Lactobacillus casei Ferments the
            <i>N</i>
            -Acetylglucosamine Moiety of Fucosyl-α-1,3-
            <i>N</i>
            -Acetylglucosamine and Excretes
            <scp>l</scp>
            -Fucose

Rodrı́guez-Dı́az, Jesús; Rubio‐del‐Campo, Antonio; Yebra, Marı́a J.

doi:10.1128/aem.00474-12

Cited by 41 publications

(42 citation statements)

References 48 publications

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“…A previous work has shown that Lactobacillus casei BL23 is able to utilize the fucosyl-disaccharide fucosyl-␣1,3-N-acetylglucosamine (Fuc-␣1,3-GlcNAc), which is transported by a specific permease of the PTS class and split into L-fucose and N-acetylglucosamine by the ␣-L-fucosidase AlfB (29). However, this strain is not able to use L-fucose and only the N-acetylglucosamine moiety of Fuc-␣1,3-GlcNAc is catabolized, while L-fucose is quantitatively expelled to the growth medium.…”

Section: Resultsmentioning

confidence: 99%

An l -Fucose Operon in the Probiotic Lactobacillus rhamnosus GG Is Involved in Adaptation to Gastrointestinal Conditions

Becerra

Yebra

Monedero

2015

Appl Environ Microbiol

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L-Fucose is a sugar present in human secretions as part of human milk oligosaccharides, mucins, and other glycoconjugates in the intestinal epithelium. The genome of the probiotic Lactobacillus rhamnosus GG (LGG) carries a gene cluster encoding a putative L-fucose permease (fucP), L-fucose catabolic pathway (fucI, fucK, fucU, and fucA), and a transcriptional regulator (fucR). The metabolism of L-fucose in LGG results in 1,2-propanediol production, and their fucI and fucP mutants displayed a severe and mild growth defect on L-fucose, respectively. Transcriptional analysis revealed that the fuc genes are induced by L-fucose and subject to a strong carbon catabolite repression effect. This induction was triggered by FucR, which acted as a transcriptional activator necessary for growth on L-fucose. LGG utilized fucosyl-␣1,3-N-acetylglucosamine and contrarily to other lactobacilli, the presence of fuc genes allowed this strain to use the L-fucose moiety. In fucI and fucR mutants, but not in fucP mutant, L-fucose was not metabolized and it was excreted to the medium during growth on fucosyl-␣1,3-N-acetylglucosamine. The fuc genes were induced by this fucosyl-disaccharide in the wild type and the fucP mutant but not in a fucI mutant, showing that FucP does not participate in the regulation of fuc genes and that L-fucose metabolism is needed for FucR activation. The L-fucose operon characterized here constitutes a new example of the many factors found in LGG that allow this strain to adapt to the gastrointestinal conditions. O ne of the few hexoses with "L" configuration found in nature is L-fucose (6-deoxy-L-galactose). It forms part of many glycans present at the surface of eukaryotic cells such as H and Lewis antigens, which are present not only in blood cells but also in epithelial cells at different mucosal sites (1). It is also present at the highly glycosylated mucin proteins of the intestinal mucosa and in a high proportion of the oligosaccharides present in human milk (2, 3). These facts make L-fucose an important sugar in the microbial ecology of the gastrointestinal tract. The importance of the fucosylation of mucosal glycoconjugates on the intestinal ecology is reflected by the fact that the intestinal microbiota composition is dependent on the secretor status of the individuals, which is defined by mutations in the FUT2 gene coding for an ␣(1,2)-fucosyltransferase that participates in the fucosylation of mucosal glycans (4, 5). Owing to its elevated concentration found in the intestinal niche, L-fucose can be used as a carbon source and its utilization has been identified as a key factor for intestinal colonization in some bacteria. Thus, the pathogen Campylobacter jejuni, primarily thought to be an asaccharolytic microorganism, was able to use L-fucose, and this provides it with a competitive advantage, as determined in a neonatal piglet infection model (6). Also, a ⌬fucAO mutant of the probiotic Escherichia coli Nissle 1917 strain, unable to use L-fucose, showed 2 orders of magnitude lower colonization le...

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Section: Resultsmentioning

confidence: 99%

An l -Fucose Operon in the Probiotic Lactobacillus rhamnosus GG Is Involved in Adaptation to Gastrointestinal Conditions

Becerra

Yebra

Monedero

2015

Appl Environ Microbiol

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“…In the following years, the proteins forming the glucose-and mannose-specific PTSs in E. coli (202,203) and the lactose-specific PTS in S. aureus (204) were identified, and their role in transport and phosphorylation of the two hexoses and the disaccharide was established. Since then, a huge number of PTSs transporting a large variety of substrates, including hexoses, 6-deoxy-hexoses (14), amino sugars, N-acetyl-amino sugars, gluconic acids (205), pentitols (206,207), ascorbate (208), and disaccharides, have been identified. We recently obtained evidence that the E. faecalis maltose-specific EIICBA Mal (MalT) (15) also transports and phosphorylates the trisaccharide maltotriose and the tetrasaccharide maltotetraose (J. Deutscher, A. Hartke, J. Thompson, C. Magni, C. Henry, V. Blancato, G. Repizo, N. Sauvageot, A. Pikis, T. Kentache, and A. Mokhtari, unpublished results).…”

Section: Discussionmentioning

confidence: 99%

“…In the last step, PϳEIIB transfers its phosphoryl group to a carbohydrate molecule bound to the cognate EIIC. There is so far only one carbohydrate, fucosyl-␣-1,3-N-acetylglucosamine, which is transported by a PTS (that of Lactobacillus casei) without being phosphorylated (14). All other carbohydrates are phosphorylated during their transport and subsequently converted into phosphorylated intermediates of either the Embden-MeyerhofParnas, pentose phosphate, or Entner-Doudoroff pathway.…”

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confidence: 99%

The Bacterial Phosphoenolpyruvate:Carbohydrate Phosphotransferase System: Regulation by Protein Phosphorylation and Phosphorylation-Dependent Protein-Protein Interactions

Deutscher

Aké

Derkaoui

et al. 2014

Microbiol Mol Biol Rev

352

374

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SUMMARY The bacterial phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS) carries out both catalytic and regulatory functions. It catalyzes the transport and phosphorylation of a variety of sugars and sugar derivatives but also carries out numerous regulatory functions related to carbon, nitrogen, and phosphate metabolism, to chemotaxis, to potassium transport, and to the virulence of certain pathogens. For these different regulatory processes, the signal is provided by the phosphorylation state of the PTS components, which varies according to the availability of PTS substrates and the metabolic state of the cell. PEP acts as phosphoryl donor for enzyme I (EI), which, together with HPr and one of several EIIA and EIIB pairs, forms a phosphorylation cascade which allows phosphorylation of the cognate carbohydrate bound to the membrane-spanning EIIC. HPr of firmicutes and numerous proteobacteria is also phosphorylated in an ATP-dependent reaction catalyzed by the bifunctional HPr kinase/phosphorylase. PTS-mediated regulatory mechanisms are based either on direct phosphorylation of the target protein or on phosphorylation-dependent interactions. For regulation by PTS-mediated phosphorylation, the target proteins either acquired a PTS domain by fusing it to their N or C termini or integrated a specific, conserved PTS regulation domain (PRD) or, alternatively, developed their own specific sites for PTS-mediated phosphorylation. Protein-protein interactions can occur with either phosphorylated or unphosphorylated PTS components and can either stimulate or inhibit the function of the target proteins. This large variety of signal transduction mechanisms allows the PTS to regulate numerous proteins and to form a vast regulatory network responding to the phosphorylation state of various PTS components.

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“…We have recently reported that probiotic bacterium L. casei strain BL23 can use Fuc-␣-1,3-GlcNAc as a carbon source and that AlfB is necessary for this. However, Fuc-␣-1,6-GlcNAc cannot be metabolized by L. casei (25). This suggests that, in spite of the high specific activity of AlfC on Fuc-␣-1,6-GlcNAc (13.6 mol fucose liberated/min · mg protein, compared to 27.5 mol fucose liberated from Fuc-␣-1,3-GlcNAc/min · mg protein for AlfB [7]), this disaccharide is not the natural substrate of AlfC.…”

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confidence: 99%

Synthesis of Fucosyl- N -Acetylglucosamine Disaccharides by Transfucosylation Using α-l-Fucosidases from Lactobacillus casei

Rodrı́guez-Dı́az

Carbajo²,

Pineda‐Lucena³

et al. 2013

Appl Environ Microbiol

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bAlfB and AlfC ␣-L-fucosidases from Lactobacillus casei were used in transglycosylation reactions, and they showed high efficiency in synthesizing fucosyldisaccharides. AlfB and AlfC activities exclusively produced fucosyl-␣-1,3-N-acetylglucosamine and fucosyl-␣-1,6-N-acetylglucosamine, respectively. The reaction kinetics showed that AlfB can convert 23% p-nitrophenyl-␣-L-fucopyranoside into fucosyl-␣-1,3-N-acetylglucosamine and AlfC at up to 56% into fucosyl-␣-1,6-N-acetylglucosamine. Fucosyl-oligosaccharides (FUS) are coming to be of great interest due to their presence in human milk and their antiadhesive activity against pathogens. This last property is characterized by blocking the attachment of intestinal pathogens to host cells due to FUS structural similarity with cell-surface glycoconjugate receptors (1-4). In addition, genes encoding ␣-L-fucosidases (EC 3.2.1.51) have recently been found in genomes of bifidobacteria (5, 6) and lactobacilli (7), which raises issues regarding the role of these enzymes in the metabolism of fucose-containing structures and opens up the possibility of using FUS as novel prebiotics (8).To fully establish the biological function of FUS as antiadhesins and prebiotics, extensive studies are required, for which synthesis of sufficient amounts would be necessary. Chemical synthesis of FUS such as ABH and Lewis blood group antigens has long been performed, but it is a tedious and expensive process since it requires multiple protection and deprotection steps to achieve the desired selectivity (9, 10). The enzymatic synthesis of FUS is an alternative to chemical methods, and it offers the advantage of forming specific glycosidic linkages in the presence of other reactive functional groups. There are two types of enzymes that can carry out fucosylation: fucosyltransferases and fucosidases. Fucosyltransferases present high specificity toward the acceptor and do not hydrolyze the product (11, 12). However, those enzymes are generally difficult to express and they require an expensive sugar nucleotide or a complex multienzymatic system for the regeneration (11, 13). In contrast, oligosaccharides have been largely produced using the transglycosylation activity of glycosidases and engineered glycosynthases (14,15). Only a few works describe the synthesis of FUS by transglycosylation using ␣-L-fucosidases; however, they cover a wide range of sources such as mammalian tissues (16), fungi (17), and bacteria (18,19). The transglycosylation activity of ␣-L-fucosidases is generally moderate compared with the hydrolysis activity, although it is variable and depends on the origin of the enzymes (20).We have previously isolated three ␣-L-fucosidases (AlfA, AlfB, and AlfC) from Lactobacillus casei that cleaved ␣-linked L-fucose from natural FUS (7). AlfB and AlfC showed high specific activity on fucosyl-␣-1,3-N-acetylglucosamine (Fuc-␣-1,3-GlcNAc) and fucosyl-␣-1,6-N-acetylglucosamine (Fuc-␣-1,6-GlcNAc), respectively. These substrate specificities led us in the present work to explore the likely us...

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Lactobacillus casei Ferments the N -Acetylglucosamine Moiety of Fucosyl-α-1,3- N -Acetylglucosamine and Excretes l -Fucose

Abstract: ABSTRACTWe have previously characterized fromLactobacillus caseiBL23 three α-l-fucosidases, AlfA, AlfB, and AlfC, which hydrolyzein vitronatural fucosyl-oligosaccharides. In this work, we have shown thatL. caseiis able to grow in the presence of fucosyl-α-1,3-N-acetylg… Show more

Cited by 41 publications

References 48 publications

An l -Fucose Operon in the Probiotic Lactobacillus rhamnosus GG Is Involved in Adaptation to Gastrointestinal Conditions

An l -Fucose Operon in the Probiotic Lactobacillus rhamnosus GG Is Involved in Adaptation to Gastrointestinal Conditions

The Bacterial Phosphoenolpyruvate:Carbohydrate Phosphotransferase System: Regulation by Protein Phosphorylation and Phosphorylation-Dependent Protein-Protein Interactions

Synthesis of Fucosyl- N -Acetylglucosamine Disaccharides by Transfucosylation Using α-l-Fucosidases from Lactobacillus casei

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