## 2 Materials and Methods
### Evaluation of Relative Growth Rate and Doubling Time in Each Medium  
Cell responses of _S. cerevisiae_ in each medium condition were analyzed as shown below. Optical density at 600 nm (OD\({}_{600}\)) was measured to quantify the strain cells. To analyze the growth activities of _S. cerevisiae_, 200 uL of _S. cerevisiae_ broth was cultivated in each well of a 96-well plate under a strong vibration mode (5 s, strong mixing mode) in each 30 min at 23 \({}^{\circ}\)C as room temperature, and monitored at OD\({}_{600}\) with a 96-well microplate reader SH-1300 Lab (Hitachi High-Tech Science Co., Tokyo, Japan). The initial OD\({}_{600}\) was calibrated to approximately 0.1-0.3. Relative growth rate (\(\mu\)) and doubling time (\(td\)) were calculated by the formulas below (\(t_{1}\): sampling time 1 (\(h\)); \(t_{2}\): sampling time 2 (\(h\)) (\(t_{2}>t_{1}\)); \(S_{1}\): scale of concentration indicated by OD\({}_{600}\) at \(t_{1}\); \(S_{2}\): scale of concentration indicated by OD\({}_{600}\) at \(t_{2}\)). \[\mu=\frac{ln(S_{2}/S_{1})}{(t_{2}-t_{1})}\ \left(h^{-1}\right)\]  
\[td=\frac{ln2}{\mu max}\ (h)\]  
Evaluation of Concentrations of Glucose, Ethanol, and General Media Components (Carbohydrate, Lipid, and Protein)  
Glucose and ethanol concentration were quantified using a high-performance liquid chromatography (HPLC) system after cultivation and pre-preparation. For cultivation, 10 mL of _S. cerevisiae_ broth was cultivated with 130 strokes per minute (spm) at 30 \({}^{\circ}\)C, and also monitored at OD\({}_{600}\) with a U-2900 (Hitachi High-Technologies Co., Tokyo, Japan). The initial OD\({}_{600}\) was calibrated to approximately 0.1. As pre-preparation, 1 mL of each broth was collected at the middle of the exponential phase, and the supernatant was obtained by centrifugation at 5000\(\times\)_g_ for 1 min. The supernatant was filtered with a Millex(r) syringe filter (pore size: 0.45 um) (Merck KGaA), and the flow-through was analyzed with the HPLC system. The HPLC system consisted of a LC-20 AD pump (Shimadzu Co., Kyoto, Japan), a CTO-10A column oven (Shimadzu Co.), an SPD-20 AV detector (Shimadzu Co.), a RezexTM RHM-Monosaccharide H\({}^{+}\) column (300 \(\times\) 7.8 mm) (Phenomenec Inc., Tokyo, Japan), and a 7725 injector (Rheodyne Inc., Bensheim, Germany). The concentrations of glucose and ethanol were identified and quantified at 190 nm, with chromatographic data monitored by the SPD-20 AV detector. The mobile phase was water, the column oven was set at 80 \({}^{\circ}\)C, and the flow rate was 0.6 mL\(\cdot\)min\({}^{-1}\). Carbohydrate concentration, lipid concentration, and protein concentration were evaluated by the methods shown below. Evaluation of total carbohydrate quantity: the carbohydrate in the medium was quantified by the anthrone method [26; 27]. An object was added to 0.004% (\(w/v\)) anthrone in 75% H\({}_{2}\)SO\({}_{4}\) on ice and stirred well. The mixture was then placed in a block incubator at 100 \({}^{\circ}\)C for 15 min. After 10 min on ice, the mixture was stirred well via vortexing. The correlation between absorbance at 620 nm and carbohydrate content was indicated by a calibration curve that was prepared with glucose, and the supernatant in the reaction was quantitatively evaluated with the calibration curve after a 20-fold dilution. Evaluation of total lipid quantity: lipids in the medium supernatant were methyl esterified with a fatty acid methylation kit (Nacalai Tesque Inc.) [28; 29]. The fatty acid methyl esters were identified and quantified with a capillary gas chromatograph GC-2025 (Shimadzu Co., Kyoto, Japan), which was equipped with a DB-23 capillary column (60 m, 0.25 mm internal diameter, 0.15 \(\upmu\)m film thickness) (Agilent Technologies Ltd., Santa Clara, CA, USA) and the previous method was then followed [29]. Heptadecanoic acid (Sigma-Aldrich Co., St. Louis, MO, USA) was used as an internal standard, and rapeseed oil (Merck KgaA) was used as a quantitative standard.