The yeast adaptation was initiated by adding 60 mL of inoculum
(Section 2.1) in 540 mL of culture medium A, for an initial biomass
concentration of 0.1 g/L. The batch was maintained until more than
90% of the initial glucose was consumed. Then, the agitation was
stopped for 15 min to allow the biomass flocculation and the
fermented broth separation. The fermented broth (540 mL) was
removed from the vessel by pumping it out through the outlet
pipe, taking care to not disturb the biomass at the bottom of the
reactor. A volume of 60 mL of the fermented broth was maintained
inside the reactor to be used as inoculum for the subsequent batch.
Then, 540 mL of sterile medium A was loaded again to the reactor
and the agitation was resumed when the loading ended, marking
the start of a new batch. Duplicate samples were taken every 8–
12 h for biomass and glucose measurement, accounting to 2–3
samples per batch. Each batch lasted 24 h approximately, and
media replacement was made every day within a time interval of
2–3 h, reducing the possibility of starvation periods.