Transcriptomic Analysis of Candida boidinii Wild-Type Strain CICC#33029 and Its Engineered L-Lactic Acid-Producing Chassis Strain LA001-14 Cultured with Glucose or Methanol as the Sole Carbon Source

Subsequently, 3 mL of the culture was transferred to a 250 mL Erlenmeyer flask containing 100 mL of liquid YPD medium and incubated at 30°C with shaking at 250 rpm for 12 hours. Cells were then harvested by centrifugation at 6,000 × g for 5 minutes and washed twice with phosphate-buffered saline (PBS).

The cell pellets were resuspended in 100 mL of liquid YPD medium (supplemented with 2% w/v glucose) or YPM medium (supplemented with 2% w/v methanol) in 250 mL Erlenmeyer flasks. Following 6 hours of incubation at 30°C and 250 rpm, yeast cells were collected by centrifugation at 6,000 × g for 5 minutes and flash-frozen in liquid nitrogen. Total RNA was extracted from the samples using TRIzol reagent (Invitrogen, Grand Island, USA) and subjected to high-throughput RNA sequencing.