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Literature summary extracted from

  • McLeish, M.J.; Kneen, M.M.; Gopalakrishna, K.N.; Koo, C.W.; Babbitt, P.C.; Gerlt, J.A.; Kenyon, G.L.
    Identification and characterization of a mandelamide hydrolase and an NAD(P)+dependent benzaldehyde dehydrogenase from Pseudomonas putida ATCC 12633 (2003), J. Bacteriol., 185, 2451-2456.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.2.1.3 expressed in Escherichia coli coli strain BL21(DE3)pLysS Pseudomonas putida
3.5.1.86 expressed in Escherichia coli BL21(DE3)pLysS cells Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.1.3 0.0399
-
 benzaldehyde at 25°C and pH 8.5, with NADP+ as the electron acceptor Pseudomonas putida
1.2.1.3 0.0634
-
 benzaldehyde at 25°C and pH 8.5, with NAD+ as the electron acceptor Pseudomonas putida
3.5.1.86 0.0198
-
 (S)-(+)-mandelamide at 30°C and pH 7.8 Pseudomonas putida
3.5.1.86 0.0342
-
 (R)-(+)-mandelamide at 30°C and pH 7.8 Pseudomonas putida

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.2.1.3 47430
-
 electrospray mass spectrometry Pseudomonas putida
1.2.1.3 47430
-
 deduced from amino acid sequence Pseudomonas putida
3.5.1.86 52600
-
 sedimentation analysis Pseudomonas putida
3.5.1.86 53820
-
 electrospray mass spectrometry Pseudomonas putida
3.5.1.86 53820
-
 deduced from amino acid sequence Pseudomonas putida

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.3 Pseudomonas putida
-
 strain ATCC 12633
-
3.5.1.86 Pseudomonas putida
-
 strain ATCC 12633
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.2.1.3 Sephacryl S-200 gel filtration, DEAE-Sepharose anion-exchange column chromatography, and Affi-Gel blue column chromatography Pseudomonas putida
3.5.1.86 Sephacryl S-200 gel filtration, HiPrep Q anion-exchange column chromatography, and HiTrap Q column chromatography Pseudomonas putida

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.1.3 benzaldehyde + NAD(P)+ + H2O
-
 Pseudomonas putida benzoate + NAD(P)H + H+
-
 ?
1.2.1.3 additional information the enzyme is only marginally active with acetaldehyde, while longer-chain aliphatic aldehydes are poor substrates Pseudomonas putida ?
-
 ?
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 3,4-dichloroisocoumarin + H2O
-
 Pseudomonas putida ?
-
 ?
3.5.1.86 3-phenylpropionamide + H2O
-
 Pseudomonas putida 3-phenylpropionate + NH3
-
 ?
3.5.1.86 additional information benzamide is not a substrate Pseudomonas putida ?
-
 ?
3.5.1.86 phenylacetamide + H2O
-
 Pseudomonas putida phenylacetate + NH3
-
 ?
3.5.1.86 phenylmethylsulfonyl fluoride + H2O
-
 Pseudomonas putida ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.1.86 monomer 1 * 52600, sedimentation analysis Pseudomonas putida

Synonyms

EC Number Synonyms Comment Organism
1.2.1.3 BADH
-
 Pseudomonas putida
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
1.2.1.3 0.0425
-
 benzaldehyde at 25°C and pH 8.5, with NADP+ as the electron acceptor Pseudomonas putida
1.2.1.3 0.137
-
 benzaldehyde at 25°C and pH 8.5, with NAD+ as the electron acceptor Pseudomonas putida
3.5.1.86 0.031 0.51 (S)-(+)-mandelamide at 30°C and pH 7.8 Pseudomonas putida
3.5.1.86 7.73
-
 (S)-(+)-mandelamide at 30°C and pH 7.8 Pseudomonas putida
3.5.1.86 16.1
-
 (R)-(+)-mandelamide at 30°C and pH 7.8 Pseudomonas putida

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.3 NAD+ dependent, the enzyme exhibits greater affinity for NAD+ than NADP+ Pseudomonas putida
1.2.1.3 NADP+ dependent, the enzyme exhibits greater affinity for NAD+ than NADP+ Pseudomonas putida