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Literature summary for 1.14.13.196 extracted from

  • Shirey, C.; Badieyan, S.; Sobrado, P.
    Role of Ser-257 in the sliding mechanism of NADP(H) in the reaction catalyzed by the Aspergillus fumigatus flavin-dependent ornithine N5-monooxygenase SidA (2013), J. Biol. Chem., 288, 32440-32448.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Aspergillus fumigatus

Protein Variants

Protein Variants Comment Organism
S257A following oxygen consumption, the Km value of the S257A enzyme for NADPH was not significantly changed, whereas the kcat value increased. When the activity is determined by measuring the formation of N5-hydroxyornithine, the kcat value for the S257A enzyme is lower than that for the wild-type enzyme. The S257A mutation causes an increase in flexibility in two small loops, loop 253–257, which is within interacting distance of NADP+ (including Ser257 and Gln256, which are able to form hydrogen bonds with NADP during MD simulations), and loop 453–458, which resides 5 A from the FAD phosphate group. In contrast, loop 97–105 (including Gln-102, which makes a hydrogen bond with FAD) presents reduced flexibility in the mutant enzyme Aspergillus fumigatus

General Stability

General Stability Organism
the presence of NADP+ is essential for activity, as it is required for stabilization of the C4a-hydroperoxyflavin, which is the hydroxylating species Aspergillus fumigatus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.014
-
NADPH pH 7.6, 25°C, mutant enzyme S257A Aspergillus fumigatus
0.017
-
NADPH pH 7.6, 25°C, wild-type enzyme Aspergillus fumigatus
0.21
-
L-ornithine pH 7.6, 25°C, mutant enzyme S257A, cofactor: NADPH Aspergillus fumigatus
0.3
-
L-ornithine pH 7.6, 25°C, wild-type enzyme, cofactor: NADPH Aspergillus fumigatus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-ornithine + NADH + H+ + O2 Aspergillus fumigatus the enzyme is essential for virulence in Aspergillus fumigatus N5-hydroxy-L-ornithine + NAD+ + H2O
-
?
L-ornithine + NADPH + H+ + O2 Aspergillus fumigatus the enzyme is essential for virulence in Aspergillus fumigatus N5-hydroxy-L-ornithine + NADP+ + H2O
-
?

Organism

Organism UniProt Comment Textmining
Aspergillus fumigatus Q5SE95
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-ornithine + NADH + H+ + O2 the enzyme is essential for virulence in Aspergillus fumigatus Aspergillus fumigatus N5-hydroxy-L-ornithine + NAD+ + H2O
-
?
L-ornithine + NADPH + H+ + O2 the enzyme is essential for virulence in Aspergillus fumigatus Aspergillus fumigatus N5-hydroxy-L-ornithine + NADP+ + H2O
-
?
L-ornithine + NADPH + H+ + O2 NADPH reduces the flavin, and the resulting NADP+ is the last product to be released Aspergillus fumigatus N5-hydroxy-L-ornithine + NADP+ + H2O
-
?

Synonyms

Synonyms Comment Organism
SidA
-
Aspergillus fumigatus
siderophore A
-
Aspergillus fumigatus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Aspergillus fumigatus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
additional information presteady-state kinetic parameters, cofactor: NADPH Aspergillus fumigatus
0.03
-
L-ornithine pH 7.6, 25°C, mutant enzyme S257A, cofactor: NADPH Aspergillus fumigatus
0.03
-
L-ornithine pH 7.6, 25°C, wild-type enzyme, cofactor: NADPH Aspergillus fumigatus
0.6
-
NADPH pH 7.6, 25°C, wild-type enzyme Aspergillus fumigatus
0.85
-
NADPH pH 7.6, 25°C, mutant enzyme S257A Aspergillus fumigatus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Aspergillus fumigatus

Cofactor

Cofactor Comment Organism Structure
NADPH
-
Aspergillus fumigatus

General Information

General Information Comment Organism
physiological function the enzyme is essential for virulence in Aspergillus fumigatus Aspergillus fumigatus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.09
-
L-ornithine pH 7.6, 25°C, mutant enzyme S257A, cofactor: NADPH Aspergillus fumigatus
0.14
-
L-ornithine pH 7.6, 25°C, wild-type enzyme, cofactor: NADPH Aspergillus fumigatus
14
-
NADPH pH 7.6, 25°C, wild-type enzyme Aspergillus fumigatus
40
-
NADPH pH 7.6, 25°C, mutant enzyme S257A Aspergillus fumigatus