2.5. Batch cultivation

A colony was picked from YPD agar plate and grown in a culture tube containing 4–5 ml of YE media for 16–18 hrs. The culture was then used as inoculum for batch growth kinetic studies carried out in a 250 ml baffled shake flask containing 50 ml of the same media. An initial optical density (OD₆₀₀) was approximately 0.1–0.2. The cultures were grown in an incubator shaker (Model Innova44, New Brunswick Scientific, USA) at 30 °C with a shaking rate of 100 rpm. Cell samples and supernatants were collected periodically to monitor cell growth, consumed sugar and byproducts.