scholarly journals Bifidobacterium breveUCC2003 Employs Multiple Transcriptional Regulators To Control Metabolism of Particular Human Milk Oligosaccharides

2018 ◽  
Vol 84 (9) ◽  
pp. e02774-17 ◽  
Author(s):  
Kieran James ◽  
Mary O'Connell Motherway ◽  
Christophe Penno ◽  
Rebecca Louise O'Brien ◽  
Douwe van Sinderen
Keyword(s):  
Transcriptional Regulation ◽  
Human Milk ◽  
Carbohydrate Metabolism ◽  
Metabolic Pathways ◽  
Transcriptional Regulators ◽  
Prototype Strain ◽  
Regulatory Mechanisms ◽  
Human Milk Oligosaccharides ◽  
Content Type ◽  
Bifidobacterium Breve

ABSTRACTBifidobacterial carbohydrate metabolism has been studied in considerable detail for a variety of both plant- and human-derived glycans, particularly involving the bifidobacterial prototype strainBifidobacterium breveUCC2003. We recently elucidated the metabolic pathways by which thehumanmilkoligosaccharide (HMO) constituents lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT) and lacto-N-biose (LNB) are utilized byB. breveUCC2003. However, to date, no work has been carried out on the regulatory mechanisms that control the expression of the genetic loci involved in these HMO metabolic pathways. In this study, we describe the characterization of three transcriptional regulators and the corresponding operator and associated (inducible) promoter sequences, with the latter governing the transcription of the genetic elements involved in LN(n)T/LNB metabolism. The activity of these regulators is dependent on the release of specific monosaccharides, which are believed to act as allosteric effectors and which are derived from the corresponding HMOs targeted by the particular locus.IMPORTANCEHuman milk oligosaccharides (HMOs) are a key factor in the development of the breastfed-infant microbiota. They function as prebiotics, selecting for a specific range of microbes, including a number of infant-associated species of bifidobacteria, which are thought to provide a range of health benefits to the infant host. While much research has been carried out on elucidating the mechanisms of HMO metabolism in infant-associated bifidobacteria, to date there is very little understanding of the transcriptional regulation of these pathways. This study reveals a multicomponent transcriptional regulation system that controls the recently identified pathways of HMO metabolism in the infant-associatedBifidobacterium breveprototype strain UCC2003. This not only provides insight into the regulatory mechanisms present in other infant-associated bifidobacteria but also provides an example of a network of sequential steps regulating microbial carbohydrate metabolism.

scholarly journals Variation in Consumption of Human Milk Oligosaccharides by Infant Gut-Associated Strains of Bifidobacterium breve

2013 ◽  
Vol 79 (19) ◽  
pp. 6040-6049 ◽  
Author(s):  
Santiago Ruiz-Moyano ◽  
Sarah M. Totten ◽  
Daniel A. Garrido ◽  
Jennifer T. Smilowitz ◽  
J. Bruce German ◽  
...  
Keyword(s):  
Human Milk ◽  
Intestinal Microbiota ◽  
Glycosyl Hydrolase ◽  
Bifidobacterium Longum ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides ◽  
Content Type ◽  
Bifidobacterium Breve ◽  
Infant Feces ◽  
Breast Fed

ABSTRACTHuman milk contains a high concentration of complex oligosaccharides that influence the composition of the intestinal microbiota in breast-fed infants. Previous studies have indicated that select species such asBifidobacterium longumsubsp.infantisandBifidobacterium bifidumcan utilize human milk oligosaccharides (HMO)in vitroas the sole carbon source, while the relatively fewB. longumsubsp.longumandBifidobacterium breveisolates tested appear less adapted to these substrates. Considering the high frequency at whichB. breveis isolated from breast-fed infant feces, we postulated that someB. brevestrains can more vigorously consume HMO and thus are enriched in the breast-fed infant gastrointestinal tract. To examine this, a number ofB. breveisolates from breast-fed infant feces were characterized for the presence of different glycosyl hydrolases that participate in HMO utilization, as well as by their ability to grow on HMO or specific HMO species such as lacto-N-tetraose (LNT) and fucosyllactose. AllB. brevestrains showed high levels of growth on LNT and lacto-N-neotetraose (LNnT), and, in general, growth on total HMO was moderate for most of the strains, with several strain differences. Growth and consumption of fucosylated HMO were strain dependent, mostly in isolates possessing a glycosyl hydrolase family 29 α-fucosidase. Glycoprofiling of the spent supernatant after HMO fermentation by select strains revealed that allB. brevestrains can utilize sialylated HMO to a certain extent, especially sialyl-lacto-N-tetraose. Interestingly, this specific oligosaccharide was depleted before neutral LNT by strain SC95. In aggregate, this work indicates that the HMO consumption phenotype inB. breveis variable; however, some strains display specific adaptations to these substrates, enabling more vigorous consumption of fucosylated and sialylated HMO. These results provide a rationale for the predominance of this species in breast-fed infant feces and contribute to a more accurate picture of the ecology of the developing infant intestinal microbiota.

scholarly journals Oligosaccharides Released from Milk Glycoproteins Are Selective Growth Substrates for Infant-Associated Bifidobacteria

2016 ◽  
Vol 82 (12) ◽  
pp. 3622-3630 ◽  
Author(s):  
Sercan Karav ◽  
Annabelle Le Parc ◽  
Juliana Maria Leite Nobrega de Moura Bell ◽  
Steven A. Frese ◽  
Nina Kirmiz ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Human Milk ◽  
Bifidobacterium Longum ◽  
Bovine Milk ◽  
Selective Growth ◽  
Whey Protein Concentrate ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides ◽  
Content Type ◽  
Growth Substrates

ABSTRACTMilk, in addition to nourishing the neonate, provides a range of complex glycans whose construction ensures a specific enrichment of key members of the gut microbiota in the nursing infant, a consortium known as the milk-oriented microbiome. Milk glycoproteins are thought to function similarly, as specific growth substrates for bifidobacteria common to the breast-fed infant gut. Recently, a cell wall-associated endo-β-N-acetylglucosaminidase (EndoBI-1) found in various infant-borne bifidobacteria was shown to remove a range of intactN-linked glycans. We hypothesized that these released oligosaccharide structures can serve as a sole source for the selective growth of bifidobacteria. We demonstrated that EndoBI-1 releasedN-glycans from concentrated bovine colostrum at the pilot scale. EndoBI-1-releasedN-glycans supported the rapid growth ofBifidobacterium longumsubsp.infantis(B. infantis), a species that grows well on human milk oligosaccharides, but did not support growth ofBifidobacterium animalissubsp.lactis(B. lactis), a species which does not. Conversely,B. infantisATCC 15697 did not grow on the deglycosylated milk protein fraction, clearly demonstrating that the glycan portion of milk glycoproteins provided the key substrate for growth. Mass spectrometry-based profiling revealed thatB. infantisconsumed 73% of neutral and 92% of sialylatedN-glycans, whileB. lactisdegraded only 11% of neutral and virtually no (<1%) sialylatedN-glycans. These results provide mechanistic support thatN-linked glycoproteins from milk serve as selective substrates for the enrichment of infant-associated bifidobacteria capable of carrying out the initial deglycosylation. Moreover, releasedN-glycans were better growth substrates than the intact milk glycoproteins, suggesting that EndoBI-1 cleavage is a key initial step in consumption of glycoproteins. Finally, the variety ofN-glycans released from bovine milk glycoproteins suggests that they may serve as novel prebiotic substrates with selective properties similar to those of human milk oligosaccharides.IMPORTANCEIt has been previously shown that glycoproteins serve as growth substrates for bifidobacteria. However, which part of a glycoprotein (glycans or polypeptides) is responsible for this function was not known. In this study, we used a novel enzyme to cleave conjugatedN-glycans from milk glycoproteins and tested their consumption by various bifidobacteria. The results showed that the glycans selectively stimulated the growth ofB. infantis, which is a key infant gut microbe. The selectivity of consumption of individualN-glycans was determined using advanced mass spectrometry (nano-liquid chromatography chip–quadrupole time of flight mass spectrometry [nano-LC-Chip-Q-TOF MS]) to reveal thatB. infantiscan consume the range of glycan structures released from whey protein concentrate.

scholarly journals Reconstruction of the Bifidobacterial Pan-Secretome Reveals the Network of Extracellular Interactions between Bifidobacteria and the Infant Gut

2018 ◽  
Vol 84 (16) ◽  
Author(s):  
Gabriele Andrea Lugli ◽  
Walter Mancino ◽  
Christian Milani ◽  
Sabrina Duranti ◽  
Francesca Turroni ◽  
...  
Keyword(s):  
Human Milk ◽  
Protein Translocation ◽  
Bifidobacterium Longum ◽  
Extracellular Proteins ◽  
Glycoside Hydrolases ◽  
Secreted Proteins ◽  
Human Milk Oligosaccharides ◽  
Secretion Systems ◽  
Milk Oligosaccharides ◽  
Content Type

ABSTRACT The repertoire of secreted proteins decoded by a microorganism represents proteins released from or associated with the cell surface. In gut commensals, such as bifidobacteria, these proteins are perceived to be functionally relevant, as they regulate the interaction with the gut environment. In the current study, we screened the predicted proteome of over 300 bifidobacterial strains among the currently recognized bifidobacterial species to generate a comprehensive database encompassing bifidobacterial extracellular proteins. A glycobiome analysis of this predicted bifidobacterial secretome revealed that a correlation exists between particular bifidobacterial species and their capability to hydrolyze human milk oligosaccharides (HMOs) and intestinal glycoconjugates, such as mucin. Furthermore, an exploration of metatranscriptomic data sets of the infant gut microbiota allowed the evaluation of the expression of bifidobacterial genes encoding extracellular proteins, represented by ABC transporter substrate-binding proteins and glycoside hydrolases enzymes involved in the degradation of human milk oligosaccharides and mucin. Overall, this study provides insights into how bifidobacteria interact with their natural yet highly complex environment, the infant gut.IMPORTANCE The ecological success of bifidobacteria relies on the activity of extracellular proteins that are involved in the metabolism of nutrients and the interaction with the environment. To date, information on secreted proteins encoded by bifidobacteria is incomplete and just related to few species. In this study, we reconstructed the bifidobacterial pan-secretome, revealing extracellular proteins that modulate the interaction of bifidobacteria with their natural environment. Furthermore, a survey of the secretion systems between bifidobacterial genomes allowed the identification of a conserved Sec-dependent secretion machinery in all the analyzed genomes and the Tat protein translocation system in the chromosomes of 23 strains belonging to Bifidobacterium longum subsp. longum and Bifidobacterium aesculapii.

scholarly journals Bifidobacterium longum subsp. infantis ATCC 15697 α-Fucosidases Are Active on Fucosylated Human Milk Oligosaccharides

2011 ◽  
Vol 78 (3) ◽  
pp. 795-803 ◽  
Author(s):  
David A. Sela ◽  
Daniel Garrido ◽  
Larry Lerno ◽  
Shuai Wu ◽  
Kemin Tan ◽  
...  
Keyword(s):  
Human Milk ◽  
Turnover Rate ◽  
Bifidobacterium Longum ◽  
Small Mass ◽  
Human Milk Oligosaccharides ◽  
High Turnover ◽  
Milk Oligosaccharides ◽  
Content Type ◽  
Biochemical Kinetics ◽  
Time Dependent Effect

ABSTRACTBifidobacterium longumsubsp.infantisATCC 15697 utilizes several small-mass neutral human milk oligosaccharides (HMOs), several of which are fucosylated. Whereas previous studies focused on endpoint consumption, a temporal glycan consumption profile revealed a time-dependent effect. Specifically, among preferred HMOs, tetraose was favored early in fermentation, with other oligosaccharides consumed slightly later. In order to utilize fucosylated oligosaccharides, ATCC 15697 possesses several fucosidases, implicating GH29 and GH95 α-l-fucosidases in a gene cluster dedicated to HMO metabolism. Evaluation of the biochemical kinetics demonstrated that ATCC 15697 expresses three fucosidases with a high turnover rate. Moreover, several ATCC 15697 fucosidases are active on the linkages inherent to the HMO molecule. Finally, the HMO cluster GH29 α-l-fucosidase possesses a crystal structure that is similar to previously characterized fucosidases.

scholarly journals A Solution to Antifolate Resistance in Group B Streptococcus: Untargeted Metabolomics Identifies Human Milk Oligosaccharide-Induced Perturbations That Result in Potentiation of Trimethoprim

mBio ◽  
2020 ◽  
Vol 11 (2) ◽  
Author(s):  
Schuyler A. Chambers ◽  
Rebecca E. Moore ◽  
Kelly M. Craft ◽  
Harrison C. Thomas ◽  
Rishub Das ◽  
...  
Keyword(s):  
Human Milk ◽  
Metabolic Response ◽  
Group B Streptococcus ◽  
Resistance Mechanisms ◽  
Human Milk Oligosaccharides ◽  
Intrinsic Resistance ◽  
Milk Oligosaccharides ◽  
Content Type ◽  
Ultrahigh Performance Liquid Chromatography ◽  
Group B

ABSTRACT Adjuvants can be used to potentiate the function of antibiotics whose efficacy has been reduced by acquired or intrinsic resistance. In the present study, we discovered that human milk oligosaccharides (HMOs) sensitize strains of group B Streptococcus (GBS) to trimethoprim (TMP), an antibiotic to which GBS is intrinsically resistant. Reductions in the MIC of TMP reached as high as 512-fold across a diverse panel of isolates. To better understand HMOs’ mechanism of action, we characterized the metabolic response of GBS to HMO treatment using ultrahigh-performance liquid chromatography–high-resolution tandem mass spectrometry (UPLC-HRMS/MS) analysis. These data showed that when challenged by HMOs, GBS undergoes significant perturbations in metabolic pathways related to the biosynthesis and incorporation of macromolecules involved in membrane construction. This study represents reports the metabolic characterization of a cell that is perturbed by HMOs. IMPORTANCE Group B Streptococcus is an important human pathogen that causes serious infections during pregnancy which can lead to chorioamnionitis, funisitis, premature rupture of gestational membranes, preterm birth, neonatal sepsis, and death. GBS is evolving antimicrobial resistance mechanisms, and the work presented in this paper provides evidence that prebiotics such as human milk oligosaccharides can act as adjuvants to restore the utility of antibiotics.

scholarly journals Ability of Bifidobacterium breve To Grow on Different Types of Milk: Exploring the Metabolism of Milk through Genome Analysis

2011 ◽  
Vol 77 (20) ◽  
pp. 7408-7417 ◽  
Author(s):  
Francesca Turroni ◽  
Elena Foroni ◽  
Fausta Serafini ◽  
Alice Viappiani ◽  
Barbara Montanini ◽  
...  
Keyword(s):  
Human Milk ◽  
Genome Analysis ◽  
Content Type ◽  
Bifidobacterium Breve ◽  
Milk Samples ◽  
Representative Isolate ◽  
Different Types

ABSTRACTWe have investigated the occurrence of bifidobacteria in human milk samples, and we provide evidence regarding the predominance of members of theBifidobacterium brevespecies in this environment. Moreover, evaluation of the growth capabilities and transcriptomic analyses of one representative isolate of this species, i.e.,B. breve4L, on different milk types were performed.

scholarly journals Human Milk Oligosaccharides Promote the Growth of Staphylococci

2012 ◽  
Vol 78 (14) ◽  
pp. 4763-4770 ◽  
Author(s):  
K. M. Hunt ◽  
J. Preuss ◽  
C. Nissan ◽  
C. A. Davlin ◽  
J. E. Williams ◽  
...  
Keyword(s):  
Human Milk ◽  
Bacterial Communities ◽  
Bacterial Species ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides ◽  
Content Type ◽  
Nutritional Conditions ◽  
Culture Independent ◽  
Lactating Mammary Gland

ABSTRACTHuman milk oligosaccharides (HMO), which constitute a major component of human milk, promote the growth of particular bacterial species in the infant's gastrointestinal tract. We hypothesized that HMO also interact with the bacterial communities present in human milk. To test this hypothesis, two experiments were conducted. First, milk samples were collected from healthy women (n= 16); culture-independent analysis of the bacterial communities was performed, HMO content was analyzed, and the relation between these factors was investigated. A positive correlation was observed between the relative abundance ofStaphylococcusand total HMO content (r= 0.66). In a follow-up study, we conducted a series ofin vitrogrowth curve experiments utilizingStaphylococcus aureusorStaphylococcus epidermidisand HMO isolated from human milk. HMO exhibited stimulatory effects on bacterial growth under various nutritional conditions. Analysis of culture supernatants from these experiments revealed that HMO did not measurably disappear from the culture medium, indicating that the growth-enhancing effects were not a result of bacterial metabolism of the HMO. Instead, stimulation of growth caused greater utilization of amino acids in minimal medium. Collectively, the data provide evidence that HMO may promote the growth ofStaphylococcusspecies in the lactating mammary gland.

scholarly journals Metabolite Changes Signal Genetic Regulatory Mechanisms for Robust Cell Behavior

mBio ◽  
2014 ◽  
Vol 5 (1) ◽  
Author(s):  
Sang Jun Lee ◽  
Andrei Trostel ◽  
Sankar Adhya
Keyword(s):  
Metabolic Pathways ◽  
Regulatory Networks ◽  
Critical Role ◽  
Genetic Regulatory Networks ◽  
Regulatory Mechanisms ◽  
Ripple Effect ◽  
Content Type ◽  
Metabolite Pool ◽  
Gene Regulatory ◽  
Transcript Profiles

ABSTRACTExploiting mechanisms of utilizing the sugard-galactose inEscherichia colias a model system, we explored the consequences of accumulation of critical intermediates of thed-galactose metabolic pathways by monitoring cell growth, metabolites, and transcript profiles. These studies revealed both metabolic network changes far from thed-galactose pathway and changes in the global gene regulatory network. The concentration change of a critical intermediate disturbs the equilibrium state, generating a ripple effect through several metabolic pathways that ends up signaling up- or downregulation of specific sets of genes in a programmed manner to cope with the imbalance. Such long-range effects on metabolites and genetic regulatory mechanisms not only may be a common feature in bacteria but very likely operate during cellular development and differentiation in higher organisms as well as in disease cells, like cancer cells.IMPORTANCEMetabolite accumulation can create adverse intracellular conditions that are relieved by compensatory immediate changes of metabolite pools and later changes of transcript levels. It has been known that gene expression is normally regulated by added catabolic substrates (induction) or anabolic end products (repression). It is becoming apparent now that change in the concentration of metabolic intermediates also plays a critical role in genetic regulatory networks for metabolic homeostasis. Our study provides new insight into how metabolite pool changes transduce signals to global gene regulatory networks.

scholarly journals Metabolism of Sialic Acid by Bifidobacterium breve UCC2003

2014 ◽  
Vol 80 (14) ◽  
pp. 4414-4426 ◽  
Author(s):  
Muireann Egan ◽  
Mary O'Connell Motherway ◽  
Marco Ventura ◽  
Douwe van Sinderen
Keyword(s):  
Sialic Acid ◽  
Human Milk ◽  
Bifidobacterium Bifidum ◽  
Commensal Bacteria ◽  
Functional Genomic ◽  
Specific Group ◽  
Content Type ◽  
Bifidobacterium Breve ◽  
Human Milk Oligosaccharide ◽  
Uptake And Metabolism

ABSTRACTBifidobacteria constitute a specific group of commensal bacteria that inhabit the gastrointestinal tracts of humans and other mammals.Bifidobacterium breveUCC2003 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 thatB. breveUCC2003 can cross feed on sialic acid derived from the metabolism of 3′-sialyllactose, an abundant HMO, by another infant gut bifidobacterial strain,Bifidobacterium bifidumPRL2010.

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