Information on EC 5.1.1.10 - amino-acid racemase

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

EC NUMBER
COMMENTARY hide
5.1.1.10
-
RECOMMENDED NAME
GeneOntology No.
amino-acid racemase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
an L-amino acid = a D-amino acid
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
racemization
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
cysteine metabolism
-
-
serine metabolism
-
-
Glycine, serine and threonine metabolism
-
-
Cysteine and methionine metabolism
-
-
D-Glutamine and D-glutamate metabolism
-
-
D-Arginine and D-ornithine metabolism
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
amino-acid racemase
A pyridoxal-phosphate protein.
CAS REGISTRY NUMBER
COMMENTARY hide
9068-61-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
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
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
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
D-Arg
L-Arg
show the reaction diagram
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
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
6.2% activity, compared to D-Lys
-
-
?
D-Ile
L-Ile
show the reaction diagram
-
-
-
-
r
D-Leu
L-Leu
show the reaction diagram
D-Lys
L-Lys
show the reaction diagram
100% activity
-
-
?
D-Met
L-Met
show the reaction diagram
D-norvaline
L-norvaline
show the reaction diagram
6.5% activity, compared to D-Lys
-
-
?
D-ornithine
L-ornithine
show the reaction diagram
42% activity, compared to D-Lys
-
-
?
D-Phe
L-Phe
show the reaction diagram
D-serine
L-serine
show the reaction diagram
D-serine
pyruvate + NH3
show the reaction diagram
-
alpha,beta-elimination reaction
-
-
?
D-Trp
L-Trp
show the reaction diagram
-
-
-
-
r
D-Tyr
L-Tyr
show the reaction diagram
-
-
-
-
r
D-Val
L-Val
show the reaction diagram
-
-
-
-
r
L-2-Aminobutyrate
D-2-Aminobutyrate
show the reaction diagram
L-Ala
D-Ala
show the reaction diagram
L-Asn
D-Asn
show the reaction diagram
-
-
-
-
r
L-Asn
L-Asn
show the reaction diagram
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
L-Ethionine
D-Ethionine
show the reaction diagram
L-Glu
D-Glu
show the reaction diagram
L-His
D-His
show the reaction diagram
L-Homoarginine
L-Homoarginine
show the reaction diagram
L-Homocitrulline
D-Homocitrulline
show the reaction diagram
L-Homocysteine
D-Homocysteine
show the reaction diagram
-
55% of the activity relative to L-Lys
-
-
L-Homoserine
L-Homoserine
show the reaction diagram
L-Ile
D-Ile
show the reaction diagram
-
-
-
-
r
L-Leu
D-Leu
show the reaction diagram
L-Lys
D-Lys
show the reaction diagram
L-Met
D-Met
show the reaction diagram
L-Methionine
D-Methionine
show the reaction diagram
-
-
-
-
r
L-Norleucine
D-Norleucine
show the reaction diagram
L-Norvaline
D-Norvaline
show the reaction diagram
-
-
-
-
L-Orn
D-Orn
show the reaction diagram
L-Phe
D-Phe
show the reaction diagram
-
-
-
-
r
L-Selenohomocysteine
D-Selenohomocysteine
show the reaction diagram
-
27% of the activity relative to L-Lys
-
-
L-Ser
D-Ser
show the reaction diagram
L-serine
D-serine
show the reaction diagram
L-serine
pyruvate + NH3
show the reaction diagram
-
alpha,beta-elimination reaction
-
-
?
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
L-threonine
3-hydroxy-2-butenoic acid + NH3
show the reaction diagram
-
alpha,beta-elimination reaction
-
-
?
L-Trp
D-Trp
show the reaction diagram
L-Tyr
D-Tyr
show the reaction diagram
-
-
-
-
r
L-Val
D-Val
show the reaction diagram
-
-
-
-
r
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
N6-Acetyl-L-Lys
N6-Acetyl-D-Lys
show the reaction diagram
S-Methyl-L-Cys
S-Methyl-D-Cys
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
D-Leu
L-Leu
show the reaction diagram
O57878
the enzyme is likely responsible for utilization of d-amino acids for growth
-
-
?
D-Met
L-Met
show the reaction diagram
D-Phe
L-Phe
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyridoxal 5'-phosphate
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ni2+
-
enhances activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
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-cycloserine
-
-
D-penicillamine
-
-
dithiothreitol
-
-
EDTA
-
inhibition of both activities
Fe2+
-
slight inhibition of both activities
hydroxylamine
Ni2+
-
slight inhibition of both activities
O-acetylserine
S-(N-Methylthiocarbamoyl)-D-Cys
-
-
S-(N-Methylthiocarbamoyl)-L-Cys
-
-
Zn2+
-
inhibition of both activities
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
7.4 - 35.6
D-Ala
18 - 103
D-Asn
94
D-Aspartate
-
in 50 mM Tris-HCl buffer, pH 8.0, at 30°C
7.8
D-Gln
-
-
9.54
D-Ile
-
pH 7.0, 80°C
4.19
D-Leu
-
pH 7.0, 80°C
0.42 - 30
D-Lys
1.6 - 42
D-Met
4.8 - 24.3
D-methionine
2.78 - 26
D-Phe
1 - 18
D-Ser
14.5 - 49
D-serine
9.8
D-Thr
-
-
1.68
D-Trp
-
pH 7.0, 80°C
2.77
D-Tyr
-
pH 7.0, 80°C
9.82
D-Val
-
pH 7.0, 80°C
36
L-2-aminobutyrate
7 - 56
L-Ala
30 - 101
L-Asn
8
L-aspartate
-
in 50 mM Tris-HCl buffer, pH 8.0, at 30°C
35
L-ethionine
-
-
3.94
L-Ile
-
pH 7.0, 80°C
4.95 - 33
L-Leu
0.55 - 2.5
L-Lys
1.2 - 39
L-Met
4.1 - 22.3
L-methionine
0.9
L-Orn
8.23
L-Phe
-
pH 7.0, 80°C
3.8 - 30
L-serine
31
L-Thr
-
-
5.84
L-Trp
-
pH 7.0, 80°C
2.23
L-Tyr
-
pH 7.0, 80°C
3.41
L-Val
-
pH 7.0, 80°C
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
-
-
0.12
N-succinyl-L-alanine
-
-
3.8
N-succinyl-L-His
-
-
3.7
N-succinyl-L-Ile
-
-
1.3
N-Succinyl-L-Leu
-
-
3.6
N-Succinyl-L-Met
-
-
0.8
N-succinyl-L-Phe
-
-
0.13
N-succinyl-L-phenylalanine
-
-
3
N-succinyl-L-Ser
-
-
7.1
N-succinyl-L-Trp
-
-
2.6
N-succinyl-L-Tyr
-
-
2.5
N-succinyl-L-Val
-
-
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
14.5
D-serine
-
pH 8.0, 37°C, presence of 1 mM ATP, racemization reaction
3.8
L-serine
-
pH 8.0, 37°C, 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
-
-
additional information
additional information
-
no activity with N-Succinyl-L-Pro; N-Succinyl-L-Cys, low specific rotation prevented kinetic analysis; N-Succinyl-L-Thr, low specific rotation prevented kinetic analysis
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.95
D-Ala
-
pH 7.0, 80°C
4.12
D-Ile
-
pH 7.0, 80°C
18.4
D-Leu
-
pH 7.0, 80°C
12.7
D-Met
-
pH 7.0, 80°C
19.9
D-Phe
-
pH 7.0, 80°C
1.31
D-Trp
-
pH 7.0, 80°C
5.69
D-Tyr
-
pH 7.0, 80°C
2.67
D-Val
-
pH 7.0, 80°C
0.159
L-Ala
-
pH 7.0, 80°C
5.5
L-Ile
-
pH 7.0, 80°C
11.9
L-Leu
-
pH 7.0, 80°C
8.26
L-Met
-
pH 7.0, 80°C
11
L-Phe
-
pH 7.0, 80°C
2.39
L-Trp
-
pH 7.0, 80°C
11.4
L-Tyr
-
pH 7.0, 80°C
1.97
L-Val
-
pH 7.0, 80°C
0.047
N-acetyl-D-alanine
-
-
0.004
N-acetyl-D-asparagine
-
-
3
N-acetyl-D-methionine
-
-
0.467
N-acetyl-D-phenylalanine
-
-
0.061
N-acetyl-D-tryptophan
-
-
0.039
N-acetyl-L-alanine
-
-
0.002
N-acetyl-L-asparagine
-
-
2.8
N-acetyl-L-methionine
-
-
0.37
N-acetyl-L-phenylalanine
-
-
0.06
N-acetyl-L-tryptophan
-
-
1.2
N-carbamoyl-D-methionine
-
-
1.1
N-carbamoyl-L-methionine
-
-
119
N-succinyl-D-alanine
-
-
38.1
N-succinyl-D-phenylalanine
-
-
352
N-succinyl-L-alanine
-
-
35.1
N-succinyl-L-phenylalanine
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
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
2
-
substrate: D-Ala, pH 8.0, 80°C
3.9
-
substrate: D-Val, pH 8.0, 80°C
10.5
-
substrate: D-Met, pH 8.0, 80°C
15.6
-
substrate: D-Leu, pH 8.0, 80°C
22.7
-
substrate: D-Phe, pH 8.0, 80°C
920
-
L-Lys as substrate
1120
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
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 7.5
-
at a temperature of 60°C, the highest activity is detected at around pH 6.5-7.5
7.5 - 9
-
-
7.5 - 10
-
-
7.5 - 8.5
-
-
8 - 9.5
-
-
8.5
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5
-
negligible activity below
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20
-
assay at
50 - 60
-
-
65
maximum activity
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50 - 80
-
the relative activities at 50°C and 80°C are 12% and 70%, respectively, as compared with that at 95°C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
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
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35400
-
2 * 35400, calculated from amino acid sequence
36000
-
2 * 36000, SDS-PAGE
36121
-
x * 36121, MALDI-MS, x * 36123, calculated
36123
-
x * 36121, MALDI-MS, x * 36123, calculated
37000
-
2 * 37000, SDS-PAGE
40000
-
2 * 40000, SDS-PAGE
41000
-
2 * 41000, SDS-PAGE
41500
calculated from cDNA
42000
-
x * 42000, SDS-PAGE
42300
-
molecular weight of monomeric subunit, SDS-PAGE
43000
-
SDS-PAGE
52392
-
calculated from sequence
55000
-
isoforms A and B, gel filtration
57000
-
gel filtration
62000 - 65000
-
gel filtration
73000
-
gel filtration
76000
-
sedimentation equilibrium method
78000
-
gel filtration
82000
-
gel filtration
84000
gel filtration
91000
-
gel filtration
110000
150000
-
gel filtration, tetramer
170000
-
tetramer, analytical ultracentrifugation
177300
-
tetramer, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
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
COMMENTARY hide
LITERATURE
proteolytic modification
N-terminal signal sequence is cleaved off
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
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
715591
7 - 11
-
at 30°C, 1 h, stable
2049
10
-
at 50°C, 10 min, stable
2049
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
14 - 50
-
the enzyme activity increases between 14-45°C and starts to decrease at 50°C for both the Ser and Asp conversion
50
-
pH 10.0, 10 min, stable
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
buffer without organic content provides excellent stability at moderate temperatures (20–35°C) while addition of 20% acetonitrile or methanol drastically reduces the half-life of the racemase
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
0-5°C, 0.01 M potassium phosphate buffer, pH 7.0, 2 mM pyridoxal 5'-phosphate, 1 month, stable
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
both isoforms A and B
-
expression in Escherichia coli with N-terminal His-tag, purification protocol from inclusion bodies
-
His-tag is removed by thrombin cleavage
-
His6-tagged BAR is applied onto a His-Trap HP 1-ml column
-
Ni-NTA column chromatography and Sepharcryl 200 gel filtration
-
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
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recombinant enzyme expressed in insect cells
-
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
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
amino acid racemase of Pseudomonas putida DSM 3263 is overexpressed in Escherichia coli and delivered cell free extract with easily sufficient activity (20–50 U/mg total protein) for application in an enzyme membrane reactor (EMR) setting
-
enzyme is produced mainly as an inclusion body in Escherichia coli. In this study, expression of the recombinant protein into the soluble fraction is markedly improved by coexpression with chaperone molecules
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expressed as a His-taged fusion protein
-
expressed in Escherichia coli as a His-tagged fusion protein
expressed in Escherichia coli BL21(DE3) cells
-
expression in Escherichia coli
-
over-expressed in Escherichia coli
protein is expressed in Escherichia coli strain BL21
-
recombinant BAR is cloned in Escherichia coli BL-21 as His-tagged BAR
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
activity is enhanced in cells grown in the medium supplemented with D-amino acids especially D-allo-Ile
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
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)
L126H/Y396C
-
specific activity: 0.0524 (substrate Trp), 0.0436 (substrate Phe), 0.477 (substrate Lys), 0.0144 (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)
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
COMMENTARY hide
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
synthesis
-
enzymatic production of L-Trp from DL-Ser and indole by a coupled reaction of tryptophan synthase and amino acid racemase