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Literature summary for 3.2.1.151 extracted from

  • Yoshizawa, T.; Shimizu, T.; Hirano, H.; Sato, M.; Hashimoto, H.
    Structural basis for inhibition of xyloglucan-specific endo-beta-1,4-glucanase (XEG) by XEG-protein inhibitor (2012), J. Biol. Chem., 287, 18710-18716.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of GST-tagged enzyme in Escherichia coli BL21 Daucus carota

Crystallization (Commentary)

Crystallization (Comment) Organism
purified enzyme free or in complex with substrate xyloglucan or inhibitor extracellular dermal glycoprotein, hanging drop vapor diffusion method, from 0.1 M sodium acetate, pH 5.5, and 1.5 M ammonium sulfate, or 0.1 M sodium acetate, pH 4.6, 25% PEG 3000, and 5 mg/ml digested xyloglucan, 20°C, X-ray diffraction structure determination and analysis at 0.95-2.70 A resolution Daucus carota

Protein Variants

Protein Variants Comment Organism
W13A site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Daucus carota
W13A/W28A site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Daucus carota
W28A site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Daucus carota
Y24A site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme Daucus carota

Inhibitors

Inhibitors Comment Organism Structure
extracellular dermal glycoprotein a glycoside hydrolase inhibitor protein inhibitor of the enzyme, it closely mimics the XEG-xyloglucan interaction Daucus carota

Organism

Organism UniProt Comment Textmining
Daucus carota Q05929
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant GST-tagged enzyme from Escherichia coli BL21 by glutathione affinity chromatography and gel filtration Daucus carota

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
tamarind seed xyloglucan + H2O
-
Daucus carota ?
-
?

Synonyms

Synonyms Comment Organism
XEG
-
Daucus carota
xyloglucanase
-
Daucus carota

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
assay at room temperature Daucus carota

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
4.6
-
assay at Daucus carota

General Information

General Information Comment Organism
evolution the enzyme is a member of GH family 12, GH12 Daucus carota
physiological function xyloglucan is a major component of the cell wall in most plants, and xyloglucan-specific endo-beta-1,4-glucanase digests the polysaccharide chains of plant cell walls. To protect the cell wall from these enzymes, plants secrete glycoside hydrolase inhibitor proteins Daucus carota