Literature summary for 3.2.1.4 extracted from

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

Search for more literature for 3.2.1.4

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

Protein Variants	Comment	Organism
E201A	site-directed mutagenesis, crystal structure determination with bound ligands	Pyrococcus horikoshii
E342A	site-directed mutagenesis, crystal structure determination with bound ligands	Pyrococcus horikoshii
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

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

Metals/Ions

Metals/Ions	Comment	Organism	Structure
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

Organism	UniProt	Comment	Textmining
Pyrococcus horikoshii	O58925	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
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

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

Temperature Optimum [°C]

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

pH Optimum

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

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Release 2023.1 (January 2023)