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Literature summary for 1.13.12.3 extracted from

  • Gaweska, H.M.; Taylor, A.B.; Hart, P.J.; Fitzpatrick, P.F.
    Structure of the flavoprotein tryptophan 2-monooxygenase, a key enzyme in the formation of galls in plants (2013), Biochemistry, 52, 2620-2626.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as a His-tagged fusion protein Pseudomonas savastanoi

Crystallization (Commentary)

Crystallization (Comment) Organism
determined by X-ray diffraction methods to a resolution of 1.95 A. The overall structure of the protein shows that it has the same fold as members of the monoamine oxidase family of flavoproteins, with the greatest similarities to the L-amino acid oxidases Pseudomonas savastanoi

Protein Variants

Protein Variants Comment Organism
K365M mutation decreases the kcat and kcat/Km (L-tryptophan) values by 60000- and 2 millionfold compared to wild-type, respectively Pseudomonas savastanoi
W466F mutation decreases the kcat value less than 2fold and the kcat/Km (L-tryptophan) value only 5fold compared to wild-type Pseudomonas savastanoi
W466M mutation results in an enzyme lacking flavin and detectable activity Pseudomonas savastanoi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.04
-
L-tryptophan pH 8.3, 25°C, wild-type Pseudomonas savastanoi
0.13
-
L-tryptophan pH 8.3, 25°C, mutant W466F Pseudomonas savastanoi
1.2
-
L-tryptophan pH 8.3, 25°C, mutant K365M Pseudomonas savastanoi
90
-
O2 pH 8.3, 25°C, wild-type Pseudomonas savastanoi
140
-
O2 pH 8.3, 25°C, mutant W466F Pseudomonas savastanoi

Organism

Organism UniProt Comment Textmining
Pseudomonas savastanoi P06617
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-tryptophan + O2
-
Pseudomonas savastanoi indole-3-acetamide + CO2 + H2O
-
?

Synonyms

Synonyms Comment Organism
tryptophan 2-monooxygenase
-
Pseudomonas savastanoi

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Pseudomonas savastanoi

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.00022
-
L-tryptophan pH 8.3, 25°C, cosubstrate: O2, mutant K365M Pseudomonas savastanoi
9.2
-
L-tryptophan pH 8.3, 25°C, cosubstrate: O2, mutant W466F Pseudomonas savastanoi
13.2
-
L-tryptophan pH 8.3, 25°C, cosubstrate: O2, wild-type Pseudomonas savastanoi

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.3
-
assay at Pseudomonas savastanoi

Cofactor

Cofactor Comment Organism Structure
flavin
-
Pseudomonas savastanoi

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.00019
-
L-tryptophan pH 8.3, 25°C, mutant K365M Pseudomonas savastanoi
70
-
O2 pH 8.3, 25°C, mutant W466F Pseudomonas savastanoi
72
-
L-tryptophan pH 8.3, 25°C, mutant W466F Pseudomonas savastanoi
140
-
O2 pH 8.3, 25°C, wild-type Pseudomonas savastanoi
360
-
L-tryptophan pH 8.3, 25°C, wild-type Pseudomonas savastanoi