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Douwe Van Sinderen - One of the best experts on this subject based on the ideXlab platform.
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ecological importance of cross feeding of the intermediate metabolite 1 2 propanediol between bacterial gut symbionts
Applied and Environmental Microbiology, 2020Co-Authors: Christopher C Cheng, Douwe Van Sinderen, Rebbeca M Duar, Xiaoxi Lin, Maria Elisa Perezmunoz, Stephanie Tollenaar, Janpeter Van Pijkeren, Michael G Ganzle, Jens WalterAbstract:Cross-feeding based on the metabolite 1,2-propanediol has been proposed to have an important role in the establishment of trophic interactions among gut symbionts, but its ecological importance has not been empirically established. Here, we show that in vitro growth of Lactobacillus reuteri (syn. Limosilactobacillus reuteri) ATCC PTA 6475 is enhanced through 1,2-propanediol produced by Bifidobacterium Breve UCC2003 and Escherichia coli MG1655 from the metabolization of fucose and rhamnose, respectively. Work with isogenic mutants showed that the trophic interaction is dependent on the pduCDE operon in L. reuteri, which encodes the ability to use 1,2-propanediol, and the l-fucose permease (fucP) gene in B. Breve, which is required for 1,2-propanediol formation from fucose. Experiments in gnotobiotic mice revealed that, although the pduCDE operon bestows a fitness burden on L. reuteri ATCC PTA 6475 in the mouse digestive tract, the ecological performance of the strain was enhanced in the presence of B. Breve UCC2003 and the mucus-degrading species Bifidobacterium bifidum The use of the respective pduCDE and fucP mutants of L. reuteri and B. Breve in the mouse experiments indicated that the trophic interaction was specifically based on 1,2-propanediol. Overall, our work established the ecological importance of cross-feeding relationships based on 1,2-propanediol for the fitness of a bacterial symbiont in the vertebrate gut.IMPORTANCE Through experiments in gnotobiotic mice that employed isogenic mutants of bacterial strains that produce (Bifidobacterium Breve) and utilize (Lactobacillus reuteri) 1,2-propanediol, this study provides mechanistic insight into the ecological ramifications of a trophic interaction between gut symbionts. The findings improve our understanding on how cross-feeding influences the competitive fitness of L. reuteri in the vertebrate gut and revealed a putative selective force that shaped the evolution of the species. The findings are relevant since they provide a basis to design rational microbial-based strategies to modulate gut ecosystems, which could employ mixtures of bacterial strains that establish trophic interactions or a personalized approach based on the ability of a resident microbiota to provide resources for the incoming microbe.
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cross feeding by Bifidobacterium Breve ucc2003 during co cultivation with Bifidobacterium bifidum prl2010 in a mucin based medium
BMC Microbiology, 2014Co-Authors: Muireann Egan, Marco Ventura, Mary Oconnell Motherway, Michelle Kilcoyne, Marian Kane, Lokesh Joshi, Douwe Van SinderenAbstract:Background Bifidobacteria constitute a specific group of commensal bacteria that commonly inhabit the mammalian gastrointestinal tract. Bifidobacterium Breve UCC2003 was previously shown to utilize a variety of plant/diet/host-derived carbohydrates, including cellodextrin, starch and galactan, as well as the mucin and HMO-derived monosaccharide, sialic acid. In the current study, we investigated the ability of this strain to utilize parts of a host-derived source of carbohydrate, namely the mucin glycoprotein, when grown in co-culture with the mucin-degrading Bifidobacterium bifidum PRL2010.
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a bile inducible membrane protein mediates bifidobacterial bile resistance
Microbial Biotechnology, 2012Co-Authors: Lorena Ruiz, Abelardo Margolles, Clara G De Los Reyesgavilan, Aldert Zomer, Mary Oconnellmotherway, Douwe Van SinderenAbstract:Bbr_0838 from Bifidobacterium Breve UCC2003 is predicted to encode a 683 residue membrane protein, containing both a permease domain that displays similarity to transporters belonging to the major facilitator superfamily, as well as a CBS (cystathionine beta synthase) domain. The high level of similarity to bile efflux pumps from other bifidobacteria suggests a significant and general role for Bbr_0838 in bile tolerance. Bbr_0838 transcription was shown to be monocistronic and strongly induced upon exposure to bile. Further analysis delineated the transcriptional start site and the minimal region required for promoter activity and bile regulation. Insertional inactivation of Bbr_0838 in B. Breve UCC2003 resulted in a strain, UCC2003:838(800) , which exhibited reduced survival upon cholate exposure as compared with the parent strain, a phenotype that was reversed when a functional, plasmid-encoded Bbr_0838 gene was introduced into UCC2003:838(800) . Transcriptome analysis of UCC2003:838(800) grown in the presence or absence of bile demonstrated that transcription of Bbr_0832, which is predicted to encode a macrolide efflux transporter gene, was significantly increased in the presence of bile, representing a likely compensatory mechanism for bile removal in the absence of Bbr_0838. This study represents the first in-depth analysis of a bile-inducible locus in bifidobacteria, identifying a key gene relevant for bifidobacterial bile tolerance.
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Bifidobacterium Breve ucc2003 surface exopolysaccharide production is a beneficial trait mediating commensal host interaction through immune modulation and pathogen protection
Gut microbes, 2012Co-Authors: Saranna Fanning, Lindsay J Hall, Douwe Van SinderenAbstract:Bifidobacteria constitute a substantial proportion of the human gut microbiota. There are currently many bifidobacterial strains with claimed probiotic attributes. The mechanism through which these strains reside within their host and exert benefits to the host is far from fully understood. We have shown in the case of Bifidobacterium Breve UCC2003 that a cell surface exopolysaccharide (EPS) plays a role in in vivo persistence. Biosynthesis of two possible EPSs is controlled by a bidirectional gene cluster which guides alternate EPS synthesis by means of a reorienting promoter. The presence of EPS impacts on host immune response: the wild type, EPS-positive B. Breve UCC2003 efficiently evades the adaptive B-cell host response, while its isogenic, EPS-deficient equivalent elicits a strong adaptive immune response. Functionally, EPS positive strains were more resilient to presence of acid and bile and were responsible for reduced colonization levels of Citrobacter rodentium, a gut pathogen. In conclusion, we have found that EPS is important in host interactions and pathogen protection, the latter indicative of a probiotic ability for the EPS of B. Breve UCC2003.
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bile inducible efflux transporter from Bifidobacterium longum ncc2705 conferring bile resistance
Applied and Environmental Microbiology, 2009Co-Authors: Miguel Gueimonde, Douwe Van Sinderen, Christel Garrigues, Clara G De Los Reyesgavilan, Abelardo MargollesAbstract:Bifidobacteria are normal inhabitants of the human gut. Some strains of this genus are considered health promoting or probiotic, being included in numerous food products. In order to exert their health benefits, these bacteria must overcome biological barriers, including bile salts, to colonize and survive in specific parts of the intestinal tract. The role of multidrug resistance (MDR) transporters in bile resistance of probiotic bacteria and the effect of bile on probiotic gene expression are not fully understood. In the present study, the effect of subinhibitory concentrations of bile on the expression levels of predicted MDR genes from three different bifidobacterial strains, belonging to Bifidobacterium longum subsp. longum, Bifidobacterium Breve, and Bifidobacterium animalis subsp. lactis, was tested. In this way, two putative MDR genes whose expression was induced by bile, BL0920 from B. longum and its homolog, Bbr0838, from B. Breve, were identified. The expression of the BL0920 gene in Escherichia coli was shown to confer resistance to bile, likely to be mediated by active efflux from the cells. To the best of our knowledge, this represents the first identified bifidobacterial bile efflux pump whose expression is induced by bile.
Catherine Stanton - One of the best experts on this subject based on the ideXlab platform.
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myosin cross reactive antigen mcra protein from Bifidobacterium Breve is a fad dependent fatty acid hydratase which has a function in stress protection
BMC Biochemistry, 2011Co-Authors: Gerald F. Fitzgerald, Eva Rosbergcody, Alena Liavonchanka, Cornelia Gobel, Paul R Ross, Orla Osullivan, Ivo Feussner, Catherine StantonAbstract:Background: The aim of this study was to determine the catalytic activity and physiological role of myosin-crossreactive antigen (MCRA) from Bifidobacterium Breve NCIMB 702258. MCRA from B. Breve NCIMB 702258 was cloned, sequenced and expressed in heterologous hosts (Lactococcus and Corynebacterium) and the recombinant proteins assessed for enzymatic activity against fatty acid substrates. Results: MCRA catalysed the conversion of palmitoleic, oleic and linoleic acids to the corresponding 10-hydroxy fatty acids, but shorter chain fatty acids were not used as substrates, while the presence of trans-double bonds and double bonds beyond the position C12 abolished hydratase activity. The hydroxy fatty acids produced were not metabolised further. We also found that heterologous Lactococcus and Corynebacterium expressing MCRA accumulated increasing amounts of 10-HOA and 10-HOE in the culture medium. Furthermore, the heterologous cultures exhibited less sensitivity to heat and solvent stresses compared to corresponding controls. Conclusions: MCRA protein in B. Breve can be classified as a FAD-containing double bond hydratase, within the carbon-oxygen lyase family, which may be catalysing the first step in conjugated linoleic acid (CLA) production, and this protein has an additional function in bacterial stress protection.
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intestinal bifidobacteria that produce trans 9 trans 11 conjugated linoleic acid a fatty acid with antiproliferative activity against human colon sw480 and ht 29 cancer cells
Nutrition and Cancer, 2006Co-Authors: M Coakley, Gerald F. Fitzgerald, Rosaleen Devery, Paul R Ross, Mark C Johnson, Emma Mcgrath, Shafiqur Rahman, Catherine StantonAbstract:Bifidobacterium Breve species of human intestinal origin have the ability to synthesize cis-9, trans-11 (c9, t11) conjugated linoleic acid (CLA) from free linoleic acid. In this study, the ability of Bifidobacterium species to isomerize C(18) polyunsaturated fatty acids was investigated, and the antiproliferative activities of the two main microbially produced CLA isomers were assessed. Linoleic acid was converted principally to c9, t11 CLA and lesser amounts of t9, t11 CLA, whereas c9, t11 CLA was converted mainly to t9, t11 CLA. Likewise, t10, c12 CLA was converted principally to t9, t11 CLA, which was incorporated into the bacterial cell membranes. To examine the antiproliferative effect of the two main CLA isomers formed, SW480 and HT-29 human colon cancer cells were cultured in the presence of c9, t11 CLA and t9, t11 CLA. The t9, t11 CLA had a more potent antiproliferative effect than c9, t11 CLA. It is tempting to suggest that the ability of Bifidobacterium to produce such bioactive metabolites may be associated with the beneficial effects of bifidobacteria present in the human gastrointestinal tract.
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Conjugated linoleic acid biosynthesis by human-derived Bifidobacterium species
Journal of Applied Microbiology, 2002Co-Authors: Mairead Coakley, R.p. Ross, Marcus Nordgren, Gerald F. Fitzgerald, Rosaleen Devery, Catherine StantonAbstract:Aims: To assess strains of Lactobacillus, Lactococcus, Pediococcus and Bifidobacterium for their ability to produce the health-promoting fatty acid conjugated linoleic acid (CLA) from free linoleic acid. Methods and Results: In this study, strains of Lactobacillus, Lactococcus, Pediococcus and Bifidobacterium were grown in medium containing free linoleic acid. Growth of the bacteria in linoleic acid and conversion of the linoleic acid to CLA was assessed. Of the bacteria assessed, nine strains of Bifidobacterium produced the c9, t11 CLA isomer from free linoleic acid. The t9, t11 CLA isomer was also produced by some strains, but at much lower concentrations. Conclusions: The production of CLA by bifidobacteria exhibited considerable interspecies variation. Bifidobacterium Breve and B. dentium were the most efficient CLA producers among the range of strains tested, with B. Breve converting up to 65% linoleic acid to c9, t11 CLA when grown in 0·55 mg ml−1 linoleic acid. Strains also varied considerably with respect to their sensitivity to linoleic acid. Significance and Impact of the Study: The production of CLA by probiotic bifidobacteria offers a possible mechanism for some health-enhancing properties of bifidobacteria and provides novel opportunities for the development of functional foods.
Hoi Shan Kwan - One of the best experts on this subject based on the ideXlab platform.
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mechanistic study of utilization of water insoluble saccharomyces cerevisiae glucans by Bifidobacterium Breve strain jcm1192
Applied and Environmental Microbiology, 2017Co-Authors: Hoi Yee Keung, Lokto Sham, Man Kit Cheung, Peter C K Cheung, Hoi Shan KwanAbstract:Bifidobacteria exert beneficial effects on hosts and are extensively used as probiotics. However, due to their genetic inaccessibility, little is known about their mechanism of carbohydrate utilization and regulation. Bifidobacterium Breve strain JCM1192 can grow on water insoluble yeast cell wall glucans (YCWG), which are recently considered as potential prebiotics. From the result of 1 H NMR spectrometry, the YCWG were composed of highly branched (1→3)(1→6)-β-glucan and (1→4)(1→6)-α-glucan. Although the YCWG composed of 78.3% β-glucan and 21.7% α-glucan, only α-glucan was consumed by the B. Breve strain. The ABC transporter ( malEFG1 ) and pullulanase ( aapA ) genes were transcriptionally upregulated in the metabolism of insoluble yeast glucan, suggesting their potential involvement in the process. A non-sense mutation identified in the gene encoding an ABC transporter ATP-binding protein (MalK) led to growth failure of an ethyl methanesulfonate-generated mutant on yeast glucans. Co-culture of the wild type and the mutant showed that this protein was responsible for the import of yeast glucan or its breakdown products instead of export of α-glucan catabolic enzymes. Further carbohydrate utilization characterization of the mutant and three of its revertants indicated that this mutation was pleiotropic — the mutant could not grow on maltose, glycogen, dextrin, raffinose, cellobiose, melibiose, or turanose. We propose that insoluble yeast α-glucan is hydrolyzed by extracellular pullulanase into maltose/malto-oligosaccharides, which are then transported into the cell through the ABC transport system comprised of MalEFG1 and MalK. The mechanism unraveled here will facilitate the development of B. Breve and water insoluble yeast glucans as a novel synbiotics. Importance Bifidobacterium strains are in general genetically intractable. Coupling classic forward genetics with next generation sequencing, we identified here an ABC transporter ATP-binding protein (MalK) responsible for the import of insoluble yeast glucan breakdown products by B. Breve JCM1192. We demonstrated the pleiotropic effect of the ABC transporter ATP-binding protein in maltose/malto-oligosaccharides, raffinose, cellobiose, melibiose, and turanose transport. With the addition of transcriptional analysis, we propose that insoluble yeast glucan is broken down by extracellular pullulanase into maltose/malto-oligosaccharides, which are then transported into the cell through the ABC transport system comprised of MalEFG1 and MalK. The mechanism unraveled here will facilitate the development of B. Breve and water insoluble yeast glucans as a novel synbiotics.
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mechanistic study of utilization of water insoluble saccharomyces cerevisiae glucans by Bifidobacterium Breve strain jcm1192
Applied and Environmental Microbiology, 2017Co-Authors: Hoi Yee Keung, Lokto Sham, Man Kit Cheung, Peter C K Cheung, Hoi Shan KwanAbstract:Bifidobacteria exert beneficial effects on hosts and are extensively used as probiotics. However, due to the genetic inaccessibility of these bacteria, little is known about their mechanisms of carbohydrate utilization and regulation. Bifidobacterium Breve strain JCM1192 can grow on water-insoluble yeast (Saccharomyces cerevisiae) cell wall glucans (YCWG), which were recently considered as potential prebiotics. According to the results of 1H nuclear magnetic resonance (NMR) spectrometry, the YCWG were composed of highly branched (1→3,1→6)-β-glucans and (1→4,1→6)-α-glucans. Although the YCWG were composed of 78.3% β-glucans and 21.7% α-glucans, only α-glucans were consumed by the B. Breve strain. The ABC transporter (malEFG1) and pullulanase (aapA) genes were transcriptionally upregulated in the metabolism of insoluble yeast glucans, suggesting their potential involvement in the process. A nonsense mutation identified in the gene encoding an ABC transporter ATP-binding protein (MalK) led to growth failure of an ethyl methanesulfonate-generated mutant with yeast glucans. Coculture of the wild-type strain and the mutant showed that this protein was responsible for the import of yeast glucans or their breakdown products, rather than the export of α-glucan-catabolizing enzymes. Further characterization of the carbohydrate utilization of the mutant and three of its revertants indicated that this mutation was pleiotropic: the mutant could not grow with maltose, glycogen, dextrin, raffinose, cellobiose, melibiose, or turanose. We propose that insoluble yeast α-glucans are hydrolyzed by extracellular pullulanase into maltose and/or maltooligosaccharides, which are then transported into the cell by the ABC transport system composed of MalEFG1 and MalK. The mechanism elucidated here will facilitate the development of B. Breve and water-insoluble yeast glucans as novel synbiotics.IMPORTANCE In general, Bifidobacterium strains are genetically intractable. Coupling classic forward genetics with next-generation sequencing, here we identified an ABC transporter ATP-binding protein (MalK) responsible for the import of insoluble yeast glucan breakdown products by B. Breve JCM1192. We demonstrated the pleiotropic effects of the ABC transporter ATP-binding protein in maltose/maltooligosaccharide, raffinose, cellobiose, melibiose, and turanose transport. With the addition of transcriptional analysis, we propose that insoluble yeast glucans are broken down by extracellular pullulanase into maltose and/or maltooligosaccharides, which are then transported into the cell by the ABC transport system composed of MalEFG1 and MalK. The mechanism elucidated here will facilitate the development of B. Breve and water-insoluble yeast glucans as novel synbiotics.
Muireann Egan - One of the best experts on this subject based on the ideXlab platform.
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Comparative genomics and genotype-phenotype associations in Bifidobacterium Breve
Scientific Reports, 2018Co-Authors: Francesca Bottacini, Justin Van Breen, Evgenia Dikareva, Jolanda Lambert, Kees Van Limpt, Ruth Morrissey, María Esteban-torres, Kieran James, Muireann Egan, Jan KnolAbstract:Bifidobacteria are common members of the gastro-intestinal microbiota of a broad range of animal hosts. Their successful adaptation to this particular niche is linked to their saccharolytic metabolism, which is supported by a wide range of glycosyl hydrolases. In the current study a large-scale gene-trait matching (GTM) effort was performed to explore glycan degradation capabilities in B . Breve . By correlating the presence/absence of genes and associated genomic clusters with growth/no-growth patterns across a dataset of 20 Bifidobacterium Breve strains and nearly 80 different potential growth substrates, we not only validated the approach for a number of previously characterized carbohydrate utilization clusters, but we were also able to discover novel genetic clusters linked to the metabolism of salicin and sucrose. Using GTM, genetic associations were also established for antibiotic resistance and exopolysaccharide production, thereby identifying (novel) bifidobacterial antibiotic resistance markers and showing that the GTM approach is applicable to a variety of phenotypes. Overall, the GTM findings clearly expand our knowledge on members of the B . Breve species, in particular how their variable genetic features can be linked to specific phenotypes.
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cross feeding by Bifidobacterium Breve ucc2003 during co cultivation with Bifidobacterium bifidum prl2010 in a mucin based medium
BMC Microbiology, 2014Co-Authors: Muireann Egan, Marco Ventura, Mary Oconnell Motherway, Michelle Kilcoyne, Marian Kane, Lokesh Joshi, Douwe Van SinderenAbstract:Background Bifidobacteria constitute a specific group of commensal bacteria that commonly inhabit the mammalian gastrointestinal tract. Bifidobacterium Breve UCC2003 was previously shown to utilize a variety of plant/diet/host-derived carbohydrates, including cellodextrin, starch and galactan, as well as the mucin and HMO-derived monosaccharide, sialic acid. In the current study, we investigated the ability of this strain to utilize parts of a host-derived source of carbohydrate, namely the mucin glycoprotein, when grown in co-culture with the mucin-degrading Bifidobacterium bifidum PRL2010.
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metabolism of sialic acid by Bifidobacterium Breve ucc2003
Applied and Environmental Microbiology, 2014Co-Authors: Muireann Egan, Marco Ventura, Mary Oconnell Motherway, Douwe Van SinderenAbstract:Bifidobacteria constitute a specific group of commensal bacteria that inhabit the gastrointestinal tracts of humans and other mammals. Bifidobacterium Breve UCC2003 has previously been shown to utilize several plant-derived carbohydrates that include cellodextrins, starch, and galactan. In the present study, we investigated the ability of this strain to utilize the mucin- and human milk oligosaccharide (HMO)-derived carbohydrate sialic acid. Using a combination of transcriptomic and functional genomic approaches, we identified a gene cluster dedicated to the uptake and metabolism of sialic acid. Furthermore, we demonstrate that B. Breve UCC2003 can cross feed on sialic acid derived from the metabolism of 3′-sialyllactose, an abundant HMO, by another infant gut bifidobacterial strain, Bifidobacterium bifidum PRL2010.
Jens Walter - One of the best experts on this subject based on the ideXlab platform.
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ecological importance of cross feeding of the intermediate metabolite 1 2 propanediol between bacterial gut symbionts
Applied and Environmental Microbiology, 2020Co-Authors: Christopher C Cheng, Douwe Van Sinderen, Rebbeca M Duar, Xiaoxi Lin, Maria Elisa Perezmunoz, Stephanie Tollenaar, Janpeter Van Pijkeren, Michael G Ganzle, Jens WalterAbstract:Cross-feeding based on the metabolite 1,2-propanediol has been proposed to have an important role in the establishment of trophic interactions among gut symbionts, but its ecological importance has not been empirically established. Here, we show that in vitro growth of Lactobacillus reuteri (syn. Limosilactobacillus reuteri) ATCC PTA 6475 is enhanced through 1,2-propanediol produced by Bifidobacterium Breve UCC2003 and Escherichia coli MG1655 from the metabolization of fucose and rhamnose, respectively. Work with isogenic mutants showed that the trophic interaction is dependent on the pduCDE operon in L. reuteri, which encodes the ability to use 1,2-propanediol, and the l-fucose permease (fucP) gene in B. Breve, which is required for 1,2-propanediol formation from fucose. Experiments in gnotobiotic mice revealed that, although the pduCDE operon bestows a fitness burden on L. reuteri ATCC PTA 6475 in the mouse digestive tract, the ecological performance of the strain was enhanced in the presence of B. Breve UCC2003 and the mucus-degrading species Bifidobacterium bifidum The use of the respective pduCDE and fucP mutants of L. reuteri and B. Breve in the mouse experiments indicated that the trophic interaction was specifically based on 1,2-propanediol. Overall, our work established the ecological importance of cross-feeding relationships based on 1,2-propanediol for the fitness of a bacterial symbiont in the vertebrate gut.IMPORTANCE Through experiments in gnotobiotic mice that employed isogenic mutants of bacterial strains that produce (Bifidobacterium Breve) and utilize (Lactobacillus reuteri) 1,2-propanediol, this study provides mechanistic insight into the ecological ramifications of a trophic interaction between gut symbionts. The findings improve our understanding on how cross-feeding influences the competitive fitness of L. reuteri in the vertebrate gut and revealed a putative selective force that shaped the evolution of the species. The findings are relevant since they provide a basis to design rational microbial-based strategies to modulate gut ecosystems, which could employ mixtures of bacterial strains that establish trophic interactions or a personalized approach based on the ability of a resident microbiota to provide resources for the incoming microbe.