The glutathione S-transferases of the following fungal species have been affinity purified on glutathione-Sepharose; Yarrowia lipolytica (NCYC825), Sporotrichum thermophile (ATCC16479), Talaromyces emersonii (ATCC26914) and Issatchenkia orientalis (CBS5147 and an isolate provided by Prof. Kumagai, Kyoto University). 2. SDS-PAGE analysis of affinity purified fungal extracts, followed by Western blots with polyclonal antisera to rat GSTs, was performed and indicated the presence of Alpha and Mu class glutathione S-transferases in the fungi immunologically and electrophoretically similar to the rat enzymes. 3. Each extract gave a characteristic affinity-gradient elution pattern on glutathione-agarose.