevibacillus laterosporus XJ-24-3 isolate. (C) Carbohydrate-Active Enzyme (CAZy) function prediction; (D) Cluster of Orthologous Groups (COG) analysis; (E) Transporter Classification Database (TCDB) annotation
Fig. 5.
uantities of secondary metabolic gene clusters in the genome of the Brevibacillus laterosporus XJ-24-3 isolate
Fig. 6.
Prediction of bacteriocin-related genes in seven regions (A–G) of the genome of Brevibacillus laterosporus XJ-24-3 strain. (A) Lantibiotic ATP-binding cassette (ABC) transporter (ATP – adenosine triphosphate) in red and 131.2 gene (encoding laterosporulin bacteriocin) in green; (B) 223.2 (uviB) gene in green; (C) 224.2 (uviB) gene in green; (D) Lantibiotic ABC transporter (encoding putative YheS) in red and bmbF gene (encoding ribosomal RNA large subunit methyltransferase Cfr) in blue; ( ) Lantibiotic HlyD transporter (encoding efflux resistance-nodulation-division transporter permease subunit) in red and bmbF gene (encoding radical s-adenosylmethionine domain protein) in blue; (F) Lantibiotic ABC transporter in red and 11.1 (bsaA2) gene in green; lanB, lanC and lanD genes (encoding lantibiotic biosynthesis proteins) in blue; (G) lapBotD genes (encoding lantibiotic biosynthesis proteins) in blue and lantibiotic ABC transporter in red
Table 1.
Prediction of molecular sequences with antimicrobial activity in the Brevibacillus laterosporus XJ-24-3 isolate
| Antimicrobial molecule | Detected by | Amino acid sequence |
|---|---|---|
| Laterosporulin | BAGEL4 | MACVNECPDAVDDWAYGDWKCHPVEGKYYRHVFAVCMNGANLYCKTEWSKGC |
| 223.2 (UviB) | BAGEL4 | MEESVMNALLQQGPFAALFVWLLFSTKKEGRDRETRLVKQAQAREAKLMEHNERMVIQLERNTSTLQQIERSLSGLEMELQELKEKVE |
| 224.2 (UviB) | BAGEL4 | MGSFGSLLYLPQGGDKERKSMEEPLFNALLSQGPFAGLFVWLLFSTKKEGRDRETRLVEQAQQREAKLMEHSERMVIQLERNTTTLQQIERSLNGLENELEELKE KVG |
| Linear azol(in)e-containing peptide | BAGEL4 | MDDFQNELKKLRVDKFQGGDVSPWENESQQDAMLVQRRCGRCHHCSCSCSCSCSCSCSCSCSCSCVCLFINCFRCSRCSRCF |
| Sactipeptide | BAGEL4 | MKNYTTPKVKVVNPGVIDVIDSCQCGSKNGAGA |
| Class I lanthipeptide | BAGEL4 | MKKEDLFDLDVQVKEASQAQGDSVVSDLICTTFCSATFCQSNCC |
| Bottromycin | BAGEL4 | MGPVVVFDCMTADFLNDDPNNAELSALELEELESWGVWSEDNQSV |
| BM1122 | antiSMASH | MNKTELIAKVAETSELTKKDATKAVDAVLDAISDALKEGDKVQLIGFGNFEVRERAARKGRNPQTGEEIEIASSKIPAFKPGKQLKDSIK |
| Lactococcin 972-like | antiSMASH | MDKSQKFPDSPLSKEEWRQLDETIVEMARRQLVGRRFIDIYGPLGEGIQTITN DIYDESRFGNMSLRGESLELTQPSKRVSLTIPIVYKDFMLYWRDMAQARTLG MPIDLSPAANAASSCALMEDDLIFNGNPEFDLPGIMNVKGRLTHIKSDWMES GNAFADIVEARNKLLKMGHSGPYALVVSPELYSLLHRVHKGTNVLEIDHIRN LVTDGVFQSPVIKGGALVATGRHNLDLAIAEDFDSAFLGDEQMNSLMRVYEC AVLRIKRPSAICTLETTEE |
| Holin | antiSMASH | MKVLFLLHKMRQGGKNGMEESVMNALLQQGPFAALFVWLLFSTKKEGRDR ETRLVKQAQAREAKLMEHNERMVIQLERNTSTLQQIERSLSGLEMELQELKE KVE |