BRENDA - Reference to 3.2.1.145; Id = 732396

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

Hungateiclostridium thermocellum

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

Hungateiclostridium thermocellum

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

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

A3DD67

-

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

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

?

-

?

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

Ct1,3Gal43A

-

Hungateiclostridium thermocellum

Cthe_0661

-

Hungateiclostridium thermocellum

exo-beta-1,3-galactanase

-

Hungateiclostridium thermocellum

37

-

assay at

Hungateiclostridium thermocellum

6

-

assay at

Hungateiclostridium thermocellum

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

Hungateiclostridium thermocellum

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

Hungateiclostridium thermocellum

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

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

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

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

?

-

?

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

37

-

assay at

Hungateiclostridium thermocellum

6

-

assay at

Hungateiclostridium thermocellum

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

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

752571

Fujita

Degradative enzymes for type ...

Bifidobacterium longum subsp. Longum

Appl. Microbiol. Biotechnol.

103

1299-1310

2019

-

1

-

3

-

1

-

6

1

6

1

-

1

-

1

-

1

-

6

1

-

1

-

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

-

5

-

1

-

3

2

-

1

-

1

-

1

-

1

-

3

2

-

1

-

731184

Fujita

Bifidobacterium longum subsp. ...

Bifidobacterium longum, Bifidobacterium longum JCM 1217

Appl. Environ. Microbiol.

80

4577-4584

2014

-

1

-

1

-

1

-

1

-

6

-

9

-

14

-

2

1

-

1

-

1

-

1

-

1

-

1

-

9

-

14

-

1

-

1

-

1

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

-

731581

Ling

An exo-beta-(1->3)-D-galactana ...

Streptomyces sp. 19(2012), Streptomyces sp.

Carbohydr. Res.

352

70-81

2012

-

1

-

3

4

-

6

-

1

-

1

-

12

1

4

1

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2

-

1

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2

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3

4

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2

-

2

-

12

2

-

2

-

2

-

1

2

-

732396

Jiang

Crystal structure of 1,3Gal43A ...

Hungateiclostridium thermocellum

J. Struct. Biol.

180

447-457

2012

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1

2

-

2

-

4

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1

-

1

2

3

1

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1

-

1

-

1

2

-

2

-

1

-

1

2

1

-

1

-

2

-

713904

Sakamoto

Characterization of an exo-bet ...

Sphingomonas sp., Sphingomonas sp. 24T

Appl. Microbiol. Biotechnol.

90

1701-1710

2011

1

-

1

-

3

-

1

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1

2

-

12

1

2

1

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1

-

1

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1

-

1

-

1

-

1

-

1

2

-

12

1

-

1

-

1

-

1

-

703061

Ling

Preparation of a new chromogen ...

Irpex lacteus

Carbohydr. Res.

344

1941-1946

2009

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1

-

1

-

1

-

2

1

-

1

-

1

-

1

-

1

-

1

-

706965

Ishida

Crystallization of selenomethi ...

Phanerochaete chrysosporium

Acta Crystallogr. Sect. F

65

1274-1276

2009

-

1

-

2

-

1

-

1

3

-

1

-

1

-

1

-

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

-

1

-

1

-

4

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

-

1

-

1

-

1

-

1

-

1

-

5

-

2

-

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

-

10

-

2

1

-

1

-

1

-

1

-

1

-

1

-

10

-

1

-

1

-

666634

Kotake

Molecular cloning of a b-galac ...

Raphanus sativus

Plant Physiol.

138

1563-1576

2005

-

1

-

4

3

-

2

-

4

-

1

-

1

-

10

1

-

1

3

1

-

1

-

4

-

3

-

2

-

1

-

1

-

10

1

-

1

3

1

-

171335

Tsumuraya

Purification of an exo-beta-(1 ...

Irpex lacteus

J. Biol. Chem.

265

7207-7215

1990

-

2

-

2

1

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1

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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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1

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2

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2

1

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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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1

-

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

Data extracted from this reference: