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Literature summary for 1.14.16.6 extracted from

  • Bhat, S.G.; Vaidyanathan, C.S.
    Purifications and properties of L-mandelate-4-hydroxylase from Pseudomonas convexa (1976), Arch. Biochem. Biophys., 176, 314-323.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
2-amino-4-hydroxy-6,7-dimethyl-tetrahydropteridine pteridine cofactor required, 2-amino-4-hydroxy-6,7-dimethyl-tetrahydropteridine is most effective, optimal concentration is 0.000025 mM Pseudomonas putida
tetrahydrofolic acid pteridine cofactor required, 50% of the activity of 2-amino-4-hydroxy-6,7-dimethyl-tetrahydropteridine Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline 0.5 mM, 88% inhibition Pseudomonas putida
2,2'-bipyridyl 0.5 mM, 83% inhibition Pseudomonas putida
2-amino-4-hydroxy-6,7-dimethyl-tetrahydropteridine above 0.000025 mM, slight inhibition Pseudomonas putida
4-hydroxymercuribenzoate partially reversible by thiol compounds Pseudomonas putida
8-hydroxyquinoline 0.5 mM, 60% inhibition Pseudomonas putida
Ag2+ 0.0001 mM, 42% inhibition Pseudomonas putida
amethopterin
-
Pseudomonas putida
Aminopterine
-
Pseudomonas putida
Cd2+ 0.0001 mM, 66% inhibition Pseudomonas putida
Cu2+ 0.0001 mM, 40% inhibition Pseudomonas putida
diethyldithiocarbamate 0.5 mM, 16% inhibition Pseudomonas putida
EDTA 0.5 mM, 64% inhibition Pseudomonas putida
guanidine hydrochloride 2.0 mM, 95% inhibition Pseudomonas putida
Hg2+ 0.0001 mM, 56% inhibition Pseudomonas putida
iodoacetamide weak Pseudomonas putida
iodoacetate weak Pseudomonas putida
NEM
-
Pseudomonas putida
SDS 0.1 mM, complete inhibition Pseudomonas putida
Thiourea 1.0 mM, complete inhibition Pseudomonas putida
Urea 2.0 mM, complete inhibition Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.1
-
(S)-2-Hydroxy-2-phenylacetate
-
Pseudomonas putida
0.19
-
NADPH
-
Pseudomonas putida

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ required Pseudomonas putida
Fe2+ Km: 0.047 mM Pseudomonas putida

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
91000
-
gel filtration Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-2-hydroxy-2-phenylacetate + tetrahydropteridine + O2 Pseudomonas putida i.e. L-mandelate, first step of oxidative degradation of L-mandelate by Pseudomonas convexa ?
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Pseudomonas putida

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.021
-
-
Pseudomonas putida

Storage Stability

Storage Stability Organism
-20°C, partially purified enzyme is stable for 36-40 h Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-2-hydroxy-2-phenylacetate + tetrahydropteridine i.e. L-mandelate, highly substrate specific, D-mandelate is not hydroxylated Pseudomonas putida (S)-4-hydroxymandelate + dihydropteridine + H2O
-
?
(S)-2-hydroxy-2-phenylacetate + tetrahydropteridine + O2 i.e. L-mandelate, first step of oxidative degradation of L-mandelate by Pseudomonas convexa Pseudomonas putida ?
-
?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
38
-
-
Pseudomonas putida

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
55
-
5 min, no loss of activity Pseudomonas putida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.4
-
citrate-phosphate buffer Pseudomonas putida

pH Range

pH Minimum pH Maximum Comment Organism
4.6 7.4 half-maximal activity at pH 4.6 and pH 7.4 Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
NADPH required Pseudomonas putida