RESUMO
Bioethanol, is a potential alternate source of energy, renewable and safe. Ethanol production from value added food and feedstock has also not shown growth as estimated. Of late, the second generation processes of production of ethanol, such as from lignocellulosic biomass out of agricultural/domestic waste has been gaining considerable momentum. Here, we explored a new approach for optimizing the conditions of physiochemical pretreatment as well as fermentation process using peels of Ananas cosmosus as substrate and immobilized yeast Pachysolen tannophilus MTCC 1077 and Pichia stipitis NCIM 3498. We have also studied the influence of process variables such as incubation temperature, inoculum concentration and different nutrients on ethanol production. Pulverized peels of A. cosmosus recorded 25 ± 0.31% cellulose, 28 ± 0.18% hemicellulose and 8 ± 0.07% of lignin on dry solid (DS) basis. Peels of A. cosmosus delignified with 1% H2SO4 yielded 18.89% glucose, 38.81% xylose and 29.31% fructose under thermochemical pretreatment using autoclave (121°C, 20 min.), with a hydrolytic efficiency of 75.52 ± 0.45%. FTIR spectroscopy results not only indicated the penetration of H2SO4 in the amorphous region of the biomass and degradation of hemicelluloses but also showed the structural differences before and after pretreatment. The enzymes required for hydrolysis were prepared from culture supernatants of Trichoderma reesei NCIM 1052 using wheat bran as carbon source under submerged fermentation conditions on rotatory shaker incubator (at 28°C for 10 days ). Enzyme activity (U/ml) of crude cellulase produced by T. reesei NCIM 1052 was 311.1 µmole/ml/min. Delignified A. cosmosus peel yielded 51.71 ± 0.44 g/l glucose when enzymatically hydrolysed by crude cellulase at the substrate enzyme ratio of 1:5. Simultaneous saccharification and fermentation (SSF) of peels of A. cosmosus by crude cellulase and separately entrapped Pichia stipitis NCIM 3498 (now known as Scheffersomyces stipitis) and Pachysolen tannophilus MTCC 1077 cells in calcium alginate beads were also investigated in the present study. The fermentation experiments were carried out at flask level. The processing parameters setup for reaching a maximum response for ethanol production was obtained when applying the optimum values for temperature (32°C), inoculum level (6%) and fermentation medium (ammonium sulphate, KH2PO4, peptone and yeast extract) for P. tannophilus MTCC 1077 and temperature (30°C), inoculum level (2%) and fermentation medium (ammonium sulphate, KH2PO4, peptone and yeast extract) for S. stipitis NCIM 3498. Maximum ethanol concentration 10.5 g/l and 10.9 g/l was obtained from P. tannophilus MTCC 1077 and S. stipitis NCIM 3498, respectively at the optimized process conditions in anaerobic batch fermentation.
RESUMO
The use of hemicellulosic hydrolysates in bioprocesses requires supplementation as to ensure the best fermentative performance of microorganisms. However, in light of conflicting data in the literature, it is necessary to establish an inexpensive and applicable medium for the development of bioprocesses. This paper evaluates the fermentative performance of Scheffersomyces (Pichia) stipitis and Candida guilliermondii growth in sugarcane bagasse hemicellulosic hydrolysate supplemented with different nitrogen sources including rice bran extract, an important by-product of agroindustry and source of vitamins and amino acids. Experiments were carried out with hydrolysate supplemented with rice bran extract and (NH4)2SO4; peptone and yeast extract; (NH4)2SO4, peptone and yeast extract and non-supplemented hydrolysate as a control. S. stipitis produced only ethanol, while C. guilliermondii produced xylitol as the main product and ethanol as by-product. Maximum ethanol production by S. stipitis was observed when sugarcane bagasse hemicellulosic hydrolysate was supplemented with (NH4)2SO4, peptone and yeast extract. Differently, the maximum xylitol formation by C. guilliermondii was obtained by employing hydrolysate supplemented with (NH4)2SO4 and rice bran extract. Together, these findings indicate that: a) for both yeasts (NH4)2SO4 was required as an inorganic nitrogen source to supplement sugarcane bagasse hydrolysate; b) for S. stipitis, sugarcane hemicellulosic hydrolysate must be supplemented with peptone and yeast extract as organic nitrogen source; and: c) for C. guilliermondii, it must be supplemented with rice bran extract. The present study designed a fermentation medium employing hemicellulosic hydrolysate and provides a basis for studies about value-added products as ethanol and xylitol from lignocellulosic materials.