Crystal structure of the catalytic domain of a bacterial cellulase belonging to family 5

Ducros, V.M.-A.; Czjzek, Mirjam; Belaı̈ch, Anne; Gaudin, Christian; Fierobe, Henri‐Pierre; Bélaı̈ch, Jean-Pierre; Davies, G.J.; Haser, R.

doi:10.1016/s0969-2126(01)00228-3

Cited by 151 publications

(103 citation statements)

References 51 publications

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“…We therefore propose that the aromatic clamp phenylalanines not only participate in substrate recognition but also partially control the reaction specificity. An aromatic clamp with similar functions has been described in GH5 family enzymes such as the fungal exoglucanase subfamily (53) and the endocellulase CelCCA (54). The clamp is also suggested to be involved in both entry and release to/from the active site and therefore in isomerase versus hydrolytic activity of the enzyme.…”

Section: Discussionmentioning

confidence: 73%

Trehalulose Synthase Native and Carbohydrate Complexed Structures Provide Insights into Sucrose Isomerization

Ravaud

Robert

Watzlawick

et al. 2007

Journal of Biological Chemistry

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137

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Various diseases related to the overconsumption of sugar make a growing need for sugar substitutes. Because sucrose is an inexpensive and readily available D-glucose donor, the industrial potential for enzymatic synthesis of the sucrose isomers trehalulose and/or isomaltulose from sucrose is large. The product specificity of sucrose isomerases that catalyze this reaction depends essentially on the possibility for tautomerization of sucrose, which is required for trehalulose formation. For optimal use of the enzyme, targeting controlled synthesis of these functional isomers, it is necessary to minimize the side reactions. This requires an extensive analysis of substrate binding modes and of the specificity-determining sites in the structure. The 1.6 -2.2-Å resolution three-dimensional structures of native and mutant complexes of a trehalulose synthase from Pseudomonas mesoacidophila MX-45 mimic successive states of the enzyme reaction. Combined with mutagenesis studies they give for the first time thorough insights into substrate recognition and processing and reaction specificities of these enzymes. Among the important outcomes of this study is the revelation of an aromatic clamp defined by Phe 256 and Phe 280 playing an essential role in substrate recognition and in controlling the reaction specificity, which is further supported by mutagenesis studies. Furthermore, this study highlights essential residues for binding the glucosyl and fructosyl moieties. The introduction of subtle changes informed by comparative three-dimensional structural data observed within our study can lead to fundamental modifications in the mode of action of sucrose isomerases and hence provide a template for industrial catalysts.

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Section: Discussionmentioning

confidence: 73%

Trehalulose Synthase Native and Carbohydrate Complexed Structures Provide Insights into Sucrose Isomerization

Ravaud

Robert

Watzlawick

et al. 2007

Journal of Biological Chemistry

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137

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“…Thus far, structures of the catalytic domains of cellulases and xylanases from seven of the families have been published: cellobiohydrolase II (CBH-II) from Trichoderma reesei (Rouvinen, Bergfors, Teeri, Knowles & Jones, 1990) and endocellulase E2 from Thermomonospora fusca (Spezio, Wilson & Karplus, 1993), both from family 6; CelA from Clostridium thermocellum (Juy et al, 1992) a representative of family 9; the endoglucanase V from H. insolens (this paper and, Davies et al, 1993) from family 45; cellobiohydrolase I (CBH-I) from T. reesei (Divne et al, 1994), family 7; a number of family 11 xylanases (Campbell et al, 1993;Wakarchuk, Campbell, Sung, Davoodi & Yaguchi, 1994;TOrr/Snen, Harkki & Rouvinen, 1994); the family 10 xylanases (Derewenda et al, 1994;Harris et al, 1994;White, Withers, Gilkes & Rose, 1994) and most recently CelCCA, a cellulase from family 5 (Ducros et al, 1996). Although all of these structures facilitate catalysis via a similar acid/base mechanism involving two or more aspartate or glutamate residues (for reviews on these mechanisms of catalysis see Koshland, 1953;Sinnott, 1990, McCarter & Withers, 1994, the structures of the cellulases from each family may be quite distinct.…”

Section: Introductionmentioning

confidence: 98%

Structure determination and refinement of the Humicola insolens endoglucanase V at 1.5 Å Resolution

Davies¹,

Dodson²,

Moore³

et al. 1996

Acta Cryst D

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The structure of the catalytic core of the endoglucanase V (EGV) from Humicola insolens has been determined by the method of multiple isomorphous replacement at 1.5 A, resolution. The final model, refined with X-PLOR and PROLSQ, has a crystallographic R factor of 0.163 (Rfree = 0.240) with deviations from stereochemical target values of 0.012 ,~ and 0.037 ° for bonds and angles, respectively. The model was further refined with SHELXL, including anisotropic modelling of the protein-atom temperature factors, to give a final model with an R factor of 0.105 and an Rfre~ of 0.154. The initial isomorphous replacement electron-density map was poor and uninterpretable but was improved by the use of synchrotron data collected at a wavelength chosen so as to optimize the f" contribution of the anomalous scattering from the heavy atoms. The structure of H. insolens EGV consists of a six-stranded/3-barrel domain, similar to that found in a family of plant defence proteins, linked by a number of disulfide-bonded loop regions. A long open groove runs across the surface of the enzyme either side of which lie the catalytic aspartate residues. The 9/~ separation of the catalytic carboxylate groups is consistent with the observation that EGV catalyzes the hydrolysis of the cellulose ,0(144) links with inversion of configuration at the anomeric CI atom. This structure is the first representative from the glycosyl hydrolase family 45.

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“…Structures for a number of cellulases from this family are known (Ducros et al, 1995;Dominguez et al, 1995;Sakon et al, 1996;Davies et al, 1998), but none were successful as molecularreplacement search models for the T. reesei mannanase. For this reason, a search for heavy-metal derivatives was initiated.…”

Section: Resultsmentioning

confidence: 99%

Crystallization and preliminary X-ray crystallographic analysis of aTrichoderma reeseiβ-mannanase from glycoside hydrolase family 5

Sabini

Brzozowski

Dauter

et al. 1999

Acta Crystallogr D Biol Cryst

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Crystals of the catalytic core domain of a Trichoderma reesei -mannanase belonging to glycoside hydrolase family 5 have been grown by the sitting-drop method at room temperature using ammonium sulfate as precipitant. The crystals grow as thin colourless plates and belong to space group P2 1 , with unit-cell parameters a = 50.0, b = 54.3, c = 60.2 A Ê , = 111.3, and have a single monomer of mannanase in the asymmetric unit. Native data to 2.0 A Ê resolution have been collected at room temperature using synchrotron radiation. Data for a platinum derivative have been collected to 1.65 A Ê at 110 K in a very short time at the CCLRC Daresbury synchrotron source, using a charge-coupled device (CCD) as detector.

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Crystal structure of the catalytic domain of a bacterial cellulase belonging to family 5

Cited by 151 publications

References 51 publications

Trehalulose Synthase Native and Carbohydrate Complexed Structures Provide Insights into Sucrose Isomerization

Trehalulose Synthase Native and Carbohydrate Complexed Structures Provide Insights into Sucrose Isomerization

Structure determination and refinement of the Humicola insolens endoglucanase V at 1.5 Å Resolution

Crystallization and preliminary X-ray crystallographic analysis of aTrichoderma reeseiβ-mannanase from glycoside hydrolase family 5

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