Lipid production by Rhodosporidium toruloides from biodiesel-derived glycerol in shake flasks and bioreactor: Impact of initial C/N molar ratio and added onion-peel extract

Abstract

Rhodosporidium toruloides NRRL Y-27012 grown in glycerol, produced yeast biomass and secondary metabolites as lipids (L) and intra-cellular polysaccharides (IPS) in trials under nitrogen limitation (initial C/N molar ratios ≈ 50, ≈ 100, ≈ 160 and ≈ 240). The initial glycerol (Glol0) concentration for all experiments was ≈ 90 g/L. The yeast demonstrated significant dry cell weight (DCW) production irrespective of the C/N ratio employed, ranging between 19 and 29 g/L. The optimum C/N ratio medium was that of 100 moles/moles, where a DCWmax = 24.5 g/L containing c. 50% w/w of lipids was recorded. Endopolysaccharides presented indeed impressive values (≥ 50% w/w) even at the very early growth steps, while IPS/DCW values decreased as the fermentation proceeded, with simultaneous increase of L/DCW values. Experiments were also conducted in media with a hydroglycerolic extract of onion peels (at 40 mL/L), while a blank experiment (no extract) was also realized. The trial with the added extract was performed under both aseptic (previous sterilization) and non-aseptic (previous thermal treatment at T = 100 °C; 5 min) conditions. The aseptic experiment with the added extract resulted in a relative decrease in lipid production in relation to the control (L=8.6 g/L in the control vs L = 6.2 g/L with the extract). Under non-aseptic conditions, lipid production was slightly lower (L = 5.2 g/L). Finally, the optimum trial in shake flasks was scaled-up in a batch bioreactor, and higher glycerol assimilation rate, and biomass and lipid production occurred compared to the flask trial (final DCW and lipid values ≈27 g/L and 13.5 g/L respectively). Cellular lipids, rich in palmitic and oleic acid, were mainly composed of neutral fractions. Phospholipids were more saturated than the neutral lipids.