METHOD OF OBTAINING PROTEIN Bg12p, REPRESENTING HOMOLOGUE OF ANTIGEN-MARKER OF MYCOSES CAUSATIVE AGENTS IN HUMANS AND ANIMALS Russian patent published in 2015 - IPC C12P21/02 C07K14/395 C12R1/865 

FIELD: chemistry.

SUBSTANCE: invention relates to field of biotechnology. Claimed is method of obtaining protein Bgl2p - homologue of antigen-marker of causative agents of mycoses in humans and animals. Dried biomass of commercial yeasts Saccharomyces cerevisiae is soaked in culture medium for 15-30 minutes at 27-33°C. After that, cells are precipitated by centrifugation for 2-5 minutes at 1500-2500 g and at room temperature. Obtained sediment is washed under the same conditions with distilled water. Washed cells are re-suspended in distilled water and frozen at temperature from -20 to -79°C. After that, cells are defrosted at room temperature. Toluene is added to final concentration 1-5%. Obtained mixture is incubated on rocker at 120-180 rev/min and temperature 35-45°C for 45-90 hours. After that, mixture is centrifuged, sediment of cell walls, which is washed with distilled water and re-centrifuged for 0.5-5 minutes at 2000-10000 g. Obtained sediment is re-suspended in distilled water and processed by ultrasound with frequency 20-30 kilohertz with power 30-50 Wt with cooling on ice for 2-5 minutes with 30 second interval each 20-40 seconds. After that cell walls are precipitated by centrifugation at 2000-10000 g. Washings of sediment are carried out by re-suspension and following centrifugation at room temperature in solution of 0.1-1% Triton X-100 one time, in solution of 0.2-1M NaCl three times, in 1-2% solution of sucrose one time and three times with distilled water. Target product is extracted from cell walls into distilled water with heating to 70-100°C for 5-15 minutes. After that, supernatant, containing target protein, is separated by double centrifugation at 3000-10000 g for 2-10 minutes.

EFFECT: invention makes it possible to obtain protein Bgl2p with output 70-100 mcg per a gram of dry yeasts and purity degree not lower than 95%.

1 dwg, 6 ex