Information on EC 5.1.1.1 - Alanine racemase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY
5.1.1.1
-
RECOMMENDED NAME
GeneOntology No.
Alanine racemase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
L-alanine = D-alanine
show the reaction diagram
-
-
-
-
L-alanine = D-alanine
show the reaction diagram
the active site of the alanine racemase reacts asymmetrically with the enantiomers of the substrate and has a conformation which greatly favors the D-enantiomer
-
L-alanine = D-alanine
show the reaction diagram
the alanine racemase builds two different bases in the active site. The base for D-Ala may be closer to the enzyme surface, and that for L-Ala inside
-
L-alanine = D-alanine
show the reaction diagram
consensus water sites at the interface between the two enzyme monomers maintain and stabilize dimer and supply the active site continuously with water molecules to allow rapid equilibration of active site protons
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
isomerization
-
stereoisomerization
racemization
-
-
-
-
racemization
-
-
racemization
-
-
racemization
P10725, P94494
;
racemization
-
-
racemization
B9WYE8, -
-
racemization
-
-
racemization
-
-
racemization
-
-
racemization
Bacillus subtilis 168
-
;
-
racemization
Medicago sativa L.
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
alanine biosynthesis I
-
alanine degradation I
-
anaerobic energy metabolism (invertebrates, cytosol)
-
D-Alanine metabolism
-
Metabolic pathways
-
SYSTEMATIC NAME
IUBMB Comments
Alanine racemase
A pyridoxal-phosphate protein.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
alanine racemase
-
-
alanine racemase
-
-
alanine racemase
Q81VF6
-
alanine racemase
-
-
alanine racemase
B3VI72
-
alanine racemase
B3VI72
-
-
alanine racemase
P10725, P94494
-
alanine racemase
Bacillus subtilis 168
P10725, P94494
-
-
alanine racemase
Q208U6
-
alanine racemase
-
-
alanine racemase
-
-
alanine racemase
-
-
alanine racemase
Q837J0
-
alanine racemase
-
-
alanine racemase
P0A6B4
-
alanine racemase
P10724
-
alanine racemase
P45257
-
alanine racemase
Q1XG01
-
alanine racemase
-
-
alanine racemase
-
-
alanine racemase
Medicago sativa L.
-
-
-
alanine racemase
P0A4X2
-
alanine racemase
Q9HTQ2
-
alanine racemase
P0A2W8
-
AlaR
Q837J0
; gene name
AlaR
P0A6B4
-
AlaR
Q1XG01
-
ALR
Pseudomonas putida YZ-26
-
-
-
AlrBax
Q81VF6
-
BA0252
Q81VF6
-
BAS0238
Q81VF6
-
D-alanine racemase
-
-
DadX
Pseudomonas fluorescens LRB3W1, Pseudomonas fluorescens TM5-2
-
-
-
dadXOF4
-
gene name
dadXOF4
B3VI72
gene name
dadXOF4
B3VI72
gene name
-
dal1
Q81VF6
gene name of AlrBax
L-Alanine racemase
-
-
-
-
L-Alanine:D-alanine racemase
-
-
-
-
Racemase, alanine
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9024-06-0
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Acidiphilium organovorum 13H
13H
-
-
Manually annotated by BRENDA team
strain YM-1
-
-
Manually annotated by BRENDA team
strain YM-1
-
-
Manually annotated by BRENDA team
strain 168
SwissProt
Manually annotated by BRENDA team
Bacillus subtilis 168
strain 168
SwissProt
Manually annotated by BRENDA team
subsp. Kurstaki
SwissProt
Manually annotated by BRENDA team
alanine racemase isoform 1
UniProt
Manually annotated by BRENDA team
alanine racemase isoform 2
UniProt
Manually annotated by BRENDA team
alanine racemase isoform 1
UniProt
Manually annotated by BRENDA team
alanine racemase isoform 2
UniProt
Manually annotated by BRENDA team
O-carbamoyl-D-Ser-resistant mutant
-
-
Manually annotated by BRENDA team
strain B
-
-
Manually annotated by BRENDA team
NCTC 11637
SwissProt
Manually annotated by BRENDA team
Medicago sativa L.
L.
-
-
Manually annotated by BRENDA team
no activity in Bacillus subtilis
strain DN1686, D-Ala auxotrophic mutant
-
-
Manually annotated by BRENDA team
no activity in Bacillus subtilis DN1686
strain DN1686, D-Ala auxotrophic mutant
-
-
Manually annotated by BRENDA team
PAO1, two independent alanine racemases: DadX and Alr
-
-
Manually annotated by BRENDA team
strain A237
-
-
Manually annotated by BRENDA team
Pseudomonas aeruginosa A237
strain A237
-
-
Manually annotated by BRENDA team
soil-borne strain LRB3W1; strain TM5-2
-
-
Manually annotated by BRENDA team
Pseudomonas fluorescens LRB3W1
-
-
-
Manually annotated by BRENDA team
Pseudomonas fluorescens TM5-2
-
-
-
Manually annotated by BRENDA team
Pseudomonas fluorescens TM5-2
strain TM5-2
-
-
Manually annotated by BRENDA team
Pseudomonas putida YZ-26
-
-
-
Manually annotated by BRENDA team
encoded by the dadB and alr gene
-
-
Manually annotated by BRENDA team
Salmonella enterica subsp. enterica serovar Typhimurium overproducing
overproducing strain
-
-
Manually annotated by BRENDA team
Thalassiosira sp.
culture under germ-free conditions
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-, Q62JA4
during log phase growth without D-alanine, the viable counts of alanine racemase-deficient mutants of Burkholderia pseudomallei decrease within 2 h by about 10fold, and no viable bacteria are present at 24 h
malfunction
-, Q63ME6, Q63SZ2
during log phase growth without D-alanine, the viable counts of alanine racemase-deficient mutants of Burkholderia pseudomallei decrease within 2 h by about 1000fold, and no viable bacteria are present at 24 h. The alanine racemase-deficient mutant of Burkholderia pseudomallei K96243 exhibits attenuation versus its isogenic parental strain with respect to growth and survival in murine peritoneal macrophages; during log phase growth without D-alanine, the viable counts of alanine racemase-deficient mutants of Burkholderia pseudomallei decrease within 2 h by about 1000fold, and no viable bacteria are present at 24 h. The alanine racemase-deficient mutant of Burkholderia pseudomallei K96243 exhibits attenuation versus its isogenic parental strain with respect to growth and survival in murine peritoneal macrophages
malfunction
-
during log phase growth without D-alanine, the viable counts of alanine racemase-deficient mutants of Burkholderia pseudomallei decrease within 2 h by about 10fold, and no viable bacteria are present at 24 h
-
malfunction
-
during log phase growth without D-alanine, the viable counts of alanine racemase-deficient mutants of Burkholderia pseudomallei decrease within 2 h by about 1000fold, and no viable bacteria are present at 24 h. The alanine racemase-deficient mutant of Burkholderia pseudomallei K96243 exhibits attenuation versus its isogenic parental strain with respect to growth and survival in murine peritoneal macrophages; during log phase growth without D-alanine, the viable counts of alanine racemase-deficient mutants of Burkholderia pseudomallei decrease within 2 h by about 1000fold, and no viable bacteria are present at 24 h. The alanine racemase-deficient mutant of Burkholderia pseudomallei K96243 exhibits attenuation versus its isogenic parental strain with respect to growth and survival in murine peritoneal macrophages
-
metabolism
-
important for cell wall biosynthesis
metabolism
-
biosynthesis of D-alanine as building blocks in the peptidoglycan layers of bacterial cell walls
metabolism
-, Q837J0
important for peptidoglycan biosynthesis
metabolism
-
important for peptidoglycan biosynthesis
metabolism
P45257
important for peptidoglycan biosynthesis
metabolism
Q1XG01
important for peptidoglycan biosynthesis
metabolism
-
important for peptidoglycan biosynthesis
metabolism
-
plays a role in spore germination and cell wall biosynthesis
physiological function
-
alanine racemase plays an essential role in cell wall synthesis as it racemizes L-alanine into D-alanine, a key building block in the biosynthesis of peptidoglycan
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
D-Ala
L-Ala
show the reaction diagram
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
-
-
-
-
?
D-alanine
L-alanine
show the reaction diagram
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
Thalassiosira sp.
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
Q65YW7, -
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
P0A2W8, -
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
Pseudomonas fluorescens TM5-2
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
Pseudomonas fluorescens LRB3W1
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
Medicago sativa L.
-
-
-
-
r
D-serine
L-serine
show the reaction diagram
-
-
-
-
r
L-2-Aminobutyrate
D-2-Aminobutyrate
show the reaction diagram
O59828, -
0.37% of the activity with L-Ala
-
r
L-2-aminobutyric acid
D-2-aminobutyric acid
show the reaction diagram
-
18% of the activity with L-Ala
-
?
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
r
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
r
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
r
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
?
L-Ala
D-Ala
show the reaction diagram
Q932V0, -
-
-
r
L-Ala
D-Ala
show the reaction diagram
P0A6B6, -
-
-
r
L-Ala
D-Ala
show the reaction diagram
P0A6B5, -
-
-
r
L-Ala
D-Ala
show the reaction diagram
Q93HP9, -
-
-
r
L-Ala
D-Ala
show the reaction diagram
-, Q9S5V6
-
-
r
L-Ala
D-Ala
show the reaction diagram
P0A4X2
-
-
r
L-Ala
D-Ala
show the reaction diagram
O59828, -
-
-
r
L-Ala
D-Ala
show the reaction diagram
-
specific for Ala
-
-
L-Ala
D-Ala
show the reaction diagram
-
specific for Ala
-
-
L-Ala
D-Ala
show the reaction diagram
-
highly specific for Ala
-
r
L-Ala
D-Ala
show the reaction diagram
-
highly specific for Ala
-
r
L-Ala
D-Ala
show the reaction diagram
-
the enzyme catalyzes transamination as a side function.The pyridoxal form of the enzyme is converted to the pyridoxamine form by incubation with its natural substrate, D-alanine or L-alanine, under acidic conditions: the enzyme loses its racemase activity concomitantly. The pyridoxamine form of the enzyme returns to the pyridoxal form by incubation with pyruvate at alkaline pH
-
r
L-Ala
D-Ala
show the reaction diagram
-
the ratio of the activity for conversion of D-alanine to L-alanine to that of the reverse conversion is constantly about 0.5 in the pH range 7-9.5
-
r
L-Ala
D-Ala
show the reaction diagram
-
Tyr265 and Lys39 are the catalytic bases removing alpha-hydrogen from L- and D-alanine
-
r
L-Ala
D-Ala
show the reaction diagram
-
enzyme is required for production of D-Ala, a necessary component of the bacterial cell wall
-
?
L-Ala
D-Ala
show the reaction diagram
-
the enzyme catalyzes the first committed step in bacterial cell wall biosynthesis
-
?
L-Ala
D-Ala
show the reaction diagram
Pseudomonas aeruginosa A237
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
Salmonella enterica subsp. enterica serovar Typhimurium overproducing
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
Acidiphilium organovorum 13H
-
highly specific for Ala
-
r
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
?
show the reaction diagram
-
-
-
-
-
L-Ala
?
show the reaction diagram
-
enzyme provides D-Ala as an essential building block for biosynthesis of the peptidoglycan layer of the cell wall
-
-
-
L-Ala
?
show the reaction diagram
-
enzyme provides D-Ala as an essential building block for biosynthesis of the peptidoglycan layer of the cell wall
-
-
-
L-Ala
?
show the reaction diagram
-
enzyme provides D-Ala as an essential building block for biosynthesis of the peptidoglycan layer of the cell wall
-
-
-
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
-
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
A1XDT8, -
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
?
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-, Q9UW18
-
-
?
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
Thalassiosira sp.
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
Q65YW7, -
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-, Q1XG01
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
Q1XG01
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-, Q208U6
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
?
L-alanine
D-alanine
show the reaction diagram
B9WYE8, -
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
-
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
?
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-, P10725, P94494
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-, Q837J0
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
P45257
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-, Q62JA4
-
-
-
?
L-alanine
D-alanine
show the reaction diagram
-, Q63ME6, Q63SZ2
-
-
-
?
L-alanine
D-alanine
show the reaction diagram
-
-
responsible for the synthesis of the d-alanine moiety present in cyclosporin A and of HC-toxin
-
-
L-alanine
D-alanine
show the reaction diagram
-, Q81VF6
-
enzyme provides D-Ala as a required compound for the synthesis of the peptidoglycan layer of the bacterial cell wall, Tolypocladium niveum requires alanine racemase for cyclosporin biosynthesis
-
r
L-alanine
D-alanine
show the reaction diagram
-, Q9UW18
TOXG encodes an alanine racemase whose function is to synthesize D-Ala for incorporation into HC-toxin, enzyme is involved in cyclic peptide biosynthesis
-
?
L-alanine
D-alanine
show the reaction diagram
-
a major component of the alanine pathway, D-alanine is a major component in cell wall synthesis
-
-
?
L-alanine
D-alanine
show the reaction diagram
-
first step in the biosynthesis of the peptidoglycan
-
-
?
L-alanine
D-alanine
show the reaction diagram
-
The bacterium utilizes D-alanine (DAla) for synthesis of the peptidoglycan cell wall.
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
responsible for the synthesis of the d-alanine moiety present in cyclosporin A and of HC-toxin
-
-
-
L-alanine
D-alanine
show the reaction diagram
-
optimal substrate for alanine racemase
-
-
?
L-alanine
D-alanine
show the reaction diagram
Bacillus subtilis 168
P10725, P94494
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
Q62JA4
-
-
-
?
L-alanine
D-alanine
show the reaction diagram
Q63ME6, Q63SZ2
-
-
-
?
L-alanine
D-alanine
show the reaction diagram
Pseudomonas fluorescens TM5-2
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
Pseudomonas fluorescens LRB3W1
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
Pseudomonas putida YZ-26
-
optimal substrate for alanine racemase
-
-
?
L-alanine
R-alanine
show the reaction diagram
P0A6B4
-
-
-
r
L-Arg
D-Arg
show the reaction diagram
-
stepwise mechanism for alanine racemase at both 25C and at 65C. The carbanionic intermediate is obligatory, and Arg219 may serve to destabilize it to avoid side reactions such as transamination, a detailed reaction mechanism is proposed that includes enzyme and substrate protonation states
-
r
L-isoleucine
D-isoleucine
show the reaction diagram
Pseudomonas putida, Pseudomonas putida YZ-26
-
-
-
-
?
L-leucine
D-leucine
show the reaction diagram
-
activity is 20% compared with L-alanine, other amino acids are not racemized
-
-
r
L-Ser
D-Ser
show the reaction diagram
Q932V0, -
2% of the activity with L-Ala
-
r
L-Ser
D-Ser
show the reaction diagram
O59828, -
3.7% of the activity with L-Ala
-
r
L-Ser
D-Ser
show the reaction diagram
-
50% of the activity with L-Ala
-
?
L-Ser
D-Ser
show the reaction diagram
-
at 0.5% of the activity with L-Ala
-
?
L-Ser
D-Ser
show the reaction diagram
-
40% of the activity with L-Ala
-
-
?
L-serine
D-serine
show the reaction diagram
-
-
-
-
r
additional information
?
-
-
exchange of the alpha-hydrogen of D-Ala and L-Ala with D2O
-
-
-
additional information
?
-
-
no racemization of L-Ser, L-Asp, L-Glu, L-Val and L-Arg
-
?
additional information
?
-
-
the enzyme catalyzes transamination as side reaction, R-isomer preference in the hydrogen abstraction from pyridoxamine 5'-phosphate
-
?
additional information
?
-
-
the epsilon-amino group of Lys39 participates in both racemization and transamination when catalyzed by the wild-type enzyme
-
?
additional information
?
-
-
key enzyme in cyclosporin A biosynthesis
-
-
-
additional information
?
-
-
alr racemase is constitutive and serves an anabolic function, dadB encoded enzyme is inducible and required for cell growth on L-Ala
-
-
-
additional information
?
-
-
two nonhomologous alanine racemase genes, one of which is associated with the catabolic function and the other of which presumably represents the biosynthetic function
-
-
-
additional information
?
-
Thalassiosira sp.
-
specific for alanine
-
-
-
additional information
?
-
Pseudomonas putida, Pseudomonas putida YZ-26
-
the enzyme is only specific to L-alanine and L-isoleucine, and does not catalyze isomerization the other amino acids
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
D-alanine
L-alanine
show the reaction diagram
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
-
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
P0A2W8, -
-
-
-
r
D-alanine
L-alanine
show the reaction diagram
Medicago sativa L.
-
-
-
-
r
L-Ala
D-Ala
show the reaction diagram
-
enzyme is required for production of D-Ala, a necessary component of the bacterial cell wall
-
?
L-Ala
D-Ala
show the reaction diagram
-
the enzyme catalyzes the first committed step in bacterial cell wall biosynthesis
-
?
L-Ala
?
show the reaction diagram
-
-
-
-
-
L-Ala
?
show the reaction diagram
-
enzyme provides D-Ala as an essential building block for biosynthesis of the peptidoglycan layer of the cell wall
-
-
-
L-Ala
?
show the reaction diagram
-
enzyme provides D-Ala as an essential building block for biosynthesis of the peptidoglycan layer of the cell wall
-
-
-
L-Ala
?
show the reaction diagram
-
enzyme provides D-Ala as an essential building block for biosynthesis of the peptidoglycan layer of the cell wall
-
-
-
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
A1XDT8, -
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
?
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-, Q1XG01
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
Q1XG01
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-, Q208U6
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
B9WYE8, -
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
-
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
?
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-, P10725, P94494
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-, Q837J0
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
P45257
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
-
responsible for the synthesis of the d-alanine moiety present in cyclosporin A and of HC-toxin
-
-
L-alanine
D-alanine
show the reaction diagram
-, Q81VF6
-
enzyme provides D-Ala as a required compound for the synthesis of the peptidoglycan layer of the bacterial cell wall, Tolypocladium niveum requires alanine racemase for cyclosporin biosynthesis
-
r
L-alanine
D-alanine
show the reaction diagram
-, Q9UW18
TOXG encodes an alanine racemase whose function is to synthesize D-Ala for incorporation into HC-toxin, enzyme is involved in cyclic peptide biosynthesis
-
?
L-alanine
D-alanine
show the reaction diagram
-
a major component of the alanine pathway, D-alanine is a major component in cell wall synthesis
-
-
?
L-alanine
D-alanine
show the reaction diagram
-
first step in the biosynthesis of the peptidoglycan
-
-
?
L-alanine
D-alanine
show the reaction diagram
-
The bacterium utilizes D-alanine (DAla) for synthesis of the peptidoglycan cell wall.
-
-
r
L-alanine
D-alanine
show the reaction diagram
-
responsible for the synthesis of the d-alanine moiety present in cyclosporin A and of HC-toxin
-
-
-
L-alanine
R-alanine
show the reaction diagram
P0A6B4
-
-
-
r
L-alanine
D-alanine
show the reaction diagram
Bacillus subtilis 168
P10725, P94494
-
-
-
r
additional information
?
-
-
key enzyme in cyclosporin A biosynthesis
-
-
-
additional information
?
-
-
alr racemase is constitutive and serves an anabolic function, dadB encoded enzyme is inducible and required for cell growth on L-Ala
-
-
-
additional information
?
-
-
two nonhomologous alanine racemase genes, one of which is associated with the catabolic function and the other of which presumably represents the biosynthetic function
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
pyridoxal 5'-phosphate
-
required as coenzyme
pyridoxal 5'-phosphate
-
1 mol of pyridoxal 5'-phosphate is bound per subunit
pyridoxal 5'-phosphate
-
1 pyridoxal 5'-phosphate per 42000 MW subunit
pyridoxal 5'-phosphate
-
contains one mol of pyridoxal 5'-phosphate per mol of enzyme; Km: 0.000033 mM; the sequence of 10 amino acid residues around the Lys residue, to which pyridoxal 5'-phosphate is bound, is identical with that of the dadB racemase
pyridoxal 5'-phosphate
-
2 mol of pyridoxal 5'-phosphate bound per mol of enzyme dimer
pyridoxal 5'-phosphate
-
pyridoxal 5'-phosphate dependent enzyme
pyridoxal 5'-phosphate
-
the monomeric inactive enzyme appears to bind the cofactor pyridoxal 5'-phosphate by a non-covalent linkage, although the native dimeric enzyme binds the cofactor through an aldimine Schiff base linkage
pyridoxal 5'-phosphate
-
2 mol of pyridoxal 5'-phosphate bound per mol of enzyme dimer
pyridoxal 5'-phosphate
-
pyridoxal 5'-phosphate binds to Lys of the enzyme protein and forms an aldimine Schiff base. The alpha-proton of the substrate is then abstracted, and the pyridoxal 5'-phosphate carbanion is generated; pyridoxal 5'-phosphate dependent enzyme
pyridoxal 5'-phosphate
-
not required as cofactor, slight activation at low concentrations, inhibition at high concentrations
pyridoxal 5'-phosphate
-
required as coenzyme
pyridoxal 5'-phosphate
-
Arg219 forms a hydrogen bond with the pyridine nitrogen of the cofactor, Arg136 donates a hydrogen bond to the phenolic oxygen of pyridoxal 5'-phosphate and may be involved in the binding of substrate as well as stabilization of intermediates
pyridoxal 5'-phosphate
Q9S5V6
Km: 0.005 mM, at 30C
pyridoxal 5'-phosphate
P0A6B6, -
cofactor
pyridoxal 5'-phosphate
Q932V0, -
cofactor
pyridoxal 5'-phosphate
P0A6B5, -
cofactor
pyridoxal 5'-phosphate
Q93HP9, -
cofactor
pyridoxal 5'-phosphate
-
both active sites of the dimer contain a pyridoxal 5'-phosphate molecule in aldimine linkage to Lys39 as a protonated Schiff base. The protonated pyridoxal 5'-phosphate-Lys39 Schiff base is the reactive form of the enzyme
pyridoxal 5'-phosphate
-
each monomer is comprised of two domains, an eight-stranded alpha/beta barrel containing the pyridoxal 5'-phosphate cofactor and a second domain primarily composed of beta-strands. The cofactor adopts two partially occupied conformational states that resemble previously reported and external aldimine complexes
pyridoxal 5'-phosphate
-
1 mol of enzyme contains 2 mol of cofactor
pyridoxal 5'-phosphate
-
enzyme is dependent on, maximal activity at 0.025 mM
pyridoxal 5'-phosphate
-
Km at 30C is 0.005 mM. Maximal activity is obtained in presence of more than 0.125 mM pyridoxal 5'-phosphate. The decrease in activity at incubation temperatures over 40C is consistent with the decrease in the amount of bound pyridoxal 5'-phosphate
pyridoxal 5'-phosphate
O59828, -
coenzyme
pyridoxal 5'-phosphate
-
the pyridoxal form of the enzyme is converted to the pyridoxamine form by incubation with its natural substrate, D-alanine or L-alanine, under acidic conditions: the enzyme loses its racemase activity concomitantly. The pyridoxamine form of the enzyme returns to the pyridoxal form by incubation with pyruvate at alkaline pH
pyridoxal 5'-phosphate
-
1 mol per mol of enzyme
pyridoxal 5'-phosphate
-
enzyme is dependent on
pyridoxal 5'-phosphate
-
covalently linked to enzyme at K42
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
-, Q1XG01
-
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
A1XDT8, -
-
pyridoxal 5'-phosphate
-
C-terminal region of 1 subdomain: Arg138 donates a hydrogen bond to the phenolic O atom of PLP, Arg224 donates a hydrogen bond to the pyridinyl N atom of PLP, Lys41 forms an aldimine linkage with the PLP, eliminating water to form the Schiff base, C-terminal atoms of second subunit Ser209, Gly226 and Ile227 stabilize the PLP phosphate with the help of Ser209 O(gamma), Tyr45 O(eta) and Tyr359 O(eta)
pyridoxal 5'-phosphate
-
stabilizes anionic intermediate after abstraction of alpha-hydrogen of the substrate amino acid by forming a quinoid intermediate
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
-
stabilizes anionic intermediate after abstraction of alpha-hydrogen of the substrate amino acid by forming a quinoid intermediate
pyridoxal 5'-phosphate
-
once per 1 million turnovers of racemization a H-atom is added to C4-atom of the substrate moiety of the anionic intermediate instead of the reprotonation of the abstracted hydrogen at C(alpha) resulting in pyridoxamine 5'-phosphate
pyridoxal 5'-phosphate
-
stabilizes anionic intermediate after abstraction of alpha-hydrogen of the substrate amino acid by forming a quinoid intermediate
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
P10725, P94494
;
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
B9WYE8, -
-
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
-
PLP is inherently bound to the enzyme, removal of PLP inactivates the enzyme, adding PLP restores the activity, addition of 10 microM PLP to native enzyme slightly enhances activity
pyridoxal 5'-phosphate
B3VI72, -
PLP is bound to the enzyme, adding PLP during developement of enzyme or to an assay is not necessary
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
-, Q62JA4
-
pyridoxal 5'-phosphate
-, Q63ME6, Q63SZ2
;
pyridoxal 5'-phosphate
-
-
FAD
-
not required as cofactor, slight activation at low concentrations, inhibition at high concentrations
additional information
-
contains a pyridoxal 5'-phosphate binding site
-
additional information
Thalassiosira sp.
-
exogenous pyridoxal 5-phosphate is not required, but enzyme may be pyridoxal 5-phosphate-dependent
-
additional information
-
not activated by pyridoxal 5'-phosphate
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
slight activation at 1 mM
Co2+
-
120% activity at 1 mM
Fe2+
-
slight activation at 1 mM
Mg2+
-
slight activation at 1 mM
Ni2+
-
slight activation at 1 mM
Sr2+
-
158% activity at 1 mM
Mn2+
-
130% activity at 1 mM
additional information
-
not influenced by 1 mM Zn2+ or Sn2+
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(1-Aminoethyl)boronic acid
-
-
(1-Aminoethyl)phosphonate
-
D- and L-(1-aminoethyl)phosphonate
(1-Aminoethyl)phosphonate
-
-
(4R)-4-amino-3-isoxazolidinone
-
-
-
(R)-1-aminoethylphosphonic acid
-
upon formation of the external aldimine the phosphonate group interacts with putative catalytic residues, thereby rendering them unavailable for catalysis
(R)-1-aminoethylphosphonic acid
-
in combination with pyridoxal 5'-phosphate
1,1'-(2-oxido-1,2,5-oxadiazole-3,4-diyl)-bis (1-(2-thienyl))-methanone
-
-
-
1-amino-cyclopropane phosphonate
-
-
2-(2-hydroxyphenoxy)-N-methylacetamide
-
-
-
2-(4,6-dimethyl-3-oxo-[1,2]thiazolo[5,4-b]pyridin-2-yl)-N-[2-(4-ethoxyphenyl)ethyl]acetamide
-
-
-
2-(4-methoxyphenyl)-1-morpholin-4-ylethanethione
-
-
-
2-(4-methylphenyl)-1-morpholin-4-ylethanethione
-
-
-
2-(hydoxyimino)-6-methyl-2H-benzopyran-3-carboxamide
-
-
-
2-(pyridin-3-ylcarbamothioyl sulfanyl)acetic acid
-
-
-
2-Amino-3-chlorobut-3-enoic acid
-
i.e. 3-chlorovinylglycine, irreversible
2-Amino-3-chlorobut-3-enoic acid
-
-
2-Amino-3-fluorobut-3-enoic acid
-
i.e. 3-fluorovinylglycine, irreversible
2-Amino-3-fluorobut-3-enoic acid
-
-
2-N',2-N',7-N',7-N'-tetramethyl-9H-fluorene-2,7-disulfonohydrazide
-
-
-
2-phenyl-1-piperidin-1-ylethanethione
-
-
-
3,3-dihydroxy-1H-quinoline-2,4-dione
-
-
-
5-chloro-N-(3-chloro-4-methoxyphenyl)-2-(methylsulfonyl)pyrimidine-4-carboxamide
-
-
-
6-O-[3-chloro-4-(6-methoxycarbonylpyridine-2-carbonyl)oxyphenyl]2-O-methyl pyridine-2,6-dicarboxylate
-
-
-
alanine phosphonate
-
-
-
Aminooxyacetate
-
1 mM, complete inhibition, both directions
aminooxyacetic acid
-
1 mM, complete inhibition
aminooxyacetic acid
-
-
beta,beta,beta-trifluoroalanine
-
nucleophilic attack of Lys38 on the electrophilic beta-difluoro-alpha,beta-unsaturated imine
beta,beta,beta-trifluoroalanine
-
-
beta-chloroalanine
-
-
beta-fluoroalanine
-
-
-
chloro-vinyl glycine
-
-
-
Cu2+
-
40% residual activity at 1 mM
Cycloserine
-
suicide inhibitor
Cycloserine
-
D-cycloserine or a racemic mixture of D- and L-cycloserine
Cycloserine
-
8 microg/ml bacterial culture extract markedly inhibits alanine racemase
D-Chloroalanine
-
Ki: 0.005 mM, competitive
D-cycloserine
-
1 mM, 95% inhibition
D-cycloserine
-
model for inactivation mechanism via geminal diamine and ketimine to isoxazole
D-cycloserine
-
time-dependent inactivation rate of enzyme from Streptomyces lavendulae is slower than for enzyme from Escherichia coli
D-cycloserine
Q65YW7, -
time-dependent inactivation rate of enzyme from Streptomyces lavendulae is slower than for enzyme from Escherichia coli. Enzyme from Streptomyces lavendulae is one of its self-resistance determinants
D-cycloserine
-
mechanism of inactivation and comparison with inactivation of Streptomyces lavendulae enzyme
D-cycloserine
-
mechanism of inactivation and comparison with inactivation of Bacillus stearothermophilus enzyme
D-cycloserine
A1XDT8, -
-
D-cycloserine
-
-
D-penicillamine
-
1 mM, 79% inhibition
ethyl 3-(pyridin-2-ylthio)propanoate
-
-
-
FAD
-
slight activation at low concentrations, inhibition at high concentrations
fluoro-alanine
-
-
-
fluoro-vinyl glycine
-
-
-
hydroxylamine
-
1 mM, 68% inhibition
hydroxylamine
-
1 mM, complete inhibition
hydroxylamine
-
1 mM, complete inhibition, both directions
hydroxylamine
-
-
hydroxylamine
-
-
hydroxylamine
-
non-competitive inhibition kinetics
L-alanine phosphonic acid
-
-
-
L-chloroalanine
-
Ki: 1.71 mM, noncompetitive
L-Cycloserine
-
1 mM, 90% inhibition
L-Cycloserine
-
model for inactivation mechanism via geminal diamine and ketimine to isoxazole
L-Cycloserine
-
-
L-Cycloserine
-
mechanism of inactivation and comparison with inactivation of Streptomyces lavendulae enzyme
L-Cycloserine
-
mechanism of inactivation and comparison with inactivation of Bacillus stearothermophilus enzyme
N',N',4-trimethylbenzenesulfonohydrazide
-
-
-
N-benzyl-5-chloro-2-methylsulfonylpyrimidine-4-carboxamide
-
-
-
N-hydroxy-2-(2-hydroxyphenoxy)acetamide
-
-
-
NaBH4
-
0.5 M, loss of activity
NaCl
-
slight inhibition above 600 mM
NaCl
-
at concentrations around seawater level
NEM
-
1 mM, 22% inhibition
O-Carbamoyl-D-Ser
-
inhibition of wild type enzyme but not of the O-carbamoyl-D-Ser mutant
o-carbamyl-D-serine
-
-
-
PCMB
-
1 mM, 91% inhibition
phenylhydrazine
-
1 mM, 72% inhibition
phenylhydrazine
-
-
phenylhydrazine
-
-
Propionate
-
propionate influences both Km (affinity for substrate) and Vmax (enzyme catalysis)
pyridoxal 5'-phosphate
-
slight activation at low concentrations, inhibition at high concentrations
pyridoxal 5'-phosphate
-
0.4 mM, 51% inhibition
Sodium borohydride
P0A6B6, -
reduction of the enzyme by dialysis with sodium borohydride, the reduced enzyme is catalytically inactive and addition of pyridoxal 5'-phosphate does not reverse the inactivation
Sodium borohydride
Q932V0, -
reduction of the enzyme by dialysis with sodium borohydride, the reduced enzyme is catalytically inactive and addition of pyridoxal 5'-phosphate does not reverse the inactivation
Sodium borohydride
P0A6B5, -
reduction of the enzyme by dialysis with sodium borohydride, the reduced enzyme is catalytically inactive and addition of pyridoxal 5'-phosphate does not reverse the inactivation
Sodium borohydride
Q93HP9, -
reduction of the enzyme by dialysis with sodium borohydride, the reduced enzyme is catalytically inactive and addition of pyridoxal 5'-phosphate does not reverse the inactivation
Sodium borohydride
-
1 mM, 30% inhibition
Sodium borohydride
O59828, -
complete inactivation after dialysis against
L-Penicillamine
-
1 mM, 28% inhibition
additional information
Q65YW7, -
not inhibitory: L-cycloserine
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
pyridoxal 5'-phosphate
-
0.02 mM, activation to 108% of control in the direction of D-Ala formation, activation to 124% of control in the direction of L-Ala formation
glutathione
-
required for maximal activity
additional information
-
activated in presence of monovalent anions including Cl-
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.1
-
D-Ala
-
23C, enzyme Alr
1.4
-
D-Ala
-
23C, enzyme DadX
1.8
-
D-Ala
-
90C, pH 9.5
2.2
-
D-Ala
-
D-Ala, dadB encoded enzyme
2.4
-
D-Ala
O59828, -
37C, pH 9.0
10
-
D-Ala
Q932V0, -
pH 8.3, 30C
12.2
-
D-Ala
Q9S5V6
30C, pH 8.3
73.5
-
D-Ala
-
30C, pH 8.5
119
-
D-Ala
-
-
179
-
D-Ala
-
pH 8.5, 37C
0.25
-
D-alanine
-
pH 8.2, 37C
0.304
-
D-alanine
-
+/-0.034, Alr P219A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
0.311
-
D-alanine
-
+/-0.008, Alr wildtype, 30C, pH 8.0, spectrophotometrically measured at 340 nm
0.4
-
D-alanine
-
pH 8.2, 37C
0.402
-
D-alanine
-
+/-0.055, Alr E221K, 30C, pH 8.0, spectrophotometrically measured at 340 nm
0.439
-
D-alanine
-
+/-0.080, Alr E221A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
0.513
-
D-alanine
-
+/-0.084, Alr E221P, 30C, pH 8.0, spectrophotometrically measured at 340 nm
0.528
-
D-alanine
-
+/-0.079, Alr E165K, 30C, pH 8.0, spectrophotometrically measured at 340 nm
0.592
-
D-alanine
-
+/-0.085, Alr E165A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
0.604
-
D-alanine
-
+/-0.070, Alr D164K, 30C, pH 8.0, spectrophotometrically measured at 340 nm
0.615
-
D-alanine
-
+/-0.032, Alr D164A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
0.7
-
D-alanine
Q65YW7, -
pH 8.2, 25C
1.008
-
D-alanine
-
+/-0.069, Alr wildtype, 30C, pH 8.0, spectrophotometrically measured at 340 nm
2.1
-
D-alanine
-
at 23C
4.2
-
D-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
4.7
-
D-alanine
-
pH 7.4, 30C
4.7
-
D-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
5.6
-
D-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
6.1
-
D-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
6.2
-
D-alanine
-, Q1XG01
Vmax 37.9 micromol/min/mg
6.9
-
D-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
7.3
-
D-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
8.7
-
D-alanine
-
pH 7.4, 30C
8.7
-
D-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
12
-
D-alanine
-
Vmax = 0.44 mol/s/kg
14.91
-
D-alanine
-
-
16.5
-
D-alanine
Thalassiosira sp.
-
pH 9.5
20.4
-
D-alanine
-
HPLC analysis
10
-
D-serine
-
pH 8.2, 37C
1.1
-
L-Ala
-
23C, enzyme Alr
1.4
-
L-Ala
-
23C, enzyme DadX
5
-
L-Ala
O59828, -
37C, pH 9.0
17.9
-
L-Ala
Q9S5V6
30C, pH 8.3
20
-
L-Ala
Q932V0, -
pH 8.3, 30C
167
-
L-Ala
-
pH 8.5, 37C
171
-
L-Ala
-
30C, pH 8.5
0.29
-
L-alanine
-
pH 8.2, 37C
0.4
-
L-alanine
-
pH 8.2, 37C
0.7
-
L-alanine
Q65YW7, -
pH 8.2, 25C
1.049
-
L-alanine
-
+/-0.131, Alr P219A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
1.401
-
L-alanine
-
+/-0.209, Alr E221P, 30C, pH 8.0, spectrophotometrically measured at 340 nm
1.516
-
L-alanine
-
+/-0.083, Alr E221A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
1.562
-
L-alanine
-
+/-0.256, Alr E165A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
1.993
-
L-alanine
-
+/-0.269, Alr E221K, 30C, pH 8.0, spectrophotometrically measured at 340 nm
2.057
-
L-alanine
-
+/-0.038, Alr E165K, 30C, pH 8.0, spectrophotometrically measured at 340 nm
2.8
-
L-alanine
-
-
3
-
L-alanine
-
-
3.03
-
L-alanine
-
+/-0.114, Alr D164A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
3.603
-
L-alanine
-
+/-0.180, Alr D164K, 30C, pH 8.0, spectrophotometrically measured at 340 nm
4.1
-
L-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
7.4
-
L-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
8.1
-
L-alanine
-
pH 7.4, 30C
8.1
-
L-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
9.3
-
L-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
9.8
-
L-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
10.34
-
L-alanine
-
at pH 9.0 and 37C
17.4
-
L-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
18.4
-
L-alanine
-
pH 7.4, 30C
18.4
-
L-alanine
-
in 50 mM potassium phosphate buffer pH 7.4, at 30C
29.4
-
L-alanine
Thalassiosira sp.
-
pH 9.5
29.6
-
L-alanine
-
Vmax = 1.02 mol/s/kg
41.79
-
L-alanine
-
-
43
-
L-alanine
-
HPLC analysis
100
-
L-alanine
-, Q1XG01
Vmax 909 micromol/min/mg
27
-
L-serine
-
pH 8.2, 37C
additional information
-
additional information
-
Km values of L-Ala in the presence of urea at various concentrations
-
additional information
-
additional information
-
effect of NaCl on Km-value
-
additional information
-
additional information
-
Vmax for the racemization (D- to L-alanine and L- to D-alanine) is 87.0 and 84.8 U/mg, respectively.
-
additional information
-
additional information
-
Vmax 110 micromol/min/mg D-alanine; Vmax 323 micromol/min/mg L-alanine
-
additional information
-
additional information
-
Vmax 306 U/mg: D-alanine to L-alanine; Vmax 345 U/mg: L-alanine to D-alanine
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
24.8
-
D-Ala
-
90C, pH 9.5
2314
-
D-Ala
-
pH 8.5, 37C
3270
-
D-Ala
-
30C, pH 8.5
3272
-
D-Ala
-
30C, pH 8.5
0.333
-
D-alanine
-
+/-0.033, Alr D164K, 30C, pH 8.0, spectrophotometrically measured at 340 nm
1.317
-
D-alanine
-
+/-0.100, Alr E165K, 30C, pH 8.0, spectrophotometrically measured at 340 nm
2
8
D-alanine
-
pH 8.2, 37C
4
-
D-alanine
-
+/-0.417, Alr E165A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
4.466
-
D-alanine
-
+/-0.200, Alr D164A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
5.267
-
D-alanine
-
+/-0.333, Alr P219A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
5.783
-
D-alanine
-
+/-0.483, Alr wildtype, 30C, pH 8.0, spectrophotometrically measured at 340 nm
6.35
-
D-alanine
-
+/-0.333, Alr E221K, 30C, pH 8.0, spectrophotometrically measured at 340 nm
6.817
-
D-alanine
-
+/-0.650, Alr E221A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
7.617
-
D-alanine
-
+/-0.750, Alr E221P, 30C, pH 8.0, spectrophotometrically measured at 340 nm
37.9
-
D-alanine
-, Q1XG01
-
63.3
-
D-alanine
-
pH 8.2, 37C
70
-
D-alanine
Q65YW7, -
pH 8.2, 25C
5.5
-
D-serine
-
pH 8.2, 37C
2589
-
L-Ala
-
pH 8.5, 37C
7500
-
L-Ala
-
30C, pH 8.5
7504
-
L-Ala
-
30C, pH 8.5
2.8
-
L-alanine
-
+/-0.250, Alr D164K, 30C, pH 8.0, spectrophotometrically measured at 340 nm
16.72
-
L-alanine
-
+/-0.850, Alr E165K, 30C, pH 8.0, spectrophotometrically measured at 340 nm
22
-
L-alanine
-
-
22.47
-
L-alanine
-
+/-2.667, Alr E165A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
27.6
-
L-alanine
-
pH 8.2, 37C
41.82
-
L-alanine
-
+/-2.600, Alr D164A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
51.77
-
L-alanine
-
+/-6.067, Alr P219A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
53.98
-
L-alanine
-
+/-3.217, Alr wildtype, 30C, pH 8.0, spectrophotometrically measured at 340 nm
55
-
L-alanine
-
pH 8.2, 37C
55
-
L-alanine
Q65YW7, -
pH 8.2, 25C
67.07
-
L-alanine
-
+/-7.633, Alr E221P, 30C, pH 8.0, spectrophotometrically measured at 340 nm
68.23
-
L-alanine
-
+/-7.550, Alr E221K, 30C, pH 8.0, spectrophotometrically measured at 340 nm
70.63
-
L-alanine
-
at pH 9.0 and 37C
70.92
-
L-alanine
-
+/-2.333, Alr E221A, 30C, pH 8.0, spectrophotometrically measured at 340 nm
632
-
L-alanine
-, Q1XG01
-
13.3
-
L-serine
-
pH 8.2, 37C
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
7.33
-
L-alanine
-
-
12048
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.086
-
(4R)-4-amino-3-isoxazolidinone
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.93
-
2-(4,6-dimethyl-3-oxo-[1,2]thiazolo[5,4-b]pyridin-2-yl)-N-[2-(4-ethoxyphenyl)ethyl]acetamide
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.08
-
2-phenyl-1-piperidin-1-ylethanethione
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.02
-
3,3-dihydroxy-1H-quinoline-2,4-dione
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.76
-
5-chloro-N-(3-chloro-4-methoxyphenyl)-2-(methylsulfonyl)pyrimidine-4-carboxamide
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.68
-
6-O-[3-chloro-4-(6-methoxycarbonylpyridine-2-carbonyl)oxyphenyl]2-O-methyl pyridine-2,6-dicarboxylate
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.33
-
D-cycloserine
-
mutant Y265F, 37C, pH 9.1
0.87
-
D-cycloserine
Q65YW7, -
pH 8.2, 25C
1.2
-
D-cycloserine
-
pH 8.2, 25C
4
-
D-cycloserine
-
or higher, wild-type, 37C, pH 9.1
0.038
-
ethyl 3-(pyridin-2-ylthio)propanoate
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.16
-
hydroxylamine
-
-
6
-
L-Cycloserine
-
wild-type, 37C, pH 9.1
8.2
-
L-Cycloserine
-
pH 8.2, 25C
0.63
-
N-benzyl-5-chloro-2-methylsulfonylpyrimidine-4-carboxamide
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
30
-
Propionate
-
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.058
-
(4R)-4-amino-3-isoxazolidinone
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.0049
-
1,1'-(2-oxido-1,2,5-oxadiazole-3,4-diyl)-bis (1-(2-thienyl))-methanone
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.0057
-
2-(2-hydroxyphenoxy)-N-methylacetamide
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.0077
-
2-(4,6-dimethyl-3-oxo-[1,2]thiazolo[5,4-b]pyridin-2-yl)-N-[2-(4-ethoxyphenyl)ethyl]acetamide
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.0033
-
2-(4-methoxyphenyl)-1-morpholin-4-ylethanethione
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.0065
-
2-(4-methylphenyl)-1-morpholin-4-ylethanethione
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.0028
-
2-(hydoxyimino)-6-methyl-2H-benzopyran-3-carboxamide
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.0131
-
2-(pyridin-3-ylcarbamothioyl sulfanyl)acetic acid
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.0016
-
2-N',2-N',7-N',7-N'-tetramethyl-9H-fluorene-2,7-disulfonohydrazide
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.006
-
2-phenyl-1-piperidin-1-ylethanethione
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.0052
-
3,3-dihydroxy-1H-quinoline-2,4-dione
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.0082
-
5-chloro-N-(3-chloro-4-methoxyphenyl)-2-(methylsulfonyl)pyrimidine-4-carboxamide
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.001
-
6-O-[3-chloro-4-(6-methoxycarbonylpyridine-2-carbonyl)oxyphenyl]2-O-methyl pyridine-2,6-dicarboxylate
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.0026
-
ethyl 3-(pyridin-2-ylthio)propanoate
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.009
-
N',N',4-trimethylbenzenesulfonohydrazide
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.0068
-
N-benzyl-5-chloro-2-methylsulfonylpyrimidine-4-carboxamide
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
0.0082
-
N-hydroxy-2-(2-hydroxyphenoxy)acetamide
-
in 100 mM Tris-Tricine, pH 8.5, temperature not specified in the publication
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3.8
-
-
L-alanine
11.5
-
-
crude extract, using L-alanine as substrate, at pH 9.0 and 37C
16.5
-
-
after 3.7fold purification, using L-isoleucine as substrate, at pH 9.0 and 37C
42.6
-
-
after 3.7fold purification, using L-alanine as substrate, at pH 9.0 and 37C
61.8
-
-, Q1XG01
in presence of 50 mM L-alanine with a 5.0% yield
209
-
Q932V0, -
L-Ala, 30C
295
-
-
at 37C
567
-
-
-
650
-
Q9S5V6
L-Ala, 0C
2807
-
-
-
3708
-
-
substrate is D-alanine
10500
-
-
substrate is L-alanine
additional information
-
-
-
additional information
-
-
assay method based on circular dichroism spectra of D- and L-alanine
additional information
-
-
288 units/mg
additional information
-
-
Vmax of racemization of L- to D-alanine is 101 micromol/min/mg
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8
10
Q932V0, -
-
8.3
-
-, Q1XG01
-
8.5
-
-
-
8.5
-
-
D-Ala as substrate
9
-
-
L-Ala as substrate
9
-
O59828, -
D-Ala as substrate
9.5
-
O59828, -
L-Ala as substrate
9.5
-
-
racemization from D-Ala to L-Ala
9.5
-
-
formation of D-Ala
9.5
-
Thalassiosira sp.
-
-
9.5
-
P10725, P94494
dal, extracts are assayed (in triplicate) by monitoring NADH production in spectrophotometric assay with L-alanine dehydrogenase
10
10.5
P10725, P94494
yncD, extracts are assayed (in triplicate) by monitoring NADH production in spectrophotometric assay with L-alanine dehydrogenase
10
-
-
racemization from L-Ala to D-Ala
10
-
-
formation of L-Ala
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
11
-
pH 7.0: about 75% of maximal activity, pH 11.0: about 75% of maximal activity
7
11
-
pH 7.0: about 40% of maximal activity in formation of D-Ala, about 50% of maximal activity in formation of L-Ala, pH 11: about 95% of maximal activity in formation of L-Ala and D-Ala
7
9.5
-
7.0: about 45% of maximal activity, 9.5: about 95% of maximal activity
7.5
12.5
-
at pH 7.5 and at pH 12.5 activity is just under 40%, at pH 7 and at pH 13 the enzyme is inactive
8.5
11
-
pH 8.5: about 55% of maximal activity, pH 11.0: about 65% of maximal activity, racemization from L-Ala to D-Ala
8.5
11
P10725, P94494
pH 8.5: 35% of max. activity, pH 11: 8% of max. activity, dal, extracts are assayed (in triplicate) by monitoring NADH production in spectrophotometric assay with L-alanine dehydrogenase; pH 8.5: 42% of max. activity, pH 11: 58% of max. activity, yncD, extracts are assayed (in triplicate) by monitoring NADH production in spectrophotometric assay with L-alanine dehydrogenase
9
11
-
pH 9: about 50% of maximal activity, pH 11.0: about 35% of maximal activity, racemization from D-Ala to L-Ala
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
35
40
-
-
35
-
Q9S5V6
in absence of pyridoxal 5'-phosphate
37
-
-
formation of L-Ala or D-Ala
37
-
-, Q1XG01
-
40
-
-
with L-Ala as substrate
50
-
Q932V0, -
in presence or absence of pyridoxal 5'-phosphate
50
-
P10725, P94494
dal, extracts are assayed (in triplicate) by monitoring NADH production in spectrophotometric assay with L-alanine dehydrogenase
55
-
P10725, P94494
yncD, extracts are assayed (in triplicate) by monitoring NADH production in spectrophotometric assay with L-alanine dehydrogenase
60
-
Q9S5V6
in presence of an excess amount of pyridoxal 5'-phosphate, 0.4 mM
additional information
-
-
no temperature dependency of the enzyme is observed with D-Ala
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0
40
-
0C: about 25% of maximal activity, 40C: about 45% of maximal activity
0
60
Q9S5V6
about 30% of maximal activity at 0C and at 60C, in absence of pyridoxal 5'-phosphate
0
70
-
between 20 to 50C the enzyme activity is 75-100%, at 0C activity is about 28% and at 70C activity is 20%
20
50
-
20C: about 60% of maximal activity, 50C, about 35% of maximal activity
25
50
-
25C: about 65% of maximal activity, formation of L-Ala or D-Ala, 50C: about 50% of maximal activity in formation of D-Ala, about 60% of maximal activity in formation of L-Ala
30
50
Q9S5V6
30C, 45% of maximal activity, 50C: about 50% of maximal activity, in presence of an excess amount of pyridoxal 5'-phosphate, 0.4 mM
30
60
P10725, P94494
30C and 60C: approx. 20% of max. activity, dal, extracts are assayed (in triplicate) by monitoring NADH production in spectrophotometric assay with L-alanine dehydrogenase
30
70
P10725, P94494
30C: approx. 25% of max. activity, 70C: approx. 8% of max. activity, yncD, extracts are assayed (in triplicate) by monitoring NADH production in spectrophotometric assay with L-alanine dehydrogenase
35
100
-
35C: about 75% of maximal activity, 100C: about 55% of maximal activity
40
70
-
40C: about 60% of maximal activity, 70C: about 50% of maximal activity
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.2
-
-
isoelectric focusing
6.5
-
-, Q9UW18
calculation from nucleotide sequence
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Acidiphilium organovorum 13H
-
-
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Actinobacillus succinogenes (strain ATCC 55618 / 130Z)
Aeromonas hydrophila subsp. hydrophila (strain ATCC 7966 / NCIB 9240)
Aeromonas hydrophila subsp. hydrophila (strain ATCC 7966 / NCIB 9240)
Bartonella henselae (strain ATCC 49882 / Houston 1)
Burkholderia phymatum (strain DSM 17167 / STM815)
Clostridium difficile (strain 630)
Clostridium difficile (strain 630)
Clostridium difficile (strain 630)
Corynebacterium glutamicum (strain ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025)
Enterococcus faecalis (strain ATCC 700802 / V583)
Enterococcus faecalis (strain ATCC 700802 / V583)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Oenococcus oeni (strain ATCC BAA-331 / PSU-1)
Pseudomonas aeruginosa (strain ATCC 15692 / PAO1 / 1C / PRS 101 / LMG 12228)
Staphylococcus aureus (strain COL)
Staphylococcus aureus (strain Mu50 / ATCC 700699)
Streptococcus pneumoniae serotype 4 (strain ATCC BAA-334 / TIGR4)
Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
39850
-
-
SDS-PAGE
40000
-
-
gel filtration
40000
-
-
SDS-PAGE
40100
-
-
SDS-PAGE
40500
-
-
SDS-PAGE
41000
-
-
SDS-PAGE
41400
-
-
gel filtration
42370
-
-
gel filtration
43650
-
P10725, P94494
precided weight from primary sequence of 394 amino acids, yncD
43660
-
-
immunoblotting
43810
-
-
mass-spectrometry
44000
-
O59828, -
gel filtration
44220
-
-
calculated from amino acid sequence
45000
-
-
enzyme including linker and His6-tag
45000
-
-
SDS-PAGE
46000
-
P0A6B6, -
gel filtration
46000
-
Q932V0, -
gel filtration
46000
-
P0A6B5, -
gel filtration
46000
-
Q93HP9, -
gel filtration
60000
-
-
gel filtration
65000
-
-
gel filtration
67000
-
-
gel filtration
68000
-
-, Q1XG01
gel-filtration analysis
72000
-
-
gel filtration
73000
-
Q9S5V6
gel filtration
76000
-
-
gel filtration in presence or absence of 2-mercaptoethanol
78000
-
-
equilibrium sedimentation method
78000
-
-
gel filtration
80000
-
-
gel filtration
80000
-
-
gel filtration
80000
-
Q65YW7, -
gel filtration
80000
-
-
gel filtration
85000
-
-
HPLC gel filtration
87000
-
-
gel filtration
90000
-
-
gel filtration
93000
-
-
dynamic light scattering (DLS)
120000
150000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 42000, SDS-PAGE
?
-
x * 43341, calculation from nucleotide sequence
?
-
3 or 4 * 37000, SDS-PAGE
?
-, Q9UW18
x * 42700, calculation from nucleotide sequence
?
-
x * 45770, calculated
dimer
-
2 * 39000, SDS-PAGE
dimer
-
2 * 38000, SDS-PAGE
dimer
Q9S5V6
2 * 42500, SDS-PAGE
dimer
-
each monomer is comprised of two domains, an eight-stranded alpha/beta barrel containing the pyridoxal 5'-phosphate cofactor and a second domain primarily composed of beta-strands
dimer
-
2 * 33000, SDS-PAGE
dimer
-
2 * 41000
dimer
-
2 * 44000, SDS-PAGE
dimer
-
formed by two crystallographically different monomers, crystallization data
dimer
-
2 * 39900, calculated, 2 * 42000, SDs-PAGE
dimer
Q65YW7, -
2 * 39900, calculated, 2 * 42000, SDs-PAGE
dimer
-
2 * 39000, SDS-PAGE, enzyme shows monomer-dimer equilibrium
dimer
-
2 * 39000, SDS-PAGE
dimer
-, Q1XG01
2 * 37000, SDS-PAGE
dimer
-
crystallography
dimer
-
4 EcAlr monomers form 2 identical dimers, crystallography
dimer
-
2 * 43700, dynamic light scattering (DLS)
dimer
-
2 * 41000, gel filtration and dynamic light scattering
dimer
Acidiphilium organovorum 13H
-
2 * 33000, SDS-PAGE
-
dimer
-
2 * 41000, gel filtration and dynamic light scattering
-
dimer
Pseudomonas fluorescens TM5-2
-
2 * 39000, SDS-PAGE, enzyme shows monomer-dimer equilibrium
-
homodimer
-
computational simulations
homodimer
-
x-ray crystallography
homodimer
-
2 * 39152, calculated from amino acid sequence; 2 * 40000, SDS-PAGE
homodimer
-
2 * 38337, calculated from amino acid sequence; 2 * 39068, calculated from amino acid sequence; 2 * 40000, SDS-PAGE
homodimer
-
2 * 38883, calculated from amino acid sequence; 2 * 38978, calculated from amino acid sequence; 2 * 40000, SDS-PAGE
homodimer
-
2 * 38765, calculated from amino acid sequence; 2 * 40000, SDS-PAGE
homodimer
-
2 * 39075, calculated from amino acid sequence; 2 * 40000, SDS-PAGE
homodimer
Pseudomonas fluorescens LRB3W1, Pseudomonas fluorescens TM5-2
-
2 * 38883, calculated from amino acid sequence; 2 * 38978, calculated from amino acid sequence; 2 * 40000, SDS-PAGE
-
monomer
-
1 * 39000, SDS-PAGE; 1 * 39044, calculation from nucleotide sequence
monomer
-
dadB enzyme and alr enzyme
monomer
P0A6B6, -
1 * 43000, SDS-PAGE
monomer
Q932V0, -
1 * 43000, SDS-PAGE
monomer
P0A6B5, -
1 * 43000, SDS-PAGE
monomer
Q93HP9, -
1 * 43000, SDS-PAGE
monomer
-
1 * 58000, SDS-PAGE
monomer
O59828, -
1 * 44000, SDS-PAGE
monomer
-
1 * 39000, SDS-PAGE, 1 * 39700, calculated, enzyme shows monomer-dimer equilibrium; 1 * 39000, SDS-PAGE, 1 * 39800, calculated, enzyme shows monomer-dimer equilibrium
monomer
-
1 * 40500, SDS-PAGE
monomer
-
gel filtration
monomer
-
1 * 43000, SDS-PAGE; 1 * 44217, calculated from amino acid sequence
monomer
Pseudomonas fluorescens TM5-2
-
1 * 39000, SDS-PAGE, 1 * 39700, calculated, enzyme shows monomer-dimer equilibrium; 1 * 39000, SDS-PAGE, 1 * 39800, calculated, enzyme shows monomer-dimer equilibrium
-
monomer
Pseudomonas putida YZ-26
-
1 * 43000, SDS-PAGE; 1 * 44217, calculated from amino acid sequence
-
monomer
Salmonella enterica subsp. enterica serovar Typhimurium overproducing
-
1 * 42000, SDS-PAGE
-
additional information
P0A6B6, -
the monomeric enzyme interacts with other monomers in the presence of substrate
additional information
Q932V0, -
the monomeric enzyme interacts with other monomers in the presence of substrate
additional information
P0A6B5, -
the monomeric enzyme interacts with other monomers in the presence of substrate
additional information
Q93HP9, -
the monomeric enzyme interacts with other monomers in the presence of substrate
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
sequence contains a signal peptide of 19 residues at the N-terminus
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
enzyme is subjected to a reductive-methylation procedure
-
using the vapor diffusion method with sitting drops
-
using the hanging-drop vapour diffusion method at 291 K
-
hanging-drop vapor-diffusion method, protein concentration: 20 mg/ml; crystal screen buffer: 0.1 M HEPES, pH 8.0, 22% (w/v) PEG 8000, 0.3 M Ca-acetate, and 1/10 (v/v) cyclohexyl-methyl-beta-D-maltoside, drop: ratio 1/2 mixed protein solution and buffer (sum 2 microl) hanging over 1 ml of buffer; cryo-protecting solution: 0.1 M HEPES, pH 8.0, 22% (w/v) PEG 8000, 0.3 M Ca-acetate, 30% (v/v) glycerol and 1/10 (v/v) cyclohexyl-methyl-beta-D-maltoside for flashcooling in liquid nitrogen, diffracted to 2.5 A
-
hanging drop method using purified enzyme concentrated to 22 mg/ml. The hanging drop contains 0.01 ml of the protein solution, 0.01 ml of 23% polyethylene glycol 4K, 200 mM sodium acetate and 100 mM Tris, pH 8.5. Drops are equilibrated against 0.7 ml of polyethylene glycol 4K solution
-
hanging drop method, determination of the crystal structure of the (R)-1-aminoethylphosphonic acid-pyridoxal 5'-phosphate aldimine in complex with alanine racemase at 1.6 A resolution
-
hanging drop method, the structure of the enzyme with the inhibitor propionate bound in the active site is determined by X-ray crystallography to a resolution of 1.9 A
-
hanging-drop vapor diffusion method. Crystal structure of the enzyme bound with reaction intermediate analogs, N-(5'-phosphopyridoxyl)-L-alanine and N-(5'-phosphopyridoxyl)-D-alanine, determined at 2.0 A resolution with the crystallographic R factor of 17.2 for PLP-L-Ala and 16.9 for PLP-D-Ala complexes
-
mutant Y265F in complex with D- and with L-cycloserine
-
study of water molecules by cluster analysis of several crystal structures
-
sitting drops equilibrated versus 1.5 M (NH4)2SO4, 2% polyethylene glycol 400 and 0.1 M HEPES, pH 7.5, crystal strcuture at 1.45 A resolution
-
sitting drop vapor diffusion method
-
sitting drop vapor diffusion method, at 4C in 1.2 M sodium citrate, 0.1 M MES, pH 7.2, and 10% (v/v) glycerol
-
free enzyme and in complex with D- or L-cycloserine
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
11
-
when the enzyme is incubated at 37C in 100 mM sodium acetate, 100 mM sodium phosphate, 100 mM Tris/HCl, and 100 mM borate/NaOH buffer for 30 min, more than 80% of the initial activity is retained at pH 7.0-11.0
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
-
-
1 h, stable
25
80
-
95% activity at 25C, 80% activity at 40C, less than 20% activity at 50C, less than 10% activity at 60 and 70C, inactive at 80C
25
85
-
between 60 and 70C 50% of the enzyme activity is lost, at 85C 50% of the protein molecules are denatured, the residual activity is 4%
30
-
-
over 30C, 1 h, quick loss of activity
30
-
-
5 min, resistant to incubation with 0.08% SDS, 1 M guanidine hydrochloride, 4 M urea, 45% ethanol or 50% dimethyl sulfoxide
30
-
-
5 min, about 50% loss of activity after incubation with 0.08% SDS, 1 M guanidine hydrochloride, 4 M urea, 45% ethanol or 50% dimethyl sulfoxide; over 30C, 1 h, quick loss of activity
35
-
Q9S5V6
pH 8.3, 1 h, the purified is extremely labile over 35C
40
50
-
pH 8.0, 5 min, sharp decrease of activity between 40C and 50C
40
-
P0A6B6, -
1 h, quick loss of activity in absence of pyridoxal 5'-phosphate
40
-
Q932V0, -
1 h, quick loss of activity in absence of pyridoxal 5'-phosphate
40
-
P0A6B5, -
1 h, quick loss of activity in absence of pyridoxal 5'-phosphate
40
-
Q93HP9, -
1 h, quick loss of activity in absence of pyridoxal 5'-phosphate
40
-
-
the decrease in the enzyme activity at incubation temperatures over 40C is consistent with the decrease in the amount of bound pyridoxal 5'-phosphate, no effect on stability at lower temperatures, 0-30C
50
-
Q9S5V6
pH 8.3, 1 h, stabilized up to 50C in presence of excess amounts of pyridoxal 5'-phosphate, 0.4 mM, complete inactivation in absence of pyridoxal 5'-phosphate
50
-
P0A6B6, -
the enzyme is stabilized up to 50C in presence of 0.4 mM pyridoxal 5'-phosphate
50
-
Q932V0, -
the enzyme is stabilized up to 50C in presence of 0.4 mM pyridoxal 5'-phosphate
50
-
P0A6B5, -
the enzyme is stabilized up to 50C in presence of 0.4 mM pyridoxal 5'-phosphate
50
-
Q93HP9, -
the enzyme is stabilized up to 50C in presence of 0.4 mM pyridoxal 5'-phosphate
60
-
-
pH 8.0, 5 min, almost complete inactivation
65
-
-
30 min, no loss of activity
70
-
-
30 min, enzyme loses most of its activity
75
-
-
1 h, inactivation over 75C
85
-
-
1 h, no denaturation
95
-
-
pH 8.0, 50 mM Tris-HCl, 0.5 M sodium phosphate, maintains more than 90% of its activity after 2 h
additional information
-
-
thermostable
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
pyridoxal 5'-phosphate present in the solvent suppresses the enzyme's unfolding
-
5 min, resistant to incubation with 0.08% SDS, 1 M guanidine hydrochloride, 4 M urea, 45% ethanol or 50% dimethyl sulfoxide
-
denatured by 3.5 M urea in one transition phase
-
in 0.6 M to 1.5 M guanidine hydrochloride the dimeric enzyme is dissociated into a monomeric form, which is catalytically inactive
-
stabilized in presence of monovalent anions including Cl-
-
5 min, about 50% loss of activity after incubation with 0.08% SDS, 1 M guanidine hydrochloride, 4 M urea, 45% ethanol or 50% dimethyl sulfoxide
-
denatured by urea in two transition phases, dissociation of pyridoxal 5'-phosphate with 4.0 M urea, unfolding with 5.5 M urea
-
NaB3H4-reduced DadB holoenzyme is resistant to alpha-chymotrypsin and trypsin and is labile only towards subtilisin
-
stable in 30% ammonium sulfate. Irreversibly diminished activity by exposure to ammonium sulfate concentrations near or above 40%, where precipitation occurs
-
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
acetonitrile
-
50%, enzyme maintains more than 90% of its activity
guanidine-HCl
-
4 M, maintains more than 60% of its activity
hexane
-
no significant change in activity
urea
-
2 M, maintains more than 60% of its activity
isopropanol
-
50%, enzyme maintains more than 30% of its activity
additional information
-
ethyl acetate destroyes the enzyme activity
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
enzyme loses about 20% of activity during storage for 20 days at -20C, 0C and 5C
-
4C, 50 mM potassium phosphate buffer, pH 7.2, 0.01 mM pyridoxal 5'-phosphate, 6 months, 20% loss of activity
-
-20C, stable for 1 month
-
4C, stable for 1 week
-
-70C, 10% glycerol, more than 90% of the activity is retained after 2 years
-
-80C, over 60 days
P0A6B6, -
4C, 3 weeks
P0A6B6, -
-80C, over 60 days
Q932V0, -
4C, 3 weeks
Q932V0, -
-80C, over 60 days
P0A6B5, -
4C, 3 weeks
P0A6B5, -
-80C, over 60 days
Q93HP9, -
4C, 3 weeks
Q93HP9, -
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant enzyme
Q9S5V6
alanine racemase is purified from the muscle of C. grata by using Toyopearl DEAE, Toyopearl Butyl, Hydroxyapatite and Sephacryl S200 column chromatographies.
-
ammonium sulfate fractionation
-
Ni2+-NTA resin (His-tagged overexpressed protein), Hitrap Q-column, Superdex S prep grad column
-
recombinant protein
-
enzyme overproduced in Escherichia coli W3110 lacIq
-
by Ether-Toyopearl, Phenyl-Toyopearl and DEAE-Toyopearl column chromatographic steps
-
partial
-
recombinant enzymes Alr and DadX
-
Ni2+-NTA column chromatography and Sephacryl S-200 gel filtration
-
recombinant enzyme
O59828, -
-
P0A6B6, -
-
Q93HP9, -
ammonium sulfate precipitation, anion-exchange chromatography, hydrophobic interaction chromatography, and gel filtration
-
purified to electrophoretic homogeneity
-
recombinant protein
Q65YW7, -
partial, from stationary growth phase
Thalassiosira sp.
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli
A1XDT8, -
expression in Escherichia coli as a His-tagged protein
-
amplified by PCR, cloned and overexpressed with pOPINB in Escherichia coli Rosetta pLysS cells, His-tagged product
-
analyzation of alanine racemase-transcripts (RT-PCR) during growth and sporulation of wild-type shows low transcription level during early and higher during late sporulation states, analyzed expression level (immunoblotting) shows only alanine racemase during late sporulating states, growth rate of alr-gene knockout mutant and wild-type are identical, mutant only produces half as many spores as wild-type, mutant and wild-type show same resistance against heat, lysozyme and organic solvents, germination takes place in mutant at lower levels of L-alanine (a suggested germination activator) than in wild-type (D-alanine ist suggested to inhibit germination when unfavourable growth conditions), conversion of phase-bright spores to phase-dark germinating cells takes already place within mother cells of the alr mutant (phase dark to phase light), not in wild-type cells resulting in non-resistant mutant-germinants and resistant wild-type-spores respectively
-
expressed as recombinant monomeric protein in Escherichia coli
-
subcloned in a D-alanine auxotrophic Escherichia coli strain MB2795 and in Escherichia coli BL21(DE3)
-
subcloned in a D-alanine auxotrophic Escherichia coli strain MB2795 and in Escherichia coli BL21(DE3)
B3VI72, -
expression in Escherichia coli SOLR with a plasmid yYOK3
Q9S5V6
recombinant expression in D-Alanine auxotrophic Escherichia coli strain MB2159 and DN1686, a D-Ala auxotrophic mutant; recombinant expression in D-Alanine auxotrophic Escherichia coli strains MB2159 and DN1686, a D-Ala auxotrophic mutant
P10725, P94494
expressed in Escherichia coli strain ALA1
-, Q62JA4
expressed in Escherichia coli strain ALA1; expressed in Escherichia coli strain ALA1
-, Q63ME6, Q63SZ2
expression in Escherichia coli
-
amplified from genomic DNA from Enterococcus faecalis via PCR, integrated into pET22b, transformation of Escherichia coli BL21 (DE3) for overexpression as His-tagged protein
-
Escherichia coli strain BL21 (DE3) was transformed with pET28aAlr (1 wild-type and 8 single point mutants)
-
expressed in Escherichia coli BL21(DE3) cells
-
31-54% sequence homologies with Bacillus subtilis and Salmonella typhimurium dadB and alr enzymes
-
expression in Escherichia coli
-
expression in Escherichia coli C600
-
expression in Escherichia coli
-, Q1XG01
from muscle and hepatopancreas
-
isolation and sequencing of cDNA clones, nucleotide sequence 1798 bp including 5- and 3-nonreading frame, poly-A-tail and open reading frame of 1263 bp, expression in E. coli - transformed with open reading frame sequence in pET32Xa/LIC, produced fusion protein with alanine racemase by Escherichia coli, predicted molecular weight of fusion protein of 62000 Da (SDS-PAGE, immunoblotting)
B9WYE8, -
amplified by PCR, cloned and overexpressed in Escherichia coli BL21, both as native and His-tagged products
-
expressed in Escherichia coli as an N-terminal polyhistidine fusion
-
cloning of two independent alanine racemases in Escherichia coli: Alr and DadX
-
expressed in Escherichia coli BL21(DE3) cells
-
strain has one single enzyme gene, expression in Escherichia coli with His-tag
-
expressed in Escherichia coli BL21 cells
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli BL21(DE3) cells
-
expression in Escherichia coli. Heterologous expression renders Saccharomyces cerevisiae capable of utilization of D-Ala as a nitrogen source but also relieves the yeast from the toxicity of D-Ala
O59828, -
expression in Escherichia coli JM109
P0A6B6, -
expression in Escherichia coli JM109
Q932V0, -
expression in Escherichia coli JM109
P0A6B5, -
expression in Escherichia coli JM109
Q93HP9, -
expressed in Escherichia coli BL21(DE3) pLysS cells
-
expression in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D48A
-
no effect on the enzyme activity
deltaalr
-
mutant with knocked out alanine racemase gene alr (2 alanine racemase-genes have been found in Bacillus anthracis jet)
K41A
-
completely inactive
Y270A
-
impaired enzyme activity
D164A
-
alanine racemase Alr from Escherichia coli with single point mutation from D to A at position 164
D164K
-
alanine racemase Alr from Escherichia coli with single point mutation from D to K at position 164
E165A
-
alanine racemase Alr from Escherichia coli with single point mutation from E to A at position 165
E165K
-
alanine racemase Alr from Escherichia coli with single point mutation from E to K at position 165
E221A
-
alanine racemase Alr from Escherichia coli with single point mutation from E to A at position 221
E221K
-
alanine racemase Alr from Escherichia coli with single point mutation from E to K at position 221
P219A
-
alanine racemase Alr from Escherichia coli with single point mutation from P to A at position 219
K39A
-
mutant enzyme is inactive as a catalyst for racemization as well as transamination
R219E
-
catalytical active mutant, the catalytic effect in the Arg219Glu mutant enzyme is due to a combined solvent and inherent stabilizing effect of the protonated cofactor, in contrast to the wild-type enzyme where the catalytic effect may be ascribed to solvent effects alone
Y265A
-
Lys39 is the catalytic residue required for the abstraction and addition of the alpha-hydrogen of D-alanine, As shown by site-directed mutagenesis (K39A mutant) and chemical rescue studies. Tyr265 is catalytic residue for L-alanine, shown by site-directed mutagenesis (Y265A mutant)
E221P
-
alanine racemase Alr from Escherichia coli with single point mutation from E to P at position 221
additional information
-
DNA shuffling of enzyme genes from Salmonella typhimurium and Escherichia coli selecting clones that exhibit higher catalytic activity toward alanine as well as serine. Specific activities of selected clones were increased up to three times more than of wild types. One mutant achieves posttranslationally a high protein level
K39A
-
Lys39 is the catalytic residue required for the abstraction and addition of the alpha-hydrogen of D-alanine, As shown by site-directed mutagenesis (K39A mutant) and chemical rescue studies. Tyr265 is catalytic residue for L-alanine, shown by site-directed mutagenesis (Y265A mutant)
additional information
-
mutant gene which tandemly encodes the two polypeptides of the enzyme subunit, fragment 1 and fragment 2, cleaved at the position corresponding to the predicted hinge region. The mutant fragmentary alanine racemase is active at about 40% of the activity of the wild type enzyme
additional information
-
-
Y265F
-
1600fold reduction of racemization
additional information
-
in-frame deletion mutation, loss of ability to grow on D-alanine
additional information
-
DNA shuffling of enzyme genes from Salmonella typhimurium and Escherichia coli selecting clones that exhibit higher catalytic activity toward alanine as well as serine. Specific activities of selected clones were increased up to three times more than of wild types. One mutant achieves posttranslationally a high protein level
additional information
-
double mutant for the alr encoded enzyme and the dad B encoded enzyme display a phenotype of requirement for exogenous D-Ala for growth
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
enzyme denatured in 6 M guanidine hydrochloride is renatured either by dialysis or dilution to reduce the guanidine hydrochloride concentration
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pharmacology
-
potential candidate for the development of a recombinant vaccine
biotechnology
-
use of enzyme gene as a promoter-screening tool for identification of conditional promoters in Lactobacillus plantarum. Screen for clones capable of complementing the D-alanine auxotroph phenotype of enzyme deletion mutant in media containing enzyme inhibitor D-cycloserine
medicine
-
drug target against Mycobacterium tuberculosis
medicine
-
the requirement for D-Ala as a necessary component of the bacterial cell wall makes the enzyme a logical target for the development of novel antibiotics
medicine
-
since it is absent in humans, this enzyme is an attractive target for the development of drugs against Streptococcus pneumoniae and other bacterial pathogens