Simple and effective protocols of cell wall disruption were elaborated for tested fungal strains: Penicillium citrinum, Aspergillus fumigatus, Rhodotorula gracilis. Several techniques of cell wall disintegration were studied, including ultrasound disintegration, homogenization in bead mill, application of chemicals of various types, and osmotic shock. The release of proteins from fungal cells and the activity of a cytosolic enzyme, glucose-6-phosphate dehydrogenase, in the crude extracts were assayed to determine and compare the efficacy of each method. The presented studies allowed adjusting the particular method to a particular strain. The mechanical methods of disintegration appeared to be the most effective for the disintegration of yeast, R. gracilis, and filamentous fungi, A. fumigatus and P. citrinum. Ultrasonication and bead milling led to obtaining fungal cell-free extracts containing high concentrations of soluble proteins and active glucose-6-phosphate dehydrogenase systems.
- MeSH
- Aspergillus fumigatus účinky léků enzymologie MeSH
- buněčná stěna účinky léků MeSH
- buněčné extrakty chemie MeSH
- detergenty farmakologie MeSH
- fungální proteiny analýza metabolismus MeSH
- glukosa-6-fosfátdehydrogenasa analýza metabolismus MeSH
- osmotický tlak MeSH
- Penicillium účinky léků enzymologie MeSH
- Rhodotorula účinky léků enzymologie MeSH
- vibrace ultrazvukové MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- MeSH
- Aspergillus fumigatus enzymologie genetika MeSH
- chromatografie na tenké vrstvě metody MeSH
- elektroforéza v polyakrylamidovém gelu metody MeSH
- genom fungální MeSH
- mannosidasy genetika izolace a purifikace metabolismus MeSH
- regulace genové exprese u hub MeSH
- sekvence aminokyselin MeSH
- Publikační typ
- techniky in vitro MeSH
