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Literature summary extracted from

  • Hirano, J.; Miyamoto, K.; Ohta, H.
    Purification and characterization of thermostable H2O2-forming NADH oxidase from 2-phenylethanol-assimilating Brevibacterium sp. KU1309 (2008), Appl. Microbiol. Biotechnol., 80, 71-78.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.6.3.3 (NH4)2SO4 250 mM, 10fold activation Brevibacterium sp.
1.6.3.3 p-chloromercuribenzoate 1 mM, 1.9fold activation Brevibacterium sp.

Application

EC Number Application Comment Organism
1.6.3.3 synthesis regeneration of NAD+ utilizing this enzyme made selective oxidation of mandelic acid or L-phenylalanine possible. This thermostable enzyme is expected to be applicable as a useful biocatalyst for NAD+ recycling Brevibacterium sp.

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.6.3.3 AgNO3 1 mM, 63% inhibition Brevibacterium sp.
1.6.3.3 CuCl2 1 mM, 58% inhibition Brevibacterium sp.
1.6.3.3 HgCl2 1 mM, 29% inhibition Brevibacterium sp.
1.6.3.3 ZnCl2 1 mM, 61% inhibition Brevibacterium sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.6.3.3 0.022
-
 NADH pH 8.8, 25°C Brevibacterium sp.

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.6.3.3 cytoplasm
-
 Brevibacterium sp. 5737
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.6.3.3 57000
-
 2 * 57000, SDS-PAGE Brevibacterium sp.
1.6.3.3 102000
-
 gel filtration Brevibacterium sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.6.3.3 NADH + H+ + O2 Brevibacterium sp. the enzyme is highly specific for NADH, no activity with NADPH NAD+ + H2O2
-
 ?
1.6.3.3 NADH + H+ + O2 Brevibacterium sp. KU1309 the enzyme is highly specific for NADH, no activity with NADPH NAD+ + H2O2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.6.3.3 Brevibacterium sp.
-
-
-
1.6.3.3 Brevibacterium sp. KU1309
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.6.3.3
-
 Brevibacterium sp.

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.6.3.3 culture condition:2-phenylethanol-grown cell
-
 Brevibacterium sp.
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.6.3.3 5.2
-
 pH 8.8, 25°C Brevibacterium sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.6.3.3 NADH + H+ + O2 the enzyme is highly specific for NADH, no activity with NADPH Brevibacterium sp. NAD+ + H2O2
-
 ?
1.6.3.3 NADH + H+ + O2 the enzyme is highly specific for NADH, no activity with NADPH Brevibacterium sp. KU1309 NAD+ + H2O2
-
 ?

Subunits

EC Number Subunits Comment Organism
1.6.3.3 dimer 2 * 57000, SDS-PAGE Brevibacterium sp.

Synonyms

EC Number Synonyms Comment Organism
1.6.3.3 H2O2-forming NADH oxidase
-
 Brevibacterium sp.
1.6.3.3 NOX
-
 Brevibacterium sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.6.3.3 25
-
 assay at Brevibacterium sp.
1.6.3.3 70
-
-
 Brevibacterium sp.

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.6.3.3 30 70 activity at 30°C is about 40% of the maximal at 70°C Brevibacterium sp.

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.6.3.3 70
-
 1 h, no significant loss of activity Brevibacterium sp.
1.6.3.3 80
-
 1 h, about 30% loss of activity Brevibacterium sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.6.3.3 8.8
-
 assay at Brevibacterium sp.
1.6.3.3 9
-
-
 Brevibacterium sp.

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.6.3.3 7.9 11 pH 7.8: about 60% of maximal activity, pH 11.0: about 90% of maximal activity Brevibacterium sp.

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.6.3.3 6 10 1 h, stable Brevibacterium sp.

Cofactor

EC Number Cofactor Comment Organism Structure
1.6.3.3 NADH the enzyme is highly specific for NADH, relative activity with NADPH is 2% compared to the activity with NADH Brevibacterium sp.