Electropulsation as an alternative method for protein extraction from yeast.
The application of series of high intensity electric pulses to a yeast suspension provoked a considerable release of some cytoplasmic proteins, glutathione reductase, 3-phosphoglycerate kinase and alcohol dehydrogenase. A maximal yield was achieved 3-8 h after pulsation. The electro-induced protein efflux was accelerated by pretreatment with the reducing agent dithiothreitol and showed a strong dependence on the growth phase and the presence of monovalent ions in the post-pulse incubation medium. The results obtained for two strains of Saccharomyces cerevisiae, PV3 (diploid) and Y47 (wild haploid), showed that electropulsation can be used for the effective extraction of cytoplasmic proteins with a preserved functional activity.
Ganeva
V
V
Department of Biophysics and Radiobiology, Faculty of Biology, Sofia University, Bulgaria. valia@biofac.uni-sofia.bg
Galutzov
B
B
D000426:Alcohol Dehydrogenase / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D018274:Electroporation / Q000379:methods*; D005656:Fungal Proteins / Q000302:isolation & purification* / Q000378:metabolism; D005980:Glutathione Reductase / Q000378:metabolism; D010735:Phosphoglycerate Kinase / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000201:enzymology
D005656:Fungal Proteins; D000426:Alcohol Dehydrogenase; D005980:Glutathione Reductase; D010735:Phosphoglycerate Kinase