3.1. Preliminary cultivations in Erlenmeyer flasks: Effect of cultivation method, sugarcane bagasse steam explosion pretreatment, and substrate particle size on enzyme production
The responses for the effects of sugarcane bagasse pretreatment, particle size (dp), and cultivation method on endoglucanase production are shown in Fig. 1. Cultivations in shake flasks were carried out at 32 °C and 200 rpm, using untreated (USB) and steam explosion pretreated (PSB) sugarcane bagasse, with particle size ranges of dp ≤ 0.5 and 1.0 ≤dp ≤ 2.0. The cultivation methods employed were conventional submerged fermentation (SmF) and the new sequential solid-state and submerged fermentation (SF). The combination of cultivation conditions investigated was planned according to a 23factorial design, with all experiments carried out in duplicate (Table 2).