Figure 3.

Total Isobutanol production of engineered P. pastoris in a mixed-carbon source medium. Engineered strains were precultured in 5-mL aliquots in MGYH minimal medium (2% glycerol) overnight and used to inoculate 5 mL yeast selective medium (2% total sugar; 1.70% glucose and 0.30% xylose) to achieve an initial optical density of 0.05 at 600 nm (OD₆₀₀). The cultures were grown at 30 °C and 250 rpm in an orbital shaking incubator. Samples were taken at 24, 48, and 72 h for isobutanol quantification and OD₆₀₀ measurement. Values are the mean of three biological replicates ± standard deviation (n = 3). Lp, Lachnoclostridium phytofermentans; Psp, Piromyces sp. E2; Pp, Pichia pastoris; Ll, Lactococcus lactis.