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Literature summary extracted from

  • Kim, H.W.; Ishikawa, K.
    Functional analysis of hyperthermophilic endocellulase from Pyrococcus horikoshii by crystallographic snapshots (2011), Biochem. J., 437, 223-230.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.4 expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Pyrococcus horikoshii

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.4 purified enzyme mutants in complex with either the substrate or product ligands, hanging drop vapour diffusion method, 20 mg/ml protein in 50 mM Tris/HCl, pH 8.0, mixing of 0.0015 ml of both protein and reservoir solution, the latter contaning 1.5 M ammonium phosphate, and 0.1 M MES, pH 6.5, 22°C, 3 days, X-ray diffraction structure determination and analysis at 1.65-2.01 A resolution Pyrococcus horikoshii

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.4 E201A site-directed mutagenesis, crystal structure determination with bound ligands Pyrococcus horikoshii
3.2.1.4 E342A site-directed mutagenesis, crystal structure determination with bound ligands Pyrococcus horikoshii
3.2.1.4 Y299F site-directed mutagenesis, crystal structure determination with bound ligands, the mutant shows reduced activity compare to the wild-type enzyme, and a rare enzyme-substrate complex structure Pyrococcus horikoshii

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.4 membrane
-
 Pyrococcus horikoshii 16020
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.4 Zn2+ zinc ions tightly bound between the two catalytic glutamate residues, which present an obstacle for the entrance of ligand in the active site Pyrococcus horikoshii

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.4 Pyrococcus horikoshii O58925
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.4 additional information with substrate cellulose, the substrate position is fixed by the alignment of one cellobiose unit between the two aromatic amino acid residues at subsites +1 and +2. During the enzyme reaction, the glucose structure of cellulose substrates is distorted at subsite -1, and the beta-1,4-glucoside bond between glucose moieties is twisted between subsites -1 and +1. Subsite -2 specifically recognizes the glucose residue, but recognition by subsites +1 and +2 is loose during the enzyme reaction. This type of recognition is important for creation of the distorted boat form of the substrate at subsite -1 Pyrococcus horikoshii ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.4 beta-1,4-endoglucanase
-
 Pyrococcus horikoshii
3.2.1.4 endocellulase
-
 Pyrococcus horikoshii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.4 85
-
 assay at Pyrococcus horikoshii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.4 5.5
-
 assay at Pyrococcus horikoshii