Abstract
This study was carried out to understand the probiotic features, ability to utilize non-digestible carbohydrates and comparative genomics of anti-inflammatory Bifidobacterium strains isolated from human infant stool samples. Bacterial strains were isolated from the stool samples using serial dilution on MRS agar plates supplemented with 0.05% l-cysteine hydrochloride and mupirocin. Molecular characterization of the strains was carried out by 16S rRNA gene sequencing. Anti-inflammatory activity was determined using TNF-α and lipopolysaccharide (LPS) induced inflammation in Caco2 cells. Probiotic attributes were determined as per the established protocols. Isomaltooligosaccharides (IMOS) utilization was determined in the broth cultures. Whole genome sequencing and analysis was carried out for three strains. Four obligate anaerobic, Gram positive Bifidobacterium strains were isolated from the infant stool samples. Strains were identified as Bifidobacterium longum Bif10, B. breve Bif11, B. longum Bif12 and B. longum Bif16. The strains were able to prevent inflammation in the Caco2 cells through lowering of IL8 production that was caused by TNF-α and LPS treatment. The strains exhibited desirable probiotic attributes such as acid and bile tolerance, mucin binding, antimicrobial activity, bile salt hydrolase activity, cholesterol lowering ability and could ferment non-digestible carbohydrates such as isomaltooligosaccharides and raffinose. Furthermore, Isomaltooligosaccharides supported the optimum growth of the strains in vitro, which was comparable to that on glucose. Strains could metabolize IMOS through cell associated α-glucosidase activity. Genomic features revealed the presence of genes responsible for the utilization of IMOS and for the probiotic attributes.
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Acknowledgements
Authors would like to acknowledge National Agri-Food Biotechnology Institute (NABI); Department of Biotechnology, Government of India and Indian Council of Medical Research for providing financial support for this research. DBT-e-Library Consortium (DeLCON) is gratefully acknowledged for providing access to electronic journals.
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This study was funded by Core Grant from NABI, DBT, Government of India and Indian council of Medical Research: File No: 5/9/1312/2020-Nut.
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SS1, SS2, KK and MB conceived the research and designed the experiments. SS2, KK and SKB isolated the Bifidobacterium strains; SS1, SS2, RM and PR conducted the experiments. VC, SM, AC, SR and RS performed the bacterial genomic characterization and in-silico data analysis. SS1, SS2, MB, SKB and KK analyzed the data. SS1, SS2, VC, SM, AC, MB and KK wrote the manuscript. All authors read and approved the manuscript.
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The data sets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request. Whole genome sequencing data presented in this study are openly available in The National Centre for Biotechnology Information (NCBI).
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Sharma, S., Singh, S., Chaudhary, V. et al. Isomaltooligosaccharides utilization and genomic characterization of human infant anti-inflammatory Bifidobacterium longum and Bifidobacterium breve strains. 3 Biotech 12, 89 (2022). https://doi.org/10.1007/s13205-022-03141-2
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DOI: https://doi.org/10.1007/s13205-022-03141-2