title: 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.
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.
authors.affiliation: Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur 784028, Assam, India.
authors.forename: Ajay K
authors.initials: AK
authors.lastname: Manhar
authors.affiliation: Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur 784028, Assam, India.
authors.forename: Yasir
authors.initials: Y
authors.lastname: Bashir
authors.affiliation: Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur 784028, Assam, India.
authors.forename: Devabrata
authors.initials: D
authors.lastname: Saikia
authors.affiliation: Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur 784028, Assam, India.
authors.forename: Dhrubajyoti
authors.initials: D
authors.lastname: Nath
authors.affiliation: Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur 784028, Assam, India.
authors.forename: Kuldeep
authors.initials: K
authors.lastname: Gupta
authors.affiliation: Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur 784028, Assam, India.
authors.forename: Bolin K
authors.initials: BK
authors.lastname: Konwar
authors.affiliation: Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur 784028, Assam, India.
authors.forename: Rahul
authors.initials: R
authors.lastname: Kumar
authors.affiliation: Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur 784028, Assam, India.
authors.forename: Nima D
authors.initials: ND
authors.lastname: Namsa
authors.affiliation: Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur 784028, Assam, India. Electronic address: mandal@tezu.ernet.in.
authors.forename: Manabendra
authors.initials: M
authors.lastname: Mandal
mesh_terms: D000821:Animal Feed; D000818:Animals; D000898:Antibiosis; D001412:Bacillus subtilis / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism* / Q000502:physiology; D001422:Bacterial Adhesion; D001647:Bile Acids and Salts / Q000378:metabolism; D018938:Caco-2 Cells; D002480:Cellulase / Q000235:genetics; D002482:Cellulose / Q000378:metabolism*; D004847:Epithelial Cells / Q000382:microbiology; D005503:Food Additives; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D010209:Paper; D018547:Plant Stems / Q000378:metabolism; D016133:Polymerase Chain Reaction; D019936:Probiotics / Q000302:isolation & purification* / Q000378:metabolism*; D012486:Salmonella typhimurium / Q000254:growth & development; D017422:Sequence Analysis, DNA; D013025:Glycine max / Q000382:microbiology; D013696:Temperature; D003313:Zea mays / Q000378:metabolism
chemical_list: D001647:Bile Acids and Salts; D005503:Food Additives; D002482:Cellulose; D002480:Cellulase
keywords: Animal feed; Bacillus subtilis; Caco-2; Cellulolytic; Probiotics