• »
• Protein extraction from yeast Pichia pastoris with Minilys

Protein extraction from yeast Pichia pastoris with Minilys

Instrument: Minilys homogenizer

Sample:  Pichia pastoris yeast

Protocol:

• 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.

Results:

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.