The liquid was then decanted and the biocatalyst was washed twice with 400 ml offresh glucose medium. Amounts of 100 g (wet weight) of the biocatalyst and 4.9 g(wet weight) of harvested yeast culture were frozen to −45◦C at a cooling rate of3◦C/min in a controlled rate freezer (BioCool, FTS Systems, NY, USA), with no additions of cryoprotecting media.