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Factors Affecting Heat Inactivation and Partial Reactivation of Peroxidase Purified by Ion‐Exchange Chromatography a, b
Author(s) -
WILDER C. J.
Publication year - 1962
Publication title -
journal of food science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.772
H-Index - 150
eISSN - 1750-3841
pISSN - 0022-1147
DOI - 10.1111/j.1365-2621.1962.tb00141.x
Subject(s) - peroxidase , chemistry , guaiacol , hydrogen peroxide , ionic strength , chromatography , ion chromatography , fractionation , enzyme , biochemistry , organic chemistry , aqueous solution
SUMMARY Diethylaminoethyl‐cellulose (DEAE) was used in an ion‐exchange chromatographic fractionation of commercially prepared horse‐radish peroxidase. One major fraction was separated from other minor fractions and used for heat inactivation and reactivation work. Heat inactivation was carried out at 250–300° F, with most of the inactivation work at 280° F. Peroxidase activity was assayed in measuring the rate of utilization of hydrogen peroxide in the formation of tetraguaiacol from guaiacol. The concentration of tetraguaiacol was measured with a recording spectrophotometer. The ionic strength of the buffer solution in which the peroxidase was heated had an effect on its inactivation. Within limits, the higher the ionic strength the greater the heat inactivation of the enzyme. Activity was maximum at pH 7. Purified peroxidase heated 30 set at 280° F and stored 24 hr at 72° F, with the heat treatment and storage time repeated three times, decreased in activity as a first‐order reaction. The storage temperature of heat‐inactivated peroxidase affected the rate and degree of reactivation.