Biotechnology and Bioengineering, Vol.44, No.6, 736-744, 1994
The Release of Virus-Like Particles from Recombinant Saccharomyces-Cerevisiae - Effect of Freezing and Thawing on Homogenization and Bead Milling
Recombinant cells of Saccharomyces cerevisiae, expressing virus-like particles (Ty-VLPs), can be readily disrupted in a high pressure homogenizer and show identical disruption kinetics to the untransformed host strain. When the cells are freeze/thawed before disruption, they become about four times more resistant to homogenization. This effect increases with the number of freeze/thaw cycles, but is independent of the time the cells remain frozen. The freeze/thaw effect is observed with cells harvested during both the logarithmic and stationary phase of growth, and occurs with the untransformed host strain as well as the transformed one. Freeze/thawed cells are twice as resistant to disruption in the bead mill as fresh cells.
Keywords:HIGH-PRESSURE HOMOGENIZER;ESCHERICHIA-COLI;BACTERIAL ENZYMES;MICROBIAL-CELLS;BAKERS-YEAST;DISRUPTION;MECHANISM;DISINTEGRATION;PROTEINS