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A thermostable lipoxygenase activity was recovered from defatted horsegram seeds. Horsegram seeds are the richest source of an acidic lipoxygenase activity reported so far with 2500 U/ g of flour. The enzyme was most active at pH 4.0 and was stable between pH 3.5 to 6.0. The enzyme was thermostable retaining greater than 75% activity when incubated at 90°C for 90 min. The half-life of the enzyme was 120 at 90°C and 150 minutes at 85°C in 0.02 M sodium phosphate buffer, pH 6.8. The enzyme was active against linoleic acid and released both 13 and 9 hydroperoxides in the ratio of 3:2.