Lp0037b

The Saccharomyces cerevisiae strains used in this study are shown in Table S5. Yeast cells were grown at 25°C in YP medium (1 % Yeast Extract, 21275, Becton Dickinson; 2 % bacteriological peptone, LP0037B, Oxoid) supplemented with 2 % Raffinose. In each case, a 12-litre exponential culture was grown to 2-3 x 10⁷ cells/ml and then induced for 6 h at 20°C by addition of Galactose to a final concentration of 2 %. Cells were collected by centrifugation and washed once with lysis buffer (indicated below for each purification) lacking protease inhibitors. Cell pellets (~ 30 g) were then resuspended in 0.3 volumes of the indicated lysis buffer containing protease inhibitors. The resulting suspensions were then frozen dropwise in liquid nitrogen and stored at – 80°C. Subsequently, the entire sample of frozen yeast cells were ground in the presence of liquid nitrogen, using a SPEX CertiPrep 6850 Freezer/Mill with 3 cycles of 2’ at a rate of 15. The resulting powders were then stored at – 80 °C.

The Saccharomyces cerevisiae strains used in this study are shown in Appendix Tables S1–S2. Yeast cells were grown at 25°C in YP medium (1% Yeast Extract, 21275, Becton Dickinson; 2% bacteriological peptone, LP0037B, Oxoid) supplemented with 2% Raffinose. In each case, a 12‐litre exponential culture was grown to 2‐3 x 10⁷ cells / ml and then induced for 6 h at 20°C by addition of galactose to a final concentration of 2%. Cells were collected by centrifugation and washed once with lysis buffer (indicated below for each purification) lacking protease inhibitors. Cell pellets (˜ 30 g) were then resuspended in 0.3 volumes of the indicated lysis buffer containing protease inhibitors. The resulting suspensions were then frozen dropwise in liquid nitrogen and stored at – 80°C. Subsequently, the entire sample of frozen yeast cells were ground in the presence of liquid nitrogen, using a SPEX CertiPrep 6850 Freezer/Mill with 6 cycles of 2’ at a rate of 15. The resulting powders were then stored at −80°C. Details for each protein are given in Appendix Materials and Methods.

The fresh sugarcane molasses, whose initial liquid concentration is 80 °Bx, was diluted to 40 °Bx, 30 °Bx, 20 °Bx, 10 °Bx, 7.5 °Bx, and 5 °Bx, respectively, using deionized water. To ensure sufficient nitrogen sources in the fermentation solution, 5 g/L of yeast extract (Oxoid, LP0021), 5 g/L of peptone (Oxoid, LP0037B), and 10 g/L of beef extract (Oxoid, LP0029B) were added to various molasses concentrations according to the MRS (De Man, Rogosa and Sharpe) basic medium formula. Then the pH value was adjusted to 6.0.