BRENDA - Enzyme Database
show all sequences of 3.2.1.145

Crystal structure of 1,3Gal43A, an exo-beta-1,3-galactanase from Clostridium thermocellum

Jiang, D.; Fan, J.; Wang, X.; Zhao, Y.; Huang, B.; Liu, J.; Zhang, X.C.; J. Struct. Biol. 180, 447-457 (2012)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
recombinant expression of N-terminally His-tagged wild-type and selenomethionine-labeled enzymes in Escherichia coli strain BL21(DE3)
Hungateiclostridium thermocellum
Crystallization (Commentary)
Crystallization (Commentary)
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,16C, 3 days, X-ray diffraction structure determination and analysis at 2.7-3.2 A resolution
Hungateiclostridium thermocellum
Engineering
Protein Variants
Commentary
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
Hungateiclostridium thermocellum
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
Hungateiclostridium thermocellum
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
58529
-
x * 64400, recombinant His-tagged enzyme, SDS-PAGE, x * 58529, sequence calculation
Hungateiclostridium thermocellum
64400
-
x * 64400, recombinant His-tagged enzyme, SDS-PAGE, x * 58529, sequence calculation
Hungateiclostridium thermocellum
Organism
Organism
UniProt
Commentary
Textmining
Hungateiclostridium thermocellum
A3DD67
-
-
Purification (Commentary)
Purification (Commentary)
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
Hungateiclostridium thermocellum
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
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
732396
Hungateiclostridium thermocellum
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
?
x * 64400, recombinant His-tagged enzyme, SDS-PAGE, x * 58529, sequence calculation
Hungateiclostridium thermocellum
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
Hungateiclostridium thermocellum
Synonyms
Synonyms
Commentary
Organism
Ct1,3Gal43A
-
Hungateiclostridium thermocellum
Cthe_0661
-
Hungateiclostridium thermocellum
exo-beta-1,3-galactanase
-
Hungateiclostridium thermocellum
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
37
-
assay at
Hungateiclostridium thermocellum
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6
-
assay at
Hungateiclostridium thermocellum
Cloned(Commentary) (protein specific)
Commentary
Organism
recombinant expression of N-terminally His-tagged wild-type and selenomethionine-labeled enzymes in Escherichia coli strain BL21(DE3)
Hungateiclostridium thermocellum
Crystallization (Commentary) (protein specific)
Crystallization
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,16C, 3 days, X-ray diffraction structure determination and analysis at 2.7-3.2 A resolution
Hungateiclostridium thermocellum
Engineering (protein specific)
Protein Variants
Commentary
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
Hungateiclostridium thermocellum
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
Hungateiclostridium thermocellum
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
58529
-
x * 64400, recombinant His-tagged enzyme, SDS-PAGE, x * 58529, sequence calculation
Hungateiclostridium thermocellum
64400
-
x * 64400, recombinant His-tagged enzyme, SDS-PAGE, x * 58529, sequence calculation
Hungateiclostridium thermocellum
Purification (Commentary) (protein specific)
Commentary
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
Hungateiclostridium thermocellum
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
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
732396
Hungateiclostridium thermocellum
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 64400, recombinant His-tagged enzyme, SDS-PAGE, x * 58529, sequence calculation
Hungateiclostridium thermocellum
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
Hungateiclostridium thermocellum
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
37
-
assay at
Hungateiclostridium thermocellum
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
6
-
assay at
Hungateiclostridium thermocellum
General Information
General Information
Commentary
Organism
evolution
the enzyme belongs to the glycoside hydrolase family 43, GH43
Hungateiclostridium thermocellum
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
Hungateiclostridium thermocellum
General Information (protein specific)
General Information
Commentary
Organism
evolution
the enzyme belongs to the glycoside hydrolase family 43, GH43
Hungateiclostridium thermocellum
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
Hungateiclostridium thermocellum
Other publictions for EC
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
752571
Fujita
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103
1299-1310
2019
-
-
1
-
-
-
-
-
-
-
-
-
-
3
-
-
1
-
-
-
-
-
6
1
6
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
6
1
1
-
-
-
1
-
-
-
-
3
3
-
-
-
752397
Godoy
Crystal structure of a putati ...
Bifidobacterium bifidum, Bifidobacterium bifidum NCIMB 41171, Bifidobacterium bifidum S17
Acta Crystallogr. Sect. F
72
288-293
2016
-
-
1
1
-
-
-
-
-
-
1
-
-
5
-
-
1
-
-
-
-
-
3
2
2
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
3
2
-
-
-
-
-
-
-
-
-
1
1
-
-
-
731184
Fujita
Bifidobacterium longum subsp. ...
Bifidobacterium longum, Bifidobacterium longum JCM 1217
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4577-4584
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-
1
-
1
-
-
1
-
-
1
-
-
6
-
-
-
-
-
-
9
-
14
-
2
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
1
-
-
1
-
-
-
-
-
-
-
9
-
14
-
1
-
-
-
1
-
-
-
1
2
2
1
-
-
731202
Okawa
Identification of an exo-beta- ...
Fusarium oxysporum 12S, Fusarium oxysporum
Appl. Microbiol. Biotechnol.
97
9685-9694
2013
-
-
1
-
-
-
-
-
-
-
-
-
-
5
-
-
1
-
-
-
1
-
12
1
2
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
12
1
1
-
-
-
1
-
-
-
-
1
1
-
-
-
731581
Ling
An exo-beta-(1->3)-D-galactana ...
Streptomyces sp. 19(2012), Streptomyces sp.
Carbohydr. Res.
352
70-81
2012
-
1
1
-
-
-
-
-
-
-
3
4
-
6
-
-
1
-
-
-
1
-
12
1
4
1
-
-
-
2
-
-
-
-
1
-
-
2
2
-
-
-
-
-
-
-
-
-
-
3
4
-
-
-
2
-
-
2
-
12
2
2
-
-
-
2
-
-
2
-
1
2
-
-
-
732396
Jiang
Crystal structure of 1,3Gal43A ...
Hungateiclostridium thermocellum
J. Struct. Biol.
180
447-457
2012
-
-
1
1
2
-
-
-
-
-
2
-
-
4
-
-
1
-
-
-
-
-
1
2
3
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
1
2
-
-
-
-
-
-
-
2
-
-
-
-
1
-
-
-
-
1
2
1
-
-
-
1
-
-
-
-
2
2
-
-
-
713904
Sakamoto
Characterization of an exo-bet ...
Sphingomonas sp., Sphingomonas sp. 24T
Appl. Microbiol. Biotechnol.
90
1701-1710
2011
1
-
-
-
-
-
-
-
-
-
1
-
-
3
-
-
1
-
-
1
2
-
12
1
2
1
-
1
-
1
-
1
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
1
-
1
2
-
12
1
1
-
1
-
1
-
1
-
-
-
-
-
-
-
703061
Ling
Preparation of a new chromogen ...
Irpex lacteus
Carbohydr. Res.
344
1941-1946
2009
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
1
-
2
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
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-
-
-
-
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-
-
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-
-
-
1
1
-
1
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
706965
Ishida
Crystallization of selenomethi ...
Phanerochaete chrysosporium
Acta Crystallogr. Sect. F
65
1274-1276
2009
-
-
1
1
-
-
-
-
-
-
-
-
-
2
-
-
1
-
-
-
-
-
-
1
3
-
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1
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1
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1
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-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
707784
Kotake
Molecular cloning and expressi ...
Irpex lacteus, Irpex lacteus NBRC 5367
Biosci. Biotechnol. Biochem.
73
2303-2309
2009
-
-
1
-
-
-
-
-
-
-
1
-
-
2
-
-
1
-
-
-
1
-
4
1
2
1
1
-
-
1
1
-
-
-
-
-
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-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
1
-
4
1
1
1
-
-
1
1
-
-
-
-
-
-
-
-
677662
Ichinose
Characterization of an exo-bet ...
Hungateiclostridium thermocellum
Appl. Environ. Microbiol.
72
3515-3523
2006
-
-
1
-
-
-
-
-
-
-
1
-
-
4
-
-
1
-
-
-
1
-
5
-
2
2
-
-
-
1
1
-
-
-
-
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-
1
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-
-
-
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-
1
-
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-
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1
-
-
1
-
5
-
2
-
-
-
1
1
-
-
-
-
-
-
-
-
678786
Ichinose
Characterization of an exo-bet ...
Streptomyces avermitilis, Streptomyces avermitilis NBRC 14893
Biosci. Biotechnol. Biochem.
70
2745-2750
2006
-
-
1
-
-
-
-
-
-
-
1
-
-
4
-
1
1
-
-
-
1
-
10
-
2
1
-
-
-
1
-
-
-
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-
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1
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1
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1
1
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1
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10
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
666634
Kotake
Molecular cloning of a b-galac ...
Raphanus sativus
Plant Physiol.
138
1563-1576
2005
-
-
1
-
-
-
4
3
-
-
2
-
-
4
-
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1
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-
-
1
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10
1
1
1
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1
3
1
1
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1
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4
-
3
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2
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1
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1
-
10
1
1
-
1
3
1
1
-
-
-
-
-
-
-
-
171335
Tsumuraya
Purification of an exo-beta-(1 ...
Irpex lacteus
J. Biol. Chem.
265
7207-7215
1990
-
-
-
-
-
-
-
2
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2
1
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1
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1
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1
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1
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1
1
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1
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1
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2
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1
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1
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4
1
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1
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1
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