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Literature summary extracted from

  • Jiang, D.; Fan, J.; Wang, X.; Zhao, Y.; Huang, B.; Liu, J.; Zhang, X.C.
    Crystal structure of 1,3Gal43A, an exo-beta-1,3-galactanase from Clostridium thermocellum (2012), J. Struct. Biol., 180, 447-457.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.145 recombinant expression of N-terminally His-tagged wild-type and selenomethionine-labeled enzymes in Escherichia coli strain BL21(DE3) Acetivibrio thermocellus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.145 purified recombinant His-tagged wild-type and selenomethionine-labeled enzymes in complex with galactose, IPTG, galactan, and lactose, hanging drop vapor diffusion method, mixing of 0.001 ml of 50 mg/ml protein in a buffer containing 20 mM Tris-HCl, pH 8.0, and 100 mM NaCl, with 0.001 ml of reservoir solution containing 100 mM sodium acetate, pH 4.5, and 2.9-3.3 M sodium chloride,16°C, 3 days, X-ray diffraction structure determination and analysis at 2.7-3.2 A resolution Acetivibrio thermocellus

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.145 E112A site-directed mutagenesis, catalytically inactive mutant, the mutation eliminates the galactan hydrolysis activity of the enzyme while it does not disrupt its ligand binding ability Acetivibrio thermocellus
3.2.1.145 E112Q site-directed mutagenesis, catalytically inactive mutant, the mutation eliminates the galactan hydrolysis activity of the enzyme while it does not disrupt its ligand binding ability Acetivibrio thermocellus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.145 58529
-
 x * 64400, recombinant His-tagged enzyme, SDS-PAGE, x * 58529, sequence calculation Acetivibrio thermocellus
3.2.1.145 64400
-
 x * 64400, recombinant His-tagged enzyme, SDS-PAGE, x * 58529, sequence calculation Acetivibrio thermocellus

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.145 Acetivibrio thermocellus A3DD67
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.145 recombinant expression of His-tagged wild-type and selenomethionine-labeled enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Acetivibrio thermocellus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.145 additional information the enzyme specifically hydrolyses beta-1,3 glycosidic bonds in galactose-based oligosaccharides or polysaccharides, and shows maximum activity towards beta-1,3-D-galactotetraose. The enzyme also can perform beta-1-3 linked main chain hydrolysis in the presence of a beta-1,6 linked branch Acetivibrio thermocellus ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.145 ? x * 64400, recombinant His-tagged enzyme, SDS-PAGE, x * 58529, sequence calculation Acetivibrio thermocellus
3.2.1.145 More the enzyme is comprised of an N-terminal glycoside hydrolase family 43 module (GH43, residues 31-350), a carbohydrate binding module 13 (CBM13, residues 351-493), and a C-terminal type I dockerin-like domain (residues 494-571), three-dimensional structure analysis, overview Acetivibrio thermocellus

Synonyms

EC Number Synonyms Comment Organism
3.2.1.145 Ct1,3Gal43A
-
 Acetivibrio thermocellus
3.2.1.145 Cthe_0661
-
 Acetivibrio thermocellus
3.2.1.145 exo-beta-1,3-galactanase
-
 Acetivibrio thermocellus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.145 37
-
 assay at Acetivibrio thermocellus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.145 6
-
 assay at Acetivibrio thermocellus

General Information

EC Number General Information Comment Organism
3.2.1.145 evolution the enzyme belongs to the glycoside hydrolase family 43, GH43 Acetivibrio thermocellus
3.2.1.145 additional information two modes of substrate binding are observed at the beta site of the CtCBM13 domain, and one galactobiose molecule is found in an L shaped pocket of the CtGH43 domain, which appears large enough to accommodate two more galactose units. Glu112 serves as the general base for substrate hydrolysis. Putative mechanism of substrate delivery via the CBM13 domain, overview Acetivibrio thermocellus