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Literature summary for 3.2.1.73 extracted from

  • Zhang, L.; Zhao, P.; Chen, C.C.; Huang, C.H.; Ko, T.P.; Zheng, Y.; Guo, R.T.
    Preliminary X-ray diffraction analysis of a thermophilic beta-1,3-1,4-glucanase from Clostridium thermocellum (2014), Acta Crystallogr. Sect. F, 70, 946-948.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of the His-tagged isolated catalytic domain of the enzyme, residues 31-251, in Escherichia coli strain BL21trxB(DE3) with an extra linker AGAGA at its N-terminus Acetivibrio thermocellus

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant catalytic domain, sitting drop vapour diffusion method, mixing of 0.002 ml of 30 mg/ml protein in 25 mM Tris pH 7.5, 150 mM NaCl, and 0.002 ml reservori solution containing 0.04 M citric acid, 0.06 M Bis-Tris propane, pH 6.4, 18% w/v PEG 3350, 25°C, 7 days, optimized method, X-ray diffraction structure determination and analysis at 1.95 A resolution, molecular replacement method Acetivibrio thermocellus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
28000
-
x * 28000, about, recombinant catalytic domain, SDS-PAGE Acetivibrio thermocellus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Acetivibrio thermocellus the enzyme catalyzes the specific hydrolysis of internal beta-1,4-glycosidic bonds adjacent to the 3-O-substituted glucose residues in mixed-linked beta-glucans. The Clostridium thermocellum enzyme exhibits a high specific activity towards barley beta-glucan and lichenan, but is not active towards laminarin, curdlan and cellulosic substrates ?
-
?
additional information Acetivibrio thermocellus DSM 1237 the enzyme catalyzes the specific hydrolysis of internal beta-1,4-glycosidic bonds adjacent to the 3-O-substituted glucose residues in mixed-linked beta-glucans. The Clostridium thermocellum enzyme exhibits a high specific activity towards barley beta-glucan and lichenan, but is not active towards laminarin, curdlan and cellulosic substrates ?
-
?

Organism

Organism UniProt Comment Textmining
Acetivibrio thermocellus A3DBX3
-
-
Acetivibrio thermocellus DSM 1237 A3DBX3
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged isolated catalytic domain of the enzyme from Escherichia coli strain BL21trxB(DE3) by nickel affinity chromatography and dialysis, tag cleavage by factor Xa digestion, and removal of the tag by another step of nickel affinity chromatography Acetivibrio thermocellus

Reaction

Reaction Comment Organism Reaction ID
beta-D-glucopyranosyl-(1-3)-beta-D-glucopyranosyl-(1-3)-beta-D-glucopyranosyl-(1-4)-beta-D-glucopyranosyl-(1-3)-beta-D-glucopyranose + 4 H2O = 5 beta-D-glucopyranose beta-1,3-1,4-glucanase catalyzes strict endo-hydrolysis of the beta-1,4-glycosidic linkage adjacent to a 3-O-substituted glucose residue in the mixed-linked beta-glucans via a double-displacement mechanism Acetivibrio thermocellus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
barley beta-glucan + H2O
-
Acetivibrio thermocellus ?
-
?
barley beta-glucan + H2O
-
Acetivibrio thermocellus DSM 1237 ?
-
?
lichenan + H2O
-
Acetivibrio thermocellus ?
-
?
lichenan + H2O
-
Acetivibrio thermocellus DSM 1237 ?
-
?
additional information the enzyme catalyzes the specific hydrolysis of internal beta-1,4-glycosidic bonds adjacent to the 3-O-substituted glucose residues in mixed-linked beta-glucans. The Clostridium thermocellum enzyme exhibits a high specific activity towards barley beta-glucan and lichenan, but is not active towards laminarin, curdlan and cellulosic substrates Acetivibrio thermocellus ?
-
?
additional information the enzyme catalyzes the specific hydrolysis of internal beta-1,4-glycosidic bonds adjacent to the 3-O-substituted glucose residues in mixed-linked beta-glucans. The Clostridium thermocellum enzyme exhibits a high specific activity towards barley beta-glucan and lichenan, but is not active towards laminarin, curdlan and cellulosic substrates Acetivibrio thermocellus DSM 1237 ?
-
?

Subunits

Subunits Comment Organism
? x * 28000, about, recombinant catalytic domain, SDS-PAGE Acetivibrio thermocellus

Synonyms

Synonyms Comment Organism
beta-1,3-1,4-glucanase
-
Acetivibrio thermocellus
CtGlu16A
-
Acetivibrio thermocellus
Lichenase
-
Acetivibrio thermocellus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
-
Acetivibrio thermocellus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8 9
-
Acetivibrio thermocellus

pH Range

pH Minimum pH Maximum Comment Organism
5 12 50% of maximal activity within this range Acetivibrio thermocellus

General Information

General Information Comment Organism
evolution beta-1,3-1,4-glucanases from various microorganisms and plants belong to glycoside hydrolase families 16 and 17, respectively Acetivibrio thermocellus