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Literature summary for 1.14.13.84 extracted from

  • Kamerbeek, N.M.; Fraaije, M.W.; Janssen, D.B.
    Identifying determinants of NADPH specificity in Baeyer-Villiger monooxygenases (2004), Eur. J. Biochem., 271, 2107-2116.
    View publication on PubMed
Search for more literature for 1.14.13.84

Cloned(Commentary)

Cloned (Comment) Organism
hapE gene, expression in Escherichia coli TOP10 Pseudomonas fluorescens

Protein Variants

Protein Variants Comment Organism
K439A mutant with 100fold decrease in catalytic efficiency with NADPH, mainly caused by increased Km, 4fold increased efficiency with NADH Pseudomonas fluorescens
K439F mutant with higher activity with NADH compared to wild-type HAPMO Pseudomonas fluorescens
K439N mutant with higher activity with NADH compared to wild-type HAPMO Pseudomonas fluorescens
K439P mutant with higher activity with NADH compared to wild-type HAPMO Pseudomonas fluorescens
R339A mutant with largely decreased affinity for NADPH and decreased catalytic efficiency with NADH Pseudomonas fluorescens
R440A totally inactive mutant, Arg-440 is crucial for completing the catalytic cycle Pseudomonas fluorescens

Inhibitors

Inhibitors Comment Organism Structure
acetyl-NADP+ 0.5 mM, 35% inhibition Pseudomonas fluorescens
amino-NADP+ very effective inhibitor, 0.005 mM, 80% inhibition Pseudomonas fluorescens
additional information not inhibited by NADP+ Pseudomonas fluorescens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.012
-
 NADPH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, wild-type HAPMO Pseudomonas fluorescens
0.017
-
 NADPH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439P mutant Pseudomonas fluorescens
0.078
-
 NADPH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439N mutant Pseudomonas fluorescens
0.08
-
 NADPH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439F mutant Pseudomonas fluorescens
0.2
-
 NADPH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439A mutant Pseudomonas fluorescens
0.3
-
 NADH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439F mutant Pseudomonas fluorescens
0.52
-
 NADH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439A mutant Pseudomonas fluorescens
0.78
-
 NADH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439N mutant Pseudomonas fluorescens
1.28
-
 NADH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439P mutant Pseudomonas fluorescens
1.44
-
 NADH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, wild-type HAPMO Pseudomonas fluorescens
3
-
 NADPH above, pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, R339A mutant Pseudomonas fluorescens
3
-
 NADH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, R339A mutant Pseudomonas fluorescens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
145000
-
-
 Pseudomonas fluorescens

Organism

Organism UniProt Comment Textmining
Pseudomonas fluorescens
-
-
-
Pseudomonas fluorescens ACB
-
-
-

Purification (Commentary)

Purification (Comment) Organism
wild-type and mutant HAPMO Pseudomonas fluorescens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-hydroxyacetophenone + NADH + H+ + O2 700fold preference for NADPH over NADH Pseudomonas fluorescens 4-hydroxyphenyl acetate + NAD+ + H2O
-
 ?
4-hydroxyacetophenone + NADH + H+ + O2 700fold preference for NADPH over NADH Pseudomonas fluorescens ACB 4-hydroxyphenyl acetate + NAD+ + H2O
-
 ?
4-hydroxyacetophenone + NADPH + H+ + O2 700fold preference for NADPH over NADH, Arg-440 plays an important role in catalysis Pseudomonas fluorescens 4-hydroxyphenyl acetate + NADP+ + H2O
-
 ?
4-hydroxyacetophenone + NADPH + H+ + O2 700fold preference for NADPH over NADH, Arg-440 plays an important role in catalysis Pseudomonas fluorescens ACB 4-hydroxyphenyl acetate + NADP+ + H2O
-
 ?
additional information catalyzes Baeyer-Villiger oxidation reactions on various ketones, oxidizes a variety of aromatic ketones and sulfides Pseudomonas fluorescens ?
-
 ?
additional information catalyzes Baeyer-Villiger oxidation reactions on various ketones, oxidizes a variety of aromatic ketones and sulfides Pseudomonas fluorescens ACB ?
-
 ?

Subunits

Subunits Comment Organism
dimer
-
 Pseudomonas fluorescens

Synonyms

Synonyms Comment Organism
HAPMO
-
 Pseudomonas fluorescens

Temperature Optimum [°C]

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.1
-
 NADPH above, pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, R339A mutant Pseudomonas fluorescens
0.19
-
 NADH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, R339A mutant Pseudomonas fluorescens
0.2
-
 NADPH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439P mutant Pseudomonas fluorescens
0.7
-
 NADH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439P mutant Pseudomonas fluorescens
1
-
 NADPH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439F mutant Pseudomonas fluorescens
1.6
-
 NADPH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439A mutant Pseudomonas fluorescens
1.7
-
 NADH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, wild-type HAPMO Pseudomonas fluorescens
2.4
-
 NADH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439A mutant Pseudomonas fluorescens
2.4
-
 NADH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439F mutant Pseudomonas fluorescens
3.5
-
 NADPH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439N mutant Pseudomonas fluorescens
5.1
-
 NADH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, K439N mutant Pseudomonas fluorescens
10.1
-
 NADPH pH 7.5, 25°C, cosubstrate 4-hydroxyacetophenone, wild-type HAPMO Pseudomonas fluorescens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Pseudomonas fluorescens

Cofactor

Cofactor Comment Organism Structure
FAD contains one FAD molecule per subunit Pseudomonas fluorescens
NADH 700fold preference for NADPH over NADH Pseudomonas fluorescens
NADPH specific for NADPH as coenzyme, Arg-339 and Lys-439 are involved in coenzyme recognition, Arg-440 not, Lys-439 plays a role in recognizing the 2’-phosphate of NADPH, 700fold preference for NADPH over NADH Pseudomonas fluorescens