Description
NADH Oxidase is a flavoprotein enzyme that catalyzes the oxidation of NADH to NAD⁺, coupled with the reduction of molecular oxygen (O₂) to either hydrogen peroxide (H₂O₂) or water (H₂O), depending on the enzyme source and reaction conditions. This enzyme plays a key role in redox homeostasis and regenerating NAD⁺ for metabolic processes such as glycolysis and fermentation.
Expression System
Escherichia coli
Detection Method
Spectrophotometry
Concentration
200 U/mg or more
Activity
Activity can be calculated by using the following formula:
Volume activity (U/mL) =ΔOD/min (ΔOD test−ΔOD blank) × df/ 1.0×Vs ＝ ΔOD/min × 20 × df
Weight activity (U/mg) ＝ (U/mL) × 1/C
Vs: Sample volume (0.05 mL)
1.0: Unit absorbance at 600 nm due to unit definition
df: Dilution factor
C: Enzyme concentration in dissolution (mg/mL)
Unit Definition
One unit causes decrease in DCPIP by one unit of absorbance (1.0) per minute under the following conditions：27 mM Tris-HCl pH 7.5, 0.2 mM NADH, 40 μM DCPIP and 33 μg/mL BSA.
Form
Lyophilized
Synonyms
NADH Oxidase, NADH:oxygen oxidoreductase, NADH oxidizing enzyme, DPNH oxidase, H₂O₂-forming NADH oxidase, FAD-NADH oxidase, Flavoprotein oxidase, NADH oxygen oxidoreductase, NADH oxidoreductase (oxygen), NADH H₂O₂ oxidase, NADH diaphorase, EC 1.6.3.x oxidase, NADH regeneration enzyme, Redox flavoenzyme
Reconstitution
It is recommended to weigh and reconstitute 10 mg of lyophilized powder in 200 μL double-distilled water directly and incubate the solution for at least 10 mins to ensure sufficient re-dissolved.
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at -20°C or lower for long term storage.
Stability & Storage
This product is stable at -20°C for long-term storage under sterile conditions.
Avoid repeated free-thaw cycles.
The effects of various chemicals on NADH Oxidase.
The enzyme was incubated with the chemicals at 25°C for 1 hour.
pH stability of NADH Oxidase.
The enzyme powder was reconstituted by double-distilled water and treated with different pH buffer condition for 3 hours at 30°C. pH 4.0-6.0, 0.1 M Sodium citrate buffer; pH 7.0-8.0, 0.1 M Potassium phosphate buffer; pH 8.5, 0.1 M Tris-HCl buffer; pH 10.0-11.0, 0.1 M Carbonate-bicarbonate buffer.
Thermal stability of NADH Oxidase.
The enzyme powder was reconstituted by double-distilled water and treated with different temperatures for 30 minutes. Final concentration: 20 U/mL
pH activity of NADH Oxidase.
The buffer conditions with various pH values were used in the reaction at 25°C. pH 5.0-6.0, 0.1 M Sodium citrate buffer; pH 7.0-8.0, 0.1 M Potassium phosphate buffer; pH 8.5, 0.1 M Tris-HCl buffer; pH 10.0-11.0, 0.1 M Carbonate-bicarbonate buffer.
Temperature activity of NADH Oxidase.
The enzyme reactions in 0.2 M Tris-HCl buffer, pH 7.5, were carried out under different temperatures.
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