BRENDA - Enzyme Database show
show all sequences of 2.6.1.30

Engineering Mesorhizobium loti pyridoxamine-pyruvate aminotransferase for production of pyridoxamine with L-glutamate as an amino donor

Yoshikane, Y.; Tamura, A.; Yokochi, N.; Ellouze, K.; Yamamura, E.; Mizunaga, H.; Fujimoto, N.; Sakamoto, K.; Sawa, Y.; Yagi, T.; J. Mol. Catal. B 67, 104-110 (2010)
No PubMed abstract available

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli BL21(DE3)
Mesorhizobium loti
Engineering
Amino acid exchange
Commentary
Organism
M2T/V70K
M2: surface residue
Mesorhizobium loti
M2T/V70K/E212G
E212: surface residue
Mesorhizobium loti
M2T/Y35H/V70K
Y35: active site residur
Mesorhizobium loti
M2T/Y35H/V70K/E212G
E212: surface residue
Mesorhizobium loti
V70K
active site residue, increase in reactivity toward 2-oxoglutarate
Mesorhizobium loti
V70K/E212G
E212: surface residue
Mesorhizobium loti
V70R
active site residue, elimination of activity
Mesorhizobium loti
V70R/F247C
F247: hydrophobic core residue
Mesorhizobium loti
Y35H/V70K
active site residues
Mesorhizobium loti
Y35H/V70R/F247C
F247: hydrophobic core residue
Mesorhizobium loti
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
8.8
-
2-oxoglutarate
Y35H/V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
12.6
-
2-oxoglutarate
Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
17.2
-
2-oxoglutarate
M2T/Y35H/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
21.1
-
2-oxoglutarate
M2T/Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
29.3
-
L-glutamate
Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
42.4
-
L-glutamate
Y35H/V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
44.2
-
L-glutamate
M2T/Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
51
-
L-glutamate
M2T/Y35H/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
61.3
-
2-oxoglutarate
V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
77.3
-
2-oxoglutarate
M2T/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
80.1
-
2-oxoglutarate
wild type protein, pH 9.0, 30°C
Mesorhizobium loti
102.6
-
2-oxoglutarate
V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
106.8
-
2-oxoglutarate
M2T/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
110.3
-
2-oxoglutarate
V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
123.4
-
L-glutamate
wild type protein, pH 9.0, 30°C
Mesorhizobium loti
133.5
-
L-glutamate
V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
142
-
L-glutamate
V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
41000
-
4 * 41000, SDS-PAGE, gel filtration
Mesorhizobium loti
154000
-
gel filtration
Mesorhizobium loti
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Mesorhizobium loti
Q988B8
-
-
Purification (Commentary)
Commentary
Organism
hydrophobic interaction chromatography, gel filtration
Mesorhizobium loti
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
pyridoxamine + 2-oxoglutarate
-
722999
Mesorhizobium loti
pyridoxal + L-glutamate
-
-
-
r
pyridoxamine + pyruvate
-
722999
Mesorhizobium loti
pyridoxal + L-alanine
-
-
-
r
Subunits
Subunits
Commentary
Organism
homotetramer
4 * 41000, SDS-PAGE, gel filtration
Mesorhizobium loti
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.5
-
L-glutamate
wild type protein, pH 9.0, 30°C
Mesorhizobium loti
1.9
-
2-oxoglutarate
wild type protein, pH 9.0, 30°C
Mesorhizobium loti
2.7
-
L-glutamate
V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
2.9
-
L-glutamate
Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
3
-
L-glutamate
M2T/Y35H/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
3.1
-
L-glutamate
M2T/Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
3.5
-
2-oxoglutarate
M2T/Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
3.9
-
2-oxoglutarate
M2T/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
4
-
2-oxoglutarate
M2T/Y35H/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
4.1
-
2-oxoglutarate
V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
4.4
-
L-glutamate
V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
4.6
-
2-oxoglutarate
M2T/V70K/E212G mutant protein, pH 9.0, 30°C; V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
5.2
-
2-oxoglutarate
Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
6.4
-
L-glutamate
Y35H/V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
8.1
-
2-oxoglutarate
Y35H/V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
9.1
-
2-oxoglutarate
V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli BL21(DE3)
Mesorhizobium loti
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
M2T/V70K
M2: surface residue
Mesorhizobium loti
M2T/V70K/E212G
E212: surface residue
Mesorhizobium loti
M2T/Y35H/V70K
Y35: active site residur
Mesorhizobium loti
M2T/Y35H/V70K/E212G
E212: surface residue
Mesorhizobium loti
V70K
active site residue, increase in reactivity toward 2-oxoglutarate
Mesorhizobium loti
V70K/E212G
E212: surface residue
Mesorhizobium loti
V70R
active site residue, elimination of activity
Mesorhizobium loti
V70R/F247C
F247: hydrophobic core residue
Mesorhizobium loti
Y35H/V70K
active site residues
Mesorhizobium loti
Y35H/V70R/F247C
F247: hydrophobic core residue
Mesorhizobium loti
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
8.8
-
2-oxoglutarate
Y35H/V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
12.6
-
2-oxoglutarate
Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
17.2
-
2-oxoglutarate
M2T/Y35H/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
21.1
-
2-oxoglutarate
M2T/Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
29.3
-
L-glutamate
Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
42.4
-
L-glutamate
Y35H/V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
44.2
-
L-glutamate
M2T/Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
51
-
L-glutamate
M2T/Y35H/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
61.3
-
2-oxoglutarate
V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
77.3
-
2-oxoglutarate
M2T/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
80.1
-
2-oxoglutarate
wild type protein, pH 9.0, 30°C
Mesorhizobium loti
102.6
-
2-oxoglutarate
V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
106.8
-
2-oxoglutarate
M2T/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
110.3
-
2-oxoglutarate
V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
123.4
-
L-glutamate
wild type protein, pH 9.0, 30°C
Mesorhizobium loti
133.5
-
L-glutamate
V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
142
-
L-glutamate
V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
41000
-
4 * 41000, SDS-PAGE, gel filtration
Mesorhizobium loti
154000
-
gel filtration
Mesorhizobium loti
Purification (Commentary) (protein specific)
Commentary
Organism
hydrophobic interaction chromatography, gel filtration
Mesorhizobium loti
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
pyridoxamine + 2-oxoglutarate
-
722999
Mesorhizobium loti
pyridoxal + L-glutamate
-
-
-
r
pyridoxamine + pyruvate
-
722999
Mesorhizobium loti
pyridoxal + L-alanine
-
-
-
r
Subunits (protein specific)
Subunits
Commentary
Organism
homotetramer
4 * 41000, SDS-PAGE, gel filtration
Mesorhizobium loti
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.5
-
L-glutamate
wild type protein, pH 9.0, 30°C
Mesorhizobium loti
1.9
-
2-oxoglutarate
wild type protein, pH 9.0, 30°C
Mesorhizobium loti
2.7
-
L-glutamate
V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
2.9
-
L-glutamate
Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
3
-
L-glutamate
M2T/Y35H/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
3.1
-
L-glutamate
M2T/Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
3.5
-
2-oxoglutarate
M2T/Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
3.9
-
2-oxoglutarate
M2T/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
4
-
2-oxoglutarate
M2T/Y35H/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
4.1
-
2-oxoglutarate
V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
4.4
-
L-glutamate
V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
4.6
-
2-oxoglutarate
M2T/V70K/E212G mutant protein, pH 9.0, 30°C; V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
5.2
-
2-oxoglutarate
Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
6.4
-
L-glutamate
Y35H/V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
8.1
-
2-oxoglutarate
Y35H/V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
9.1
-
2-oxoglutarate
V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.012
-
L-glutamate
wild type protein, pH 9.0, 30°C
Mesorhizobium loti
0.019
-
L-glutamate
V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.023
-
2-oxoglutarate
wild type protein, pH 9.0, 30°C
Mesorhizobium loti
0.033
-
L-glutamate
V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.04
-
2-oxoglutarate
V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.042
-
2-oxoglutarate
V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.043
-
2-oxoglutarate
M2T/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.05
-
2-oxoglutarate
M2T/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.058
-
L-glutamate
M2T/Y35H/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.07
-
L-glutamate
M2T/Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.1
-
L-glutamate
Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.148
-
2-oxoglutarate
V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.152
-
L-glutamate
Y35H/V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.167
-
2-oxoglutarate
M2T/Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.234
-
2-oxoglutarate
M2T/Y35H/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.408
-
2-oxoglutarate
Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.916
-
2-oxoglutarate
Y35H/V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.012
-
L-glutamate
wild type protein, pH 9.0, 30°C
Mesorhizobium loti
0.019
-
L-glutamate
V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.023
-
2-oxoglutarate
wild type protein, pH 9.0, 30°C
Mesorhizobium loti
0.033
-
L-glutamate
V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.04
-
2-oxoglutarate
V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.042
-
2-oxoglutarate
V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.043
-
2-oxoglutarate
M2T/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.05
-
2-oxoglutarate
M2T/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.058
-
L-glutamate
M2T/Y35H/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.07
-
L-glutamate
M2T/Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.1
-
L-glutamate
Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.148
-
2-oxoglutarate
V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.152
-
L-glutamate
Y35H/V70R/F247C mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.167
-
2-oxoglutarate
M2T/Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.234
-
2-oxoglutarate
M2T/Y35H/V70K/E212G mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.408
-
2-oxoglutarate
Y35H/V70K mutant protein, pH 9.0, 30°C
Mesorhizobium loti
0.916
-
2-oxoglutarate
Y35H/V70R/F247C mutant protein, pH 9.0, 30°C
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
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-
1
1
3
-
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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.
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Vitamin B6 degradation by pyri ...
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