Abstract
Bacillus clausii UBBC07 is a commercial spore probiotic known to reduce diarrhea in children and adults. In the present study, survival and germination of UBBC07 spores were investigated under fed and fasted conditions in Simulator of Human Intestinal Microbial Ecosystem. Besides this, lantibiotic production, purification, and characterization were performed. The agar plate analysis showed that spores were 100% tolerant to fed and fasted gastrointestinal tract (GIT) conditions. Simultaneously, flow cytometry revealed that at the end of small intestinal incubation, 120% (fed) and 133% (fasted) spores were in viable germinating state. The transformation of viable germinating spores into viable vegetative cells was observed at 3 h of incubation under fasted GIT conditions. In antimicrobial evaluation, UBBC07 produced low-molecular-weight (2107.94 Da) class I lantibiotic clausin. The presence of lanB, lanC, and lanD genes confirms the clausin production. Clausin is stable at proteases (pepsin, proteinase K, and trypsin), temperature (up to 100°C), and pH (up to 11). Furthermore, the antimicrobial activity toward Gram-positive bacteria including Clostridium difficile is advantageous. In conclusion, B. clausii UBBC07 spore probiotic is capable of surviving and germinating under in vitro upper GIT conditions. The clausin production justifies strain applicability in diarrhea.
Highlights
Bacillus clausii UBBC07 (MTCC 5472) is a Gram-positive, spore-forming, motile, rod-shaped, non-toxic, and safe probiotic bacterium (Lakshmi et al, 2017) available commercially in the form of spore-suspension and dry formulations in India
Fed stomach simulation in reactor was achieved by using Simulator of Human Intestinal Microbial Ecosystem (SHIME) media (g/L: starch, 4; yeast extract, 3; mucin, 3; pectin, 2; arabinogalactan, 1.2; peptone, 1; xylan, 0.5; L-cysteine hydrochloride, 0.5; glucose, 0.4) supplemented with pepsin, phosphatidylcholine (0.17 mM), NaCl, and KCl
Fed Conditions Bacillus clausii UBBC07 total viable count (TVC) enumerated by agar plate method showed no significant effect of stomach and small intestinal conditions on survival of spores under fed state
Summary
Bacillus clausii UBBC07 (MTCC 5472) is a Gram-positive, spore-forming, motile, rod-shaped, non-toxic, and safe probiotic bacterium (Lakshmi et al, 2017) available commercially in the form of spore-suspension and dry formulations in India. The consumption of UBBC07 [2 × 109 colony-forming unit (CFU)] capsule (twice a day) for 10 days was found to be safe and effective to alleviate the symptoms of acute diarrhea in adults (Sudha et al, 2013). The supplementation of UBBC07 (2 × 109 CFU/5 ml) spore suspension (twice a day) for 5 days effectively reduced the severity of diarrhea in children under 5 years of age (Sudha et al, 2019). No adverse effects and significant changes in safety parameters were reported in UBBC07 clinical investigations (Sudha et al, 2013, 2019). UBBC07 has been reported to mitigate uremic toxins in the blood of acetaminophen-induced uremia rats (Patel et al, 2019).
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