Molecular Characterization and In Vitro Analyses of a Sporogenous Bacterium with Potential Probiotic Properties

Abstract

The Gram-positive thin rods of a Bacillus species were identified and designated as Bacillus coagulans RK - 02 through the standard microbiological and biochemical characterization procedures, followed by data analysis and comparison with the characteristics given in Bergey's manual of systematic bacteriology. The culture was further characterized and confirmed as Bacillus coagulans by 16S rDNA sequence analysis wherein about 755 nucleotides of 16S rDNA was amplified and cloned in pGEM-T vector and subsequently sequenced. Sequence was blasted against the nr database of NCBI. Multiple alignments were done with some selected and related sequences using Clustal W. Phylogenetic trees were drawn with the same software after the distances were determined by neighbor-joining algorithm. The in vitro studies on the probiotic properties demonstrated that our isolate could prove to be a potential probiotic with spore-forming and lactic acid-producing abilities coupled with acid and bile tolerance properties and antimicrobial action. In addition to these characteristics, the bacterium also produced enzymes such as amylase, cellulase, lipase, protease, lactase and catalase, which can help in improving digestion and overall health, alleviate lactose intolerance and remove oxidative stresses, required for the well-being of the consumers. In our previously reported studies, an exopolysaccharide (EPS), a probioactive molecule produced by the same bacterium, showed very significant antioxidant, antihyperglycemic and emulsification activities. Thus, Bacillus coagulans RK - 02 is a well-characterized promising probiotic for its potential commercial applications to pharmaceutical, nutraceutical and functional food formulations with care-free storage.

Isolation and characterization of novel Bacillus strains with superior probiotic potential: comparative analysis and safety evaluation

“…Next, 60 µL of the cell-free supernatant of each bacterial culture was poured into the wells and the plates were incubated at 37 °C. The diameter of the growth inhibition zoon appeared around the wells was recorded after 24 h of incubation 64 – 66 .…”

Purification and Partial Elucidation of the Structure of an Antioxidant Carbohydrate Biopolymer from the Probiotic Bacterium Bacillus coagulans RK-02

“…The probiotic bacterium B. coagulans RK-2 was isolated from a soil sample and grown at 37 °C, pH 6.7, and 180 rpm in glucose mineral salt medium. EPS isolation and quantification have been described previously. − …”

“…The physicochemical and biological properties of EPSs depend mainly on the monosaccharide composition and the linkages between the monomeric units that are joined together to form the exopolymer. 10 Considering the commercial and therapeutic importance of these products, it is therefore of paramount importance to structurally characterize the EPS that was isolated and purified from a Bacillus coagulans RK-02 probiotic culture broth 13,14 and has shown significant antioxidant and emulsifying activities. 15,16 Herein we report that the B. coagulans RK-02 EPS exhibits antioxidant activity in vivo and present the first structural characterization of a microbial EPS.…”

Assessing the probiotic potential and safety of newly isolated Bacillus coagulans VHBAX‐04: in vitro and in silico evaluations

“…To date, B. coagulans strains GBI‐30, Unique IS‐2, MTCC 5856 and SNZ‐1969 are each well‐known for their clinical safety and efficacy. As the probiotic effects are strain‐specific, it is necessary to investigate newer strains with new attributes, safety and efficacy (Sen et al ., 2010; Sudha et al ., 2010; Marzorati et al ., 2021).…”