Information on EC 3.5.1.86 - mandelamide amidase

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

EC NUMBER
COMMENTARY
3.5.1.86
-
RECOMMENDED NAME
GeneOntology No.
mandelamide amidase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
(R)-mandelamide + H2O = (R)-mandelate + NH3
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of amide bond
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Aminobenzoate degradation
-
-
Microbial metabolism in diverse environments
-
-
SYSTEMATIC NAME
IUBMB Comments
mandelamide hydrolase
-
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
amidase, mandelamide
-
-
-
-
Pseudomonas mandelamide hydrolase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
198496-85-4
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Corynebacterium nitrilophilus
ATCC 21419
-
-
Manually annotated by BRENDA team
strain C5
-
-
Manually annotated by BRENDA team
Corynebacterium sp. C5
strain C5
-
-
Manually annotated by BRENDA team
strain AC777
-
-
Manually annotated by BRENDA team
Mycobacterium sp. AC777
strain AC777
-
-
Manually annotated by BRENDA team
ATCC 12633
-
-
Manually annotated by BRENDA team
strain ATCC 12633
-
-
Manually annotated by BRENDA team
strain AK32
-
-
Manually annotated by BRENDA team
Rhodococcus sp. AK32
strain AK32
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(R)-(+)-mandelamide + H2O
(R)-(+)-mandelate + NH3
show the reaction diagram
-
-
-
-
?
(R)-(-)-mandelamide + H2O
(R)-(-)mandelate + NH3
show the reaction diagram
-
-
-
?
(R)-(-)-mandelamide + H2O
(R)-(-)mandelate + NH3
show the reaction diagram
-
-
-
?
(R)-2-aminophenylacetamide + H2O
(R)-2-aminophenylacetate + NH3
show the reaction diagram
-
-
-
-
?
(R)-lactamide + H2O
(R)-lactate + NH3
show the reaction diagram
-
low efficiency
-
-
?
(R)-mandelamide + H2O
(R)-mandelic acid + ammonia
show the reaction diagram
-
-
-
-
?
(R,S)-methyl mandelate + H2O
?
show the reaction diagram
Q84DC4
-
-
?
(S)-(+)-mandelamide + H2O
(S)-(+)-mandelate + NH3
show the reaction diagram
-
-
-
?
(S)-(+)-mandelamide + H2O
(S)-(+)-mandelate + NH3
show the reaction diagram
-
-
-
?
(S)-(+)-mandelamide + H2O
(S)-(+)-mandelate + NH3
show the reaction diagram
-
-
-
-
?
(S)-2-aminophenylacetamide + H2O
(S)-2-aminophenylacetate + NH3
show the reaction diagram
-
-
-
-
?
(S)-lactamide + H2O
(S)-lactate + NH3
show the reaction diagram
-
low efficiency
-
-
?
(S)-mandelamide + H2O
(S)-mandelic acid + ammonia
show the reaction diagram
-
-
-
-
?
2-cyclohexylacetamide + H2O
2-cyclohexylacetate + NH3
show the reaction diagram
-
-
-
-
?
2-phenylacetamide + H2O
?
show the reaction diagram
Q84DC4
-
-
?
2-phenylacetamide + H2O
2-phenylacetate + NH3
show the reaction diagram
-
-
-
-
?
3,4-dichloroisocoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
3-phenylpropionamide + H2O
?
show the reaction diagram
Q84DC4
-
-
?
3-phenylpropionamide + H2O
3-phenylpropionate + NH3
show the reaction diagram
-
-
-
-
?
acetamide + H2O
acetate + NH3
show the reaction diagram
-
-
-
-
?
butanamide + H2O
butanoate + NH3
show the reaction diagram
-
-
-
-
?
caproate + H2O
caproamide + NH3
show the reaction diagram
-
-
-
-
?
cyclohexanecarboxamide + H2O
cyclohexanecarboxylate + NH3
show the reaction diagram
-
-
-
-
?
N-methyl phenylacetamide + H2O
?
show the reaction diagram
Q84DC4
-
-
?
pentanamide + H2O
pentanoate + NH3
show the reaction diagram
-
-
-
-
?
phenylacetamide + H2O
phenylacetate + NH3
show the reaction diagram
-
-
-
-
?
phenylmethylsulfonyl fluoride + H2O
?
show the reaction diagram
-
-
-
-
?
propanamide + H2O
propanoate + NH3
show the reaction diagram
-
-
-
-
?
R,S-mandelamide + H2O
mandelic acid + ammonia
show the reaction diagram
Q84DC4
-
-
?
hexanoamide + H2O
?
show the reaction diagram
Q84DC4
-
-
?
additional information
?
-
-
benzamide is not a substrate
-
-
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3-phenyl-1,1,1-trifluoro-2-propanone
Q84DC4
competitive
3-phenyl-1-chloro-2-propanone
Q84DC4
competitive
PMSF
Q84DC4
1 mM, 80% inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0342
(R)-(+)-mandelamide
-
at 30C and pH 7.8
15
(R)-(-)-mandelamide
-
-
46
(R)-(-)-mandelamide
-
-
0.005
(R)-2-aminophenylacetamide
-
-
0.35
(R)-lactamide
-
Q207H/S316N/Q382E mutant
0.596
(R)-lactamide
-
wild-type
0.75
(R)-lactamide
-
G202S/Q207H/R236C/R369M/Q382E mutant
1
(R)-lactamide
-
G202A/Q207H/Q382E mutant
2
(R)-lactamide
-
I437N mutant
2.2
(R)-lactamide
-
T31I/I437N mutant
2.4
(R)-lactamide
-
Q207H/S316N/I437N mutant
600
(R)-lactamide
-
-
0.014
(R)-mandelamide
-
Q207H/S316N/Q382E mutant
0.018
(R)-mandelamide
-
wild-type
0.02
(R)-mandelamide
-
-
0.033
(R)-mandelamide
Q84DC4
pH 7.5, 30C
0.059
(R)-mandelamide
Q84DC4
pH 7.5, 30C, mutant S181A
0.129
(R)-mandelamide
-
I437N mutant
0.168
(R)-mandelamide
-
Q207H/S316N/I437N mutant
0.215
(R)-mandelamide
-
T31I/I437N mutant
0.871
(R)-mandelamide
Q84DC4
pH 7.5, 30C, mutant S180A
8.2
(R)-mandelamide
-
G202S/Q207H/R236C/R369M/Q382E mutant
8.8
(R)-mandelamide
-
G202A/Q207H/Q382E mutant
0.17
(R,S)-methyl mandelate
Q84DC4
pH 7.5, 30C
0.0198
(S)-(+)-mandelamide
-
at 30C and pH 7.8
0.006
(S)-2-aminophenylacetamide
-
-
0.149
(S)-lactamide
-
Q207H/S316N/I437N mutant
0.219
(S)-lactamide
-
I437N mutant
0.224
(S)-lactamide
-
T31I/I437N mutant
0.397
(S)-lactamide
-
Q207H/S316N/Q382E mutant
0.43
(S)-lactamide
-
wild-type
0.59
(S)-lactamide
-
G202A/Q207H/Q382E mutant
0.65
(S)-lactamide
-
G202S/Q207H/R236C/R369M/Q382E mutant
430
(S)-lactamide
-
-
0.0035
(S)-mandelamide
-
Q207H/S316N/Q382E mutant
0.007
(S)-mandelamide
-
-
0.0072
(S)-mandelamide
-
wild-type
0.009
(S)-mandelamide
Q84DC4
pH 7.5, 30C, mutant S181A
0.014
(S)-mandelamide
-
Q207H/S316N/I437N mutant
0.02
(S)-mandelamide
Q84DC4
pH 7.5, 30C
0.023
(S)-mandelamide
-
T31I/I437N mutant
0.029
(S)-mandelamide
-
I437N mutant
0.296
(S)-mandelamide
Q84DC4
pH 7.5, 30C, mutant S180A
3.1
(S)-mandelamide
-
G202A/Q207H/Q382E mutant
7.6
(S)-mandelamide
-
G202S/Q207H/R236C/R369M/Q382E mutant
0.04
2-cyclohexylacetamide
-
-
0.0009
2-phenylacetamide
-
-
0.0038
2-phenylacetamide
Q84DC4
pH 7.5, 30C
0.049
3-phenylpropionamide
Q84DC4
pH 7.5, 30C
490
acetamide
-
-
6.8
butanamide
-
-
0.12
Caproamide
-
-
0.55
cyclohexanecarboxamide
-
-
0.288
hexanoamide
Q84DC4
pH 7.5, 30C
0.032
N-methyl phenylacetamide
Q84DC4
pH 7.5, 30C
0.54
Pentanamide
-
-
45
propanamide
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
16.1
(R)-(+)-mandelamide
-
at 30C and pH 7.8
10.2
(R)-(-)-mandelamide
-
-
25
(R)-2-aminophenylacetamide
-
-
25
(R)-lactamide
-
T31I/I437N mutant
29
(R)-lactamide
-
I437N mutant
43
(R)-lactamide
-
; wild-type
64
(R)-lactamide
-
Q207H/S316N/I437N mutant
76
(R)-lactamide
-
G202S/Q207H/R236C/R369M/Q382E mutant
78
(R)-lactamide
-
Q207H/S316N/Q382E mutant
165
(R)-lactamide
-
G202A/Q207H/Q382E mutant
20
(R)-mandelamide
-
G202S/Q207H/R236C/R369M/Q382E mutant
51
(R)-mandelamide
-
G202A/Q207H/Q382E mutant
76
(R)-mandelamide
-
Q207H/S316N/I437N mutant
78
(R)-mandelamide
-
T31I/I437N mutant
85
(R)-mandelamide
-
I437N mutant
125
(R)-mandelamide
-
Q207H/S316N/Q382E mutant
150
(R)-mandelamide
-
; wild-type
0.031 - 0.51
(S)-(+)-mandelamide
-
at 30C and pH 7.8
4.4
(S)-(+)-mandelamide
-
-
7.73
(S)-(+)-mandelamide
-
at 30C and pH 7.8
27
(S)-2-aminophenylacetamide
-
-
19
(S)-lactamide
-
; wild-type
39
(S)-lactamide
-
I437N mutant; T31I/I437N mutant
40
(S)-lactamide
-
G202S/Q207H/R236C/R369M/Q382E mutant
49
(S)-lactamide
-
Q207H/S316N/I437N mutant
54
(S)-lactamide
-
Q207H/S316N/Q382E mutant
88
(S)-lactamide
-
G202A/Q207H/Q382E mutant
8.8
(S)-mandelamide
-
G202S/Q207H/R236C/R369M/Q382E mutant
18
(S)-mandelamide
-
G202A/Q207H/Q382E mutant
43
(S)-mandelamide
-
Q207H/S316N/Q382E mutant
82
(S)-mandelamide
-
; wild-type
89
(S)-mandelamide
-
Q207H/S316N/I437N mutant
101
(S)-mandelamide
-
I437N mutant
108
(S)-mandelamide
-
T31I/I437N mutant
1.1
2-cyclohexylacetamide
-
-
46
2-phenylacetamide
-
-
1.4
acetamide
-
-
0.61
butanamide
-
-
2.8
Caproamide
-
-
0.04
cyclohexanecarboxamide
-
-
2
Pentanamide
-
-
4.6
propanamide
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.06
3-phenyl-1,1,1-trifluoro-2-propanone
Q84DC4
pH 7.5, 30C
0.13
3-phenyl-1-chloro-2-propanone
Q84DC4
pH 7.5, 30C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.8
Q84DC4
-
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
52600
-
sedimentation analysis
680440
53810 - 53830
-
electrospray ionization mass spectrometry
288943
53820
-
deduced from amino acid sequence; electrospray mass spectrometry
680440
53850
-
matrix-assisted laser desorption ionization
288943
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
Q84DC4
x * 54000, SDS-PAGE, x * 53800, mass spectrometry
monomer
-
1 * 52600, sedimentation analysis
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, no loss of activtiy during several freeze-thaw cycles
Q84DC4
4C, phosphate buffer pH 7.8, 1 mM EDTA, stable for at least 1 month
Q84DC4
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
affinity chromatography
-
recombinant enzyme
-
Sephacryl S-200 gel filtration, HiPrep Q anion-exchange column chromatography, and HiTrap Q column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
-
expressed in Escherichia coli BL21(DE3)pLysS cells
-
expression in Escherichia coli
-
expression in Escherichia coli both wild type and His6-tagged protein
Q84DC4
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
G202A
-
site-directed mutagenesis. Gly202 appears to control the preference for aromatic substrates as the G202A variant shows three orders of magnitude decrease in kcat/Km for (R)- and (S)-mandelamide
G202A/Q207H/Q382E
-
site-directed mutagenesis
G202S/Q207H/R236C/R369M/Q382E
-
site-directed mutagenesis
G202V
-
site-directed mutagenesis. Reduction in activity increases to six orders of magnitude for the G202V variant
I437N
-
site-directed mutagenesis
K100A
Q84DC4
no enzymic activity
Q207H/S316N/I437N
-
site-directed mutagenesis
Q207H/S316N/Q382E
-
site-directed mutagenesis
S180A
Q84DC4
significant decrease in kcat value
S181A
Q84DC4
significant decrease in kcat value
S204A
Q84DC4
no enzymic activity
S217A
Q84DC4
very little enzymic activity
S218A
Q84DC4
very little enzymic activity
T31I/I437N
-
site-directed mutagenesis