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Literature summary for 1.2.1.39 extracted from

  • Debnar-Daumler, C.; Seubert, A.; Schmitt, G.; Heider, J.
    Simultaneous involvement of a tungsten-containing aldehyde ferredoxin oxidoreductase and a phenylacetaldehyde dehydrogenase in anaerobic phenylalanine metabolism (2014), J. Bacteriol., 196, 483-492 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene ebA4954, recombinant expression of N-terminally Strep-tagged enzyme in Escherichia coli strain DHalpha Aromatoleum aromaticum

Inhibitors

Inhibitors Comment Organism Structure
Mg2+ at high concentrations Aromatoleum aromaticum
NAD+ cooperative substrate inhibition Aromatoleum aromaticum
NADP+ cooperative substrate inhibition Aromatoleum aromaticum
phenylacetaldehyde the enzyme is highly specific for phenylacetaldehyde, has cooperative kinetics toward the substrate, and shows considerable substrate inhibition Aromatoleum aromaticum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetic analysis, overview. The enzyme shows cooperative kinetics toward substrate phenylacetaldehyde and considerable substrate inhibition Aromatoleum aromaticum
0.018
-
NAD+ pH 8.5, 28°C Aromatoleum aromaticum
0.025
-
NADP+ pH 8.5, 28°C Aromatoleum aromaticum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
phenylacetaldehyde + NAD+ + H2O Aromatoleum aromaticum
-
phenylacetate + NADH + 2 H+
-
r
phenylacetaldehyde + NAD+ + H2O Aromatoleum aromaticum EbN1
-
phenylacetate + NADH + 2 H+
-
r

Organism

Organism UniProt Comment Textmining
Aromatoleum aromaticum Q5P171
-
-
Aromatoleum aromaticum EbN1 Q5P171
-
-

Purification (Commentary)

Purification (Comment) Organism
native enzyme 126fold by cross-flow filtration, ultracentrifugation, and anion exchange chromatography, recombinant Strep-tagged enzyme 137fold from Escherichia coli strain DHalpha by affinity chromatography on a Strep-Tactin column Aromatoleum aromaticum

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.102
-
purified native enzyme, with cofactor NADP+, pH 8.5, 28°C Aromatoleum aromaticum
0.21
-
purified recombinant Strep-tagged enzyme, pH 8.5, 28°C Aromatoleum aromaticum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information enzyme is highly specific for phenylacetaldehyde, has cooperative kinetics toward the substrate, and shows considerable substrate inhibition. No activity with benzaldehyde and indole-3-carbaldehyde, substrate specificity, overview Aromatoleum aromaticum ?
-
?
additional information enzyme is highly specific for phenylacetaldehyde, has cooperative kinetics toward the substrate, and shows considerable substrate inhibition. No activity with benzaldehyde and indole-3-carbaldehyde, substrate specificity, overview Aromatoleum aromaticum EbN1 ?
-
?
phenylacetaldehyde + NAD+ + H2O
-
Aromatoleum aromaticum phenylacetate + NADH + 2 H+
-
r
phenylacetaldehyde + NAD+ + H2O
-
Aromatoleum aromaticum EbN1 phenylacetate + NADH + 2 H+
-
r
phenylacetaldehyde + NADP+ + H2O
-
Aromatoleum aromaticum phenylacetate + NADPH + 2 H+
-
r
phenylacetaldehyde + NADP+ + H2O
-
Aromatoleum aromaticum EbN1 phenylacetate + NADPH + 2 H+
-
r

Subunits

Subunits Comment Organism
homotetramer 4 * 53500, recombinant Strep-tagged enzyme, SDS-PAGE Aromatoleum aromaticum

Synonyms

Synonyms Comment Organism
ebA4954
-
Aromatoleum aromaticum
PDH
-
Aromatoleum aromaticum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
28
-
assay at Aromatoleum aromaticum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
53
-
NAD+ pH 8.5, 28°C Aromatoleum aromaticum
63
-
NADP+ pH 8.5, 28°C Aromatoleum aromaticum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5 6.5 assay at, reverse reaction Aromatoleum aromaticum
8.5
-
assay at, forward reaction Aromatoleum aromaticum

Cofactor

Cofactor Comment Organism Structure
additional information enzyme PDH accepts both NAD+ and NADP+ as electron acceptors, but exhibits 86% of its maximal activity with NAD+ but only 5% of its maximal activity with NADP+ Aromatoleum aromaticum
NAD+
-
Aromatoleum aromaticum
NADH
-
Aromatoleum aromaticum
NADP+
-
Aromatoleum aromaticum
NADPH
-
Aromatoleum aromaticum

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.067
-
NADP+ pH 8.5, 28°C, cooperative substrate inhibition Aromatoleum aromaticum
0.069
-
NAD+ pH 8.5, 28°C, cooperative substrate inhibition Aromatoleum aromaticum

General Information

General Information Comment Organism
metabolism two distinct enzymes are involved in the oxidation of phenylacetaldehyde to phenylacetate, an aldehyde:ferredoxin oxidoreductase (AOR) and a phenylacetaldehyde dehydrogenase (PDH). PDH is the primary enzyme during anaerobic phenylalanine degradation, whereas AOR is not essential for the metabolic pathway and exhibits probably a function as a detoxifying enzyme if high aldehyde concentrations accumulate in the cytoplasm, which would lead to substrate inhibition of PDH Aromatoleum aromaticum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0025
-
NADP+ pH 8.5, 28°C Aromatoleum aromaticum
0.003
-
NAD+ pH 8.5, 28°C Aromatoleum aromaticum