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
-
-
-
-
mandelamide hydrolase
-
-
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
-
-
-
?
(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
-
-
-
?
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
-
-
-
?
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
-
-
-
?
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
-
-
?
hexanoamide + H2O
?
show the reaction diagram
-
-
-
?
additional information
?
-
-
benzamide is not a substrate
-
-
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3-phenyl-1,1,1-trifluoro-2-propanone
competitive
3-phenyl-1-chloro-2-propanone
competitive
PMSF
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
pH 7.5, 30C
0.059
(R)-mandelamide
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
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
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
pH 7.5, 30C, mutant S181A
0.014
(S)-mandelamide
-
Q207H/S316N/I437N mutant
0.02
(S)-mandelamide
pH 7.5, 30C
0.023
(S)-mandelamide
-
T31I/I437N mutant
0.029
(S)-mandelamide
-
I437N mutant
0.296
(S)-mandelamide
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
pH 7.5, 30C
0.049
3-phenylpropionamide
pH 7.5, 30C
490
acetamide
-
-
6.8
butanamide
-
-
0.12
Caproamide
-
-
0.55
cyclohexanecarboxamide
-
-
0.288
hexanoamide
pH 7.5, 30C
0.032
N-methyl phenylacetamide
pH 7.5, 30C
0.54
Pentanamide
-
-
45
propanamide
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
16.1
(R)-(+)-mandelamide
Pseudomonas putida
-
at 30C and pH 7.8
10.2
(R)-(-)-mandelamide
Pseudomonas putida
-
-
25
(R)-2-aminophenylacetamide
Pseudomonas putida
-
-
25
(R)-lactamide
Pseudomonas putida
-
T31I/I437N mutant
29
(R)-lactamide
Pseudomonas putida
-
I437N mutant
43
(R)-lactamide
Pseudomonas putida
-
; wild-type
64
(R)-lactamide
Pseudomonas putida
-
Q207H/S316N/I437N mutant
76
(R)-lactamide
Pseudomonas putida
-
G202S/Q207H/R236C/R369M/Q382E mutant
78
(R)-lactamide
Pseudomonas putida
-
Q207H/S316N/Q382E mutant
165
(R)-lactamide
Pseudomonas putida
-
G202A/Q207H/Q382E mutant
20
(R)-mandelamide
Pseudomonas putida
-
G202S/Q207H/R236C/R369M/Q382E mutant
51
(R)-mandelamide
Pseudomonas putida
-
G202A/Q207H/Q382E mutant
76
(R)-mandelamide
Pseudomonas putida
-
Q207H/S316N/I437N mutant
78
(R)-mandelamide
Pseudomonas putida
-
T31I/I437N mutant
85
(R)-mandelamide
Pseudomonas putida
-
I437N mutant
125
(R)-mandelamide
Pseudomonas putida
-
Q207H/S316N/Q382E mutant
150
(R)-mandelamide
Pseudomonas putida
-
; wild-type
0.031 - 0.51
(S)-(+)-mandelamide
Pseudomonas putida
-
at 30C and pH 7.8
4.4
(S)-(+)-mandelamide
Pseudomonas putida
-
-
7.73
(S)-(+)-mandelamide
Pseudomonas putida
-
at 30C and pH 7.8
27
(S)-2-aminophenylacetamide
Pseudomonas putida
-
-
19
(S)-lactamide
Pseudomonas putida
-
; wild-type
39
(S)-lactamide
Pseudomonas putida
-
I437N mutant; T31I/I437N mutant
40
(S)-lactamide
Pseudomonas putida
-
G202S/Q207H/R236C/R369M/Q382E mutant
49
(S)-lactamide
Pseudomonas putida
-
Q207H/S316N/I437N mutant
54
(S)-lactamide
Pseudomonas putida
-
Q207H/S316N/Q382E mutant
88
(S)-lactamide
Pseudomonas putida
-
G202A/Q207H/Q382E mutant
8.8
(S)-mandelamide
Pseudomonas putida
-
G202S/Q207H/R236C/R369M/Q382E mutant
18
(S)-mandelamide
Pseudomonas putida
-
G202A/Q207H/Q382E mutant
43
(S)-mandelamide
Pseudomonas putida
-
Q207H/S316N/Q382E mutant
82
(S)-mandelamide
Pseudomonas putida
-
; wild-type
89
(S)-mandelamide
Pseudomonas putida
-
Q207H/S316N/I437N mutant
101
(S)-mandelamide
Pseudomonas putida
-
I437N mutant
108
(S)-mandelamide
Pseudomonas putida
-
T31I/I437N mutant
1.1
2-cyclohexylacetamide
Pseudomonas putida
-
-
46
2-phenylacetamide
Pseudomonas putida
-
-
1.4
acetamide
Pseudomonas putida
-
-
0.61
butanamide
Pseudomonas putida
-
-
2.8
Caproamide
Pseudomonas putida
-
-
0.04
cyclohexanecarboxamide
Pseudomonas putida
-
-
2
Pentanamide
Pseudomonas putida
-
-
4.6
propanamide
Pseudomonas putida
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.06
3-phenyl-1,1,1-trifluoro-2-propanone
pH 7.5, 30C
0.13
3-phenyl-1-chloro-2-propanone
pH 7.5, 30C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
?
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
4C, phosphate buffer pH 7.8, 1 mM EDTA, stable for at least 1 month
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
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
no enzymic activity
Q207H/S316N/I437N
-
site-directed mutagenesis
Q207H/S316N/Q382E
-
site-directed mutagenesis
S180A
significant decrease in kcat value
S181A
significant decrease in kcat value
S204A
no enzymic activity
S217A
very little enzymic activity
S218A
very little enzymic activity
T31I/I437N
-
site-directed mutagenesis