1. Cellulolytic potential of probiotic Bacillus Subtilis AMS6 isolated from traditional fermented soybean (Churpi): An in-vitro study with regards to application as an animal feed additive
- Author
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Dhrubajyoti Nath, Bolin Kumar Konwar, Manabendra Mandal, Rahul Kumar, Devabrata Saikia, Ajay Kumar Manhar, Yasir Bashir, Kuldeep Gupta, and Nima D. Namsa
- Subjects
0301 basic medicine ,Paper ,Salmonella typhimurium ,Cellulase ,Bacillus subtilis ,Microbiology ,Polymerase Chain Reaction ,Zea mays ,Bacterial Adhesion ,law.invention ,Bile Acids and Salts ,03 medical and health sciences ,Probiotic ,chemistry.chemical_compound ,law ,Hydrolase ,Antibiosis ,Animals ,Humans ,Glycosyl ,Cellulose ,biology ,Plant Stems ,Hydrolysis ,Probiotics ,Temperature ,Epithelial Cells ,Sequence Analysis, DNA ,Hydrogen-Ion Concentration ,Cellulose binding ,biology.organism_classification ,Animal Feed ,030104 developmental biology ,chemistry ,Biochemistry ,biology.protein ,Food Additives ,Soybeans ,Caco-2 Cells ,Antibacterial activity ,Digestion - Abstract
The aim of the present study is to evaluate the probiotic attributes of Bacillus subtilis AMS6 isolated from fermented soybean (Churpi). This isolate exhibited tolerance to low pH (pH 2.0) and bile salt (0.3%), capability to autoaggregate and coaggregate. AMS6 also showed highest antibacterial activity against the pathogenic indicator strain Salmonella enterica typhimurium (MTCC 1252) and susceptibility towards different antibiotics tested. The isolate was effective in inhibiting the adherence of food borne pathogens to Caco-2 epithelial cell lines, and was also found to be non-hemolytic which further strengthen the candidature of the isolate as a potential probiotic. Further studies revealed B. subtilis AMS6 showed cellulolytic activity (0.54±0.05 filter paper units mL(-1)) at 37°C. The isolate was found to hydrolyze carboxymethyl cellulose, filter paper and maize (Zea mays) straw. The maize straw digestion was confirmed by scanning electron microscopy studies. The isolate was able to degrade filter paper within 96h of incubation. A full length cellulase gene of AMS6 was amplified using degenerate primers consisting of 1499 nucleotides. The ORF encoded for a protein of 499 amino acids residues with a predicted molecular mass of 55.04kDa. The amino acids sequence consisted of a glycosyl hydrolase family 5 domain at N-terminal; Glycosyl hydrolase catalytic core and a CBM-3 cellulose binding domain at its C terminal. The study suggests potential probiotic B. subtilis AMS6 as a promising candidate envisaging its application as an animal feed additive for enhanced fiber digestion and gut health of animal.
- Published
- 2015