Fed‑batch cultures subject to different oxygenation
conditions and initiated with different inoculum densities
Studies examining lipid production by Y. lipolytica using
fed-batch or repeated-batch cultures are scarce. Moreover,
only glycerol has been used as a substrate for cell
growth and lipid synthesis [25]. This study utilized a
two-step process: biomass was produced using molasses
for 48 h, and then lipids were produced using glycerol as
the main carbon source. Biomass yield and lipid production
were analyzed at two different inoculum densities
(low density and high density) and under two sets of oxygenation
conditions (unregulated and regulated). In the
unregulated strategy “Oxy-const”, dissolved oxygen (DO)
was not regulated; in the regulated strategy “Oxy-regul”,
DO was regulated at 50% saturation (see “Methods”).
When Oxy-const strategy and low-density inoculum
were used, the biomass reached 50 g CDW/L and citric
acid production was 36.8 g/L after 55 h of culture
(Figure 1a, b). During the glycerol-feeding phase, cells
converted the citric acid produced into lipids. In those
conditions, total lipid concentration increased from 11 to
15.5 g/L (Figure 1c). Yeast lipid content reached 31% of
CDW, which corresponds to a volumetric lipid productivity
(QL) of 0.18 g/L/h and a coefficient of lipid yield to
glycerol consumption (YL/gly) of 0.083 g/g (Table 1). This
condition also produced a small amount of mycelial cells
(Figure 2a). Indeed, low DO levels have been shown to
induce the yeast-to-mycelium transition in Y. lipolytica.
Bellou and colleagues demonstrated that mycelial and/or
pseudomycelial forms predominated over the yeast form
when DO was low, regardless of the carbon and nitrogen
sources used [26].
When oxygenation was regulated and a low-density
inoculum was used, cell growth was surprisingly very
slow (rx = 0.24 gCDW/h) and the culture duration (the
time required for complete glycerol consumption) was
190 h. Consequently, when crude glycerol was fed into
the bioreactor at 48 h, the fructose concentration was
still high (30 g/L). In this condition, the biomass yield was