Ultrasonic treatment releases thioglucoside glucohydrolase (thioglucosidase) from insoluble particles of Crambe abyssinica seed meal. The crude enzyme is optimally activated by 10−2 M ascorbate, requires a reducing agent for stability, and is inhibited by 10−3 M p-chloromercuribenzenesulfonate.Chromatography of a meal extract on cross-linked dextran separates two enzyme fractions. About 80% of the activity elutes at an apparent molecular weight of 110 000; the remainder elutes at the void volume. Both crude soluble enzyme and the fractions from the dextran column produce goitrin from epi-progoitrin, but in the presence of ferrous ion the chief aglucon product becomes 1-cyano-2-hydroxy-3-butene. The Michaelis constant (of the major peak) at pH 5 is 0.004 M. Ferrous ion produces substrate inhibition at high epi-progoitrin levels. Sequential fractionation of meal extract by ammonium sulfate precipitation and by chromatography on cross-linked dextran effects an 80-fold purification.