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Literature summary for 1.2.1.58 extracted from

  • Hirsch, W.; Schägger, H.; Fuchs, G.
    Phenylglyoxylate:NAD+ oxidoreductase (CoA benzoylating), a new enzyme of anaerobic phenylalanine metabolism in the denitrifying bacterium Azoarcus evansii (1998), Eur. J. Biochem., 251, 907-915.
    View publication on PubMed

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.045
-
phenylglyoxylate
-
Aromatoleum evansii
0.055
-
CoA
-
Aromatoleum evansii
0.074
-
NAD+
-
Aromatoleum evansii

Metals/Ions

Metals/Ions Comment Organism Structure
Iron iron-sulfur protein, contains 35 mol Fe and 36 mol acid-labile sulfur per mol of enzyme Aromatoleum evansii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
11500
-
2 * 50000 + 2 * 48000 + 2 * 43000 + 2 * 24000 + 2 * 11500, SDS-PAGE. The enzyme complex consists of a core enzyme of 4 subunits with the composition alpha2beta2gamma2delta2, this complex is able to reduce viologen dyes. The holoenzyme contains in addition an epsilon2 unit that catalyzes the transfer of electrons from a small ferredoxin-like subunit of the core complex to NAD, this unit also catalyzes the transhydrogenase reaction, carries FAD and resembles ferredoxin:NAD(P)+-oxidoreductase Aromatoleum evansii
24000
-
2 * 50000 + 2 * 48000 + 2 * 43000 + 2 * 24000 + 2 * 11500, SDS-PAGE. The enzyme complex consists of a core enzyme of 4 subunits with the composition alpha2beta2gamma2delta2, this complex is able to reduce viologen dyes. The holoenzyme contains in addition an epsilon2 unit that catalyzes the transfer of electrons from a small ferredoxin-like subunit of the core complex to NAD, this unit also catalyzes the transhydrogenase reaction, carries FAD and resembles ferredoxin:NAD(P)+-oxidoreductase Aromatoleum evansii
43000
-
2 * 50000 + 2 * 48000 + 2 * 43000 + 2 * 24000 + 2 * 11500, SDS-PAGE. The enzyme complex consists of a core enzyme of 4 subunits with the composition alpha2beta2gamma2delta2, this complex is able to reduce viologen dyes. The holoenzyme contains in addition an epsilon2 unit that catalyzes the transfer of electrons from a small ferredoxin-like subunit of the core complex to NAD, this unit also catalyzes the transhydrogenase reaction, carries FAD and resembles ferredoxin:NAD(P)+-oxidoreductase Aromatoleum evansii
48000
-
2 * 50000 + 2 * 48000 + 2 * 43000 + 2 * 24000 + 2 * 11500, SDS-PAGE. The enzyme complex consists of a core enzyme of 4 subunits with the composition alpha2beta2gamma2delta2, this complex is able to reduce viologen dyes. The holoenzyme contains in addition an epsilon2 unit that catalyzes the transfer of electrons from a small ferredoxin-like subunit of the core complex to NAD, this unit also catalyzes the transhydrogenase reaction, carries FAD and resembles ferredoxin:NAD(P)+-oxidoreductase Aromatoleum evansii
50000
-
2 * 50000 + 2 * 48000 + 2 * 43000 + 2 * 24000 + 2 * 11500, SDS-PAGE. The enzyme complex consists of a core enzyme of 4 subunits with the composition alpha2beta2gamma2delta2, this complex is able to reduce viologen dyes. The holoenzyme contains in addition an epsilon2 unit that catalyzes the transfer of electrons from a small ferredoxin-like subunit of the core complex to NAD, this unit also catalyzes the transhydrogenase reaction, carries FAD and resembles ferredoxin:NAD(P)+-oxidoreductase Aromatoleum evansii
370000
-
gel filtration Aromatoleum evansii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
phenylglyoxylate + NAD+ + CoA-SH Aromatoleum evansii enzyme activity is induced during anaerobic growth with Phe, phenylacetate and phenylglyoxylate, but not with benzoate benzoyl-S-CoA + CO2 + NADH
-
?
phenylglyoxylate + NAD+ + CoA-SH Aromatoleum evansii enzyme of anaerobic Phe metabolism benzoyl-S-CoA + CO2 + NADH
-
?

Organism

Organism UniProt Comment Textmining
Aromatoleum evansii
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Aromatoleum evansii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
5.1
-
NAD+-dependent reaction Aromatoleum evansii
7.4
-
benzyl viologen-dependent reaction Aromatoleum evansii

Storage Stability

Storage Stability Organism
6°C, 1 mM phenylglyoxal, half-life: 2 days Aromatoleum evansii
6°C, 10% glycerol, half-life: 4 days Aromatoleum evansii
6°C, without glycerol or substrate, half-life: 10 h Aromatoleum evansii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxoisovalerate + NAD+ + CoA-SH at 15% of the activity relative to phenylglyoxylate Aromatoleum evansii ?
-
?
additional information enzyme catalyzes a NAD(H):viologen dye transhydrogenation Aromatoleum evansii ?
-
?
phenylglyoxylate + benzyl viologen + CoA-SH
-
Aromatoleum evansii benzoyl-S-CoA + CO2 + reduced benzyl viologen
-
?
phenylglyoxylate + FAD + CoA-SH
-
Aromatoleum evansii benzoyl-S-CoA + CO2 + FADH2
-
?
phenylglyoxylate + FMN + CoA-SH
-
Aromatoleum evansii benzoyl-S-CoA + CO2 + FMNH
-
?
phenylglyoxylate + methyl viologen + CoA-SH
-
Aromatoleum evansii benzoyl-S-CoA + CO2 + reduced methyl viologen
-
?
phenylglyoxylate + NAD+ + CoA-SH reaction is strictly dependent on CoA Aromatoleum evansii benzoyl-S-CoA + CO2 + NADH
-
?
phenylglyoxylate + NAD+ + CoA-SH enzyme activity is induced during anaerobic growth with Phe, phenylacetate and phenylglyoxylate, but not with benzoate Aromatoleum evansii benzoyl-S-CoA + CO2 + NADH
-
?
phenylglyoxylate + NAD+ + CoA-SH enzyme of anaerobic Phe metabolism Aromatoleum evansii benzoyl-S-CoA + CO2 + NADH
-
?

Subunits

Subunits Comment Organism
decamer 2 * 50000 + 2 * 48000 + 2 * 43000 + 2 * 24000 + 2 * 11500, SDS-PAGE. The enzyme complex consists of a core enzyme of 4 subunits with the composition alpha2beta2gamma2delta2, this complex is able to reduce viologen dyes. The holoenzyme contains in addition an epsilon2 unit that catalyzes the transfer of electrons from a small ferredoxin-like subunit of the core complex to NAD, this unit also catalyzes the transhydrogenase reaction, carries FAD and resembles ferredoxin:NAD(P)+-oxidoreductase Aromatoleum evansii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
46
-
benzyl viologen
-
Aromatoleum evansii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
Aromatoleum evansii

Cofactor

Cofactor Comment Organism Structure
FAD contains 1.1 mol FAD per mol of enzyme Aromatoleum evansii
thiamine diphosphate reaction is strictly dependent on Aromatoleum evansii