Fig. 3.

Cultivation of N. crassa in small scale stirred tank reactors. a Growth characteristics of the fungus monitored by online and offline data. Strain DHN-201 (Pvvd-glat-ht186-13×myc, exo-1, Δvib-1, Δvvd) was cultivated in a controlled 1 L bioreactor system in minimal Vogel’s medium with maltose as the carbon source at 30 °C. The dissolved oxygen level was maintained at 20% by controlling the stirrer speed. The pH value was maintained above 4.5 by the controlled addition of ammonia. b Macroscopical phenotype after 69 h of cultivation. Left culture in the bioreactor; Right Stirrer after cultivation. c Analysis of heterologous fusion protein production. Proteins in the supernatant were concentrated and separated with SDS-PAGE (S). After the Western blot transfer (WB), the tag was detected and signals were developed by electrochemiluminescence (development time of 15 min). The arrows indicate the expected molecular mass of the fusion protein. The protease assay (P) was performed as described above