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1.
Mol Microbiol ; 112(1): 114-130, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30947380

RESUMO

Human gut bifidobacteria rely on ATP-binding cassette (ABC) transporters for oligosaccharide uptake. Multiple oligosaccharide-specific solute-binding protein (SBP) genes are occasionally associated with a single ABC transporter, but the significance of this multiplicity remains unclear. Here, we characterize BlMnBP1 and BlMnBP2, the two SBPs associated to the ß-manno-oligosaccharide (MnOS) ABC transporter in Bifidobacterium animalis subsp. lactis. Despite similar overall specificity and preference to mannotriose (Kd ≈80 nM), affinity of BlMnBP1 is up to 2570-fold higher for disaccharides than BlMnBP2. Structural analysis revealed a substitution of an asparagine that recognizes the mannosyl at position 2 in BlMnBP1, by a glycine in BlMnBP2, which affects substrate affinity. Both substitution types occur in bifidobacterial SBPs, but BlMnBP1-like variants prevail in human gut isolates. B. animalis subsp. lactis ATCC27673 showed growth on gluco and galactomannans and was able to outcompete a mannan-degrading Bacteroides ovatus strain in co-cultures, attesting the efficiency of this ABC uptake system. By contrast, a strain that lacks this transporter failed to grow on mannan. This study highlights SBP diversification as a possible strategy to modulate oligosaccharide uptake preferences of bifidobacterial ABC-transporters during adaptation to specific ecological niches. Efficient metabolism of galactomannan by distinct bifidobacteria, merits evaluating this plant glycan as a potential prebiotic.


Assuntos
Transportadores de Cassetes de Ligação de ATP/metabolismo , Bifidobacterium animalis/metabolismo , Mananas/metabolismo , Transportadores de Cassetes de Ligação de ATP/fisiologia , Proteínas de Bactérias/metabolismo , Bifidobacterium/genética , Bifidobacterium/metabolismo , Bifidobacterium animalis/genética , Proteínas de Ligação a DNA/metabolismo , Galactose/análogos & derivados , Oligossacarídeos/metabolismo
2.
Benef Microbes ; 9(6): 963-974, 2018 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-30208728

RESUMO

A feeding study using rats was conducted to evaluate the utility of lablab bean husk and soya bean husk as sources of potential prebiotic fibre. Twenty 5-week-old Sprague Dawley rats were divided into 4 groups and fed one of the following diets for 3 weeks: purified diet (AIN93 G) containing 5% cellulose (CEL), or the same diet in which cellulose was replaced by corn starch (STA), lablab bean husk (LBH), or soya bean husk (SBH). Rats were sacrificed at 8 weeks of age and caecal digesta were collected. Feed intake, body weight, anatomical parameters, and caecal ammonia level did not differ significantly among diets. Rats on LBH and SBH showed higher concentrations of caecal short-chain fatty acid and lactate than those on CEL. Rats on CEL, SBH, and LBH exhibited lower caecal indole and skatole levels. LBH yielded increased caecal abundance of Akkermansia muciniphila and Oscillibacter relatives, as demonstrated by either qPCR, MiSeq, or clone library analysis. SBH favoured the growth of lactobacilli as assessed by both qPCR and MiSeq, and favoured the growth of bifidobacteria as assessed by MiSeq. In comparison with STA, LBH and SBH yielded lower caecal abundance of bacteria related to Dorea massiliensis, as demonstrated by qPCR, MiSeq, and clone library analysis. Both types of bean husk were found to contain oligosaccharides that might selectively stimulate the growth of beneficial bacteria. Based on these results, the two species of bean husk tested are considered potentially functional for promoting the gut health of monogastric animals.


Assuntos
Ração Animal , Ceco/metabolismo , Ceco/microbiologia , Fibras na Dieta/administração & dosagem , Fermentação/efeitos dos fármacos , Microbioma Gastrointestinal/efeitos dos fármacos , Prebióticos/administração & dosagem , Animais , Conteúdo Gastrointestinal/química , Metagenômica , Compostos Orgânicos/análise , Phaseolus , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNA , Glycine max
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