Cunninghamella elegans grown on Sabouraud dextrose broth had glutathione S-transferase (GST) activity. The enzyme was purified 172-fold from the cytosolic fraction (120 000Ug) of the extract from a culture of C. elegans, using Q-Sepharose ion exchange chromatography and glutathione affinity chromatography. The GST showed activity against 1-chloro-2,4-dinitrobenzene, 1,2-dichloro-4-nitrobenzene, 4-nitrobenzyl chloride, and ethacrynic acid. Sodium dodecyl sulfate^polyacrylamide gel electrophoresis gel filtration chromatography revealed that the native enzyme was homodimeric with a subunit of M r 27 000. Comparison by Western blot analysis implied that this fungal GST had no relationship with mammalian K-, W-, and Z-class GSTs, although it showed a small degree of crossreactivity with a a-class GST. The N-terminal amino acid sequence of the purified enzyme showed no significant homology with other known GSTs. ß