Protein extraction from yeast Pichia pastoris with Minilys
Instrument: Minilys homogenizer
Sample: Pichia pastoris yeast
- Precellys kit: 03961-1-004 (0.5 mm glass beads).
- Add Samples: 5.108 to 1.109 of Pichia pastoris (pellets).
- Add Buffer: 500 μL of Pichia breaking buffer (pH7.4, 50mM Na-Phosphate, 1mM EDTA, 5%(v/v) glycerol).
- Minilys: Two different speeds and different durations of cycle were tested to lyse yeast cells. Duplicate (5000 rpm) or triplicate (4000 rpm) by condition.
- 5000 rpm 20 sec x 2, 4, or 6 with 60 sec break on ice between each cycle.
- 4000 rpm 20 sec x 2 with 60 sec break on ice between each cycle.
While treatment at 4000 rpm 20 sec x 2 already gave good extraction of protein. There were no sign of the excess disintegration of the proteins even after 5000 rpm 20 sec x 6 treatment. Even the 5000 rpm 20 sec x 6 treatment gave no excess heat and the tube was less than 30°C after each treatment.
SDS-PAGE of the protein extracted from Pichia pastoris