Open AccessPurification and characterization of a glutathione S ‐transferase from the fungus Cunninghamella elegans
Author(s) -
Cha ChangJun,
Coles Brian F,
Cerniglia Carl E
Publication year - 2001
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2001.tb10850.x
Subject(s) - affinity chromatography , glutathione s transferase , gel electrophoresis , biochemistry , enzyme , polyacrylamide gel electrophoresis , glutathione , size exclusion chromatography , western blot , microbiology and biotechnology , biology , chemistry , chromatography , gene
Cunninghamella elegans grown on Sabouraud dextrose broth had glutathione S ‐transferase (GST) activity. The enzyme was purified 172‐fold from the cytosolic fraction (120 000× g ) 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 α‐, μ‐, and π‐class GSTs, although it showed a small degree of cross‐reactivity with a θ‐class GST. The N‐terminal amino acid sequence of the purified enzyme showed no significant homology with other known GSTs.