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Phenotypic and genetic characterization of differential galacto-oligosaccharide utilization in Lactobacillus plantarum.

ABSTRACT: Several Lactobacillus plantarum strains are marketed as probiotics for their potential health benefits. Prebiotics, e.g., galacto-oligosaccharides (GOS), have the potential to selectively stimulate the growth of L. plantarum probiotic strains based on their phenotypic diversity in carbohydrate utilization, and thereby enhance their health promoting effects in the host in a strain-specific manner. Previously, we have shown that GOS variably promotes the strain-specific growth of L. plantarum. In this study we investigated this variation by molecular analysis of GOS utilization by L. plantarum. HPAEC-PAD analysis revealed two distinct GOS utilization phenotypes in L. plantarum. Linking these phenotypes to the strain-specific genotypes led to the identification of a lac operon encoding a ?-galactosidase (lacA), a permease (lacS), and a divergently oriented regulator (lacR), that are predicted to be involved in the utilization of higher degree of polymerization (DP) constituents present in GOS (specifically DP of 3-4). Mutation of lacA and lacS in L. plantarum NC8 resulted in reduced growth on GOS, and HPAEC analysis confirmed the role of these genes in the import and utilization of higher-DP GOS constituents. Overall, the results enable the design of highly-selective synbiotic combinations of L. plantarum strain-specific probiotics and specific GOS-prebiotic fractions.

SUBMITTER: Fuhren J

PROVIDER: S-EPMC7728778 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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Phenotypic and genetic characterization of differential galacto-oligosaccharide utilization in Lactobacillus plantarum.

Fuhren Jori J Schwalbe Markus M Peralta-Marzal Lucía L Rösch Christiane C Schols Henk A HA Kleerebezem Michiel M

Scientific reports 20201210 1

Several Lactobacillus plantarum strains are marketed as probiotics for their potential health benefits. Prebiotics, e.g., galacto-oligosaccharides (GOS), have the potential to selectively stimulate the growth of L. plantarum probiotic strains based on their phenotypic diversity in carbohydrate utilization, and thereby enhance their health promoting effects in the host in a strain-specific manner. Previously, we have shown that GOS variably promotes the strain-specific growth of L. plantarum. In ...[more]

PMID: 33303847

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Project description:Ecological approaches can help to correct oral microbial dysbiosis and drive the advent and persistence of a symbiotic oral microbiome, which benefits long-term dental caries control. The aim of this study was to investigate the impact of the prebiotic Galacto-oligosaccharide (GOS) on the growth of probiotics L. plantarum 14,917 and its effect on the inhibitory ability of L. plantarum 14,917 against the growth of Streptococcus mutans and Candida albicans in an in vitro model. Single-species growth screenings were conducted in TSBYE broth with 1% glucose and 1-5% GOS. Interaction experiments were performed using duo- and multi-species models with inoculation of 105 CFU/mL S. mutans, 103 CFU/mL C. albicans, and 108 CFU/mL L. plantarum 14,917 under 1%, 5% GOS or 1% glucose. Viable cells and pH changes were measured. Real-time PCR was utilized to assess expression of C. albicans and S. mutans virulence genes. Six replicates were used for each group. Student's t-test, one-way ANOVA, and Kruskal-Wallis were employed to compare the outcomes of different groups. GOS significantly inhibited the growth of C. albicans and S. mutans in terms of growth quantity and speed when the two strains were grown individually. However, GOS did not affect the growth of L. plantarum 14,917. Moreover, 1% and 5% GOS enhanced the anti-fungal performance of L. plantarum 14,917 in comparison to 1% glucose. GOS as the carbon source resulted in a less acidic environment in the C. albicans and S. mutans duo-species model and multispecies model where L. plantarum 14,917 was added. When GOS was utilized as the carbohydrate substrate, S. mutans and C. albicans had a significant reduction in the expression of the HWP1, ECE1, atpD, and eno genes (p < 0.05). To our knowledge, this is the first study that reported the ability of GOS to neutralize S. mutans-C. albicans high caries of medium pH and to disrupt virulence gene expression. Moreover, as a prebiotic, GOS augmented the inhibitory ability of L. plantarum against C. albicans in vitro. The current study revealed the anti-caries potential of prebiotics GOS and shed light on novel caries prevention strategies from the perspective of prebiotics and probiotics. These findings provide a rationale for future biofilm or clinical studies to elucidate the effect of GOS on modulating oral microbiota and caries control.

Project description:BackgroundThe safety of an infant formula containing a new mixture of the prebiotics galacto-oligosaccharides (GOS) and fructo-oligosaccharide (FOS) and the probiotic Lactobacillus reuteri needs to be evaluated.MethodsHealthy term infants in Singapore were randomly assigned (using computer-generated allocation sequences) to receive exclusively an experimental infant formula containing L. reuteri, GOS (5.50 g/L), and FOS (0.36 g/L) or a control formula containing only L. reuteri from enrollment (7-14 days of age) to 4 months of age. The primary objective of this trial was to demonstrate that weight change between birth and 4 months of age in infants fed the experimental formula was not inferior to World Health Organization (WHO) Child Growth standards. The non-inferiority margin was -0.5 standard deviations (SD). The secondary objectives were to compare changes in anthropometric measurements (weight, length, body mass index, and head circumference), digestive tolerance, stool bacterial counts, urinary D- and L- lactate concentrations, and adverse events in the two formula groups.ResultsThe intention-to-treat (ITT) population included all randomized infants stratified by gender, (experimental group, N = 68 and control group, N = 72). The per-protocol (PP) population included 61 infants in the experimental and 62 infants in the control groups. The change in weight-for-age z-score between birth and 4 months was +0.93 (95% confidence interval [CI]: +0.63 to +1.23) SD in the experimental group and +0.92 (95% CI: +0.62 to +1.22) SD in the control group in the PP population, indicating non-inferior weight gain in both formulas groups compared with WHO standards. The ITT population had similar results. Liquid stools occurred more frequently in the experimental compared with the control group and median bifidobacteria, lactobacilli, and enterococci counts were higher in the experimental group (p < 0.05). Other secondary outcomes were not significantly different between groups.ConclusionsInfant formula containing L. reuteri + GOS/FOS supports normal growth and is safe.Trial registrationClinicalTrial.gov: NCT01010113.

Dataset Information

Phenotypic and genetic characterization of differential galacto-oligosaccharide utilization in Lactobacillus plantarum.

Publications

Phenotypic and genetic characterization of differential galacto-oligosaccharide utilization in Lactobacillus plantarum.

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