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(H2O2: H2O2 oxidoreductase; EC

Catalase is a heme-containing enzyme which catalyzes the following reaction:

2H2O2 -------------------> 2H2O + O2
Catalase is present in a wide variety of tissues from animals, plants and microorganisms. Mammalian liver contains high concentrations of the enzyme. Bovine liver catalase has a molecular weight of 250,000 with four subunits of equal size. Optimum pH is 7.0 and isoelectric point is 5.4.

Encapsulated or immobilized catalase is used in the food industry whenever hydrogen peroxide needs to be destroyed, for example, in the manufacture of cheese (Chu, H.D., Leeder, J.G. and S.C. Gilbert, J. Food Sci, 40, 641, 1975). Catalase can also be used in coupled systems for the determination of metabolites in biological fluids.

1.Source: Aspergillus Niger
   Form: Freeze-dried powder 
   Solubility: Distilled water or dilute buffer 
   Stability: Store at -20 C (-4 F) 
   Activity (approx.): 2000 U/mg protein 
   Protein (approx.): 95% 
   Catalog No.: 191A2000 
2.Source: Bovine Liver
   Form: Freeze-dried powder 
   Solubility: Distilled water or dilute buffer 
   Stability: Store at -20 C (-4 F) 
   Activity (approx.): 5,000 U/mg protein 
   Protein (approx.): 95% 
   Catalog No.: 194A5000 

The amount of enzyme which catalyzes the decomposition of one micromole of H2O2 per minute at 25C and pH 7.00.

The decomposition of hydrogen peroxide, which is a measure of catalase activity, can be followed by measuring the absorbance at 240 nm (Methods of Enzymatic Analysis, Bergmeyer, H.U., ed., Vol 1, p 438, 1974, Academic Press, New York).

  1. 0.05 M Potassium phosphate buffer, pH 7.0.
  2. 0.05% Hydrogen Peroxide (substrate) solution. Dilute 0.2 ml of 30% H2O2 to 100 ml with phosphate buffer. The absorbance at 240 nm should be in the range of 0.50-0.55.
  3. Catalase (enzyme) solution. Dilute in buffer to yield a concentration of 5-10 U/ml. Prepare fresh prior to assay.

  1. Set spectrophotometer (equipped with a strip chart recorder and temperature control) at 240 nm and 25C.
  2. In a quartz cuvette pipette 2.9 ml of diluted H2O2 solution (substrate). Incubate in spectrophotometer at 25C for 5 min. to attain temperature equilibration. Record absorbance at 240 nm (blank).
  3. Initiate reaction by adding 0.1 ml diluted enzyme (catalase) solution to the cuvette. Record decrease in absorbance at 240 nm for 2-3 minutes.
  4. Calculate (delta)E240 nm/min

Activity (U/mg) = (ΔE240nm/min)(Total Vol.)(Enz. Diln.)
(0.043)(Enz. Vol.)(mg Enz./ml)

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