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 ACCESSION NO.
LITERATURE
(R)-mandelamide + H2O = (R)-mandelate + NH3
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of amide bond
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
mandelamide hydrolase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
amidase, mandelamide
-
-
-
-
MAH
Q84DC4
-
mandelamide hydrolase
-
-
Pseudomonas mandelamide hydrolase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
198496-85-4
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
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 ACCESSION NO.
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 ACCESSION NO.
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]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
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]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
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]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
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]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.8
-
-, Q84DC4
-
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
52600
-
-
sedimentation analysis
53810
53830
-
electrospray ionization mass spectrometry
53820
-
-
deduced from amino acid sequence; electrospray mass spectrometry
53850
-
-
matrix-assisted laser desorption ionization
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-, Q84DC4
x * 54000, SDS-PAGE, x * 53800, mass spectrometry
monomer
-
1 * 52600, sedimentation analysis
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
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 ACCESSION NO.
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 ACCESSION NO.
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 ACCESSION NO.
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