Efficient isolation of membrane-associated exopolysaccharides of four commercial bifidobacterial strains

Ferrari, Michela; Hameleers, Lisanne; Stuart, Marc C. A.; Oerlemans, Marjolein M. P.; Vos, Paul de; Jurak, Edita; Walvoort, Marthe T. C.

doi:10.1016/j.carbpol.2021.118913

Cited by 12 publications

(7 citation statements)

References 31 publications

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“…We also determined that the mCGB was able to support the growth of the co-culture and monocultures. The EPS extraction was performed using the methods of Ferrari et al (2022) with modifications. Briefly, 9 mL of fresh medium was inoculated with 1 mL of co-culture and monoculture suspensions (OD 600 = 0.1) and incubated aerobically at 30°C.…”

Section: Methodsmentioning

confidence: 99%

Enhanced probiotic potential of Lactobacillus kefiranofaciens OSU-BDGOA1 through co-culture with Kluyveromyces marxianus bdgo-ym6

et al. 2023

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IntroductionDue to the increasing consumer demand for the development and improvement of functional foods containing probiotics, new probiotic candidates need to be explored as well as novel means to enhance their beneficial effects. Lactobacillus kefiranofaciens OSU-BDGOA1 is a strain isolated from kefir grains that has demonstrated probiotic traits. This species is the main inhabitant of kefir grains and is responsible for the production of an exopolysaccharide (EPS) whit vast technological applications and potential bioactivities. Research has shown that interkingdom interactions of yeast and lactic acid bacteria can enhance metabolic activities and promote resistance to environmental stressors.MethodsComparative genomic analyses were performed to distinguish OSU-BDGOA1 from other strains of the same species, and the genome was mined to provide molecular evidence for relevant probiotic properties. We further assessed the cumulative effect on the probiotic properties of OSU-BDGOA1 and Kluyveromyces marxianus bdgo-ym6 yeast co-culture compared to monocultures.ResultsSurvival during simulated digestion assessed by the INFOGEST digestion model showed higher survival of OSU-BDGOA1 and bdgo-ym6 in co-culture. The adhesion to intestinal cells assessed with the Caco-2 intestinal cell model revealed enhanced adhesion of OSU-BDGOA1 in co-culture. The observed increase in survival during digestion could be associated with the increased production of EPS during the late exponential and early stationary phases of co-culture that, by enhancing co-aggregation between the yeast and the bacterium, protects the microorganisms from severe gastrointestinal conditions as observed by SEM images. Immune modulation and barrier function for recovery and prevention of flagellin-mediated inflammation by Salmonella Typhimurium heat-killed cells (HKSC) in Caco-2 cells were also measured. OSU-BDGOA1 in mono- and co-culture regulated inflammation through downregulation of pro-inflammatory cytokine expression and increased membrane barrier integrity assessed by TEER, FD4 permeability, and expression of tight junctions.DiscussionThe results of the study warrant further research into the application of co-cultures of yeast and LAB in functional probiotic products and the potential to increase EPS production by co-culture strategies.

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Section: Methodsmentioning

confidence: 99%

Enhanced probiotic potential of Lactobacillus kefiranofaciens OSU-BDGOA1 through co-culture with Kluyveromyces marxianus bdgo-ym6

et al. 2023

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“…A cheap medium with a high concentration of glucose, yeast extract and soy peptone was investigated to achieve suitable biomass yields and productivities, thereby shortening process time. However, the presence of polysaccharide contaminants in complex nitrogen sources complicates the purification of potentially bioactive compounds, such as exopolysaccharides produced by lactobacilli, and the characterization of their structure (Ferrari et al, 2022). To define a simpler medium and an industrially feasible purification strategy, the effect of lower concentrations of complex nitrogen sources (and glucose) was evaluated in the Frontiers in Bioengineering and Biotechnology frontiersin.org same experimental set-up demonstrating, unexpectedly, that the reduction did not affect the titer of viable cells and of EPS produced.…”

Section: Discussionmentioning

confidence: 99%

Optimization of growth of Levilactobacillus brevis SP 48 and in vitro evaluation of the effect of viable cells and high molecular weight potential postbiotics on Helicobacter pylori

Cimini

D’ambrosio²,

Stellavato³

et al. 2022

Front. Bioeng. Biotechnol.

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Several Levilactobacillus brevis strains have the potential to be used as probiotics since they provide health benefits due to the interaction of live cells, and of their secreted products, with the host (tissues). Therefore, the development of simple fermentation processes that improve cell viability to reduce industrial production costs, and at the same time the characterization and biological evaluation of cell-free postbiotics that can further promote application, are of great interest. In the present study, small scale batch fermentations on semi defined media, deprived of animal derived raw materials, were used to optimize growth of L. brevis SP48, reaching 1.2 ± 0.4 × 1010 CFU/ml of viable cells after 16 h of growth. Displacement, competition, and inhibition assays compared the effect, on Helicobacter pylori, of L. brevis cells to that of its partially purified potentially postbiotic fraction rich in exopolysaccharides and proteins. The expression of pro and anti-inflammatory biochemical markers indicated that both samples activated antimicrobial defenses and innate immunity in a gastric model. Moreover, these compounds also acted as modulators of the inflammatory response in a gut in vitro model. These data demonstrate that the high molecular weight compounds secreted by L. brevis SP48 can contrast H. pylori and reduce inflammation related to intestinal bowel disease, potentially overcoming issues related to the preservation of probiotic viability.

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“…At the end of incubation, the tubes were centrifuged (3,500 × g, 4°C, 10 min) and the pellets were washed with saline. Each pellet was subject to alkaline treatment using 2 mL of 2 M NaOH and agitated overnight at 25°C on a rocking platform to remove the bound polysaccharides while rendering the bacterial cells intact (26). The samples were centrifuged (7,000 × g, 4°C, 30 min) and the polysaccharides (supernatant) were precipitated with 2 volumes of EtOH with overnight incubation at −20°C.…”

Section: Growth and Bound Polysaccharide Production Curvesmentioning

confidence: 99%

“…Bound polysaccharides were extracted from bacterial pellets of a 200 mL culture in the presence or absence of 0.5% MPL following the procedure of Ferrari et al (26). Hydrolysis of extracted sugars was executed by resuspending the freeze-dried powder to a concentration of 12.5 mg/mL using sonication (10 min, 37°C).…”

Section: Bound Polysaccharide Composition and Quantification Using Hp...mentioning

confidence: 99%

Milk phospholipids protect Bifidobacterium longum subsp. infantis during in vitro digestion and enhance polysaccharide production

Kosmerl,

González-Orozco,

García-Cano

et al. 2023

Front. Nutr.

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Bifidobacterium longum subsp. infantis is associated with the gut microbiota of breast-fed infants. Bifidobacterium infantis promotes intestinal barrier and immune function through several proposed mechanisms, including interactions between their surface polysaccharides, the host, and other gut microorganisms. Dairy foods and ingredients are some of the most conspicuous food-based niches for this species and may provide benefits for their delivery and efficacy in the gut. Milk phospholipid (MPL)-rich ingredients have been increasingly recognized for their versatile benefits to health, including interactions with the gut microbiota and intestinal cells. Therefore, our objective was to investigate the capacity for MPL to promote survival of B. infantis during simulated digestion and to modulate bacterial polysaccharide production. To achieve these aims, B. infantis was incubated with or without 0.5% MPL in de Man, Rogosa, and Sharpe (MRS) media at 37°C under anaerobiosis. Survival across the oral, gastric, and intestinal phases using in vitro digestion was measured using plate count, along with adhesion to goblet-like intestinal cells. MPL increased B. infantis survival at the end of the intestinal phase by at least 7% and decreased adhesion to intestinal cells. The bacterial surface characteristics, which may contribute to these effects, were assessed by ζ-potential, changes in surface proteins using comparative proteomics, and production of bound polysaccharides. MPL decreased the surface charge of the bifidobacteria from −17 to −24 mV and increased a 50 kDa protein (3-fold) that appears to be involved in protection from stress. The production of bound polysaccharides was measured using FTIR, HPLC, and TEM imaging. These techniques all suggest an increase in bound polysaccharide production at least 1.7-fold in the presence of MPL. Our results show that MPL treatment increases B. infantis survival during simulated digestion, induces a stress resistance surface protein, and yields greater bound polysaccharide production, suggesting its use as a functional ingredient to enhance probiotic and postbiotic effects.

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Efficient isolation of membrane-associated exopolysaccharides of four commercial bifidobacterial strains

Cited by 12 publications

References 31 publications

Enhanced probiotic potential of Lactobacillus kefiranofaciens OSU-BDGOA1 through co-culture with Kluyveromyces marxianus bdgo-ym6

Enhanced probiotic potential of Lactobacillus kefiranofaciens OSU-BDGOA1 through co-culture with Kluyveromyces marxianus bdgo-ym6

Optimization of growth of Levilactobacillus brevis SP 48 and in vitro evaluation of the effect of viable cells and high molecular weight potential postbiotics on Helicobacter pylori

Milk phospholipids protect Bifidobacterium longum subsp. infantis during in vitro digestion and enhance polysaccharide production

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