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Comparative accounts of probiotic properties of spore and vegetative cells of Bacillus clausii UBBC07 and in silico analysis of probiotic function.

ABSTRACT: In this study, the spores and vegetative cells of B. clausii were independently evaluated for probiotic properties such as acid, gastric juice, bile, and intestinal fluid tolerance, adhesion to solvents/mucin and zeta potential. In addition, in silico identification of genome features contributing to probiotic properties were investigated. The results showed that spores were highly stable at gastric acidity and capable to germinate and multiply under intestinal conditions as compared to vegetative cells. The higher hydrophobicity of spores, compared to vegetative cells, is advantageous for colonization and persistence in the intestine. Furthermore, the presence of F ₀ F ₁ ATP synthase, amino acid decarboxylase, bile acid symporter, mucin/collagen/fibronectin-binding proteins, heat/cold shock proteins, and universal stress proteins suggests that the strain is able to survive stress. In conclusion, the results demonstrate that B. clausii UBBC07 spores show significantly higher survival and adhesion in in vitro gastrointestinal conditions as compared to vegetative cells. Besides, this study provides a comparative analysis of the in vitro probiotic properties of spores and vegetative cells of Bacillus clausii UBBC07.

SUBMITTER: Ahire JJ

PROVIDER: S-EPMC7867671 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature

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Comparative accounts of probiotic properties of spore and vegetative cells of <i>Bacillus clausii</i> UBBC07 and in silico analysis of probiotic function.

Ahire J J JJ Kashikar M S MS Madempudi R S RS

3 Biotech 20210206 3

In this study, the spores and vegetative cells of <i>B. clausii</i> were independently evaluated for probiotic properties such as acid, gastric juice, bile, and intestinal fluid tolerance, adhesion to solvents/mucin and zeta potential. In addition, in silico identification of genome features contributing to probiotic properties were investigated. The results showed that spores were highly stable at gastric acidity and capable to germinate and multiply under intestinal conditions as compared to v ...[more]

PMID: 33604232

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Project description:BackgroundClonorchiasis, a food-borne zoonosis, is caused by Clonorchis sinensis. The intestinal tract and bile ducts are crucial places for C. sinensis metacercariae to develop into adult worms. The endospore of Bacillus subtilis is an ideal oral immunization vehicle for delivery of heterologous antigens to intestine. Cysteine protease of C. sinensis (CsCP) is an endogenous key component in the excystment of metacercariae and other physiological or pathological processes.MethodsWe constructed a fusion gene of CotC (a coat protein)-CsCP and obtained B. subtilis spores with recombinant plasmid of pEB03-CotC-CsCP (B.s-CotC-CsCP). CotC-CsCP expressed on spores' surface was detected by Western blotting and immunofluorescence. Immunological characteristics of recombinant spore coat protein were evaluated in a mouse model. The levels of CsCP-specific antibodies were detected by ELISA. Effects of recombinant spores on mouse intestine were evaluated by histological staining. The activities of biochemical enzymes in serum were assayed by microplate. Liver sections of infected mice were evaluated by Ishak score after Masson's trichrome.ResultsThe B.s-CotC-CsCP spores displayed CsCP on their coat. Specific IgG and isotypes were significantly induced by coat proteins of B.s-CotC-CsCP spores after subcutaneous immunization. IgA levels in intestinal mucus and bile of B.s-CotC-CsCP orally treated mice significantly increased. Additionally, more IgA-secreting cells were observed in enteraden and lamina propria regions of the mouse jejunum, and an increased amount of acidic mucins in intestines were also observed. There were no significant differences in enzyme levels of serum among groups. No inflammatory injury was observed in the intestinal tissues of each group. The degree of liver fibrosis was significantly reduced after oral immunization with B.s-CotC-CsCP spores.ConclusionsBacillus subtilis spores maintained the original excellent immunogenicity of CsCP expressed on their surface. Both local and systemic specific immune responses were elicited by oral administration of B.s-CotC-CsCP spores. The spores effectively promoted intestinal health by inducing secretion of acidic mucins, with no other side effects to the liver or intestine. Oral administration of spores expressing CsCP could provide effective protection against C. sinensis. This study may be a cornerstone for development of antiparasitic agents or vaccines against clonorchiasis based on B. subtilis spore expressing CsCP on the surface.

Project description:UnlabelledBacillus anthracis, a spore-forming pathogen, replicates as chains of vegetative cells by regulating the separation of septal peptidoglycan. Surface (S)-layer proteins and B. anthracis S-layer-associated proteins (BSLs) function as chain length determinants and are assembled in the envelope by binding to the secondary cell wall polysaccharide (SCWP). B. anthracis expresses six different genes encoding LytR-CpsA-Psr (LCP) enzymes (lcpB1 to -4, lcpC, and lcpD), which when expressed in Staphylococcus aureus promote attachment of wall teichoic acid to peptidoglycan. Mutations in B. anthracis lcpB3 and lcpD cause aberrations in cell size and chain length that can be explained as discrete defects in SCWP assembly; however, the function of the other lcp genes is not known. By deleting combinations of lcp genes from the B. anthracis genome, we generated variants with single lcp genes. B. anthracis expressing lcpB3 alone displayed physiological cell size, vegetative growth, spore formation, and S-layer assembly. Strains expressing lcpB1 or lcpB4 displayed defects in cell size and shape, S-layer assembly, and spore formation yet sustained vegetative growth. In contrast, the lcpB2 strain was unable to grow unless the gene was expressed from a multicopy plasmid (lcpB2(++)), and variants expressing lcpC or lcpD displayed severe defects in growth and cell shape. The lcpB2(++), lcpC, or lcpD strains supported neither S-layer assembly nor spore formation. We propose a model whereby LCP enzymes fulfill partially overlapping functions in transferring SCWP molecules to discrete sites within the bacterial envelope.ImportanceProducts of genes essential for bacterial envelope assembly represent targets for antibiotic development. The LytR-CpsA-Psr (LCP) enzymes tether bactoprenol-linked intermediates of secondary cell wall polymers to the C6 hydroxyl of N-acetylmuramic acid in peptidoglycan; however, the role of LCPs as a target for antibiotic therapy is not defined. We show here that LCP enzymes are essential for the cell cycle, vegetative growth, and spore formation of Bacillus anthracis, the causative agent of anthrax disease. Furthermore, we assign functions for each of the six LCP enzymes, including cell size and shape, vegetative growth and sporulation, and S-layer and S-layer-associated protein assembly.

Dataset Information

Comparative accounts of probiotic properties of spore and vegetative cells of Bacillus clausii UBBC07 and in silico analysis of probiotic function.

Publications

Comparative accounts of probiotic properties of spore and vegetative cells of <i>Bacillus clausii</i> UBBC07 and in silico analysis of probiotic function.

Similar Datasets

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