2.5.1. Pre-culture for submerged fermentation (SmF)
In the conventional submerged fermentation experiments, the pre-culture was initiated with the addition of 107 spores per mL of nutrient medium enriched with 30 g L−1 of glucose. The incubation was carried out for 50 h in an orbital shaker incubator at 32 °C and 200 rpm. A portion of 10% (v/v) was then transferred to the culture medium in order to initiate enzyme production in a 4 L stirred tank bioreactor or in shake flasks