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
-
-
-
-
an L-amino acid = a D-amino acid
show the reaction diagram
significant internal return of the alpha-hydrogen; single base mechanism
-
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
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
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
racemization
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Cysteine and methionine metabolism
-
D-Arginine and D-ornithine metabolism
-
D-Glutamine and D-glutamate metabolism
-
Glycine, serine and threonine metabolism
-
Metabolic pathways
-
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
-
-
13488
0.004
-
N-acetyl-D-asparagine
-
-
13490
3
-
N-acetyl-D-methionine
-
-
13530
0.467
-
N-acetyl-D-phenylalanine
-
-
13533
0.061
-
N-acetyl-D-tryptophan
-
-
22136
0.039
-
N-acetyl-L-alanine
-
-
13547
0.002
-
N-acetyl-L-asparagine
-
-
86295
2.8
-
N-acetyl-L-methionine
-
-
13566
0.37
-
N-acetyl-L-phenylalanine
-
-
13570
0.06
-
N-acetyl-L-tryptophan
-
-
13574
1.2
-
N-carbamoyl-D-methionine
-
-
86358
1.1
-
N-carbamoyl-L-methionine
-
-
86374
119
-
N-succinyl-D-alanine
-
-
293334
38.1
-
N-succinyl-D-phenylalanine
-
-
293335
352
-
N-succinyl-L-alanine
-
-
293333
35.1
-
N-succinyl-L-phenylalanine
-
-
236648
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 * 41000, SDS-PAGE
dimer
-
2 * 40000, SDS-PAGE
dimer
-
2 * 37000, SDS-PAGE
dimer
I0J1I6, -
2 * 41500 Da
homodimer
-
2 * 45000 Da, SDS-PAGE
homodimer
-
2 * 35400, calculated from amino acid sequence; 2 * 36000, SDS-PAGE
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