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Literature summary extracted from

  • McDonald, D.W.; Coddington, A.
    Properties of the assimilatory nitrate reductase from Aspergillus nidulans (1974), Eur. J. Biochem., 46, 169-178.
    View publication on PubMed

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.7.1.3 8-hydroxyquinoline pH 7.5, 20% inhibition at 0.1 mM Aspergillus nidulans
1.7.1.3 azide pH 7.5, 90% inhibition at 0.01 mM Aspergillus nidulans
1.7.1.3 cyanide pH 7.5, 100% inhibition at 0.01 mM Aspergillus nidulans
1.7.1.3 iodoacetamide pH 7.5, 40% inhibition at 1 mM Aspergillus nidulans
1.7.1.3 additional information cysteine or dithiothreitol relieve or prevent inhibition. Not inhibited by urea, glutamic acid, aspartic acid and ammonia at 10 mM Aspergillus nidulans
1.7.1.3 NADP+ inhibits NADPH oxidation, non-competitive with respect to nitrate Aspergillus nidulans
1.7.1.3 nitrite competitive with respect to nitrate and non-competitive to NADPH Aspergillus nidulans
1.7.1.3 p-hydroxymercuribenzoate pH 7.5, 85% inhibition at 0.001 mM Aspergillus nidulans
1.7.1.3 phenanthroline hydrate pH 7.5, 1:10, 20% inhibition at 0.1 mM Aspergillus nidulans
1.7.1.3 potassium chlorate pH 7.5, slight, 20% inhibition at 5 mM Aspergillus nidulans
1.7.1.3 Sodium nitrite pH 7.5, slight, 50% inhibition at 5 mM Aspergillus nidulans

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.7.1.3 0.009
-
 NADPH + nitrate, pH 6.5 and + cytochrome c, pH 7.5 Aspergillus nidulans
1.7.1.3 0.01
-
 NADPH + nitrate, pH 7.5 Aspergillus nidulans
1.7.1.3 0.015
-
 NADPH + cytochrome c, pH 8.5 Aspergillus nidulans
1.7.1.3 0.024
-
 cytochrome c cosubstrate NADPH, pH 8.5 Aspergillus nidulans
1.7.1.3 0.029
-
 cytochrome c cosubstrate NADPH, pH 7.5 Aspergillus nidulans
1.7.1.3 0.06
-
 nitrate cosubstrate NADPH, pH 7.5 Aspergillus nidulans
1.7.1.3 0.08
-
 nitrate cosubstrate NADPH, pH 6.5 Aspergillus nidulans

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.7.1.3 Molybdenum
-
 Aspergillus nidulans
1.7.1.3 phosphate stimulates, required for maximal activity Aspergillus nidulans

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.7.1.3 180000 200000 sucrose density gradient, gel-filtration Aspergillus nidulans

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.7.1.3 nitrate + NADPH Aspergillus nidulans
-
 nitrite + NADP+ + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.7.1.3 Aspergillus nidulans
-
 bi-1 a, translocation-free biotin auxotroph
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.7.1.3 ion-exchange, ammonium sulfate precipitation, gel filtration Aspergillus nidulans

Reaction

EC Number Reaction Comment Organism Reaction ID
1.7.1.3 nitrite + NADP+ + H2O = nitrate + NADPH + H+ random order rapid-equilibrium mechanism, two functional parts: 1. heat labile, FAD and haemoprotein containing, binds NADPH and transfers electrons from NADPH via FAD and perhaps cytochrome b to acceptors such as cytochrome c, 2. heat stable, molybdenum containing, accepts electrons from reduced viologen dyes and transfers them to nitrate Aspergillus nidulans
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.7.1.3 2.5
-
-
 Aspergillus nidulans
1.7.1.3 4 16.1 at different stages of purification Aspergillus nidulans

Storage Stability

EC Number Storage Stability Organism
1.7.1.3 4°C, half life of 3-5 days Aspergillus nidulans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.7.1.3 chlorate + NADPH lower affinity than to nitrate, appears to be toxic or its product Aspergillus nidulans chlorite + NADP+
-
 ?
1.7.1.3 ferricytochrome c + NADPH + H+
-
 Aspergillus nidulans ferrocytochrome c + NADP+ + H2O
-
 r
1.7.1.3 nitrate + NADPH
-
 Aspergillus nidulans nitrite + NADP+ + H2O
-
 ?
1.7.1.3 nitrate + NADPH two associated activities: cytochrome c reductase and reduced viologen dye:nitrate reductase Aspergillus nidulans nitrite + NADP+ + H2O
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.7.1.3 25
-
 assay at Aspergillus nidulans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.7.1.3 7.5
-
 assay at Aspergillus nidulans
1.7.1.3 7.5 8
-
 Aspergillus nidulans

Cofactor

EC Number Cofactor Comment Organism Structure
1.7.1.3 FAD stimulated by flavin Aspergillus nidulans
1.7.1.3 heme
-
 Aspergillus nidulans

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.7.1.3 0.04 0.048 NADP+ NADPH as substrate in saturating and non-saturating concentrations, competitive Aspergillus nidulans
1.7.1.3 0.065
-
 NADP+ nitrate as substrate in non-saturating concentrations, non-competitive Aspergillus nidulans
1.7.1.3 0.15
-
 nitrite NADPH as substrate in non-saturation concentration, non-competitive Aspergillus nidulans
1.7.1.3 0.18
-
 nitrite nitrate as substrate in saturating and non-saturating concentrations, competitive Aspergillus nidulans