BRENDA - Enzyme Database
show all sequences of 3.2.1.86

Structural insights into the substrate specificity of a 6-phospho-beta-glucosidase BglA-2 from Streptococcus pneumoniae TIGR4

Yu, W.L.; Jiang, Y.L.; Pikis, A.; Cheng, W.; Bai, X.H.; Ren, Y.M.; Thompson, J.; Zhou, C.Z.; Chen, Y.; J. Biol. Chem. 288, 14949-14958 (2013)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
gene SP_0578 or bglA-2, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
Streptococcus pneumoniae
Crystallization (Commentary)
Crystallization (Commentary)
Organism
purified recombinant enzyme in apoform or complexed with thiocellobiose 6-phosphate, sitting drop vapour diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 20 mM Tris-Cl, pH 8.0, 100 mM NaCl with 0.001 ml of reservoir solution containing 15% PEG 5000MME, and 0.1 M sodium citrate, pH 5.6, 16°C, X-ray diffraction structure determination and analysis at 2.0-2.4 A resolution, molecular replacement method
Streptococcus pneumoniae
Engineering
Protein Variants
Commentary
Organism
E171A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
E171Q
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
E364A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
E364Q
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
K430A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
M423A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Streptococcus pneumoniae
S424A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
W338A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
Y126A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
Y126F
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Streptococcus pneumoniae
Y303A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
Y303F
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
Y432F
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.478
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged wild-type enzyme
Streptococcus pneumoniae
0.512
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant M423A
Streptococcus pneumoniae
0.598
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant Y126F
Streptococcus pneumoniae
1.135
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged wild-type enzyme
Streptococcus pneumoniae
1.32
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant M423A
Streptococcus pneumoniae
1.515
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant Y126F
Streptococcus pneumoniae
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
cellobiose 6-phosphate + H2O
Streptococcus pneumoniae
-
beta-D-glucose + glucose 6-phosphate
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Streptococcus pneumoniae
A0A0H2UP35
gene SP_0578 or bglA-2
-
Purification (Commentary)
Purification (Commentary)
Organism
recombinant tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, gel filtration, and ultrafiltration
Streptococcus pneumoniae
Reaction
Reaction
Commentary
Organism
Reaction ID
6-phospho-beta-D-glucosyl-(1->4)-D-glucose + H2O = D-glucose + D-glucose 6-phosphate
active site structure and double displacement mechanism, overview
Streptococcus pneumoniae
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
4-nitrophenyl beta-D-glucopyranoside 6-phosphate + H2O
-
732101
Streptococcus pneumoniae
4-nitrophenol + beta-D-glucose 6-phosphate
-
-
-
?
cellobiose 6-phosphate + H2O
-
732101
Streptococcus pneumoniae
beta-D-glucose + glucose 6-phosphate
-
-
-
?
additional information
no activity with beta-(1,4)-thiocellobiose 6-phosphate
732101
Streptococcus pneumoniae
?
-
-
-
-
Synonyms
Synonyms
Commentary
Organism
BglA-2
-
Streptococcus pneumoniae
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Streptococcus pneumoniae
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
138
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant Y126F
Streptococcus pneumoniae
145
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant M423A
Streptococcus pneumoniae
146
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant Y126F
Streptococcus pneumoniae
168
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant M423A
Streptococcus pneumoniae
170
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged wild-type enzyme
Streptococcus pneumoniae
195
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged wild-type enzyme
Streptococcus pneumoniae
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Streptococcus pneumoniae
Cloned(Commentary) (protein specific)
Commentary
Organism
gene SP_0578 or bglA-2, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
Streptococcus pneumoniae
Crystallization (Commentary) (protein specific)
Crystallization
Organism
purified recombinant enzyme in apoform or complexed with thiocellobiose 6-phosphate, sitting drop vapour diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 20 mM Tris-Cl, pH 8.0, 100 mM NaCl with 0.001 ml of reservoir solution containing 15% PEG 5000MME, and 0.1 M sodium citrate, pH 5.6, 16°C, X-ray diffraction structure determination and analysis at 2.0-2.4 A resolution, molecular replacement method
Streptococcus pneumoniae
Engineering (protein specific)
Protein Variants
Commentary
Organism
E171A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
E171Q
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
E364A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
E364Q
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
K430A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
M423A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Streptococcus pneumoniae
S424A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
W338A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
Y126A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
Y126F
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Streptococcus pneumoniae
Y303A
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
Y303F
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
Y432F
site-directed mutagenesis, inactive mutant
Streptococcus pneumoniae
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.478
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged wild-type enzyme
Streptococcus pneumoniae
0.512
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant M423A
Streptococcus pneumoniae
0.598
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant Y126F
Streptococcus pneumoniae
1.135
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged wild-type enzyme
Streptococcus pneumoniae
1.32
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant M423A
Streptococcus pneumoniae
1.515
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant Y126F
Streptococcus pneumoniae
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
cellobiose 6-phosphate + H2O
Streptococcus pneumoniae
-
beta-D-glucose + glucose 6-phosphate
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, gel filtration, and ultrafiltration
Streptococcus pneumoniae
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
4-nitrophenyl beta-D-glucopyranoside 6-phosphate + H2O
-
732101
Streptococcus pneumoniae
4-nitrophenol + beta-D-glucose 6-phosphate
-
-
-
?
cellobiose 6-phosphate + H2O
-
732101
Streptococcus pneumoniae
beta-D-glucose + glucose 6-phosphate
-
-
-
?
additional information
no activity with beta-(1,4)-thiocellobiose 6-phosphate
732101
Streptococcus pneumoniae
?
-
-
-
-
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Streptococcus pneumoniae
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
138
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant Y126F
Streptococcus pneumoniae
145
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant M423A
Streptococcus pneumoniae
146
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant Y126F
Streptococcus pneumoniae
168
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant M423A
Streptococcus pneumoniae
170
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged wild-type enzyme
Streptococcus pneumoniae
195
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged wild-type enzyme
Streptococcus pneumoniae
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Streptococcus pneumoniae
General Information
General Information
Commentary
Organism
evolution
the enzyme belongs to the glycosyl hydrolase family 1, GH1. Members of the GH-1 family share a common catalytic mechanism and exhibit similar structural folds, including a (beta/alpha)8 TIM-barrel
Streptococcus pneumoniae
additional information
the overall structure of enzyme BglA-2 adopts a typical (beta/alpha)8 TIM-barrel, with the active site located at the center of the convex surface of the beta-barrel. Residues Tyr126, Tyr303, and Trp338, at subsite +1 of BglA-2 determine substrate specificity with respect to 1,4-linked 6-phospho-beta-glucosides. Residues Ser424, Lys430, and Tyr432 of BglA-2 play important roles in the hydrolytic selectivity toward phosphorylated rather than non-phosphorylated compounds, comparative structural analysis. Tryptophan versus a methionine/alanine residue at subsite -1 may contribute to the catalytic and substrate selectivity with respect to structurally similar 6-phospho-beta-galactosidases and 6-phospho-beta-glucosidases assigned to the GH-1 family
Streptococcus pneumoniae
General Information (protein specific)
General Information
Commentary
Organism
evolution
the enzyme belongs to the glycosyl hydrolase family 1, GH1. Members of the GH-1 family share a common catalytic mechanism and exhibit similar structural folds, including a (beta/alpha)8 TIM-barrel
Streptococcus pneumoniae
additional information
the overall structure of enzyme BglA-2 adopts a typical (beta/alpha)8 TIM-barrel, with the active site located at the center of the convex surface of the beta-barrel. Residues Tyr126, Tyr303, and Trp338, at subsite +1 of BglA-2 determine substrate specificity with respect to 1,4-linked 6-phospho-beta-glucosides. Residues Ser424, Lys430, and Tyr432 of BglA-2 play important roles in the hydrolytic selectivity toward phosphorylated rather than non-phosphorylated compounds, comparative structural analysis. Tryptophan versus a methionine/alanine residue at subsite -1 may contribute to the catalytic and substrate selectivity with respect to structurally similar 6-phospho-beta-galactosidases and 6-phospho-beta-glucosidases assigned to the GH-1 family
Streptococcus pneumoniae
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
90
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant Y126F
Streptococcus pneumoniae
110
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant M423A
Streptococcus pneumoniae
150
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged wild-type enzyme
Streptococcus pneumoniae
240
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant Y126F
Streptococcus pneumoniae
330
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant M423A
Streptococcus pneumoniae
410
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged wild-type enzyme
Streptococcus pneumoniae
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
90
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant Y126F
Streptococcus pneumoniae
110
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant M423A
Streptococcus pneumoniae
150
-
cellobiose 6-phosphate
pH 7.5, 37°C, recombinant His-tagged wild-type enzyme
Streptococcus pneumoniae
240
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant Y126F
Streptococcus pneumoniae
330
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged mutant M423A
Streptococcus pneumoniae
410
-
4-nitrophenyl-beta-D-glucopyranoside 6-phosphate
pH 7.5, 37°C, recombinant His-tagged wild-type enzyme
Streptococcus pneumoniae
Other publictions for EC 3.2.1.86
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)
750522
Lansky
Structural basis for enzyme b ...
Geobacillus stearothermophilus, Geobacillus stearothermophilus T-1
FEBS J.
284
3931-3953
2017
-
-
-
1
3
-
-
-
-
-
1
-
-
3
-
-
1
-
-
-
-
-
2
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
3
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
2
1
-
-
-
-
-
-
-
-
1
-
-
1
-
-
751192
Yang
-
Coloring and bioinformatic an ...
Oenococcus oeni
J. Chin. Inst. Food Sci. Technol.
17
197-205
2017
-
-
1
-
-
-
-
-
1
-
-
1
-
1
-
-
-
-
-
-
-
-
1
1
2
-
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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
-
-
-
-
-
-
-
1
-
-
-
-
-
-
750546
Kwan
Chemoenzymatic synthesis of 6 ...
Streptococcus pyogenes, Streptococcus pyogenes M1
FEBS Lett.
590
461-468
2016
-
-
1
1
-
-
1
1
-
-
-
-
-
2
-
-
1
-
-
-
-
-
2
-
1
-
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1
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1
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1
-
1
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1
1
1
-
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1
-
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-
-
2
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
750733
Terra
Pneumococcal 6-phospho-beta-g ...
Streptococcus pneumoniae
Infect. Immun.
84
286-292
2016
-
-
1
-
-
-
-
1
1
-
-
2
-
4
-
-
1
-
-
-
1
-
2
1
3
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
1
1
-
-
2
-
-
-
1
-
-
1
-
2
1
-
-
-
-
-
-
-
-
-
1
1
-
-
-
750963
Zangoui
Evolution of aromatic beta-gl ...
Escherichia coli
J. Bacteriol.
197
710-716
2015
-
-
-
-
1
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
4
-
2
-
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-
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1
-
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1
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-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
731055
Stepper
Structure and activity of the ...
Streptococcus pyogenes, Streptococcus pyogenes M1 GAS SF370
Acta Crystallogr. Sect. D
69
16-23
2013
-
-
1
1
-
1
1
2
-
-
-
-
-
5
-
-
1
-
-
-
-
-
4
1
1
1
-
-
1
1
-
-
-
-
-
-
-
-
1
-
1
-
1
-
1
-
2
-
-
-
-
-
-
-
1
-
-
-
-
4
1
1
-
-
1
1
-
-
-
-
3
3
-
1
1
731178
Yang
Characterization of a novel me ...
uncultured organism
Appl. Environ. Microbiol.
79
2121-2127
2013
-
-
1
-
11
-
4
1
-
3
-
-
-
1
-
-
1
-
-
-
-
-
2
-
1
1
-
-
-
1
-
-
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-
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-
1
-
-
11
-
-
4
-
1
-
3
-
-
-
-
-
1
-
-
-
-
2
-
1
-
-
-
1
-
-
-
-
2
2
-
-
-
732101
Yu
Structural insights into the s ...
Streptococcus pneumoniae
J. Biol. Chem.
288
14949-14958
2013
-
-
1
1
13
-
-
6
-
-
-
1
-
4
-
-
1
1
-
-
-
-
3
-
1
1
-
-
6
1
-
-
-
-
-
-
-
-
1
-
1
13
-
-
-
-
6
-
-
-
1
-
-
-
1
-
-
-
-
3
-
1
-
-
6
1
-
-
-
-
2
2
-
6
6
732252
Honda
Purification and characterizat ...
Lactobacillus gasseri, Lactobacillus gasseri ATCC 33323
J. Gen. Appl. Microbiol.
58
11-17
2012
-
-
1
-
-
-
-
2
-
-
1
-
-
4
-
-
1
-
-
-
-
-
4
1
2
2
-
-
2
2
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
2
-
-
1
-
-
-
-
1
-
-
-
-
4
1
2
-
-
2
2
-
-
-
-
1
1
-
2
2
715955
Capaldo
-
beta-Glucoside metabolism in O ...
Oenococcus oeni
J. Mol. Catal. B
69
27-34
2011
-
-
1
-
-
-
-
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-
-
1
-
-
1
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1
-
-
-
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-
2
1
3
-
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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2
1
-
1
-
-
1
-
-
-
-
-
-
-
-
-
705622
Hall
Evolution and biochemistry of ...
Erwinia rhapontici, Erwinia rhapontici DSM 448
Mol. Biol. Evol.
26
2487-2497
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The beta-glucoside system of E ...
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136926
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Genetic determination of the c ...
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Cellobiose metabolism in Aerob ...
Klebsiella aerogenes
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