Influência da linhagem da levedura e das condições de cultivo no processo de isomerização e fermentação simultâneas da xilose

Abstract

The conversion of the hemicellulosic fraction in ethanol is a factor that impacts on the economic viability of the second generation ethanol production process from sugar cane bagasse. Hemicellulose from bagasse is a heteropolymer constituted by pentoses and glucose, being xylose the predominant sugar (~ 21 %). Among the available technological alternatives for ethanol production from xylose, SIF process (Simultaneous Isomerization and Fermentation), consisting of xylose conversion to xylulose by glucose isomerase (GI) enzyme and xylulose fermentation by the yeast S. cerevisiae, is considered a promising alternative. The main objectives of the present work were: i) evaluate the performance of different S. cerevisiae strains towards xylulose intake and ethanol productivity; ii) assess the influence of cultivation conditions (temperature, oxygen availability and initial xylose concentration) upon ethanol and xylitol production by the selected strains; iii) define the operation conditions for the continuous SIF process, using a system of fixed bed reactors associated in series. Preliminary experiments were conducted in 50 mL flasks, containing 4 g of pellets with a load of 20 % of immobilized GI, co-immobilized with yeast (load of 10 %) in alginate gel. For the screening of yeasts showing better performance on ethanol production from xylose, two commercial baker´s yeast strains (Itaiquara® e Fleischmann®), three industrial strains (BG-1, CAT-1 e PE-2) and one lab strain (CEN.PK113-7D) were evaluated. These experiments were performed at 35 oC, using a medium composed by xylose (60 g/L), urea (5 g/L), CaCl2 (1.9g/L) and several salts, at initial pH of 5.6. Additional SIF studies were carried out with the selected yeasts Itaiquara®, BG-1 or CEN.PK113-7D under different temperature conditions (40 oC), aeration (15 mL flasks) and initial xylose concentration (130 g/L) for comparison with the results obtained at the standard conditions. For SIF cultures, samples were withdrawal and the concentrations of reducing sugars were determined by DNS method while xylose, xylulose, ethanol and by-products (xylitol, glycerol etc) concentrations were assessed by liquid chromatography. Cell viability was also measured at the beginning and end of the experiment. When comparing the different yeasts, Itaiquara® strain presented the best performance, reaching ethanol concentrations of 22.4 g/L, with a productivity of 2.1 g/Lh. The conversion of xylose was similar for all studied industrial strains as well as among the baker s yeast and lab strains. Concerning the group of additional experiments, at 40 oC, a decrease of viability and ethanol selectivity was observed for Itaiquara®, whereas productivity and selectivity for CEN.PK113-7D. was improved. For the studies conducted under semianaerobic conditions, the yeast BG-1 showed an increase in selectivity and yield. However, the reaction time increased to app. 45 hours. On the other hand, the performance of strain Itaiquara® was not altered by the lower level of oxygen tested. In the experiment with 120 g/L of xylose, more than 40 g/L of ethanol was obtained in 24 hours of cultivation. Thus, we conclude that the SIF process proposed in the present work is a viable alternative for the production of ethanol from xylose or lignocellulosic residues. For the operation of the continuous system composed by fixed bed reactors associated in series, the recommended conditions include the Itaiquara® yeast with a temperature no higher than 35 oC, keeping the total residence time around 10 hours for a feeding supply containing 60 g/L of xylose.