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- A new combined approach to improved lipid production using a strictly aerobic and oleaginous yeastPublication . Guerreiro, Fábio; Constantino, Ana; Emília Lima-Costa, Maria; Raposo, SaraMicrobial lipids have potential applications in energy, and food industry, because most of those lipids are triacylglycerol with long-chain fatty-acids that are comparable to conventional vegetable oils and can be obtained without arable land requirement. Rhodosporidium toruloides is a strictly aerobic strain, where oxygen plays a crucial role in growth, maintenance, and metabolite production, such as lipids and carotenoids. Dissolved oxygen concentration is one of the major factors affecting yeast physiological and biochemical characteristics. In this context, different approaches have been developed to increase available oxygen by the increasing the aeration and the addition of an oxygen-vector. The growth of R. toruloides in 2-L mechanical stirred tank reactor equipped with 1 or 2 porous spargers and a 70 C/N ratio, revealed a lipid content of 0.47 and 0.52 g/g and a lipidic productivity of 0.16 and 0.17 g/L day, respectively. The oxygen-vector addition, increased the lipidic productivity for 0.20 g/L day and a lipid contend of 0.51 g of lipids/g of biomass. The combined approach, combining high aeration (AA), and 1% of n-dodecane addition (DA), produced a significant improvement in the lipid accumulation (62%, w/w), when compared with the DA (51%, w/w) and the AA (52%, w/w) approaches. The increasing of lipids accumulation and smaller culture time are key factors for the success of scale-up and profitability of a bioprocess.
- Kinetic and energetic parameters of carob wastes fermentation by Saccharomyces cerevisiae: crabtree effect, ethanol toxicity, and invertase repressionPublication . Rodrigues, Brígida; Peinado, J. M.; Raposo, Sara; Constantino, Ana; Quintas, Célia; Emília Lima-Costa, MariaCarob waste is a useful raw material for the second-generation ethanol because 50% of its dry weight is sucrose, glucose, and fructose. To optimize the process, we have studied the influence of the initial concentration of sugars on the fermentation performance of Saccharomyces cerevisiae. With initial sugar concentrations (S0) of 20 g/l, the yeasts were derepressed and the ethanol produced during the exponential phase was consumed in a diauxic phase. The rate of ethanol consumption decreased with increasing S0 and disappeared at 250 g/l when the Crabtree effect was complete and almost all the sugar consumed was transformed into ethanol with a yield factor of 0.42 g/g. Sucrose hydrolysis was delayed at high S0 because of glucose repression of invertase synthesis, which was triggered at concentrations above 40 g/l. At S0 higher than 250 g/l, even when glucose had been exhausted, sucrose was hydrolyzed very slowly, probably due to an inhibition at this low water activity. Although with lower metabolic rates and longer times of fermentation, 250 g/l is considered the optimal initial concentration because it avoids the diauxic consumption of ethanol and maintains enough invertase activity to consume all the sucrose, and also avoids the inhibitions due to lower water activities at higher S0.