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The pH and stirrer speed in the fermentation broth were controlled to 4.0 5.0 (by adding 1 N NaOH if required, i.e., it was added manually only if pH dropped to less than four) and 300 500 rpm, respectively (Richter et al. 2013; Mohammadi et al. 2012; Younesi et al. 2006).
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The pH of the culture broth was controlled at between 6.5 and 7.0 by adding anion-exchange resins to remove lactate.
For P. pastoris and L. paracasei cultures, the pH of broth was controlled at 5.5 and 6.0, respectively, with 25% (w/w) NH4OH solution.
The pH of the broth was controlled at 5.5.
The pH of fermentation broth was controlled by ammonia (5% W/W) at 4.5.
Broth viscosity was controlled in case of immobilized cells.
Bacterial viability in the broth containers was controlled after inoculation by microscopy and cultivation.
To this end, 3 L of basal salt medium broth (PFPG, Invitrogen) was used, pH was controlled at 4.5 5.0, and DO was maintained between 30 and 80% during the whole fermentation process (Additional file 1: Figure S6).
The broth was pumped from the reservoir to the central compartment through a peristaltic pump, which is controlled digitally.
Approximately 30 μg of target or control proteins and 300 μl of M17 broth were added to the cell mixture.
As a control, the bacterial growth curves in enriched broth were found to be nearly identical for wt Alab49 and the rofA::aad9 construct (data not shown).
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