Sugar and ethanol dosing and total cell count
Ethanol and total sugars were quantified by high performance
liquid chromatography (HPLC) using a Shimadzu LC-20A Prominence
instrument with a SUPELCOGEL Ca column, deionized water
as the mobile phase, a flow rate of 0.5 mL/min, an 80 ◦C oven
temperature and a 20 mL injection volume. The concentrations of
ethanol and sugars were determined from previously obtained calibration
curves. The concentration of total sugars (TS) was calculated
from the sum of the concentrations of stachyose, raffinose, sucrose,
glucose and fructose. To monitor cell growth during fermentation,
we counted the cells in a Neubauer chamber.