Diluphotometer

E. coli strains were grown in 5 mL of Luria Bertani medium (5 g L⁻¹ yeast extract, 10 g L⁻¹ tryptone, and 5 g L⁻¹ NaCl) in 14 mL round bottom culture tubes, at 37 °C and 250 rpm. 100 μg mL⁻¹ of ampicillin was supplemented when necessary for selection.
Yeast peptone medium (10 g L⁻¹ yeast extract, 20 g L⁻¹ peptone) containing 20 g L⁻¹ of glucose (YPD) was prepared for routine S. boulardii culture, genome editing, and plasmid transformation. Nourseothricin (100 μg mL⁻¹), geneticin (300 μg mL⁻¹), and hygromycin B (200 μg mL⁻¹) were added to YPD medium when required for selection. Synthetic complete medium, which contains yeast nitrogen base without ammonium sulfate and complete supplement mixture (MP Biomedicals), was used for the inducible transactivation experiments together with 50 mM pH 5.5 potassium hydrogen phthalate buffer. 20 g L⁻¹ of glucose or galactose was added as a carbon source. 200 μM CuSO₄ was supplemented when required.
Microbial cell growth was monitored by absorbance at 600 nm (A600) using DiluPhotometer™ (IMPLEN). Consumption of carbon sources were measured by HP 1050 HPLC system equipped with Agilent 1100 Refractive Index Detector (G1362). 0.005 N H₂SO₄ was eluted through Rezex ROA-Organic Acid H+ (8%) column (Phenomenex) at 0.6 mL min⁻¹ and 50 °C.