Figure 5
Expression of recombinant Tb TIM in 96-well plates with EnBase at 25°C and induction at different cell densities. Expression levels of Tb TIM are shown as a ratio of insoluble protein (light green) and the target, soluble protein (dark green). (a) Growth curves after induction; (b) Total volumetric amounts of Tb TIM and (c) product amounts expressed per cell unit. The protein amounts after induction are shown for four different OD600 levels in the novel cultivation system and for the reference culture (M9ZB, induction at OD600 = 0.3). Cultures were performed with 3 AGU l-1 of glucoamylase. Product synthesis was induced by addition of 0.5 mM IPTG at the indicated times and cell densities, and the cultivation was continued for up to 18 hours after induction. Additional ammonia (at 26.2 h) and magnesium (at 36.85 h) were added during the cultivations as described in the Materials section.