Biochemical and crystallographic characterization of a glucansucrase from<i>Lactobacillus reuteri</i>180

Pijning, Tjaard; Vujicic-Zagar, A.; Kralj, Slavko; Eeuwema, Wieger; Dijkhuizen, Lubbert; Dijkstra, Bauke W.

doi:10.1080/10242420701789163

Cited by 34 publications

(29 citation statements)

References 20 publications

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“…For example, small positional differences of domain V were seen in the elongated glucansucrase structures of GTF180-DN I [9] and GTFA-DN [10]. A much larger positional difference for domain V was observed in DN 123 -GBD-CD2, yielding a much more compact structure [12].…”

Section: Discussionmentioning

confidence: 94%

“…In GTF180-DN II, as opposed to the GTF180-DN I structure, most of the C-terminal residues are visible in the electron density map. These residues (1753-1771) form a long extended loop protruding away from the core of domain V; residues 1755-1759 form a short 3 10 helix. The C-terminal~10 residues are near the active site of the enzyme in domain A, although they make only a few direct hydrogen-bonding interactions.…”

Section: Crystal Structure Of Gtf180-dn IImentioning

confidence: 99%

“…Previously, we reported the crystal structures of N-terminally truncated forms of Lactobacillus reuteri 180 glucansucrase GTF180 (which will be referred to as GTF180-DN I) [8,9] and Lactobacillus reuteri 121 GTFA (referred to as GTFA-DN) [10]. In addition, crystal structures have been reported for Streptococcus mutans GTF-SI [11] and Leuconostoc mesenteroides NRRL B-1299 DN 123 -GBD-CD2 from DSR-E [12].…”

Section: Introductionmentioning

confidence: 99%

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Flexibility of truncated and full‐length glucansucrase GTF180 enzymes from Lactobacillus reuteri 180

et al. 2014

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Glucansucrase enzymes synthesize high-molecular-mass extracellular aglucan polysaccharides from sucrose. Previously, the crystal structure of truncated glucansucrase glucosyltransferase (GTF)180-DN from Lactobacillus reuteri 180 (lacking the N-terminal domain) revealed an elongated overall structure with two remote domains (IV and V) extending away from the core. By contrast, a new crystal form of the a-1,6/a-1,3 specific glucansucrase GTF180-DN shows an approximate 120 o rotation of domain V about a hinge located between domains IV and V, giving a much more compact structure than before. Positional variability of domain V in solution is confirmed by small angle X-ray scattering experiments and rigid-body ensemble calculations. In addition, small angle Xray scattering measurements of full-length GTF180 also provide the first structural data for a full-length glucansucrase, showing that the enzyme has an almost symmetric boomerang-like molecular shape, with a bend likely located between domains IV and V. The~700-residue N-terminal domain, which is not present in the crystal structures, extends away from domain V and the catalytic core of the enzyme. We conclude that, as a result of the hinge region, in solution, GTF180-DN (and likely also the full-length GTF180) shows conformational flexibility; this may be a general feature of GH70 glucansucrases.

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Section: Discussionmentioning

confidence: 94%

Section: Crystal Structure Of Gtf180-dn IImentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Flexibility of truncated and full‐length glucansucrase GTF180 enzymes from Lactobacillus reuteri 180

et al. 2014

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“…For cloning and mutagenesis of gtf180 see SI text. Purification and crystallization of native L. reuteri GTF180-ΔN were performed as described previously (17). Selenomethionine (SeMet) labeled GTF180-ΔN was produced as formerly reported (36).…”

Section: Methodsmentioning

confidence: 99%

“…Here we describe the crystal structure of a fully active GH70 glucansucrase of Lactobacillus reuteri 180, encompassing the catalytic and C-terminal domains, but not its ∼740 residues N-terminal domain (17). This 117 kDa GTF180-ΔN (residues 742-1,772) produces a high molecular mass (∼36 MDa) glucose polymer, with both αð1 → 6Þ and αð1 → 3Þ glucosidic linkages (18).…”

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confidence: 99%

Crystal structure of a 117 kDa glucansucrase fragment provides insight into evolution and product specificity of GH70 enzymes

Vujicic-Zagar

Pijning

Kralj

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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Glucansucrases are large enzymes belonging to glycoside hydrolase family 70, which catalyze the cleavage of sucrose into fructose and glucose, with the concomitant transfer of the glucose residue to a growing α-glucan polymer. Among others, plaque-forming oral bacteria secrete these enzymes to produce α-glucans, which facilitate the adhesion of the bacteria to the tooth enamel. We determined the crystal structure of a fully active, 1,031-residue fragment encompassing the catalytic and C-terminal domains of GTF180 from Lactobacillus reuteri 180, both in the native state, and in complexes with sucrose and maltose. These structures show that the enzyme has an α-amylase-like ðβ∕αÞ 8 -barrel catalytic domain that is circularly permuted compared to the catalytic domains of members of glycoside hydrolase families 13 and 77, which belong to the same GH-H superfamily. In contrast to previous suggestions, the enzyme has only one active site and one nucleophilic residue. Surprisingly, in GTF180 the peptide chain follows a "U"-path, such that four of the five domains are made up from discontiguous N-and C-terminal stretches of the peptide chain. Finally, the structures give insight into the factors that determine the different linkage types in the polymeric product.crystal structure complexes | exopolysaccharide | Lactobacillus reuteri | dental caries

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Microbial and Enzymatic Polysaccharides

Kralj

Khandige

Guan

et al. 2020

Kirk-Othmer Encyclopedia of Chemical Technology

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Polysaccharides are ubiquitous and the largest substantive material class in nature (eg, structural in plants (cellulose, starch), algae (alginate), and (microbes) and throughout history polysaccharides have found extensive use in many industrial applications, for example in packaging (paper and board), fiber and textiles (eg, cotton and viscose), adhesives, and even in early engineered thermoplastic materials (eg, cellulose esters). Sourced from nature and in general biocompatible, many polysaccharides have applications across various food and food additive applications. The broad functionality and general good compatibility are attractive properties increasingly desired as focus on sustainability is increasing. This article will discuss the established product range of microbial polysaccharides utilized across a range of specialty applications such as food, personal care, and industrials. Product categories such as alginates, bacterial cellulose, curdlan, dextran, diutan, emulsan, fructan, gellan, pullulan, sceroglucan, succinoglycan, welan, and xanthan gum will be reviewed. In addition, the emerging platform technology enzymatic polymerization is discussed with special focus on the transformational promise for this approach toward engineered polysaccharide structures. This approach allows for the rational and systematic design of polysaccharide materials controlling aspects such as molecular weight, branching, and particle morphology.

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Biochemical and crystallographic characterization of a glucansucrase fromLactobacillus reuteri180

Cited by 34 publications

References 20 publications

Flexibility of truncated and full‐length glucansucrase GTF180 enzymes from Lactobacillus reuteri 180

Flexibility of truncated and full‐length glucansucrase GTF180 enzymes from Lactobacillus reuteri 180

Crystal structure of a 117 kDa glucansucrase fragment provides insight into evolution and product specificity of GH70 enzymes

Microbial and Enzymatic Polysaccharides

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