Purification and partial characterization of Cu/Zn superoxide dismutase from Kluyveromyces marxianus yeast.
A new thermostable Cu/Zn SOD from a thermotolerant yeast strain Kluyveromyces marxianus NBIMCC 1984 has been purified and characterized. The purification procedure comprises thermal treatment and dialysis, ion-exchange chromatography and chromatofocusing. The methodology is a rapid, efficient and highly specific, generating pure preparation (specific activity 996 U mg of protein(-1)) with a yield of 53%. The purified enzyme is a homodimer with Mw of 34,034 Da and has high N-terminal homology with other yeasts' Cu/Zn SOD enzymes. The protein is characterized with some unique features such as-thermostability (t(1/2) at 70 degrees C=30 min), pH stability in the alkaline range (7.5-8.5) and resistance to inhibitors and variety of chemicals. These characteristics reveal possibilities for wide practical application of K. marxianus Cu/Zn SOD enzyme.
Nedeva
Trayana
T
The Sofia University St. Kliment Ohridski, Faculty of Biology, 8 Dragan Tsankov Str., 1164 Sofia, Bulgaria.
Dolashka-Angelova
Pavlina
P
Moshtanska
Vesela
V
Voelter
Wolfgang
W
Petrova
Ventzislava
V
Kujumdzieva
Anna
A
D002852:Chromatography, Ion Exchange; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007716:Kluyveromyces / Q000201:enzymology*; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D013696:Temperature
D005656:Fungal Proteins; D006023:Glycoproteins; D013482:Superoxide Dismutase