Abstract
(1) Crystalline catalase prepared by Sirakawa's method was separated into two components, by application of the column chromatography with tricalcium phosphate gel of Tiselius. By stepwise elution, changing ionic concentration of phosphate buffer of pH 6.8, a main and a minor components eluted from the column with 0.1 and 0.2M.
Protein recovery was 95% and 5%, and Kat. f. was 30, 000 and 10, 000, respectively, in accordance with main and minor components.
(2) These two components behaved independently when rechromatography was performed, indicating that no denaturation occurred during chromatography.
(3) Sedimentation constant was s20W=11.0 and 8.5, and the iron content was 0.11 and 0.08%, respectively, in accordance with the main and the minor fractions.
(4) The optical absorption at hematin moiety showed no significant difference between the two components, whereas at 405mμ the minor component characteristically decreased in its absorbancy.
