Supplemental Figure 1 Fermentation results for different initial cell concentrations for 11 h RaBIT fermentation cycles performed in shake flasks. Results for a) xylose and b) ethanol are shown. Error bars represent standard deviations and are present for all data points but may be hidden by the symbol.

Supplemental Figure 2 Fraction of viable cells after RaBIT Cycles 1, 3, & 5 performed in shake flasks and separated using a separatory funnel. Error bars represent standard deviations.

Supplemental Table 1 Traditional shake flask fermentation performance after 24 h using RaBIT cycle separatory funnel settled cell fractions

Fraction* / Glucose (g/L) / Xylose (g/L) / Ethanol (g/L) / OD600
Cycle 1
1 / 1.78 / ± / 0.18 / 23.07 / ± / 1.28 / 37.37 / ± / 0.30 / 3.68 / ± / 0.10
2 / 1.88 / ± / 0.15 / 24.14 / ± / 0.92 / 36.88 / ± / 0.19 / 3.63 / ± / 0.19
3 / 1.97 / ± / 0.16 / 24.07 / ± / 1.15 / 36.49 / ± / 1.23 / 3.51 / ± / 0.10
4 / 2.04 / ± / 0.15 / 24.00 / ± / 0.95 / 36.79 / ± / 0.22 / 3.61 / ± / 0.12
5 / 1.99 / ± / 0.07 / 24.08 / ± / 1.16 / 37.04 / ± / 0.25 / 3.71 / ± / 0.07
6 / 1.98 / ± / 0.10 / 25.12 / ± / 0.83 / 36.39 / ± / 0.52 / 3.89 / ± / 0.15
Cycle 3
1 / 1.81 / ± / 0.35 / 26.70 / ± / 0.78 / 31.50 / ± / 1.75 / 2.80 / ± / 0.11
2 / 1.76 / ± / 0.36 / 26.44 / ± / 1.17 / 32.12 / ± / 1.08 / 2.72 / ± / 0.03
3 / 1.55 / ± / 0.12 / 27.27 / ± / 1.36 / 33.06 / ± / 0.73 / 2.86 / ± / 0.21
4 / 1.58 / ± / 0.12 / 26.82 / ± / 0.44 / 32.55 / ± / 0.59 / 2.89 / ± / 0.09
5 / 1.59 / ± / 0.06 / 26.57 / ± / 0.38 / 33.04 / ± / 0.65 / 2.75 / ± / 0.00
6 / 1.56 / ± / 0.07 / 27.01 / ± / 1.01 / 33.25 / ± / 0.49 / 2.79 / ± / 0.22
Cycle 5
1 / 33.91 / ± / 7.01 / 32.50 / ± / 0.82 / 12.11 / ± / 3.73 / 1.00 / ± / 0.03
2 / 35.18 / ± / 5.01 / 32.38 / ± / 0.77 / 11.29 / ± / 2.70 / 0.87 / ± / 0.03
3 / 34.44 / ± / 5.81 / 32.45 / ± / 0.66 / 11.16 / ± / 2.83 / 0.92 / ± / 0.04
4 / 32.22 / ± / 7.65 / 32.40 / ± / 0.93 / 12.79 / ± / 4.03 / 0.97 / ± / 0.05
5 / 32.87 / ± / 5.85 / 32.48 / ± / 0.84 / 12.43 / ± / 2.86 / 0.92 / ± / 0.03
6 / 38.80 / ± / 5.06 / 32.53 / ± / 0.73 / 9.20 / ± / 2.55 / 0.89 / ± / 0.08

* Fractions were ordered from bottom (1) to top (6)
Error values represent standard deviations

Supplemental Figure 3 RaBIT cycle performance comparison using end of cycle cells for 23 h traditional fermentations (1 g/L DCW inoculum) using shake flasks. Error bars represent standard deviations and are present for all data points but may be hidden by the symbol.