o-Dihydroxyphenoloxidase (E.C. 188.8.131.52) (o-DPOase) was extracted from acetone-extracted freeze-dried yam tubers, and fractionated by (NH4)2SO4 precipitation and chromatography on DEAE-cellulose to give two forms from Dioscorea alta L. (cv. UM680) and three forms from D. rotundata Poir. (cv. Nwopoke). Polyacrylamide gel electrophoresis revealed one of the forms from each species to contain two protein bands with o-DPOase activity. o-DPOase from D. alata showed activity with catechol, (+)-catechin, (−)-epicatechin and chlorogenic acid, but not with tyrosine, as substrate, o-DPOase from D. rotundata showed activity only with catechol and (+)-catechin. Km values for D. rotundata enzymes, calculated assuming a two-substrate reaction, were between 0.2 and 0.8 mM for oxygen and 70 and 120 mM for catechol. The enzyme was most active between pH 5.5 and pH 7.0, and showed slight activation after holding for 2 minutes at 40 or 50°C. After heating to above 60°C the enzyme showed evidence of irreversible denaturation. Theo-DPOase activity extracted from ten cultivars of five species of D. alata L., D. bulbifera L., D. cayenensis Lam., D. dumetorum Pax, and D. rotundata Poir. were compared.