New screening methods for probiotics with adhesion properties to sialic acid and sulphate residues in human colonic mucin using the Biacore assay

Abstract

Aims: To determine the relationship between adhesive ability of probiotics and acidic residues in human colonic mucin, we developed a new screening method using Biacore to evaluate adherence of bacteria before and after sialic acid or sulphate residues were blocked or removed from mucin. Methods and Results: Ten strains of lactobacilli and three strains of bifidobacteria isolated from human faeces were evaluated for their adhesive properties to soluble human colonic mucin (sHCM) using the Biacore binding assay. Three strains (Lactobacillus strain ME-522, Lact. gasseri ME-527 and Bifidobacterium bifidum MCC1092) showing significant adherence were selected. Decreased binding activities were observed after removing sialic acid of sHCM using sialidase. However, after removing the sulphate residue using sulphatase, the adhesion of ME-527 decreased; whereas the remaining two strains had increased adhesion. The adhesion of three probiotics significantly decreased after the sulphate residue was blocked by elution with barium chloride. Conclusions: A new evaluation method using the Biacore assay was developed to observe binding properties to the acidic residues of sHCM. Results indicated that there was a strong relationship between probiotic adhesion and acidic residues of sHCM. Significance and Impact of the Study: This is the first report showing a screening method that quantitatively measures the binding between bacteria and acidic residues in sHCM using the Biacore binding assay; and provides a new method for the selection of probiotics in the future.

Evaluation of bifidobacterial adhesion to acidic sugar chains of porcine colonic mucins

“…Interestingly, adhesion of B. animalis subsp. animalis MCC-1489 was significantly increased after sulfatase treatment, suggesting that the binding sites may be exposed after sulfatase treatment, as described by Huang et al, 14) who showed similar interactions in the adhesion of Lactobacillus ME-522 and B. bifidum MCC-1092 to mucin. However, additional studies are needed to characterize the binding properties of B. animalis subsp.…”

“…12) Thus, for probiotics, including bifidobacteria, adhesion to acidic mucins is thought to be an advantage for survival during transit and colonization in the intestinal tract. [13][14][15] The interactions between Bifidobacterium strains and carbohydrate moieties of glycoconjugates, including glycolipids and mucins, have been described previously; for example, Bifidobacterium pseudolongum ATCC 25526 and Bifidobacterium adolescentis VTT E-001561 specifically adhere to α-1,4-linked glucose sugars, 16) while Bifidobacterium bifidum EB102 and Bifidobacterium longum SBT2928 bind to gangliotetraosylceramide. 17,18) A recent study showed that B. bifidum MCC-1092 binds to both sialic acid and sulfated carbohydrates of human colonic mucins because adhesion of the bacterium to mucins was reduced by treatment with sialidase and sulfatase and blocked by barium chloride.…”

Probiotic roles ofLactobacillussp. in swine: insights from gut microbiota

“…Probiotic lactobacilli are further selected for their ability to adhere to mucus and epithelial cells, as these characteristics are essential for their ability to colonize the gut's mucosal and epithelial layer and enhance their competitiveness against pathogens (Huang et al . ; Valeriano et al . ).…”

“…Autochthonous lactobacilli are frequently selected as potential probiotics because of their natural ability to withstand the harsh physiological conditions caused by stomach acid, bile salts and pancreatic enzymes during their passage through the GIT (Walter 2008). Probiotic lactobacilli are further selected for their ability to adhere to mucus and epithelial cells, as these characteristics are essential for their ability to colonize the gut's mucosal and epithelial layer and enhance their competitiveness against pathogens (Huang et al 2013;Valeriano et al 2014). This competitive profile is most likely present in autochthonous lactobacilli, which are used as biomarkers of health and are important components of the gut microflora of pigs.…”

Use of Atomic Force Microscopy to Study the Multi-Modular Interaction of Bacterial Adhesins to Mucins

“…Methodologies to screen for bacterial adhesion to mucins have previously employed thin layer chromatography overlay [26], enzyme-linked immunosorbent assay [15], micro-titre plate assays [8,25,27], surface plasmon resonance [16,28,29,30,31], fluorescence spectroscopy [20], mucin microarrays [32], flow cytometry [33], and cell-based assays [26,34,35,36]. However, due to the complexity and diversity of mucin glycosylation, these methods typically provide qualitative binding data indicating only presence or absence of interaction.…”