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- Chemo-enzymatic saccharification strategy of microalgae chlorella sorokinianaPublication . Constantino, Ana; Rodrigues, Brígida; Raposo, SaraBiofuel production using microalgae attracted much attention because it can be cultured using CO2 and sunlight. With high carbohydrate content, microalgae have the potential to be used as a fermentation feedstock for bioethanol production. In present work, chemo-enzymatic saccharification of Chlorella sorokiniana microalgae were investigated. Chemical hydrolysis of the biomass followed by enzymatic hydrolysis and was also evaluated the effect of combining the two enzymes and the sequential addition. The effect of α-amylase concentrations was analyzed in ranged between 50 and 8000 U/g of biomass and for amyloglucosidase between 90 and 600 U/g of biomass. The higher concentrations showed the highest conversion of reducing sugars. The α-amylase concentration 8000 U/g of biomass presented a conversion of 43.06 ± 2.92% (w/w), while amyloglucosidase with 600 U/g of biomass obtained 76.57 ± 6.42% (w/w). The combination of two enzymes simultaneously was more efficient than the sequential addition for low enzyme concentrations (α-amylase 50 U/g and amyloglucosidase 90 U/g) with a total reducing sugar of 22.78 ± 3.06 and 16.92 ± 2.06% (w/w), respectively. On the other hand, using the higher enzymes concentrations, no difference was observed between the two addition strategies, 58.9 ± 3.55 and 57.05 ± 2.33% (w/w) for the sequential and simultaneous, respectively. Both strategies didn’t present advantage, since the amyloglucosidase enzyme alone produced slightly higher results. Even thought, the obtained results showed successfully performed saccharification of microalgal biomass and clearly point to microalgae use for saccharification and subsequent bioethanol production.
- 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.
- Chemical and physical pretreatments of microalgal biomassPublication . Rodrigues, Brígida; Borges, Rodrigo; Castro, Maria; Constantino, Ana; Raposo, SaraNon-axenic microalga Chlorella sorokiniana was cultivated in batch cultures, and its total sugar composition was determined. The microalga under study showed a total sugar concentration of 21.44 ± 0.46% (w/w). The effects of freeze-drying, oven-drying, freezing and thawing, chemical and the combination of hydrothermal and chemical pretreatments were evaluated. In the combined pretreatment different concentrations of H2SO4 and reaction times were also optimized. It was possible to determine that the sugar extraction yields more significant were 59.5% for the lyophilization, 6.2% with 6 cycles of freeze and thawing and around 100% for 2 and 4% (v/v) of H2SO4 at 121 °C for 30 min. Some of the methods that were described in this study are interesting to facilitate cost-efficient conversion of microalgal biomass into biofuels.