Figure 3.
Modification of arfA expression impacts colony macromorphology and protein secretion in submerged cultures. 20 ml of MM supplemented with 5% glucose and various doxycycline concentrations (0.25, 0.5, and 10 μg/ml) were inoculated with a final concentration of 5 × 106 spores/ml and incubated at 30°C for 72 h. Each cultivation was performed in biological triplicates. Cultures were analyzed for their dry biomass production 72 h after cultivation (A) and pellet diameter (B). The amount of secreted proteins/g dry biomass (C) was analyzed using a Bradford assay. The amount of secreted GlaA (D) was monitored via Western analysis and normalized firstly against the dry biomass for each corresponding strain/condition. A second normalization was conducted where GlaA was reported relative to the control strain MF31.2 at each respective condition. Students t-test was used for significance determination and p-values are reported.