Freezing and thawing of a microbial cell paste will inevitably cause ice crystals to form and their expansion followed by thawing will lead to some subsequent disruption of cells. It is slow, with limited release of cellular materials, and has not often been used as a technique on its own, although it is often used in combination with other techniques. /3-Glucosidase has been obtained from S. cerevisiae by this method (Honig and Kula, 1976). A sample of 360 g of frozen yeast
paste was thawed at 5° for 10 hours. This cycle was repeated twice before further processing.
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