ABSTRACITomato (Lycopersicon esculentum Mill) anionic peroxidase was found to catalyze oxidase reactions with NADH, glutathione, dithiothreitol, oxaloacetate, and hydroquinone as substrates with a mean activity 30% that of horseradish peroxidase; this is in contrast to the negligible activity of the tomato enzyme as compared to the horseradish enzyme in catalyzing an indoleacetic acid-oxidase reaction with only Mn2' and a phenol as cofactors. Substitution of Ce3' for Mn2' produced an 18-fold larger response with the tomato enzyme than with the horseradish enzyme, suggesting a significant difference in the autocatalytic indoleacetic acidoxidase reactions with these two enzymes. In attempting to compare enzyme activities with 2,4-dichlorophenol as a cofactor, it was found that reaction rates increased exponentially with both increasing cofactor concentration and increasing enzyme concentration. While the former response may be analogous to allosteric control of enzyme activity, the latter response is contrary to the principle that reaction rate is proportional to enzyme concentration, and additionally makes any comparison of enzyme activity difficult.Tomato ripening, like that of pears, bananas, and avocadoes, appears to be at least partly controlled by auxin levels (12); free IAA decreases and peroxides accumulate in tomato ripening.An isoelectric point of less than 5.5 for tomato anionic peroxidase, as suggested by its column chromatographic behavior, is lower than those of the major horseradish peroxidase isozymes (1 1) and is in particular lower than the value of 8.9 for horseradish peroxidase isozyme C, which predominates in commercial horseradish peroxidase preparations. This is of interest because of the statement (2) that cationic peroxidases serve as IAAoxidases in vivo. The more cationic members of isozyme sets often show the highest specific activity in the IAA-oxidase reaction or the highest ratio of IAA-oxidase to peroxidase activity (1 1, 20, 22, 24), though this is not consistently the case (5, 1 1). In considering the results of in vitro comparisons of peroxidases, it should be recalled that IAA-oxidase activity is more closely related to the oxidation-reduction potential of the enzyme than to the isoelectric point (24), that peroxidase isozymes may differ greatly in their peroxidase activity (15), and that relative IAAoxidase activities may depend greatly on the conditions of assay used (1, 22).The major peroxidase oftomato fruits, termed tomato anionic peroxidase, differs markedly from horseradish peroxidase in its ability to oxidize IAA (17). The horseradish enzyme catalyzes an autocatalytic IAA-oxidase2 reaction, which is stimulated by phenols and Mn2". The IAA-oxidase reaction of the tomato enzyme, however, was found to be completely dependent on the presence of catalytic amounts of H202 in the assay mixtures (18) MATERIALS AND METHODS Enzymes. Tomato anionic peroxidase was prepared from tomato pericarp using extraction, (NH4)2SO4 fractionation, and QAE-Sephadex column chromatography with grad...