Literature summary for 3.2.1.145 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.

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Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

Protein Variants	Comment	Organism
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
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]

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

Organism

Organism	UniProt	Comment	Textmining
Acetivibrio thermocellus	A3DD67	-	-

Purification (Commentary)

Purification (Comment)	Organism
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)

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

Subunits	Comment	Organism
?	x * 64400, recombinant His-tagged enzyme, SDS-PAGE, x * 58529, sequence calculation	Acetivibrio thermocellus
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

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

Temperature Optimum [°C]

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

pH Optimum

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

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the glycoside hydrolase family 43, GH43	Acetivibrio thermocellus
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