A thermostable manganese-containing superoxide dismutase from the thermophilic fungus Thermomyces lanuginosus.
A thermostable superoxide dismutase (SOD) from a Thermomyces lanuginosus strain (P134) was purified to homogeneity by fractional ammonium sulfate precipitation, ion-exchange chromatography on DEAE-Sepharose, Phenyl-Sepharose hydrophobic interaction chromatography, and gel filtration on Sephacryl S-100. The molecular mass of a single band of the enzyme was estimated to be 22.4 kDa, using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Using gel filtration on Sephacryl S-100, the molecular mass was estimated to be 89.1 kDa, indicating that this enzyme was composed of four identical subunits of 22.4 kDa each. The SOD was found to be inhibited by NaN3, but not by KCN or H2O2, suggesting that the SOD in T. lanuginosus was of the manganese superoxide dismutase type. The SOD exhibited maximal activity at pH 7.5. The optimum temperature for the activity was 55 degrees C. It was thermostable at 50 and 60 degrees C and retained 55% activity after 60 min at 70 degrees C. The half-life of the SOD at 80 degrees C was approximately 28 min and even retained 20% activity after 20 min at 90 degrees C.
Department of Environmental Biology, Shandong Agricultural University, Taian, Shandong 271018, China. lidc20@sdau.edu.cn
Duo-Chuan
DC
Li
Jing
J
Gao
Ya-Ling
YL
Li
Jing
J
Lu
D000645:Ammonium Sulfate / Q000494:pharmacology; D001203:Ascomycota / Q000201:enzymology*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004983:Ethanolamines / Q000737:chemistry; D006358:Hot Temperature; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008345:Manganese / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry; D015398:Signal Transduction; D013482:Superoxide Dismutase / Q000737:chemistry*; D013696:Temperature; D013997:Time Factors
D004983:Ethanolamines; D008345:Manganese; D012685:Sepharose; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase; C007369:2-diethylaminoethanol; D000645:Ammonium Sulfate