Literature summary for 3.2.1.136 extracted from

Sainz-Polo, M.A.; Valenzuela, S.V.; Gonzalez, B.; Pastor, F.I.; Sanz-Aparicio, J.
Structural analysis of glucuronoxylan-specific Xyn30D and its attached CBM35 domain gives insights into the role of modularity in specificity (2014), J. Biol. Chem., 289, 31088-31101.

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Cloned(Commentary)

Cloned (Comment)	Organism
expression of His-tagged full-length enzyme and isolated catalytic domain in Escherichia coli strain BL21Star (DE3)	Paenibacillus barcinonensis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant catalytic domain GH30 or extended domain CBM35, free or in complex with glucuronic acid, crystallization by mixing of 0.001 ml of 22 mg/ml protein in 20 mM Tris, pH 8.0, and 150 mM NaCl with 0.001 ml of a solution containing 20% w/v PEG 6000, 0.2 M Ca2Cl, and either 0.1 M MES, pH 6.0, or 0.1 M Hepes, pH 7.0, and equilibration by vapor diffusion at room temperature, complexes are obtained by the soaking technique using glucuronic acid or a mixture of aldotriouronic, aldotetraouronic, and aldopentaouronic acids, in a ratio of 2:2:1, cryoprotection by 20% v/v glycerol, X-ray diffraction structure determination and analysis at 2.4 A resolution, molecular replacement, modelling	Paenibacillus barcinonensis

Crystallization (Comment)

Organism

purified recombinant catalytic domain GH30 or extended domain CBM35, free or in complex with glucuronic acid, crystallization by mixing of 0.001 ml of 22 mg/ml protein in 20 mM Tris, pH 8.0, and 150 mM NaCl with 0.001 ml of a solution containing 20% w/v PEG 6000, 0.2 M Ca2Cl, and either 0.1 M MES, pH 6.0, or 0.1 M Hepes, pH 7.0, and equilibration by vapor diffusion at room temperature, complexes are obtained by the soaking technique using glucuronic acid or a mixture of aldotriouronic, aldotetraouronic, and aldopentaouronic acids, in a ratio of 2:2:1, cryoprotection by 20% v/v glycerol, X-ray diffraction structure determination and analysis at 2.4 A resolution, molecular replacement, modelling

Paenibacillus barcinonensis

Inhibitors

Inhibitors	Comment	Organism	Structure
EDTA	partial inhibition	Paenibacillus barcinonensis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Ca2+	the enzyme's catalytic CBM35 domain displays a jellyroll beta-sandwich including two calcium ions, structure overview	Paenibacillus barcinonensis

Organism

Organism	UniProt	Comment	Textmining
Paenibacillus barcinonensis	H6WCZ0	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged full-length enzyme and isolated catalytic domain from Escherichia coli strain BL21Star (DE3) by immobilized metal affinity chromatography	Paenibacillus barcinonensis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
rye glucuronoarabinoxylan + H2O	the enzyme shows calcium-dependent glucuronic acid binding and also some binding to arabinose in presence of calcium chelating EDTA. Recognition of uronic acid by the CBM35 domains is always dependent on calcium, which interacts with the carboxylate of the ligand. Glu129, from the loop linking beta10-beta11, is a determinant of substrate specificity	Paenibacillus barcinonensis	?	-	?

Subunits

Subunits	Comment	Organism
More	Xyn30D is a modular enzyme	Paenibacillus barcinonensis

Synonyms

Synonyms	Comment	Organism
glucuronoxylan-specific Xyn30D	-	Paenibacillus barcinonensis
glucuronoxylanase	-	Paenibacillus barcinonensis
Xyn30D	-	Paenibacillus barcinonensis

General Information

General Information	Comment	Organism
evolution	glucuronoxylanase Xyn30D is a modular enzyme containing a family 30 glycoside hydrolase catalytic domain and an attached carbohydrate binding module of the CBM35 family. GH30 family enzymes structure comparisons, overview	Paenibacillus barcinonensis
metabolism	the enzyme plays a role in depolymerization of highly substituted chemically complex xylans, it belongs to the secretome of Paenibacillus barcinonensis, a powerful xylan degrading microorganism that shows a complex set of carbohydratases, including several GH10 and GH11 xylanases	Paenibacillus barcinonensis
additional information	the catalytic domain folds into an (alpha/beta)8 barrel with an associated beta-structure, whereas the attached CBM35 domain displays a jellyroll beta-sandwich including two calcium ions, molecular surface of Xyn30D-CBM35, atomic interactions between Xyn30D-CBM35, the calcium ion and the GlcA ligand, overview. An extended 12-residue segment links the CBM35 to the catalytic GH30 domain. Although both domains fold in an independent manner, the linker region makes polar interactions with the catalytic domain, allowing a moderate flexibility	Paenibacillus barcinonensis