EC Number | Cloned (Comment) | Organism |
---|---|---|
3.5.1.86 | recombinant expression of C-terminally His6-tagged wild-type and mutant enzymes | Pseudomonas putida |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.5.1.86 | F150L | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type | Pseudomonas putida |
3.5.1.86 | F433A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type | Pseudomonas putida |
3.5.1.86 | F433L | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type | Pseudomonas putida |
3.5.1.86 | K100A | site-directed mutagenesis, mutation of a catalytic residue, inactive mutant | Pseudomonas putida |
3.5.1.86 | S180A | site-directed mutagenesis, mutation of a catalytic residue, 4200fold decrease in the kcat/Km value | Pseudomonas putida |
3.5.1.86 | S204A | site-directed mutagenesis, mutation of a catalytic residue, inactive mutant | Pseudomonas putida |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
3.5.1.86 | benzyl (carbamoyloxy)carbamate | weak inhibition | Pseudomonas putida | |
3.5.1.86 | additional information | O-acyl hydroxamates are no irreversible inactivators of MAH but some are found to be transient inhibitors. Reactivity of MAH with some recently studied beta-lactamase inhibitors, overview | Pseudomonas putida | |
3.5.1.86 | N-(carbamoyloxy)benzamide | - |
Pseudomonas putida | |
3.5.1.86 | O-methoxycarbonylbenzohydroxamate | - |
Pseudomonas putida | |
3.5.1.86 | phenyl carbamate | - |
Pseudomonas putida | |
3.5.1.86 | phenylmethylboronic acid | a potent inhibitor of enzyme MAH that forms a transition state analogue structure with the enzyme | Pseudomonas putida |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.5.1.86 | 0.009 | - |
(S)-mandelamide | recombinant mutant F150W, pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 0.018 | - |
(S)-mandelamide | recombinant wild-type enzyme, pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 0.043 | - |
(S)-mandelamide | recombinant mutant F433W, pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 0.135 | - |
(S)-mandelamide | recombinant mutant F433L, pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 1.53 | - |
(S)-mandelamide | recombinant mutant F433A, pH 7.8, 25°C | Pseudomonas putida |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.5.1.86 | (R)-mandelamide + H2O | Pseudomonas putida | - |
(R)-mandelate + NH3 | - |
? | |
3.5.1.86 | (S)-mandelamide + H2O | Pseudomonas putida | - |
(S)-mandelate + NH3 | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.5.1.86 | Pseudomonas putida | Q84DC4 | i.e. Arthrobacter siderocapsulatus | - |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.5.1.86 | (R)-mandelamide + H2O | - |
Pseudomonas putida | (R)-mandelate + NH3 | - |
? | |
3.5.1.86 | (S)-mandelamide + H2O | - |
Pseudomonas putida | (S)-mandelate + NH3 | - |
? | |
3.5.1.86 | 4-nitroacetanilide + H2O | - |
Pseudomonas putida | ? | - |
? | |
3.5.1.86 | 4-nitrophenylacetamide + H2O | - |
Pseudomonas putida | 4-nitrophenol + acetamide | - |
? | |
3.5.1.86 | 4-nitrophenylacetate + H2O | - |
Pseudomonas putida | 4-nitrophenol + acetate | - |
? | |
3.5.1.86 | 4-nitrophenylcarbonate + H2O | - |
Pseudomonas putida | 4-nitrophenol + CO2 | - |
? | |
3.5.1.86 | methyl 4-nitrophenylcarbonate + H2O | - |
Pseudomonas putida | ? | - |
? | |
3.5.1.86 | methyl mandelate + H2O | - |
Pseudomonas putida | ? | - |
? | |
3.5.1.86 | additional information | the lack of reactivity of MAH with the corresponding ethyl ester confirms the very limited size of the MAH leaving group site. The reactivity of MAH with 4-nitrophenyl acetate and methyl 4-nitrophenyl carbonate, therefore, suggests formation of an inverse acyl-enzyme where the small acyl-group occupies the normal leaving group site. Both (R)- and (S)-mandelamide are excellent substrates for MAH, while the enzyme can hydrolyze phenylacetamide with even higher catalytic efficiency. Reaction scheme for interaction of substrates with MAH. Substrate specificity, overview. No activity with ethyl 4-nitrophenylacetate, phenyl carbamate, and N-benzyloxycarbonyl-glycinamide, poor activity with phenyl acetate, 4-nitrophenyl n-butyrate, and ethyl 4-nitrophenyl carbonate | Pseudomonas putida | ? | - |
- |
|
3.5.1.86 | N-benzyloxycarbonylglycinamide + H2O | - |
Pseudomonas putida | ? | - |
? | |
3.5.1.86 | phenyl carbamate + H2O | - |
Pseudomonas putida | ? | - |
? | |
3.5.1.86 | phenylacetamide + H2O | - |
Pseudomonas putida | phenylacetate + NH3 | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.5.1.86 | MAH | - |
Pseudomonas putida |
3.5.1.86 | mandelamide hydrolase | - |
Pseudomonas putida |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.5.1.86 | 25 | - |
assay at | Pseudomonas putida |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.5.1.86 | 20 | - |
(S)-mandelamide | recombinant mutant F433A, pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 43 | - |
(S)-mandelamide | recombinant mutant F150W, pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 90 | - |
(S)-mandelamide | recombinant mutant F433W, pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 105 | - |
(S)-mandelamide | recombinant mutant F433L, pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 150 | - |
(S)-mandelamide | recombinant wild-type enzyme, pH 7.8, 25°C | Pseudomonas putida |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
3.5.1.86 | 7.8 | - |
assay at | Pseudomonas putida |
EC Number | Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.5.1.86 | 0.000025 | - |
O-methoxycarbonylbenzohydroxamate | pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 0.000025 | - |
phenyl carbamate | pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 0.000027 | - |
phenylmethylboronic acid | pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 1.4 | - |
benzyl (carbamoyloxy)carbamate | pH 7.8, 25°C | Pseudomonas putida |
EC Number | General Information | Comment | Organism |
---|---|---|---|
3.5.1.86 | additional information | the catalytic triad of MAH is composed of Ser204, Ser180, and Lys100. Modeling of substrate binding. A model of the deacylation tetrahedral intermediate is derived from reaction of MAH with phenylacetamide, overview | Pseudomonas putida |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.5.1.86 | 13.1 | - |
(S)-mandelamide | recombinant mutant F433A, pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 777.8 | - |
(S)-mandelamide | recombinant mutant F433L, pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 2093 | - |
(S)-mandelamide | recombinant mutant F433W, pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 4778 | - |
(S)-mandelamide | recombinant mutant F150W, pH 7.8, 25°C | Pseudomonas putida | |
3.5.1.86 | 8333 | - |
(S)-mandelamide | recombinant wild-type enzyme, pH 7.8, 25°C | Pseudomonas putida |