After cooling to room temperature, 25 ml of 6 × 106 spore/ml of M. hiemalis or a loop full of S. cerevisiae from petri dishes was added for inoculation. The flask was incubated at 32 ± 0.5 ◦C and 130 rpm for 24 h. The biomass was separated by centrifugation (4000 rpm for 20 min) under sterile conditions and used for SSF