Information on EC 2.6.1.30 - pyridoxamine-pyruvate transaminase

for references in articles please use BRENDA:EC2.6.1.30
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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
2.6.1.30
-
RECOMMENDED NAME
GeneOntology No.
pyridoxamine-pyruvate transaminase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
pyridoxamine + pyruvate = pyridoxal + L-alanine
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
amino group transfer
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-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
vitamin B6 degradation
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Vitamin B6 metabolism
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Microbial metabolism in diverse environments
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SYSTEMATIC NAME
IUBMB Comments
pyridoxamine:pyruvate aminotransferase
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CAS REGISTRY NUMBER
COMMENTARY hide
9023-38-5
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
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Manually annotated by BRENDA team
Gram-negative bacillus isolate T2, 99.9% related to Ochrobactrum anthropi
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Manually annotated by BRENDA team
Soil bacterium
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-
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Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(S)-2-aminobutyrate + pyruvate
?
show the reaction diagram
2-norpyridoxal + L-alanine
2-norpyridoxamine + pyruvate
show the reaction diagram
-
-
-
r
3-hydroxypyridine-4-aldehyde + L-alanine
3-hydroxy-4-aminomethylpyridine + pyruvate
show the reaction diagram
-
-
-
r
5-deoxypyridoxal + L-alanine
5-deoxypyridoxamine + pyruvate
show the reaction diagram
omega-methylpyridoxal + L-alanine
omega-methylpyridoxamine + pyruvate
show the reaction diagram
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i.e. 3-hydroxy-5-hydroxymethyl-2-ethylpyridine-4-carboxaldehyde
-
r
pyridoxal + L-alanine
pyridoxamine + pyruvate
show the reaction diagram
-
-
-
-
?
pyridoxamine + 2-oxobutyrate
?
show the reaction diagram
pyridoxamine + 2-oxobutyrate
pyridoxal + (S)-2-aminobutyrate
show the reaction diagram
poor substrate
-
-
r
pyridoxamine + 2-oxoglutarate
pyridoxal + L-glutamate
show the reaction diagram
-
-
-
-
r
pyridoxamine + pyruvate
pyridoxal + L-alanine
show the reaction diagram
pyridoxamine 5'-phosphate + pyruvate
pyridoxal 5'-phosphate + L-alanine
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
pyridoxamine + pyruvate
pyridoxal + L-alanine
show the reaction diagram
additional information
?
-
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enzyme is part of the degradative pathway for vitamin B6 compounds utilized by Pseudomonas sp. MA-1
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyridoxal 5'-phosphate
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K+
-
activates
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-Deoxypyridoxal
-
inhibits pyridoxamine formation from pyridoxal + alanine
4-Nitrosalicylaldehyde
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inhibits pyridoxamine formation from pyridoxal + alanine
N-Methylpyridoxal
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inhibits pyridoxamine formation from pyridoxal + alanine
O-Methylpyridoxal
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inhibits pyridoxamine formation from pyridoxal + alanine
Pyridine-4-aldehyde
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inhibits pyridoxamine formation from pyridoxal + alanine
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
phosphate is neither essential nor inhibitory
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
20
(S)-2-aminobutyrate
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; 30C, pH 9.0
0.59
2-norpyridoxal
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pH 8.9, 25C
0.16
2-Norpyridoxamine
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pH 8.9, 25C
7.1
2-oxobutyrate
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; 30C, pH 9.0
8.8 - 110.3
2-oxoglutarate
0.89
3-hydroxypyridine-4-aldehyde
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pH 8.9, 25C
0.009
5-Deoxypyridoxal
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pH 8.9, 25C
0.014
5-Deoxypyridoxamine
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pH 8.9, 25C
0.58 - 64.5
L-alanine
29.3 - 142
L-glutamate
1.3
omega-methylpyridoxal
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pH 8.9, 25C
0.012 - 0.059
pyridoxal
0.013 - 2.5
pyridoxamine
3.4
pyridoxamine 5'-phosphate
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mutant E68A, pH 9.0, 30C
0.1 - 7.4
pyruvate
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
9
(S)-2-aminobutyrate
-
; 30C, pH 9.0
24
2-oxobutyrate
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; 30C, pH 9.0
1.9 - 9.1
2-oxoglutarate
41
L-alanine
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; 30C, pH 9.0
1.5 - 6.4
L-glutamate
41
pyridoxal
-
; 30C, pH 9.0
0.77 - 8
pyridoxamine
0.062 - 0.22
pyridoxamine 5'-phosphate
29
pyruvate
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; 30C, pH 9.0
additional information
additional information
-
-
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.023 - 0.916
2-oxoglutarate
0.012 - 0.152
L-glutamate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00631
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at 30C
7.3
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purified enzyme
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
9.5
-
estimated based on the Km and kcat values
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 10
Soil bacterium
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
assay at
37
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assay at
70
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at pH 9, 100 mM borate/KOH buffer; in 100 mM borate/KOH buffer pH 9.0
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 80
-
30: 17% of maximal activity, 80C: 3% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Mesorhizobium japonicum (strain LMG 29417 / CECT 9101 / MAFF 303099)
Mesorhizobium japonicum (strain LMG 29417 / CECT 9101 / MAFF 303099)
Mesorhizobium japonicum (strain LMG 29417 / CECT 9101 / MAFF 303099)
Mesorhizobium japonicum (strain LMG 29417 / CECT 9101 / MAFF 303099)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
32000
-
x * 32000, SDS-PAGE
41589
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4 * 41589, calculated from amino acid sequence; 4 * 41589, calculated from sequence
154000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 32000, SDS-PAGE
homotetramer
tetramer
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4 * 41000, SDS-PAGE; 4 * 41589, calculated from sequence
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme, space group P43212, diffraction to 2.0 A resolution. Complexes with pyridoxamine, pyridoxal, and pyridoxyl-L-alanine at 1.7 A, 1.7 A, and 2.0 A resolution, respectively. Enzyme is a homotetramer and each subunit is composed of a large N-terminal domain, consisting of seven beta-sheets and eight alpha-helices, and a smaller C-terminal domain, consisting of three beta-sheets and four alpha-helices. The substrate pyridoxal is bound through an aldimine linkage to Lys197 in the active site. The carboxylate group of the substrate amino/keto acid is hydrogen-bonded to Arg336 and Arg345
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addition of saturated ammonium sulfate solution to the concentrated protein solution until the first permanent turbidity appears, several days at 5C
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 80
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17% and 3% of the maximal activity is observed at 30C and 80C, respectively
70
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10 min, stable
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
K+ stabilizes
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
loss of activity within 12 h at 25C in sodium phosphate buffer, but is stable in potassium buffer under the same conditions
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
from tobacco leaf
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hydrophobic interaction chromatography, gel filtration
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partial; partial, QA52 column chromatography and butyl-Toyopearl column chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3)
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expressed in Escherichia coli strains JM109 and BL21(DE3); expression in Escherichia coli
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expression in Escherichia coli
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C198A
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enzyme shows the same specific activity as that of the wild type enzyme; shows activity of the wild type enzyme
E68A
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low pyridoxamine 5'-phosphate:pyruvate aminotransferase activity, decrease in activity towards pyridoxamine
E68G
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low pyridoxamine 5'-phosphate:pyruvate aminotransferase activity, decrease in activity towards pyridoxamine
K197L
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mutant enzyme shows no activity; shows no activity
M2T/V70K
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M2: surface residue
M2T/V70K/E212G
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E212: surface residue
M2T/Y35H/V70K
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Y35: active site residur
M2T/Y35H/V70K/E212G
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E212: surface residue
R336A
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significant decrease in affinity for pyruvate
V70K
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active site residue, increase in reactivity toward 2-oxoglutarate
V70K/E212G
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E212: surface residue
V70R
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active site residue, elimination of activity
V70R/F247C
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F247: hydrophobic core residue
Y35H/V70K
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active site residues
Y35H/V70R/F247C
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F247: hydrophobic core residue
C198A
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shows activity of the wild type enzyme
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E68A
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low pyridoxamine 5'-phosphate:pyruvate aminotransferase activity, decrease in activity towards pyridoxamine
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E68G
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low pyridoxamine 5'-phosphate:pyruvate aminotransferase activity, decrease in activity towards pyridoxamine
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K197L
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shows no activity
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R336A
-
significant decrease in affinity for pyruvate
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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determination method for individual natural vitamin B6 compounds. Compounds are specifically converted into 4-pyridoxolactone by pyridoxal 4-dehydrogenase and coupling reactions involving pyridoxine 4-oxidase, pyridoxal 4-dehydrogenase, or pyridoxamine-pyruvate aminotransferase and pyridoxal 4-dehydrogenase. Application of method for analysis of food samples