Fig4

Fig 4. Fermentative capacity and the survival of OsMDHAR-expressing yeast cells during batch fermentation. (A) Fermentative capacity was analyzed by measuring the alcohol (AC) and residual glucose (RG) concentrations in YG medium after fermentation for 72 h at 30°C. Upward triangles, AC of WT cells; circles, AC of TC cells; squares, RG of WT cells; diamonds, RG of TC cells. (B) Time-dependent OsMDHAR expression during batch fermentation was evaluated by western blotting. Tubulin (Tub) was used as a loading control. (C) Growth kinetics during fermentation was assessed by measuring the OD600 at 2-h intervals for the indicated time. Squares, WT cells; circles, TC cells. (D) Cell viability during fermentation at 30°C was assessed by a spotting assay. Cells were harvested after 24 h (upper panel), 48 h (middle panel), and 72 h (lower panel) of fermentation and serially diluted to 10−9. A 5-µL aliquot of each diluted solution was spotted onto YPD agar plates. After incubation for 3 days, the plates were photographed. (E) Stress response to ethanol. Mid-log phase yeast cells (OD600 ≈ 2.0) were exposed to different concentrations of ethanol (0, 15%, and 20%) for 1 h, serially diluted, and spotted onto YPD agar plates. WT, yeast cells with an empty vector; TC, OsMDHAR-expressing yeast cells.