Hi Cory, I am afraid to say that Pichia and Saccharomyces are tough cells to break compared to bacteria. We also purifiy membrane proteins in yeast in my lab so we have gone through this process. With regular yeast we can break in an emulsiflex but it takes some hard work and this is not really viable option on a large scale once you process several liters of cell culture. So we use a bead-beater ( from BIOSPEC) and glass beads (0.5 mm for yeast, the diameter depends on the kind of organism you want to disrupt). it works well and is quite fast , however you have to deal with the bead cleaning part and the losses due to the mass of liquid trapped in the beads (rinsing of beads with buffer is fine but it results in an increase burden at the membrane centrifugation step) It takes some optimization (mass of beads/mass of cells processed), but once this is is set this is probably the way to go, Bead beaters come in various sizes depending on the volumes you want to process. It is just a blender after all.
Hope this helps. Best regards -- Pascal F. Egea, PhD Assistant Professor UCLA, David Geffen School of Medicine Department of Biological Chemistry 314 Biomedical Sciences Research Building office (310)-983-3515 lab (310)-983-3516 email pe...@mednet.ucla.edu
