BRENDA - Enzyme Database show
show all sequences of 2.6.1.30

Molecular cloning, expression and characterization of pyridoxamine-pyruvate aminotransferase

Yoshikane, Y.; Yokochi, N.; Ohnishi, K.; Hayashi, H.; Yagi, T.; Biochem. J. 396, 499-507 (2006)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli strains JM109 and BL21(DE3); expression in Escherichia coli
Mesorhizobium loti
Engineering
Amino acid exchange
Commentary
Organism
C198A
enzyme shows the same specific activity as that of the wild type enzyme; shows activity of the wild type enzyme
Mesorhizobium loti
K197L
mutant enzyme shows no activity; shows no activity
Mesorhizobium loti
Inhibitors
Inhibitors
Commentary
Organism
Structure
additional information
hydroxylamine (10 mM), phenylhydrazine (1 mM) and sodium borohydride (1 mM) do not inactivate the enzyme
Mesorhizobium loti
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.044
-
pyridoxamine
; 30°C, pH 9.0
Mesorhizobium loti
0.059
-
pyridoxal
; 30°C, pH 9.0
Mesorhizobium loti
0.34
-
pyruvate
; 30°C, pH 9.0
Mesorhizobium loti
7.1
-
2-oxobutyrate
; 30°C, pH 9.0
Mesorhizobium loti
11
-
L-alanine
; 30°C, pH 9.0
Mesorhizobium loti
20
-
(S)-2-aminobutyrate
; 30°C, pH 9.0
Mesorhizobium loti
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
41000
-
4 * 41000, SDS-PAGE; 4 * 41000, SDS-PAGE
Mesorhizobium loti
41589
-
4 * 41589, calculated from amino acid sequence; 4 * 41589, calculated from sequence
Mesorhizobium loti
154000
-
gel filtration; gel filtration
Mesorhizobium loti
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Mesorhizobium loti
-
-
-
Mesorhizobium loti MAFF303099
-
-
-
Mesorhizobium loti
Q988B8
-
-
Purification (Commentary)
Commentary
Organism
partial; partial, QA52 column chromatography and butyl-Toyopearl column chromatography
Mesorhizobium loti
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(S)-2-aminobutyrate + pyruvate
(S)-2-aminobutyrate is a poor substrate
671864
Mesorhizobium loti
?
-
-
-
?
(S)-2-aminobutyrate + pyruvate
(S)-2-aminobutyrate is a poor substrate
671864
Mesorhizobium loti MAFF303099
?
-
-
-
?
additional information
the enzyme shows less than 0.5% of the activity of pyridoxamine towards pyridoxamine 5'-phosphate (3.3 mM) with pyruvate as the amino acceptor, the enzyme shows less than 0.5% of the activity of pyruvate towards 2-oxoglutarate, 3-hydroxypyruvate, indol-pyruvate, phenylpyruvate, 3-methyl-2-oxobutyrate and 4-hydroxyphenylpyruvate when they are used at 10 mM
671864
Mesorhizobium loti
?
-
-
-
-
additional information
the enzyme shows less than 0.5% of the activity of pyridoxamine towards pyridoxamine 5'-phosphate (3.3 mM) with pyruvate as the amino acceptor, the enzyme shows less than 0.5% of the activity of pyruvate towards 2-oxoglutarate, 3-hydroxypyruvate, indol-pyruvate, phenylpyruvate, 3-methyl-2-oxobutyrate and 4-hydroxyphenylpyruvate when they are used at 10 mM
671864
Mesorhizobium loti MAFF303099
?
-
-
-
-
pyridoxamine + 2-oxobutyrate
-
671864
Mesorhizobium loti
?
-
-
-
?
pyridoxamine + 2-oxobutyrate
-
671864
Mesorhizobium loti MAFF303099
?
-
-
-
?
pyridoxamine + 2-oxobutyrate
poor substrate
671864
Mesorhizobium loti
pyridoxal + (S)-2-aminobutyrate
-
-
-
r
pyridoxamine + pyruvate
-
671864
Mesorhizobium loti
pyridoxal + L-alanine
-
-
-
?
pyridoxamine + pyruvate
-
671864
Mesorhizobium loti
pyridoxal + L-alanine
-
-
-
r
pyridoxamine + pyruvate
-
671864
Mesorhizobium loti MAFF303099
pyridoxal + L-alanine
-
-
-
?
Subunits
Subunits
Commentary
Organism
homotetramer
4 * 41000, SDS-PAGE; 4 * 41589, calculated from amino acid sequence
Mesorhizobium loti
tetramer
4 * 41000, SDS-PAGE; 4 * 41589, calculated from sequence
Mesorhizobium loti
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
70
-
at pH 9, 100 mM borate/KOH buffer; in 100 mM borate/KOH buffer pH 9.0
Mesorhizobium loti
Temperature Range [°C]
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
30
80
30: 17% of maximal activity, 80°C: 3% of maximal activity
Mesorhizobium loti
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
30
80
17% and 3% of the maximal activity is observed at 30°C and 80°C, respectively
Mesorhizobium loti
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2
8
pyridoxamine
; 30°C, pH 9.0
Mesorhizobium loti
9
-
(S)-2-aminobutyrate
; 30°C, pH 9.0
Mesorhizobium loti
24
-
2-oxobutyrate
; 30°C, pH 9.0
Mesorhizobium loti
29
-
pyruvate
; 30°C, pH 9.0
Mesorhizobium loti
41
-
L-alanine
; 30°C, pH 9.0
Mesorhizobium loti
41
-
pyridoxal
; 30°C, pH 9.0
Mesorhizobium loti
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
9.5
-
estimated based on the Km and kcat values
Mesorhizobium loti
Cofactor
Cofactor
Commentary
Organism
Structure
additional information
pyridoxal 5'-phosphate-independent enzyme
Mesorhizobium loti
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli strains JM109 and BL21(DE3)
Mesorhizobium loti
expression in Escherichia coli
Mesorhizobium loti
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
additional information
pyridoxal 5'-phosphate-independent enzyme
Mesorhizobium loti
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
C198A
enzyme shows the same specific activity as that of the wild type enzyme
Mesorhizobium loti
C198A
shows activity of the wild type enzyme
Mesorhizobium loti
K197L
mutant enzyme shows no activity
Mesorhizobium loti
K197L
shows no activity
Mesorhizobium loti
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
additional information
hydroxylamine (10 mM), phenylhydrazine (1 mM) and sodium borohydride (1 mM) do not inactivate the enzyme
Mesorhizobium loti
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.044
-
pyridoxamine
30°C, pH 9.0
Mesorhizobium loti
0.044
-
pyridoxamine
-
Mesorhizobium loti
0.059
-
pyridoxal
30°C, pH 9.0
Mesorhizobium loti
0.059
-
pyridoxal
-
Mesorhizobium loti
0.34
-
pyruvate
30°C, pH 9.0
Mesorhizobium loti
0.34
-
pyruvate
-
Mesorhizobium loti
7.1
-
2-oxobutyrate
30°C, pH 9.0
Mesorhizobium loti
7.1
-
2-oxobutyrate
-
Mesorhizobium loti
11
-
L-alanine
30°C, pH 9.0
Mesorhizobium loti
11
-
L-alanine
-
Mesorhizobium loti
20
-
(S)-2-aminobutyrate
30°C, pH 9.0
Mesorhizobium loti
20
-
(S)-2-aminobutyrate
-
Mesorhizobium loti
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
41000
-
4 * 41000, SDS-PAGE
Mesorhizobium loti
41589
-
4 * 41589, calculated from sequence
Mesorhizobium loti
41589
-
4 * 41589, calculated from amino acid sequence
Mesorhizobium loti
154000
-
gel filtration
Mesorhizobium loti
Purification (Commentary) (protein specific)
Commentary
Organism
partial
Mesorhizobium loti
partial, QA52 column chromatography and butyl-Toyopearl column chromatography
Mesorhizobium loti
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(S)-2-aminobutyrate + pyruvate
(S)-2-aminobutyrate is a poor substrate
671864
Mesorhizobium loti
?
-
-
-
?
(S)-2-aminobutyrate + pyruvate
(S)-2-aminobutyrate is a poor substrate
671864
Mesorhizobium loti MAFF303099
?
-
-
-
?
additional information
the enzyme shows less than 0.5% of the activity of pyridoxamine towards pyridoxamine 5'-phosphate (3.3 mM) with pyruvate as the amino acceptor, the enzyme shows less than 0.5% of the activity of pyruvate towards 2-oxoglutarate, 3-hydroxypyruvate, indol-pyruvate, phenylpyruvate, 3-methyl-2-oxobutyrate and 4-hydroxyphenylpyruvate when they are used at 10 mM
671864
Mesorhizobium loti
?
-
-
-
-
additional information
the enzyme shows less than 0.5% of the activity of pyridoxamine towards pyridoxamine 5'-phosphate (3.3 mM) with pyruvate as the amino acceptor, the enzyme shows less than 0.5% of the activity of pyruvate towards 2-oxoglutarate, 3-hydroxypyruvate, indol-pyruvate, phenylpyruvate, 3-methyl-2-oxobutyrate and 4-hydroxyphenylpyruvate when they are used at 10 mM
671864
Mesorhizobium loti MAFF303099
?
-
-
-
-
pyridoxamine + 2-oxobutyrate
-
671864
Mesorhizobium loti
?
-
-
-
?
pyridoxamine + 2-oxobutyrate
-
671864
Mesorhizobium loti MAFF303099
?
-
-
-
?
pyridoxamine + 2-oxobutyrate
poor substrate
671864
Mesorhizobium loti
pyridoxal + (S)-2-aminobutyrate
-
-
-
r
pyridoxamine + pyruvate
-
671864
Mesorhizobium loti
pyridoxal + L-alanine
-
-
-
?
pyridoxamine + pyruvate
-
671864
Mesorhizobium loti
pyridoxal + L-alanine
-
-
-
r
pyridoxamine + pyruvate
-
671864
Mesorhizobium loti MAFF303099
pyridoxal + L-alanine
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
homotetramer
4 * 41000, SDS-PAGE; 4 * 41589, calculated from amino acid sequence
Mesorhizobium loti
tetramer
4 * 41000, SDS-PAGE; 4 * 41589, calculated from sequence
Mesorhizobium loti
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
70
-
at pH 9, 100 mM borate/KOH buffer
Mesorhizobium loti
70
-
in 100 mM borate/KOH buffer pH 9.0
Mesorhizobium loti
Temperature Range [°C] (protein specific)
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
30
80
30: 17% of maximal activity, 80°C: 3% of maximal activity
Mesorhizobium loti
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
30
80
17% and 3% of the maximal activity is observed at 30°C and 80°C, respectively
Mesorhizobium loti
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2
8
pyridoxamine
30°C, pH 9.0
Mesorhizobium loti
2
8
pyridoxamine
-
Mesorhizobium loti
9
-
(S)-2-aminobutyrate
30°C, pH 9.0
Mesorhizobium loti
9
-
(S)-2-aminobutyrate
-
Mesorhizobium loti
24
-
2-oxobutyrate
30°C, pH 9.0
Mesorhizobium loti
24
-
2-oxobutyrate
-
Mesorhizobium loti
29
-
pyruvate
30°C, pH 9.0
Mesorhizobium loti
29
-
pyruvate
-
Mesorhizobium loti
41
-
L-alanine
30°C, pH 9.0
Mesorhizobium loti
41
-
L-alanine
-
Mesorhizobium loti
41
-
pyridoxal
30°C, pH 9.0
Mesorhizobium loti
41
-
pyridoxal
-
Mesorhizobium loti
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
9.5
-
estimated based on the Km and kcat values
Mesorhizobium loti
Other publictions for EC 2.6.1.30
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
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)
739366
Huang
Enzymatic transamination of py ...
Nicotiana tabacum
Plant Sci.
212
55-59
2013
-
-
-
-
-
-
1
2
-
-
1
-
-
1
-
-
1
-
-
1
-
-
3
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
2
-
-
1
-
-
-
-
1
-
1
-
-
3
1
1
-
-
-
1
-
-
-
-
-
-
-
-
-
722999
Yoshikane
-
Engineering Mesorhizobium loti ...
Mesorhizobium loti
J. Mol. Catal. B
67
104-110
2010
-
-
1
-
10
-
-
17
-
-
2
-
-
1
-
-
1
-
-
-
-
-
2
1
-
-
-
16
-
-
-
-
-
-
-
-
-
1
-
-
10
-
-
-
-
17
-
-
2
-
-
-
-
1
-
-
-
-
2
1
-
-
-
16
-
-
-
-
-
-
-
-
17
17
687694
Yoshikane
Crystal structure of pyridoxam ...
Mesorhizobium loti, Mesorhizobium loti MAFF303099
J. Biol. Chem.
283
1120-1127
2008
-
-
1
1
3
-
-
7
-
-
-
-
-
4
-
-
-
1
-
-
-
-
4
-
-
-
-
5
-
-
-
1
-
-
-
-
-
1
1
1
3
-
-
-
-
7
-
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
5
-
-
-
-
-
-
-
-
-
-
688568
Nishimura
Determination of individual vi ...
Mesorhizobium loti
J. Nutr. Sci. Vitaminol.
54
18-24
2008
-
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
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1
-
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-
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-
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-
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-
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-
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-
-
-
-
671864
Yoshikane
Molecular cloning, expression ...
Mesorhizobium loti, Mesorhizobium loti MAFF303099
Biochem. J.
396
499-507
2006
-
-
1
-
2
-
1
6
-
-
3
-
-
14
-
-
1
-
-
-
-
-
10
2
1
1
1
6
1
-
-
1
-
-
-
-
-
2
1
-
4
-
-
1
-
12
-
-
6
-
-
-
-
2
-
-
-
-
10
2
2
1
1
12
1
-
-
-
-
-
-
-
-
-
670891
Yanee
-
Vitamin B6 degradation by pyri ...
Ochrobactrum anthropi
Science Asia
31
307-311
2005
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
639958
Ford
-
Effect of pH and temperature o ...
Soil bacterium
J. Ala. Acad. Sci.
49
16-30
1978
-
-
-
-
-
-
-
1
-
-
-
1
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
1
-
-
-
-
-
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-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
639959
Gilmer
Pyridoxamine-pyruvate transami ...
Pseudomonas sp.
Biochemistry
16
5241-5246
1977
-
-
-
-
-
1
-
1
-
1
-
2
-
1
-
-
-
-
-
-
-
1
3
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
1
-
1
-
2
-
-
-
-
-
-
-
1
3
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
639960
Gilmer
Pyridoxamine-pyruvate transami ...
Pseudomonas sp.
Biochemistry
16
5246-5253
1977
-
-
-
-
-
-
-
1
-
-
-
1
-
1
-
-
-
1
-
-
-
-
2
-
1
-
-
-
-
-
-
-
-
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-
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-
-
-
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-
1
-
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-
1
-
-
-
-
-
-
-
-
2
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
639957
Ayling
Relation of structure to activ ...
Pseudomonas sp.
Biochemistry
7
1626-1636
1968
-
-
-
-
-
-
5
20
-
-
-
1
-
1
-
-
-
-
-
-
-
-
6
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
5
-
20
-
-
-
1
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639956
Wada
Enzymatic transamination pf py ...
Pseudomonas sp.
J. Biol. Chem.
237
133-137
1962
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