Shake-flask cultures in 250 mL Erlenmeyer flasks contained
90 mL of the wastes based medium. The pH was adjusted to 6.0.
The wastes medium was sterilized by autoclaving before use. The
cultures were initiated with 10% of a 24-h-old seed culture
(approximately 107cells/mL) and incubated at room temperature
(30 ± 2 C) on a rotary shaker at 140 rpm for 72 h. A two-phase process
was attempted, in which cell growth was promoted in the first
phase and the lipid accumulation was then enhanced during the
second phase. The cells from the first phase were harvested by centrifugation
at 1585g for 15 min and resuspended in the fresh
medium.