BRENDA - Enzyme Database
show all sequences of 4.1.2.52

Purification and biochemical characterization of a pyruvate-specific class II aldolase, HpaI

Wang, W.; Seah, S.Y.; Biochemistry 44, 9447-9455 (2005)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
HpaI wild type and R70A mutant expressed in Escherichia coli BL21(lambda DE3) under the control of the T7 promoter from expression plasmid pT7-7
Escherichia coli
Engineering
Protein Variants
Commentary
Organism
R70A
replacement by site-specific mutagenesis results in an enzyme that lacks both aldolase and decarboxylase activities. The mutant enzyme is also unable to catalyze pyruvate proton exchange
Escherichia coli
Inhibitors
Inhibitors
Commentary
Organism
Structure
pyruvate
-
Escherichia coli
sodium oxalate
-
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.006
-
4-hydroxy-2-oxopentanoate
Km (app) with Co2+, pH 8.0, 25C
Escherichia coli
0.0175
-
4-hydroxy-2-oxopentanoate
Km (app) with Mn2+, pH 8.0, 25C
Escherichia coli
0.16
-
4-hydroxy-2-oxohexanoate
pH 8.0, 25C
Escherichia coli
0.289
-
4-hydroxy-2-oxopentanoate
Km (app) with Mg2+, pH 8.0, 25C
Escherichia coli
0.38
-
4-hydroxy-2-oxopentanoate
pH 8.0, 25C
Escherichia coli
14.22
-
3-deoxy-D-manno-oct-2-ulosonic acid
i.e. KDO, pH 8.0, 25C
Escherichia coli
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Cd2+
0.5 mM chloride salt, 4% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Co2+
0.5 mM chloride salt, 100% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Cu2+
0.5 mM chloride salt, 1% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Fe2+
0.5 mM chloride salt, 57% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Mg2+
0.5 mM chloride salt, 49% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Mn2+
0.5 mM chloride salt, 99% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
additional information
no activity with Ca2+, Fe3+, Cr3+ and Al3+
Escherichia coli
Ni2+
0.5 mM chloride salt, 11% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Zn2+
0.5 mM chloride salt, 73% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
28000
-
subunit, SDS-PAGE
Escherichia coli
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
4-hydroxy-2-oxoheptanedioate
Escherichia coli
-
pyruvate + succinate semialdehyde
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Escherichia coli
Q47098
-
-
Purification (Commentary)
Purification (Commentary)
Organism
wild type protein and R70A mutant protein purified to homogeneity using anion exchange, hydrophobic interaction, and gel filtration chormatography
Escherichia coli
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
3-deoxy-D-manno-oct-2-ulosonic acid
3-deoxy-D-manno-oct-2-ulosonic acid i.e. KDO
721601
Escherichia coli
?
-
-
-
?
4-hydroxy-2-oxoheptanedioate
-
721601
Escherichia coli
pyruvate + succinate semialdehyde
-
-
-
?
4-hydroxy-2-oxohexanoate
-
721601
Escherichia coli
?
-
-
-
?
4-hydroxy-2-oxopentanoate
-
721601
Escherichia coli
?
-
-
-
?
additional information
the enzyme exhibits significant oxaloacetate decarboxylase activity, with a kcat value 2.4fold higher than the corresponding value for the aldol cleavage of 4-hydroxy-2-oxopentanoate
721601
Escherichia coli
?
-
-
-
?
Synonyms
Synonyms
Commentary
Organism
HpaI
-
Escherichia coli
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.55
-
3-deoxy-D-manno-oct-2-ulosonic acid
i.e. KDO, pH 8.0, 25C
Escherichia coli
229
-
4-hydroxy-2-oxohexanoate
pH 8.0, 25C
Escherichia coli
353
-
4-hydroxy-2-oxopentanoate
pH 8.0, 25C
Escherichia coli
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.0055
-
sodium oxalate
pH 8.0, 25C
Escherichia coli
0.53
-
pyruvate
pH 8.0, 25C
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
HpaI wild type and R70A mutant expressed in Escherichia coli BL21(lambda DE3) under the control of the T7 promoter from expression plasmid pT7-7
Escherichia coli
Engineering (protein specific)
Protein Variants
Commentary
Organism
R70A
replacement by site-specific mutagenesis results in an enzyme that lacks both aldolase and decarboxylase activities. The mutant enzyme is also unable to catalyze pyruvate proton exchange
Escherichia coli
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
pyruvate
-
Escherichia coli
sodium oxalate
-
Escherichia coli
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.0055
-
sodium oxalate
pH 8.0, 25C
Escherichia coli
0.53
-
pyruvate
pH 8.0, 25C
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.006
-
4-hydroxy-2-oxopentanoate
Km (app) with Co2+, pH 8.0, 25C
Escherichia coli
0.0175
-
4-hydroxy-2-oxopentanoate
Km (app) with Mn2+, pH 8.0, 25C
Escherichia coli
0.16
-
4-hydroxy-2-oxohexanoate
pH 8.0, 25C
Escherichia coli
0.289
-
4-hydroxy-2-oxopentanoate
Km (app) with Mg2+, pH 8.0, 25C
Escherichia coli
0.38
-
4-hydroxy-2-oxopentanoate
pH 8.0, 25C
Escherichia coli
14.22
-
3-deoxy-D-manno-oct-2-ulosonic acid
i.e. KDO, pH 8.0, 25C
Escherichia coli
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Cd2+
0.5 mM chloride salt, 4% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Co2+
0.5 mM chloride salt, 100% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Cu2+
0.5 mM chloride salt, 1% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Fe2+
0.5 mM chloride salt, 57% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Mg2+
0.5 mM chloride salt, 49% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Mn2+
0.5 mM chloride salt, 99% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
additional information
no activity with Ca2+, Fe3+, Cr3+ and Al3+
Escherichia coli
Ni2+
0.5 mM chloride salt, 11% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Zn2+
0.5 mM chloride salt, 73% relative activity, with 4-hydroxy-2-oxopentanoate as substrate
Escherichia coli
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
28000
-
subunit, SDS-PAGE
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
4-hydroxy-2-oxoheptanedioate
Escherichia coli
-
pyruvate + succinate semialdehyde
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
wild type protein and R70A mutant protein purified to homogeneity using anion exchange, hydrophobic interaction, and gel filtration chormatography
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
3-deoxy-D-manno-oct-2-ulosonic acid
3-deoxy-D-manno-oct-2-ulosonic acid i.e. KDO
721601
Escherichia coli
?
-
-
-
?
4-hydroxy-2-oxoheptanedioate
-
721601
Escherichia coli
pyruvate + succinate semialdehyde
-
-
-
?
4-hydroxy-2-oxohexanoate
-
721601
Escherichia coli
?
-
-
-
?
4-hydroxy-2-oxopentanoate
-
721601
Escherichia coli
?
-
-
-
?
additional information
the enzyme exhibits significant oxaloacetate decarboxylase activity, with a kcat value 2.4fold higher than the corresponding value for the aldol cleavage of 4-hydroxy-2-oxopentanoate
721601
Escherichia coli
?
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.55
-
3-deoxy-D-manno-oct-2-ulosonic acid
i.e. KDO, pH 8.0, 25C
Escherichia coli
229
-
4-hydroxy-2-oxohexanoate
pH 8.0, 25C
Escherichia coli
353
-
4-hydroxy-2-oxopentanoate
pH 8.0, 25C
Escherichia coli
General Information
General Information
Commentary
Organism
metabolism
enzyme is involved in the catabolic pathway of hydroxyphenylacetate
Escherichia coli
General Information (protein specific)
General Information
Commentary
Organism
metabolism
enzyme is involved in the catabolic pathway of hydroxyphenylacetate
Escherichia coli
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.03868
-
3-deoxy-D-manno-oct-2-ulosonic acid
i.e. KDO, pH 8.0, 25C
Escherichia coli
940
-
4-hydroxy-2-oxopentanoate
pH 8.0, 25C
Escherichia coli
1460
-
4-hydroxy-2-oxohexanoate
pH 8.0, 25C
Escherichia coli
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.03868
-
3-deoxy-D-manno-oct-2-ulosonic acid
i.e. KDO, pH 8.0, 25C
Escherichia coli
940
-
4-hydroxy-2-oxopentanoate
pH 8.0, 25C
Escherichia coli
1460
-
4-hydroxy-2-oxohexanoate
pH 8.0, 25C
Escherichia coli
Other publictions for EC 4.1.2.52
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)
725493
Coincon
Crystal structure of reaction ...
Escherichia coli
J. Biol. Chem.
287
36208-36221
2012
-
-
-
1
5
-
-
4
-
1
-
1
-
1
-
-
-
-
-
-
-
-
3
1
2
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
1
5
-
-
-
-
4
-
1
-
1
-
-
-
-
-
-
-
-
3
1
-
-
-
4
-
-
-
-
-
-
-
-
3
3
702377
Wang
Comparison of two metal-depend ...
Escherichia coli
Biochemistry
49
3774-3782
2010
-
-
-
1
-
-
5
10
-
1
-
1
-
1
-
-
-
1
-
-
-
-
9
-
1
-
-
-
9
-
-
-
-
6
-
-
-
-
-
-
1
-
-
-
5
6
10
-
1
-
1
-
-
-
-
-
-
-
-
9
-
-
-
-
9
-
-
-
-
-
-
-
-
9
9
722258
Wang
The role of a conserved histid ...
Escherichia coli
FEBS Lett.
582
3385-3388
2008
-
-
1
-
2
1
-
4
-
1
1
-
-
1
-
-
1
-
-
-
-
-
1
-
1
-
-
-
3
-
1
-
-
-
-
-
-
-
1
-
-
2
1
-
-
-
4
-
1
1
-
-
-
-
1
-
-
-
-
1
-
-
-
-
3
-
1
-
-
-
-
-
-
3
3
722955
Rea
Structure and mechanism of Hpc ...
Escherichia coli
J. Mol. Biol.
373
866-876
2007
-
-
1
1
-
-
-
-
-
-
1
1
-
2
-
-
1
-
-
-
-
-
1
1
2
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
1
1
-
-
-
1
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
1
1
-
-
-
721601
Wang
Purification and biochemical c ...
Escherichia coli
Biochemistry
44
9447-9455
2005
-
-
1
-
1
-
2
6
-
9
1
1
-
1
-
-
1
-
-
-
-
-
5
-
1
-
-
-
3
-
-
-
-
2
-
-
-
-
1
-
-
1
-
-
2
2
6
-
9
1
1
-
-
-
1
-
-
-
-
5
-
-
-
-
3
-
-
-
-
-
1
1
-
3
3