Yeast liquid cultures were grown in Falcon flasks containing
30 mL of yeast extract peptone dextrose (YEPD) media (10 g/L yeast
extract, 20 g/L peptone, 20 g/L glucose). The cultures were incubated
for 24 h at 28 ◦C. Medium-free suspension cells were obtained
by cell culture centrifugation at 1.5 g for 10 min followed by successive
washing (twice) with sterile distilled water. The pellets
were resuspended in sterile distilled water up to a concentration of
1.0
×
108 cells/mL. The cells were counted using a Neubauer chamber
under optical microscope (Nikon E-200) and concentration
adjusted to 1.0
×
108 cells/mL (Zhao et al., 2008).