Structural analysis of the α-d-glucan (EPS35-5) produced by the Lactobacillus reuteri strain 35-5 glucansucrase GTFA enzyme

Leeuwen, Sander S. van; Kralj, Slavko; Geel-Schutten, Ineke H. van; Gerwig, Gerrit J.; Dijkhuizen, Lubbert; Kamerling, Johannis P.

doi:10.1016/j.carres.2008.01.044

Cited by 63 publications

(112 citation statements)

References 16 publications

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“…The 10,11,22 This is further corroborated by the R H-2 signal at δ 3.258, indicating that residue R is not 3-or 4-substituted. 22 The split B H-1 signal at δ 5.353/5.343 fits best with the occurrence of an (-)R-D-Glcp-(1f3)-unit.…”

Section: Fraction 8 Maldi-tof-ms Analysis Of Fraction 8 Showed [M + Na]mentioning

confidence: 78%

“…This observation adds a new structural-reporter-group signal to the concept. 10,11,22 These data lead to a structural element F1f4B1f3E1f6R. The final residue D has to be positioned to cause a branched structure, which can only occur at residue E, resulting in an…”

Section: Fraction 11 Maldi-tof-ms Analysis Of Fraction 11 Showed [M mentioning

confidence: 99%

“…Structural analysis of the homopolysaccharides produced by the glucansucrases GTF180 and GTFA revealed highly complex structures, and composite models have been proposed. 10,11 Previous studies on the site-directed mutagenesis near the catalytic Asp1133 (putative transition state stabilizing residue) of GTFA have shown that specific amino acid mutations in this glucansucrase gave rise to large changes in the EPS structure and its properties. 12 Similar modifications in the homologous region of the gtf180 gene have been performed (unpublished data).…”

Section: Introductionmentioning

confidence: 99%

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Structural Analysis of Bioengineered α-d-Glucan Produced by a Triple Mutant of the Glucansucrase GTF180 Enzyme from Lactobacillus reuteri Strain 180: Generation of (α1→4) Linkages in a Native (1→3)(1→6)-α-d-Glucan

et al. 2008

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Site-directed mutagenesis of the glucansucrase gtf180 gene from Lactobacillus reuteri strain 180 was used to transform the active site region. The R-D-glucan (mEPS-PNNS) produced by the triple mutant V1027P:S1137N: A1139S differed in structure from that of the wild-type R-D-glucan (EPS180). Besides (R1f3) and (R1f6) linkages, as present in EPS180, mEPS-PNNS also contained (R1f4) linkages. Linkage analysis, periodate oxidation, and 1D/2D 1 H NMR spectroscopy of the intact mEPS-PNNS, as well as MS and NMR analysis of oligosaccharides obtained by partial acid hydrolysis of mEPS-PNNS afforded a composite model, which includes all identified structural features.

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Section: Fraction 8 Maldi-tof-ms Analysis Of Fraction 8 Showed [M + Na]mentioning

confidence: 78%

Section: Fraction 11 Maldi-tof-ms Analysis Of Fraction 11 Showed [M mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Structural Analysis of Bioengineered α-d-Glucan Produced by a Triple Mutant of the Glucansucrase GTF180 Enzyme from Lactobacillus reuteri Strain 180: Generation of (α1→4) Linkages in a Native (1→3)(1→6)-α-d-Glucan

et al. 2008

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“…Nuclear magnetic resonance (NMR) was carried out as described previously (25). Samples were exchanged twice with 200 l of 99.9 atom% D 2 O (Cambridge Isotope Laboratories Ltd., United Kingdom) with intermediate lyophilization and finally dissolved in 650 l of 99.9 atom% D 2 O containing 25 ppm acetone as an internal standard (␦1H 2.225).…”

Section: Methodsmentioning

confidence: 99%

Differential Metabolism of Exopolysaccharides from Probiotic Lactobacilli by the Human Gut Symbiont Bacteroides thetaiotaomicron

Bueren

Saraf

Martens

et al. 2015

Appl Environ Microbiol

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bProbiotic microorganisms are ingested as food or supplements and impart positive health benefits to consumers. Previous studies have indicated that probiotics transiently reside in the gastrointestinal tract and, in addition to modulating commensal species diversity, increase the expression of genes for carbohydrate metabolism in resident commensal bacterial species. In this study, it is demonstrated that the human gut commensal species Bacteroides thetaiotaomicron efficiently metabolizes fructan exopolysaccharide (EPS) synthesized by probiotic Lactobacillus reuteri strain 121 while only partially degrading reuteran and isomalto/malto-polysaccharide (IMMP) ␣-glucan EPS polymers. B. thetaiotaomicron metabolized these EPS molecules via the activation of enzymes and transport systems encoded by dedicated polysaccharide utilization loci specific for ␤-fructans and ␣-glucans. Reduced metabolism of reuteran and IMMP ␣-glucan EPS molecules may be due to reduced substrate binding by components of the starch utilization system (sus). This study reveals that microbial EPS substrates activate genes for carbohydrate metabolism in B. thetaiotaomicron and suggests that microbially derived carbohydrates provide a carbohydrate-rich reservoir for B. thetaiotaomicron nutrient acquisition in the gastrointestinal tract.T rillions of bacteria inhabit the gut, imparting symbiotic effects that benefit the overall health and well-being of individuals (1). While the human genome has approximately 30,000 genes, the human microbiome contributes an additional 3 to 9 million gene products (termed the gut metagenome) that contribute to functionalities in human lifestyle (2). One prominent example is the human utilization of carbohydrates. Carbohydrates provide a substantial proportion of the daily energy intake of an individual. However, humans have the ability to metabolize only very few dietary carbohydrate compounds, such as lactose, starch, and sucrose, by enzymes either present in saliva (amylases) or anchored to the epithelial wall of the small intestine with glycosylphosphatidylinositol (GPI) anchors (such as invertases and lactases). All other carbohydrates that traverse the gastrointestinal (GI) tract have the potential to be metabolized by commensal gut bacteria, where it is estimated that gut microbiota provide an additional 30,000 enzymes which facilitate the breakdown of carbohydrates in the gut (3).Approximately 30% of resident human gut microbiota are from the Bacteriodetes phylum. These Gram-negative, obligate anaerobes are well armed with a repertoire of carbohydrate-degrading enzymes for harvesting carbohydrate nutritional resources. One species, Bacteroides thetaiotaomicron, devotes approximately 18% of its genome to carbohydrate foraging (4). Discrete polysaccharide utilization loci (PULs) within its genome are upregulated in response to carbohydrates dependent on the source. Most carbohydrate sources for B. thetaiotaomicron nutrient acquisition originate from dietary sources (such as nondigestible plant polysaccharides)...

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“…In the present study, glucans from different strains of Lactobacillus reuteri (ATCC-55730, 180, ML1 and 121) of varying molecular weight, linkage types, and degree of branching 7,[21][22][23][24] were analysed for binding to native β-lactoglobulin, denatured β-lactoglobulin and κ-casein in the range pH 4.0-5.5 by using SPR. The extent of individual interactions was correlated with the molecular properties of the glucans.…”

Section: Introductionmentioning

confidence: 99%

Binding Interactions Between α-glucans from Lactobacillus reuteri and Milk Proteins Characterised by Surface Plasmon Resonance

et al. 2012

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Structural analysis of the α-d-glucan (EPS35-5) produced by the Lactobacillus reuteri strain 35-5 glucansucrase GTFA enzyme

Cited by 63 publications

References 16 publications

Structural Analysis of Bioengineered α-d-Glucan Produced by a Triple Mutant of the Glucansucrase GTF180 Enzyme from Lactobacillus reuteri Strain 180: Generation of (α1→4) Linkages in a Native (1→3)(1→6)-α-d-Glucan

Structural Analysis of Bioengineered α-d-Glucan Produced by a Triple Mutant of the Glucansucrase GTF180 Enzyme from Lactobacillus reuteri Strain 180: Generation of (α1→4) Linkages in a Native (1→3)(1→6)-α-d-Glucan

Differential Metabolism of Exopolysaccharides from Probiotic Lactobacilli by the Human Gut Symbiont Bacteroides thetaiotaomicron

Binding Interactions Between α-glucans from Lactobacillus reuteri and Milk Proteins Characterised by Surface Plasmon Resonance

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