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Literature summary for 1.14.13.59 extracted from

  • Dick, S.; Siemann, S.; Frey, H.E.; Lepock, J.R.; Viswanatha, T.
    Recombinant lysine:N(6)-hydroxylase: effect of cysteine-->alanine replacements on structural integrity and catalytic competence (2002), Biochim. Biophys. Acta, 1594, 219-233.
    View publication on PubMed
Search for more literature for 1.14.13.59

Cloned(Commentary)

Cloned (Comment) Organism
gene iucD, expression of wild-type and mutants in strain DH5alpha Escherichia coli

Protein Variants

Protein Variants Comment Organism
C146A site-directed mutagenesis, unaffected thermal stability, and affinity for L-lysine and FAD compared to the wild-type enzyme Escherichia coli
C158A site-directed mutagenesis, unaffected thermal stability, and affinity for L-lysine and FAD compared to the wild-type enzyme Escherichia coli
C166A site-directed mutagenesis, unaffected thermal stability, and affinity for L-lysine and FAD compared to the wild-type enzyme Escherichia coli
C31A site-directed mutagenesis, unaffected thermal stability, and affinity for L-lysine and FAD compared to the wild-type enzyme Escherichia coli
C31A/C51A site-directed mutagenesis, unaffected thermal stability, and affinity for L-lysine and FAD compared to the wild-type enzyme Escherichia coli
C51A site-directed mutagenesis, about 2fold increased activity, unaffected thermal stability, and affinity for L-lysine and FAD compared to the wild-type enzyme Escherichia coli
C51A/C158A site-directed mutagenesis, about 2fold increased activity, unaffected thermal stability, and affinity for L-lysine and FAD compared to the wild-type enzyme Escherichia coli
C51A/C166A site-directed mutagenesis, about 2fold increased activity, unaffected thermal stability, and affinity for L-lysine and FAD compared to the wild-type enzyme Escherichia coli
additional information a covalent C51A-dichloropehno indophenol conjugate accomodates FAD in its catalytic function Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
additional information number of cysteine residues in wild-type and mutant enzymes accessible for modification with DTNB, overview Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0016
-
 FAD mutant C51A, pH 7.0, 37°C Escherichia coli
0.0019
-
 FAD mutant C146A, pH 7.0, 37°C Escherichia coli
0.0021
-
 FAD mutant C51A/C158A, pH 7.0, 37°C Escherichia coli
0.0022
-
 FAD mutant C166A, pH 7.0, 37°C Escherichia coli
0.0025
-
 FAD mutant C31A, pH 7.0, 37°C Escherichia coli
0.0033
-
 FAD wild-type enzyme, pH 7.0, 37°C Escherichia coli
0.0047
-
 FAD mutant C51A/C166A, pH 7.0, 37°C Escherichia coli
0.0056
-
 FAD mutant C31A/C51A, pH 7.0, 37°C Escherichia coli
0.0057
-
 FAD mutant C158A, pH 7.0, 37°C Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-lysine + NADPH + O2 Escherichia coli
-
 N6-hydroxy-L-lysine + NADP+ + H2O
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutants from strain DH5alpha Escherichia coli

Reaction

Reaction Comment Organism Reaction ID
L-lysine + NADPH + H+ + O2 = N6-hydroxy-L-lysine + NADP+ + H2O reaction mechanism, Cys51 is involved, while Cys31, Cys146, Cys158, and Cys166 are not Escherichia coli
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-lysine + NADPH + O2
-
 Escherichia coli N6-hydroxy-L-lysine + NADP+ + H2O
-
 ?

Synonyms

Synonyms Comment Organism
IucD
-
 Escherichia coli
lysine:N(6)-hydroxylase
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.243
-
 FAD mutant C146A, pH 7.0, 37°C Escherichia coli
0.305
-
 FAD mutant C166A, pH 7.0, 37°C Escherichia coli
0.351
-
 FAD mutant C31A, pH 7.0, 37°C Escherichia coli
0.405
-
 FAD wild-type enzyme, pH 7.0, 37°C Escherichia coli
0.425
-
 FAD mutant C158A, pH 7.0, 37°C Escherichia coli
0.543
-
 FAD mutant C51A/C158A, pH 7.0, 37°C Escherichia coli
0.614
-
 FAD mutant C31A/C51A, pH 7.0, 37°C Escherichia coli
0.667
-
 FAD mutant C51A, pH 7.0, 37°C Escherichia coli
0.731
-
 FAD mutant C51A/C166A, pH 7.0, 37°C Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
FAD required for activity Escherichia coli
NADPH required for activity Escherichia coli