<P><B>Highlights</B></P><P>► New bioprocesses for the production of cellulolytic enzymes are needed. ► A novel combined cultivation method for cellulase production is being proposed. ► The new method resulted in a 3-fold improvement over conventional fermentation.</P> <P><B>Abstract</B></P><P>Sequential solid-state and submerged cultivation with sugarcane bagasse as substrate for cellulase production by <I>Aspergillus niger</I> A12 was assessed by measuring endoglucanase activity. An unconventional pre-culture with an initial fungal growth phase under solid-state cultivation was followed by a transition to submerged fermentation by adding the liquid culture medium to the mycelium grown on solid substrate. For comparison, control experiments were conducted using conventional submerged cultivation. The cultures were carried out in shake flasks and in a 5-L bubble column bioreactor. An endoglucanase productivity of 57±13IU/L/h was achieved in bubble column cultivations prepared using the new method, representing an approximately 3-fold improvement compared to conventional submerged fermentation. Therefore, the methodology proposed here of a sequential fermentation process offers a promising alternative for cellulase production.</P>