Information on EC 5.1.1.10 - amino-acid racemase

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

EC NUMBER
COMMENTARY
5.1.1.10
-
RECOMMENDED NAME
GeneOntology No.
amino-acid racemase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
an L-amino acid = a D-amino acid
show the reaction diagram
the results of the mechanistic studies argue against a single base mechanism and are best rationalized in terms of a double base model where only one of the bases undergoes proton exchange with the solvent while the amino acid is enzyme-bound
-
an L-amino acid = a D-amino acid
show the reaction diagram
enzyme abstracts a hydrogen nonstereospecifically from C-4' of the coenzyme
-
an L-amino acid = a D-amino acid
show the reaction diagram
single base mechanism, significant internal return of the alpha-hydrogen
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
racemization
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Cysteine and methionine metabolism
-
cysteine metabolism
BRENDA
BRENDA
BRENDA
D-Arginine and D-ornithine metabolism
-
D-Glutamine and D-glutamate metabolism
-
Glycine, serine and threonine metabolism
-
Metabolic pathways
-
serine metabolism
BRENDA
BRENDA
BRENDA
SYSTEMATIC NAME
IUBMB Comments
amino-acid racemase
A pyridoxal-phosphate protein.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
amino acid racemase
-
-
amino acid racemase
I0J1I6
-
amino-acid racemase
-
-
broad specificity amino acid racemase
-
-
D-amino acid racemase 1
-
-
L-Amino acid racemase
-
-
-
-
N-succinylamino acid racemase
-
-
Racemase, amino acid
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9068-61-5
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Aeromonas punctata subsp. caviae
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
bifunctional enzyme, racemization of serine and elimination of L-serine and L-serine-O-sulfate to form pyruvate
-
-
Manually annotated by BRENDA team
bifunctional enzyme, racemization of serine and elimination of L-serine and L-serine-O-sulfate to form pyruvate
-
-
Manually annotated by BRENDA team
bifunctional enzymes, racemization of serine and alpha,beta-elimination of L-serine
-
-
Manually annotated by BRENDA team
isoforms A and B, bifunctional enzymes, racemization of serine and elimination of L-serine and L-serine-O-sulfate to form pyruvate
-
-
Manually annotated by BRENDA team
DSM 3263
-
-
Manually annotated by BRENDA team
IFO 12996
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-oxoglutarate + L-ornithine
D-glutamate + L-glutamate
show the reaction diagram
-
-
-
?
D-2,4-Diaminobutyrate
L-2,4-Diaminobutyrate
show the reaction diagram
-
10% of the activity relative to D-Lys
-
-
D-Ala
L-Ala
show the reaction diagram
I0J1I6, -
12% activity, compared to D-Lys
-
-
?
D-Arg
L-Arg
show the reaction diagram
-
-
-
-
D-Arg
L-Arg
show the reaction diagram
-
-
-
-
-
D-Arg
L-Arg
show the reaction diagram
-
65% of the activity relative to L-Lys
-
-
-
D-Arg
L-Arg
show the reaction diagram
-
60% of the activity relative to D-Lys
-
-
-
D-Arg
L-Arg
show the reaction diagram
-
21% of the activity relative to D-Gln
-
-
-
D-Arg
L-Arg
show the reaction diagram
-
80% of the activity relative to L-Lys
-
-
-
D-Arg
L-Arg
show the reaction diagram
I0J1I6, -
95% activity, compared to D-Lys
-
-
?
D-Asn
L-Asn
show the reaction diagram
-
-
-
-
r
D-Asp
L-Asp
show the reaction diagram
-
6% of the activity relative to D-Gln
-
-
D-aspartate
L-aspartate
show the reaction diagram
-
lower substrate affinity for D-aspartate compared to L-aspartate
-
-
?
D-ethionine
L-ethionine
show the reaction diagram
I0J1I6, -
11% activity, compared to D-Lys
-
-
?
D-Gln
L-Gln
show the reaction diagram
-
D-Glu is the best substrate
-
-
D-homoarginine
L-homoarginine
show the reaction diagram
I0J1I6, -
6.2% activity, compared to D-Lys
-
-
?
D-Lys
L-Lys
show the reaction diagram
I0J1I6, -
100% activity
-
-
?
D-Met
L-Met
show the reaction diagram
-
-
-
-
r
D-Met
L-Met
show the reaction diagram
I0J1I6, -
12% activity, compared to D-Lys
-
-
?
D-norvaline
L-norvaline
show the reaction diagram
I0J1I6, -
6.5% activity, compared to D-Lys
-
-
?
D-ornithine
L-ornithine
show the reaction diagram
I0J1I6, -
42% activity, compared to D-Lys
-
-
?
D-Phe
L-Phe
show the reaction diagram
-
1% of the velocity relative to D-Glu
-
-
D-serine
pyruvate + NH3
show the reaction diagram
-
alpha,beta-elimination reaction
-
-
?
D-serine
L-serine
show the reaction diagram
-
-
-
-
r
D-serine
L-serine
show the reaction diagram
-
racemization reaction
-
-
r
D-serine
L-serine
show the reaction diagram
-
similar substrate affinity for both L-serine and D-serine
-
-
?
L-2-Aminobutyrate
D-2-Aminobutyrate
show the reaction diagram
-
-
-
-
L-2-Aminobutyrate
D-2-Aminobutyrate
show the reaction diagram
-
4.7% of the activity relative to D-Lys
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
-
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
6% of the activity relative to L-Lys
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
D-Ala, 15% of the activity relative to D-Gln
-
-
-
L-Ala
D-Ala
show the reaction diagram
-
9% of the activity relative to D-Lys
-
-
-
L-Asn
L-Asn
show the reaction diagram
-
12% of the activity relative to L-Lys
-
-
-
L-Asn
L-Asn
show the reaction diagram
-
9% of the activity relative to L-Lys
-
-
L-Asn
D-Asn
show the reaction diagram
-
-
-
-
r
L-aspartate
D-aspartate
show the reaction diagram
-
higher substrate affinity for L-aspartate compared to D-aspartate
-
-
?
L-Citrulline
D-Citrulline
show the reaction diagram
-
45% of the activity relative to L-Lys
-
-
-
L-Citrulline
D-Citrulline
show the reaction diagram
-
30% of the activity relative to L-Lys
-
-
-
L-Citrulline
D-Citrulline
show the reaction diagram
-
16% of the activity relative to D-Lys
-
-
L-Ethionine
D-Ethionine
show the reaction diagram
-
-
-
-
L-Ethionine
D-Ethionine
show the reaction diagram
-
-
-
-
-
L-Ethionine
D-Ethionine
show the reaction diagram
-
67% of the activity relative to L-Lys
-
-
-
L-Ethionine
D-Ethionine
show the reaction diagram
-
83% of the activity relative to L-Lys
-
-
-
L-Ethionine
D-Ethionine
show the reaction diagram
-
76% of the activity relative to D-Lys
-
-
-
L-Glu
D-Glu
show the reaction diagram
-
-
-
-
-
L-Glu
D-Glu
show the reaction diagram
-
80% of the activity relative to L-Lys
-
-
-
L-Glu
D-Glu
show the reaction diagram
-
no activity with D-Glu
-
-
-
L-Glu
D-Glu
show the reaction diagram
-
57% of the activity relative to L-Lys
-
-
L-His
D-His
show the reaction diagram
-
6% of the activity relative to L-Lys
-
-
-
L-His
D-His
show the reaction diagram
-
1.8% of the activity relative to D-Lys
-
-
L-Homoarginine
L-Homoarginine
show the reaction diagram
-
80% of the activity relative to L-Lys
-
-
L-Homoarginine
L-Homoarginine
show the reaction diagram
-
67% of the activity relative to L-Lys
-
-
L-Homocitrulline
D-Homocitrulline
show the reaction diagram
-
45% of the activity relative to L-Lys
-
-
-
L-Homocitrulline
D-Homocitrulline
show the reaction diagram
-
34% of the activity relative to L-Lys
-
-
L-Homocysteine
D-Homocysteine
show the reaction diagram
-
55% of the activity relative to L-Lys
-
-
L-Homoserine
L-Homoserine
show the reaction diagram
-
12% of the activity relative to L-Lys
-
-
L-Homoserine
L-Homoserine
show the reaction diagram
-
22% of the activity relative to L-Lys
-
-
-
L-Leu
D-Leu
show the reaction diagram
-
-
-
-
L-Leu
D-Leu
show the reaction diagram
-
12% of the activity relative to L-Lys
-
-
-
L-Leu
D-Leu
show the reaction diagram
-
D-Leu, 13% of the activity relative to D-Gln
-
-
-
L-Leu
D-Leu
show the reaction diagram
-
10% of the activity relative to L-Lys
-
-
-
L-Leu
D-Leu
show the reaction diagram
-
3% of the activity relative to D-Lys
-
-
-
L-Lys
D-Lys
show the reaction diagram
-
-
-
-
L-Lys
D-Lys
show the reaction diagram
-
-
-
-
-
L-Lys
D-Lys
show the reaction diagram
-
-
-
-
-
L-Lys
D-Lys
show the reaction diagram
-
D-Lys at 22% of the activity relative to D-Gln
-
-
-
L-Met
D-Met
show the reaction diagram
-
-
-
-
L-Met
D-Met
show the reaction diagram
-
-
-
-
-
L-Met
D-Met
show the reaction diagram
-
-
-
-
r
L-Met
D-Met
show the reaction diagram
-
-
-
-
-
L-Met
D-Met
show the reaction diagram
-
67% of the activity relative to L-Lys
-
-
-
L-Met
D-Met
show the reaction diagram
-
D-Met, 42% of the activity relative to D-Gln
-
-
-
L-Met
D-Met
show the reaction diagram
-
66% of the activity relative to L-Lys
-
-
-
L-Met
D-Met
show the reaction diagram
-
48% of the activity relative to D-Lys
-
-
-
L-Methionine
D-Methionine
show the reaction diagram
-
-
-
-
r
L-Norleucine
D-Norleucine
show the reaction diagram
-
-
-
-
L-Norleucine
D-Norleucine
show the reaction diagram
-
45% of the activity relative to L-Lys
-
-
-
L-Norleucine
D-Norleucine
show the reaction diagram
-
28% of the activity relative to L-Lys
-
-
-
L-Norvaline
D-Norvaline
show the reaction diagram
-
-
-
-
L-Orn
D-Orn
show the reaction diagram
-
-
-
-
-
L-Orn
D-Orn
show the reaction diagram
-
80% of the activity relative to L-Lys
-
-
-
L-Orn
D-Orn
show the reaction diagram
-
79% of the activity relative to L-Lys
-
-
L-Selenohomocysteine
D-Selenohomocysteine
show the reaction diagram
-
27% of the activity relative to L-Lys
-
-
L-Ser
D-Ser
show the reaction diagram
-
10% of the activity relative to L-Lys
-
-
-
L-Ser
D-Ser
show the reaction diagram
-
8.7% of the activity relative to D-Lys
-
-
L-Ser
D-Ser
show the reaction diagram
-
D-Ser, 31% of the activity relative to D-Gln
-
-
-
L-serine
pyruvate + NH3
show the reaction diagram
-
alpha,beta-elimination reaction
-
-
?
L-serine
D-serine
show the reaction diagram
-
-
-
-
r
L-serine
D-serine
show the reaction diagram
-
-
-
-
r
L-serine
D-serine
show the reaction diagram
-
racemization reaction
-
-
r
L-serine
S-serine
show the reaction diagram
-
similar substrate affinity for both L-serine and D-serine
-
-
?
L-Thr
L-Thr
show the reaction diagram
-
no activity
-
-
-
L-Thr
L-Thr
show the reaction diagram
-
3% of the activity relative to L-Lys
-
-
L-threonine
3-hydroxy-2-butenoic acid + NH3
show the reaction diagram
-
alpha,beta-elimination reaction
-
-
?
N-acetyl-D-alanine
N-acetyl-L-alanine
show the reaction diagram
-
-
-
-
r
N-acetyl-D-asparagine
N-acetyl-L-asparagine
show the reaction diagram
-
-
-
-
r
N-acetyl-D-methionine
N-acetyl-L-methionine
show the reaction diagram
-
-
-
-
r
N-acetyl-D-phenylalanine
N-acetyl-L-phenylalanine
show the reaction diagram
-
-
-
-
r
N-acetyl-D-tryptophan
N-acetyl-L-tryptophan
show the reaction diagram
-
-
-
-
r
N-carbamoyl-L-methionine
N-carbamoyl-D-methionine
show the reaction diagram
-
-
-
-
r
N-succinyl-L-alanine
N-succinyl-D-alanine
show the reaction diagram
-
-
-
-
r
N-succinyl-L-phenylalanine
N-succinyl-D-phenylalanine
show the reaction diagram
-
-
-
-
-
N-succinyl-L-phenylalanine
N-succinyl-D-phenylalanine
show the reaction diagram
-
-
-
-
r
N6-Acetyl-L-Lys
N6-Acetyl-D-Lys
show the reaction diagram
-
80% of the activity relative to L-Lys
-
-
-
N6-Acetyl-L-Lys
N6-Acetyl-D-Lys
show the reaction diagram
-
37% of the activity relative to L-Lys
-
-
S-Methyl-L-Cys
S-Methyl-D-Cys
show the reaction diagram
-
3% of the activity relative to L-Lys
-
-
-
S-Methyl-L-Cys
S-Methyl-D-Cys
show the reaction diagram
-
43% of the activity relative to L-Lys
-
-
L-Trp
D-Trp
show the reaction diagram
-
-
-
-
r
additional information
?
-
-
catalyzes nonstereospecific transamination: production of D-Glu and L-Glu from 2-oxoglutarate through transamination with L-Orn
-
-
-
additional information
?
-
-
the relative activity is 1 for Leu, 1.8 for L-2-aminobutyrate, 3.3 for L-Ala, 5.2 for norvaline, 5.9 for L-norleucine, 7.5 for D-Arg, 10.6 for L-Met, 13.2 for D-Lys, and 15.9 for L-ethionine
-
-
-
additional information
?
-
-
no activity with: D-Glu, L-Ile, L-Phe, L-Thr, L-Val
-
-
-
additional information
?
-
-
racemization and elimination activities reside at the same active site of enzyme. Racemization activity is specific to serine, elimination activity has a broader specificity for L-amino acids with a suitable leaving group at the beta-carbon
-
-
-
additional information
?
-
-
ration of elimination reaction/racemization reaction for substrate L-serine is 3.7
-
-
-
additional information
?
-
-
DAR1 has no detectable activity with 100 mM L-glutamate or 100 mM L-alanine
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
pyridoxal 5'-phosphate
-
required for maximal activity
pyridoxal 5'-phosphate
-
holoenzyme contains 2 mol of pyridoxal 5'-phosphate per mol of enzyme; Km: 0.2 mM
pyridoxal 5'-phosphate
-
holoenzyme contains 2 mol of pyridoxal 5'-phosphate per mol of enzyme; Km: 0.0026 mM; required for maximal activity
pyridoxal 5'-phosphate
-
pyridoxal phosphate enzyme
pyridoxal 5'-phosphate
-
contains 1 mol of pyridoxal 5'-phosphate per mol of subunit; holoenzyme contains 2 mol of pyridoxal 5'-phosphate per mol of enzyme
pyridoxal 5'-phosphate
-
dependent on; pyridoxal phosphate enzyme
pyridoxal 5'-phosphate
-
holoenzyme contains 2 mol of pyridoxal 5'-phosphate per mol of enzyme; Km: 0.0026 mM
pyridoxal 5'-phosphate
-
holoenzyme contains 2 mol of pyridoxal 5'-phosphate per mol of enzyme; pyridoxal phosphate enzyme
pyridoxal 5'-phosphate
I0J1I6, -
-
pyridoxal 5'-phosphate
-
-
pyridoxal 5'-phosphate
-
dependent on
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
-
Ca2+
-
may partially replace Mg2+
Co2+
-
enzyme requires a divalent cation for activity. Maximal enzymatic activity with Co2+
Co2+
-
enhances activity, highest activity with Co2+
Mg2+
-
required, both activities
Mg2+
-
required by both isoforms A and B
Mg2+
-
MgCl2 alone (2 mM) has only a small effect on enzyme racemase activity, but when Mg2+ and ATP are present, the enzyme shows the largest activity for the D-Asp and D-Ser conversions
Mn2+
-
-
Mn2+
-
may partially replace Mg2+
Mn2+
-
assay with 0.1 mM MnCl2
Mn2+
-
enhances activity
Ni2+
-
enhances activity
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
3-chloroalanine
-
inactivator binds to the same Lys residue which binds pyridoxyl 5'-phosphate
3-Fluoroalanine
-
inactivator binds to the same Lys residue which binds pyridoxyl 5'-phosphate
8-hydroxy-5-quinolinesulfonic acid
-
-
aminooxyacetic acid
-
complete inhibition at 0.002 mM
ATP
-
inhibitory to L-serine O-sulfate dehydration reaction, activating for racemization reraction
beta-mercaptoethanol
-
-
Co2+
-
inhibition of both activities
Cu2+
-
inhibition of both activities
D-penicillamine
-
-
EDTA
-
inhibition of both activities
Fe2+
-
slight inhibition of both activities
hydroxylamine
-
-
Ni2+
-
slight inhibition of both activities
O-acetylserine
-
inactivator binds to the same Lys residue which binds pyridoxyl 5'-phosphate
O-acetylserine
-
2S,3R, 2R,3R, 2S,3S, 2R,3R
S-(N-Methylthiocarbamoyl)-D-Cys
-
-
S-(N-Methylthiocarbamoyl)-L-Cys
-
-
Zn2+
-
inhibition of both activities
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ATP
-
ATP alone (4 mM) significantly increases enzyme activity, but when the combination of ATP and Mg2+ is present, the enzyme shows the largest activity for the D-Asp and D-Ser conversions
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
7.4
-
D-Ala
-
-
18
-
D-Asn
-
pure lyophilisates, 20C, pH 7, Vmax: 770 micromol/min/g
50
-
D-Asn
-
pure lyophilisates, 25C, pH 7, Vmax: 1299 micromol/min/g
62
-
D-Asn
-
pure lyophilisates, 30C, pH 7, Vmax: 1881 micromol/min/g
66
-
D-Asn
-
pure lyophilisates, 35C, pH 7, Vmax: 2585 micromol/min/g
103
-
D-Asn
-
pure lyophilisates, 40C, pH 7, Vmax: 4032 micromol/min/g
94
-
D-Aspartate
-
in 50 mM Tris-HCl buffer, pH 8.0, at 30C
7.8
-
D-Gln
-
-
0.42
-
D-Lys
-
-
0.83
-
D-Lys
-
-
1.6
-
D-Met
-
-
2
3
D-Met
-
crude lyophilisates, 30C, pH 7, Vmax: 2075 micromol/min/g
9.5
-
D-Met
-
-
22
-
D-Met
-
crude lyophilisates, 25C, pH 7, Vmax: 1892 micromol/min/g
25
-
D-Met
-
crude lyophilisates, 20C, pH 7, Vmax: 1696 micromol/min/g
28
-
D-Met
-
crude lyophilisates, 25C, pH 8, Vmax: 3627 micromol/min/g
30
-
D-Met
-
crude lyophilisates, 35C, pH 7, Vmax: 2512 micromol/min/g
32
-
D-Met
-
crude lyophilisates, 40C, pH 7, Vmax: 2668 micromol/min/g
42
-
D-Met
-
crude lyophilisates, 25C, pH 6, Vmax: 1160 micromol/min/g
4.8
-
D-methionine
-
100% aqueous
9
-
D-methionine
-
10% methanol
11.1
-
D-methionine
-
10% acetonitril
11.9
-
D-methionine
-
40% acetonitril
13.2
-
D-methionine
-
20% acetonitril
15.8
-
D-methionine
-
20% methanol
24.3
-
D-methionine
-
40% methanol
25.7
-
D-Phe
-
-
1
-
D-Ser
-
L-Arg; L-Lys
14.5
-
D-serine
-
pH 8.0, 37C, presence of 1 mM ATP, racemization reaction
49
-
D-serine
-
pH 8.0, 37C, racemization reaction
9.8
-
D-Thr
-
-
36
-
L-2-aminobutyrate
-
-
36
-
L-2-aminobutyrate
-
L-Ala
30
-
L-Asn
-
pure lyophilisates, 20C, pH 7, Vmax: 1626 micromol/min/g
47
-
L-Asn
-
pure lyophilisates, 25C, pH 7, Vmax: 2427 micromol/min/g
76
-
L-Asn
-
pure lyophilisates, 30C, pH 7, Vmax: 3930 micromol/min/g
101
-
L-Asn
-
pure lyophilisates, 35C, pH 7, Vmax: 5683 micromol/min/g
8
-
L-aspartate
-
in 50 mM Tris-HCl buffer, pH 8.0, at 30C
35
-
L-ethionine
-
-
33
-
L-Leu
-
L-2-aminobutyrate
0.55
-
L-Lys
-
-
1.05
-
L-Lys
-
-
2.5
-
L-Lys
-
D-Lys
1.2
-
L-Met
-
-
2
3
L-Met
-
crude lyophilisates, 25C, pH 7, Vmax: 2150 micromol/min/g
20
-
L-Met
-
crude lyophilisates, 25C, pH 8, Vmax: 2951 micromol/min/g
30
-
L-Met
-
crude lyophilisates, 25C, pH 6, Vmax: 1174 micromol/min/g; crude lyophilisates, 30C, pH 7, Vmax: 2296 micromol/min/g
31
-
L-Met
-
crude lyophilisates, 20C, pH 7, Vmax: 1889 micromol/min/g
35
-
L-Met
-
crude lyophilisates, 40C, pH 7, Vmax: 2847 micromol/min/g
39
-
L-Met
-
crude lyophilisates, 35C, pH 7, Vmax: 2833 micromol/min/g
4.1
-
L-methionine
-
100% aqueous
7.8
-
L-methionine
-
10% methanol
9.4
-
L-methionine
-
20% methanol
11.5
-
L-methionine
-
10% acetonitril
13.2
-
L-methionine
-
20% acetonitril
16.1
-
L-methionine
-
40% acetonitril
22.3
-
L-methionine
-
40% methanol
0.9
-
L-Orn
-
-
0.9
-
L-Orn
-
D-Lys; L-Orn
3.8
-
L-serine
-
pH 8.0, 37C, presence of 1 mM ATP, racemization reaction
15
-
L-serine
-
in 50 mM Tris-HCl buffer, pH 8.5, for Ser reactions, at 30C
16
-
L-serine
-
in 50 mM Tris-HCl buffer, pH 8.5, for Ser reactions, at 30C
30
-
L-serine
-
pH 8.0, 37C, racemization reaction
17
-
N-acetyl-D-alanine
-
-
18
-
N-acetyl-D-asparagine
-
-
7
-
N-acetyl-D-methionine
-
-
2
3
N-acetyl-D-phenylalanine
-
-
2
-
N-acetyl-D-tryptophan
-
-
41
-
N-acetyl-L-alanine
-
-
27
-
N-acetyl-L-asparagine
-
-
8
-
N-acetyl-L-methionine
-
-
43
-
N-acetyl-L-phenylalanine
-
-
2
-
N-acetyl-L-tryptophan
-
-
2
-
N-carbamoyl-D-methionine
-
-
5
-
N-carbamoyl-L-methionine
-
-
0.13
-
N-succinyl-D-alanine
-
-
0.04
-
N-succinyl-D-phenylalanine
-
-
2.6
-
N-Succinyl-L-Ala
-
-
3.8
-
N-Succinyl-L-His
-
-
3.7
-
N-Succinyl-L-Ile
-
-
3.6
-
N-Succinyl-L-Met
-
-
0.8
-
N-succinyl-L-Phe
-
-
0.13
-
N-succinyl-L-phenylalanine
-
-
7.1
-
N-Succinyl-L-Trp
-
-
2.6
-
N-Succinyl-L-Tyr
-
-
2.5
-
N-Succinyl-L-Val
-
-
0.12
-
N-succinyl-L-alanine
-
-
additional information
-
N-Succinyl-L-Arg
-
low specific rotation prevented kinetic analysis
additional information
-
N-Succinyl-L-Asn
-
low specific rotation prevented kinetic analysis
additional information
-
N-Succinyl-L-Asp
-
low specific rotation prevented kinetic analysis
additional information
-
N-Succinyl-L-Cys
-
low specific rotation prevented kinetic analysis
additional information
-
N-Succinyl-L-Gln
-
low specific rotation prevented kinetic analysis
additional information
-
N-Succinyl-L-Glu
-
low specific rotation prevented kinetic analysis
1.3
-
N-Succinyl-L-Leu
-
-
additional information
-
N-Succinyl-L-Lys
-
low specific rotation prevented kinetic analysis
3
-
N-Succinyl-L-Ser
-
-
additional information
-
N-Succinyl-L-Thr
-
low specific rotation prevented kinetic analysis
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
14.5
-
D-serine
-
pH 8.0, 37C, presence of 1 mM ATP, racemization reaction
3.8
-
L-serine
-
pH 8.0, 37C, presence of 1 mM ATP, racemization reaction
0.8
-
N-acetyl-D-alanine
-
-
0.07
-
N-acetyl-D-asparagine
-
-
20
-
N-acetyl-D-methionine
-
-
10
-
N-acetyl-D-phenylalanine
-
-
0.09
-
N-acetyl-D-tryptophan
-
-
2
-
N-acetyl-L-alanine
-
-
0.06
-
N-acetyl-L-asparagine
-
-
22
-
N-acetyl-L-methionine
-
-
16
-
N-acetyl-L-phenylalanine
-
-
0.15
-
N-acetyl-L-tryptophan
-
-
2
-
N-carbamoyl-D-methionine
-
-
2
-
N-carbamoyl-L-methionine
-
-
15
-
N-succinyl-D-alanine
-
-
2
-
N-succinyl-D-phenylalanine
-
-
89
-
N-Succinyl-L-Ala
-
-
43
-
N-succinyl-L-alanine
-
-
0.014
-
N-Succinyl-L-Arg
-
-
2.5
-
N-Succinyl-L-Asn
-
-
0.071
-
N-Succinyl-L-Asp
-
-
9
-
N-Succinyl-L-Gln
-
-
0.073
-
N-Succinyl-L-Glu
-
-
2
-
N-Succinyl-L-His
-
-
35
-
N-Succinyl-L-Ile
-
-
17
-
N-Succinyl-L-Leu
-
-
2.1
-
N-Succinyl-L-Lys
-
-
53
-
N-Succinyl-L-Met
-
-
19
-
N-succinyl-L-Phe
-
-
5
-
N-succinyl-L-phenylalanine
-
-
45
-
N-Succinyl-L-Ser
-
-
7.9
-
N-Succinyl-L-Trp
-
-
52
-
N-Succinyl-L-Tyr
-
-
92
-
N-Succinyl-L-Val
-
-
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.047
-
N-acetyl-D-alanine
-
-
15855
0.004
-
N-acetyl-D-asparagine
-
-
20393
3
-
N-acetyl-D-methionine
-
-
10717
0.467
-
N-acetyl-D-phenylalanine
-
-
15857
0.061
-
N-acetyl-D-tryptophan
-
-
16118
0.039
-
N-acetyl-L-alanine
-
-
2972
0.002
-
N-acetyl-L-asparagine
-
-
6135
2.8
-
N-acetyl-L-methionine
-
-
1165
0.37
-
N-acetyl-L-phenylalanine
-
-
2855
0.06
-
N-acetyl-L-tryptophan
-
-
4835
1.2
-
N-carbamoyl-D-methionine
-
-
16690
1.1
-
N-carbamoyl-L-methionine
-
-
4105
119
-
N-succinyl-D-alanine
-
-
40130
38.1
-
N-succinyl-D-phenylalanine
-
-
40131
352
-
N-succinyl-L-alanine
-
-
40129
35.1
-
N-succinyl-L-phenylalanine
-
-
14015
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.00588
-
-
substrate Trp, wildtype BAR
0.00706
-
-
substrate Phe, mutant Y396C
0.00782
-
-
substrate Ala, wildtype BAR
0.00838
-
-
substrate Phe, wildtype BAR
0.0121
-
-
substrate Ala, mutant I83L, D361V, Y396C
0.0141
-
-
substrate Ala, mutant Y293A, Y301S, Y396C
0.0144
-
-
substrate Ala, mutant L126H, Y396C
0.0249
-
-
substrate Phe, mutant Y293A, Y301S, Y396C
0.0257
-
-
substrate Phe, mutant I83L, D361V, Y396C
0.026
-
-
substrate Ala, mutant I384M
0.0267
-
-
substrate Trp, mutant Y293A, Y301S, Y396C
0.0303
-
-
substrate Trp, mutant I83L, D361V, Y396C
0.03215
-
-
substrate Lys, mutant Y396C
0.0436
-
-
substrate Phe, mutant L126H, Y396C
0.0487
-
-
substrate Trp, mutant Y396C
0.0524
-
-
substrate Trp, mutant L126H, Y396C
0.0744
-
-
substrate Phe, mutant I384M
0.124
-
-
substrate Trp, mutant I384M
0.132
-
-
substrate Lys, wildtype BAR
0.223
-
-
substrate Lys, mutant I384M
0.458
-
-
substrate Lys, mutant Y293A, Y301S, Y396C
0.473
-
-
substrate Lys, mutant I83L, D361V, Y396C
0.477
-
-
substrate Lys, mutant L126H, Y396C
0.825
-
-
substrate Ala, mutant Y396C
708.4
-
-
L-Met as substrate
708.4
-
-
-
920
-
-
L-Lys as substrate
1120
-
I0J1I6, -
recombinant protein
1250
-
-
L-Lys as substrate
additional information
-
-
wildtype BAR, specific activity for L-Lys 100%, L-Arg 65%, L-Ala 33%, L-Ser 20%, L-Met 14%, L-Cys 14%, L-Leu 3.3%, L-His 1.6%, L-Phe 0.29%, L-Pro 0.1.3%, L-Thr 0.069%, L-Asn 0.054%, L-Asp 0.01%, L-Trp 0.006%, L-Val 0.003%
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.5
8.5
-
-
7.9
-
-
assay at
8
9.5
-
-
8
-
I0J1I6, -
optimal activity
8.5
-
-
assay at
10
-
I0J1I6, -
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.5
-
-
negligible activity below
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
-
-
assay at
37
-
-
assay at
37
-
I0J1I6, -
assay at
65
-
I0J1I6, -
maximum activity
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
DAR1 is localized to the medial region of the cerebral ganglion where the F- and C-clusters are situated
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
Deinococcus radiodurans (strain ATCC 13939 / DSM 20539 / JCM 16871 / LMG 4051 / NBRC 15346 / NCIMB 9279 / R1 / VKM B-1422)
Deinococcus radiodurans (strain ATCC 13939 / DSM 20539 / JCM 16871 / LMG 4051 / NBRC 15346 / NCIMB 9279 / R1 / VKM B-1422)
Deinococcus radiodurans (strain ATCC 13939 / DSM 20539 / JCM 16871 / LMG 4051 / NBRC 15346 / NCIMB 9279 / R1 / VKM B-1422)
Deinococcus radiodurans (strain ATCC 13939 / DSM 20539 / JCM 16871 / LMG 4051 / NBRC 15346 / NCIMB 9279 / R1 / VKM B-1422)
Deinococcus radiodurans (strain ATCC 13939 / DSM 20539 / JCM 16871 / LMG 4051 / NBRC 15346 / NCIMB 9279 / R1 / VKM B-1422)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
41500
-
I0J1I6, -
calculated from cDNA
42300
-
-
molecular weight of monomeric subunit, SDS-PAGE
43000
-
-
SDS-PAGE
45000
-
-
SDS-PAGE
55000
-
-
isoforms A and B, gel filtration
57000
-
-
gel filtration
62000
65000
-
gel filtration
73000
-
-
gel filtration
76000
-
-
sedimentation equilibrium method
78000
-
-
gel filtration
80000
-
-
-
82000
-
-
gel filtration
84000
-
I0J1I6, -
gel filtration
110000
-
-
sedimentation equilibrium method
110000
-
-
gel filtration
110000
-
-
-
150000
-
-
gel filtration, tetramer
170000
-
-
tetramer, analytical ultracentrifugation
177300
-
-
tetramer, gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 42000, SDS-PAGE
?
-
x * 45000, SDS-PAGE
?
-
x * 36121, MALDI-MS, x * 36123, calculated
dimer
-
2 * 40000, SDS-PAGE
dimer
-
2 * 41000, SDS-PAGE
dimer
-
2 * 37000, SDS-PAGE
dimer
I0J1I6, -
2 * 41500 Da
homodimer
-
2 * 36000, SDS-PAGE
homodimer
-
2 * 45000 Da, SDS-PAGE
homodimer
-
2 * 35400, calculated from amino acid sequence
homotetramer
-
4 * 43000, SDS-PAGE
tetramer
-
gel filtration and analytical ultracentrifugation. Tetrameric structure in the absence or presence of Co2+, 4 * 42300 Da
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
I0J1I6, -
N-terminal signal sequence is cleaved off
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.5
8.5
-
the enzyme shows little racemase activity below pH 6.5 and an increasing activity between pH 7.0-8.5 in 50 mM Tris-HCl buffer
7
11
-
at 30C, 1 h, stable
10
-
-
at 50C, 10 min, stable
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
14
50
-
the enzyme activity increases between 14-45C and starts to decrease at 50C for both the Ser and Asp conversion
50
-
-
pH 10.0, 10 min, stable
60
-
-
stable up to 60C, pH 8.0, 10 min, in 0.1 M potassium phosphate
60
-
-
GkNSAAR shows a gradual loss of activity at preincubation temperatures over 60C
80
-
-
pH 10.0, 10 min, most of the activity is lost
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
buffer without organic content provides excellent stability at moderate temperatures (2035C) while addition of 20% acetonitrile or methanol drastically reduces the half-life of the racemase
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
0-5C, 0.01 M potassium phosphate buffer, pH 7.0, 2 mM pyridoxal 5'-phosphate, 1 month, stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Ni-NTA column chromatography and Sepharcryl 200 gel filtration
-
His-tag is removed by thrombin cleavage
-
protein is purified by applying to a diethylaminoethyl sepharose fast flow column and a phenyl sepharose 6 fast flow column
-
purified in a one-step procedure by immobilized cobalt affinity chromatography
-
expression in Escherichia coli with N-terminal His-tag, purification protocol from inclusion bodies
-
both isoforms A and B
-
recombinant enzyme expressed in insect cells
-
His6-tagged BAR is applied onto a His-Trap HP 1-ml column
-
two enzyme lyophilisates of different purity are obtained from which the crude is sufficient for the racemization of methionine and the pure is used for asparagine
-
using Ni-NTA chromatography
I0J1I6, -
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
-
expressed as a His-taged fusion protein
-
over-expressed in Escherichia coli
-
protein is expressed in Escherichia coli strain BL21
-
amino acid racemase of Pseudomonas putida DSM 3263 is overexpressed in Escherichia coli and delivered cell free extract with easily sufficient activity (2050 U/mg total protein) for application in an enzyme membrane reactor (EMR) setting
-
over-expressed in Escherichia coli
-
recombinant BAR is cloned in Escherichia coli BL-21 as His-tagged BAR
-
expressed in Escherichia coli as a His-tagged fusion protein
I0J1I6, -
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
H152S
-
ratio of elimination reaction to racemization is 1.4 compared to 3.7 in wild-type
N154F
-
ratio of elimination reaction to racemization is 0.33 compared to 3.7 in wild-type
P153S
-
ratio of elimination reaction to racemization is 0.24 compared to 3.7 in wild-type
Q155D
-
ratio of elimination reaction to racemization is 0.25 compared to 3.7 in wild-type
I384M
-
I384M mutant BAR shows the highest activity for Trp. Using I384M mutant BAR the proportion of D-Trp reaches 43% while wildtype BAR racemizes only 6% of initial L-Trp, specific activity: 0.124 (substrate Trp), 0.0744 (substrate Phe), 0.223 (substrate Lys), 0.026 (substrate Ala)
I83L/D361V/Y396C
-
specific activity: 0.0303 (substrate Trp), 0.0257 (substrate Phe), 0.473 (substrate Lys), 0.0121 (substrate Ala)
Y293A/Y301S/Y396C
-
specific activity: 0.0267 (substrate Trp), 0.0249 (substrate Phe), 0.458 (substrate Lys), 0.0141 (substrate Ala)
Y396C
-
specific activity: 0.0487 (substrate Trp), 0.00706 (substrate Phe), 0.03215 (substrate Lys), 0.825 (substrate Ala)
L126H/Y396C
-
specific activity: 0.0524 (substrate Trp), 0.0436 (substrate Phe), 0.477 (substrate Lys), 0.0144 (substrate Ala)
additional information
-
random mutagenesis on bar gene is performed to obtain mutant BAR derivates with high activity for Trp. Five positive mutant are isolated after the two-step-screening of the randomly mutated BAR, substitutions at Y396 and I384 increases the Trp specific racemization activity and the racemazation activity for overall amino acids, respectively
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
analysis
-
simple procedure for in situ analysis of stereospecificity of C-4 hydrogen transfer of NADH by an NAD-dependent dehydrogenase by combination with amino acid racemase, EC 5.1.1.10, and L-leucine dehydrogenase, EC 1.4.1.9
biotechnology
-
since commercially available D-Trp is chemically synthesized and expensive a mutant BAR protein might represent an effective method to synthesize D-Trp
biotechnology
-
an enzyme lyophilisate of amino acid racemase from Pseudomonas putida is used for in situ racemization. Crystallization experiments accompanied by enzymatic racemization lead to a significant increase of crystallized L-Asn
synthesis
-
enzymatic production of L-Trp from DL-Ser and indole by a coupled reaction of tryptophan synthase and amino acid racemase