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Literature summary extracted from

  • Wang, P.F.; Yep, A.; Kenyon, G.L.; McLeish, M.J.
    Using directed evolution to probe the substrate specificity of mandelamide hydrolase (2009), Protein Eng. Des. Sel., 22, 103-110.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.86 expressed in Escherichia coli Pseudomonas putida

Protein Variants

EC Number Protein Variants Comment Organism
3.5.1.86 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 Pseudomonas putida
3.5.1.86 G202A/Q207H/Q382E site-directed mutagenesis Pseudomonas putida
3.5.1.86 G202S/Q207H/R236C/R369M/Q382E site-directed mutagenesis Pseudomonas putida
3.5.1.86 G202V site-directed mutagenesis. Reduction in activity increases to six orders of magnitude for the G202V variant Pseudomonas putida
3.5.1.86 I437N site-directed mutagenesis Pseudomonas putida
3.5.1.86 Q207H/S316N/I437N site-directed mutagenesis Pseudomonas putida
3.5.1.86 Q207H/S316N/Q382E site-directed mutagenesis Pseudomonas putida
3.5.1.86 T31I/I437N site-directed mutagenesis Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.86 0.0009
-
 2-phenylacetamide
-
 Pseudomonas putida
3.5.1.86 0.0035
-
 (S)-mandelamide Q207H/S316N/Q382E mutant Pseudomonas putida
3.5.1.86 0.005
-
 (R)-2-aminophenylacetamide
-
 Pseudomonas putida
3.5.1.86 0.006
-
 (S)-2-aminophenylacetamide
-
 Pseudomonas putida
3.5.1.86 0.007
-
 (S)-mandelamide
-
 Pseudomonas putida
3.5.1.86 0.0072
-
 (S)-mandelamide wild-type Pseudomonas putida
3.5.1.86 0.014
-
 (S)-mandelamide Q207H/S316N/I437N mutant Pseudomonas putida
3.5.1.86 0.014
-
 (R)-mandelamide Q207H/S316N/Q382E mutant Pseudomonas putida
3.5.1.86 0.018
-
 (R)-mandelamide wild-type Pseudomonas putida
3.5.1.86 0.02
-
 (R)-mandelamide
-
 Pseudomonas putida
3.5.1.86 0.023
-
 (S)-mandelamide T31I/I437N mutant Pseudomonas putida
3.5.1.86 0.029
-
 (S)-mandelamide I437N mutant Pseudomonas putida
3.5.1.86 0.04
-
 2-cyclohexylacetamide
-
 Pseudomonas putida
3.5.1.86 0.12
-
 Caproamide
-
 Pseudomonas putida
3.5.1.86 0.129
-
 (R)-mandelamide I437N mutant Pseudomonas putida
3.5.1.86 0.149
-
 (S)-lactamide Q207H/S316N/I437N mutant Pseudomonas putida
3.5.1.86 0.168
-
 (R)-mandelamide Q207H/S316N/I437N mutant Pseudomonas putida
3.5.1.86 0.215
-
 (R)-mandelamide T31I/I437N mutant Pseudomonas putida
3.5.1.86 0.219
-
 (S)-lactamide I437N mutant Pseudomonas putida
3.5.1.86 0.224
-
 (S)-lactamide T31I/I437N mutant Pseudomonas putida
3.5.1.86 0.35
-
 (R)-lactamide Q207H/S316N/Q382E mutant Pseudomonas putida
3.5.1.86 0.397
-
 (S)-lactamide Q207H/S316N/Q382E mutant Pseudomonas putida
3.5.1.86 0.43
-
 (S)-lactamide wild-type Pseudomonas putida
3.5.1.86 0.54
-
 Pentanamide
-
 Pseudomonas putida
3.5.1.86 0.55
-
 cyclohexanecarboxamide
-
 Pseudomonas putida
3.5.1.86 0.59
-
 (S)-lactamide G202A/Q207H/Q382E mutant Pseudomonas putida
3.5.1.86 0.596
-
 (R)-lactamide wild-type Pseudomonas putida
3.5.1.86 0.65
-
 (S)-lactamide G202S/Q207H/R236C/R369M/Q382E mutant Pseudomonas putida
3.5.1.86 0.75
-
 (R)-lactamide G202S/Q207H/R236C/R369M/Q382E mutant Pseudomonas putida
3.5.1.86 1
-
 (R)-lactamide G202A/Q207H/Q382E mutant Pseudomonas putida
3.5.1.86 2
-
 (R)-lactamide I437N mutant Pseudomonas putida
3.5.1.86 2.2
-
 (R)-lactamide T31I/I437N mutant Pseudomonas putida
3.5.1.86 2.4
-
 (R)-lactamide Q207H/S316N/I437N mutant Pseudomonas putida
3.5.1.86 3.1
-
 (S)-mandelamide G202A/Q207H/Q382E mutant Pseudomonas putida
3.5.1.86 6.8
-
 butanamide
-
 Pseudomonas putida
3.5.1.86 7.6
-
 (S)-mandelamide G202S/Q207H/R236C/R369M/Q382E mutant Pseudomonas putida
3.5.1.86 8.2
-
 (R)-mandelamide G202S/Q207H/R236C/R369M/Q382E mutant Pseudomonas putida
3.5.1.86 8.8
-
 (R)-mandelamide G202A/Q207H/Q382E mutant Pseudomonas putida
3.5.1.86 45
-
 propanamide
-
 Pseudomonas putida
3.5.1.86 430
-
 (S)-lactamide
-
 Pseudomonas putida
3.5.1.86 490
-
 acetamide
-
 Pseudomonas putida
3.5.1.86 600
-
 (R)-lactamide
-
 Pseudomonas putida

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.86 Pseudomonas putida
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.86 affinity chromatography Pseudomonas putida

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.86 (R)-2-aminophenylacetamide + H2O
-
 Pseudomonas putida (R)-2-aminophenylacetate + NH3
-
 ?
3.5.1.86 (R)-lactamide + H2O low efficiency Pseudomonas putida (R)-lactate + NH3
-
 ?
3.5.1.86 (R)-mandelamide + H2O
-
 Pseudomonas putida (R)-mandelic acid + ammonia
-
 ?
3.5.1.86 (S)-2-aminophenylacetamide + H2O
-
 Pseudomonas putida (S)-2-aminophenylacetate + NH3
-
 ?
3.5.1.86 (S)-lactamide + H2O low efficiency Pseudomonas putida (S)-lactate + NH3
-
 ?
3.5.1.86 (S)-mandelamide + H2O
-
 Pseudomonas putida (S)-mandelic acid + ammonia
-
 ?
3.5.1.86 2-cyclohexylacetamide + H2O
-
 Pseudomonas putida 2-cyclohexylacetate + NH3
-
 ?
3.5.1.86 2-phenylacetamide + H2O
-
 Pseudomonas putida 2-phenylacetate + NH3
-
 ?
3.5.1.86 acetamide + H2O
-
 Pseudomonas putida acetate + NH3
-
 ?
3.5.1.86 butanamide + H2O
-
 Pseudomonas putida butanoate + NH3
-
 ?
3.5.1.86 caproate + H2O
-
 Pseudomonas putida caproamide + NH3
-
 ?
3.5.1.86 cyclohexanecarboxamide + H2O
-
 Pseudomonas putida cyclohexanecarboxylate + NH3
-
 ?
3.5.1.86 pentanamide + H2O
-
 Pseudomonas putida pentanoate + NH3
-
 ?
3.5.1.86 propanamide + H2O
-
 Pseudomonas putida propanoate + NH3
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.5.1.86 MAH
-
 Pseudomonas putida
3.5.1.86 mandelamide hydrolase
-
 Pseudomonas putida

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.1.86 0.04
-
 cyclohexanecarboxamide
-
 Pseudomonas putida
3.5.1.86 0.61
-
 butanamide
-
 Pseudomonas putida
3.5.1.86 1.1
-
 2-cyclohexylacetamide
-
 Pseudomonas putida
3.5.1.86 1.4
-
 acetamide
-
 Pseudomonas putida
3.5.1.86 2
-
 Pentanamide
-
 Pseudomonas putida
3.5.1.86 2.8
-
 Caproamide
-
 Pseudomonas putida
3.5.1.86 4.6
-
 propanamide
-
 Pseudomonas putida
3.5.1.86 8.8
-
 (S)-mandelamide G202S/Q207H/R236C/R369M/Q382E mutant Pseudomonas putida
3.5.1.86 18
-
 (S)-mandelamide G202A/Q207H/Q382E mutant Pseudomonas putida
3.5.1.86 19
-
 (S)-lactamide
-
 Pseudomonas putida
3.5.1.86 19
-
 (S)-lactamide wild-type Pseudomonas putida
3.5.1.86 20
-
 (R)-mandelamide G202S/Q207H/R236C/R369M/Q382E mutant Pseudomonas putida
3.5.1.86 25
-
 (R)-2-aminophenylacetamide
-
 Pseudomonas putida
3.5.1.86 25
-
 (R)-lactamide T31I/I437N mutant Pseudomonas putida
3.5.1.86 27
-
 (S)-2-aminophenylacetamide
-
 Pseudomonas putida
3.5.1.86 29
-
 (R)-lactamide I437N mutant Pseudomonas putida
3.5.1.86 39
-
 (S)-lactamide I437N mutant Pseudomonas putida
3.5.1.86 39
-
 (S)-lactamide T31I/I437N mutant Pseudomonas putida
3.5.1.86 40
-
 (S)-lactamide G202S/Q207H/R236C/R369M/Q382E mutant Pseudomonas putida
3.5.1.86 43
-
 (R)-lactamide
-
 Pseudomonas putida
3.5.1.86 43
-
 (R)-lactamide wild-type Pseudomonas putida
3.5.1.86 43
-
 (S)-mandelamide Q207H/S316N/Q382E mutant Pseudomonas putida
3.5.1.86 46
-
 2-phenylacetamide
-
 Pseudomonas putida
3.5.1.86 49
-
 (S)-lactamide Q207H/S316N/I437N mutant Pseudomonas putida
3.5.1.86 51
-
 (R)-mandelamide G202A/Q207H/Q382E mutant Pseudomonas putida
3.5.1.86 54
-
 (S)-lactamide Q207H/S316N/Q382E mutant Pseudomonas putida
3.5.1.86 64
-
 (R)-lactamide Q207H/S316N/I437N mutant Pseudomonas putida
3.5.1.86 76
-
 (R)-lactamide G202S/Q207H/R236C/R369M/Q382E mutant Pseudomonas putida
3.5.1.86 76
-
 (R)-mandelamide Q207H/S316N/I437N mutant Pseudomonas putida
3.5.1.86 78
-
 (R)-lactamide Q207H/S316N/Q382E mutant Pseudomonas putida
3.5.1.86 78
-
 (R)-mandelamide T31I/I437N mutant Pseudomonas putida
3.5.1.86 82
-
 (S)-mandelamide
-
 Pseudomonas putida
3.5.1.86 82
-
 (S)-mandelamide wild-type Pseudomonas putida
3.5.1.86 85
-
 (R)-mandelamide I437N mutant Pseudomonas putida
3.5.1.86 88
-
 (S)-lactamide G202A/Q207H/Q382E mutant Pseudomonas putida
3.5.1.86 89
-
 (S)-mandelamide Q207H/S316N/I437N mutant Pseudomonas putida
3.5.1.86 101
-
 (S)-mandelamide I437N mutant Pseudomonas putida
3.5.1.86 108
-
 (S)-mandelamide T31I/I437N mutant Pseudomonas putida
3.5.1.86 125
-
 (R)-mandelamide Q207H/S316N/Q382E mutant Pseudomonas putida
3.5.1.86 150
-
 (R)-mandelamide
-
 Pseudomonas putida
3.5.1.86 150
-
 (R)-mandelamide wild-type Pseudomonas putida
3.5.1.86 165
-
 (R)-lactamide G202A/Q207H/Q382E mutant Pseudomonas putida