Abstract
BACKGROUND: Current invention in the field of biotechnology includes research into the probiotic potentials of enzyme-producing bacteria species. OBJECTIVE: This study evaluated the probiotic ability of cellulolytic Bacillus subtilis SBMP4 isolated from waste dump site. METHODS: Isolated Bacillus species was screened for cellulolytic ability on carboxyl methylcellulose agar (CMCA), and hydrolysis efficiency was calculated. Probiotic ability of cellulolytic Bacillus species was carried out as follows: pathogenic potentials, bacteriocin producing ability, antibiotic susceptibility testing, gastric acid, and bile tolerance. Molecular identification was done by means of 16S rRNA gene sequencing, blast search analysis and phylogenetic analysis. RESULTS: Zone of clearance of 30 ± 2.08 mm on CMCA with hydrolysis efficiency of 114 % depicted the isolate’s cellulolytic potential. Bacillus subtilis SBMP4 showed γ-hemolytic phenotype with negative lecithinase activity. Antibacterial activity of bacteriocin observed against Staphylococcus aureus ATCC 25923 reference culture, had maximum inhibition zone of 14±1.0 mm at 72 hour of incubation. Substantial susceptibility towards 9 out of 10 commonly used antibiotics further depicted the isolate’s probiotic ability. Bacillus subtilis SBMP4 exhibited good gastric acid tolerance (pH 2.5) of 5.857 Log cycle/ml and good tolerance to 5 % and 10 % bile salt concentrations with 6.134 and 6.301 Log cycle/ml of viable bacteria cells. Molecular identification of the selected bacteria species revealed it to be Bacillus subtilis SBMP4 with RefSeq accession number NR_118383, showing maximum sequence similarity of 98%. CONCLUSION: Cellulolytic Bacillus subtilis SBMP4 possesses good probiotic ability and has the potentials of being applied as additive in livestock feeding to enhance overall productivity. KEYWORDS: Cellulolytic, Probiotic, CelA gene, Bacteriocin, Bacillus subtilis
Published Version
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