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Literature summary for 1.14.16.1 extracted from

  • Ronau, J.A.; Paul, L.N.; Fuchs, J.E.; Corn, I.R.; Wagner, K.T.; Liedl, K.R.; Abu-Omar, M.M.; Das, C.
    An additional substrate binding site in a bacterial phenylalanine hydroxylase (2013), Eur. Biophys. J., 42, 691-708.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.14.16.1

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3)pLysS cells Chromobacterium violaceum

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapor diffusion method, using 0.1 M Na-HEPES, pH 7.0, 0.01 M magnesium chloride hexahydrate, 0.005 M nickel(II) chloride hexahydrate, and 15% (w/v) PEG 3350, or sitting drop vapor diffusion method, using 0.1 M Na-HEPES pH 7.0, 0.01 M magnesium chloride hexahydrate, and 15% (w/v) PEG Chromobacterium violaceum

Protein Variants

Protein Variants Comment Organism
F258A the mutant shows decreased activity and a marked decrease in the affinity for L-phenylalanine Chromobacterium violaceum
S230P the mutant shows strongly decreased activity and a marked decrease in the affinity for L-phenylalanine Chromobacterium violaceum
T254A the mutant shows decreased activity and a marked decrease in the affinity for L-phenylalanine Chromobacterium violaceum
Y155A the mutant shows decreased activity and a marked decrease in the affinity for L-phenylalanine Chromobacterium violaceum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.1365
-
 L-phenylalanine mutant enzyme Y155A, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum
0.244
-
 L-phenylalanine wild type enzyme, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum
0.414
-
 L-phenylalanine mutant enzyme T254A, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum
0.4675
-
 L-phenylalanine mutant enzyme F258A, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum
0.6872
-
 L-phenylalanine mutant enzyme S230P, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-phenylalanine + tetrahydrobiopterin + O2 Chromobacterium violaceum
-
 L-tyrosine + 4a-hydroxytetrahydrobiopterin
-
 ?
L-phenylalanine + tetrahydrobiopterin + O2 Chromobacterium violaceum ATCC 12472
-
 L-tyrosine + 4a-hydroxytetrahydrobiopterin
-
 ?

Organism

Organism UniProt Comment Textmining
Chromobacterium violaceum P30967
-
-
Chromobacterium violaceum ATCC 12472 P30967
-
-

Purification (Commentary)

Purification (Comment) Organism
glutathione-Sepharose column chromatography and Superdex S75 gel filtration Chromobacterium violaceum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-phenylalanine + tetrahydrobiopterin + O2
-
 Chromobacterium violaceum L-tyrosine + 4a-hydroxytetrahydrobiopterin
-
 ?
L-phenylalanine + tetrahydrobiopterin + O2
-
 Chromobacterium violaceum ATCC 12472 L-tyrosine + 4a-hydroxytetrahydrobiopterin
-
 ?

Synonyms

Synonyms Comment Organism
PAH
-
 Chromobacterium violaceum
phenylalanine hydroxylase
-
 Chromobacterium violaceum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.9
-
 L-phenylalanine mutant enzyme S230P, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum
8.81
-
 L-phenylalanine mutant enzyme Y155A, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum
10.9
-
 L-phenylalanine mutant enzyme F258A, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum
18
-
 L-phenylalanine wild type enzyme, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum
21.5
-
 L-phenylalanine mutant enzyme T254A, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5.6
-
 L-phenylalanine mutant enzyme S230P, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum
14
-
 L-phenylalanine mutant enzyme F258A, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum
25
-
 L-phenylalanine mutant enzyme Y155A, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum
32
-
 L-phenylalanine mutant enzyme T254A, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum
48
-
 L-phenylalanine wild type enzyme, in 0.1 M Na-HEPES (pH 7.4) at 30°C Chromobacterium violaceum