Produção de hidrolisado enzimático de farinha de batata-doce para fermentação etanólica

Recently there is a great quest of producing alcohol from starchy resources, replacing the sugar cane. The most common starchy sources are cassava, maize and sweet potatoes and a lot of research are been realized with excellent results. In this work it was evaluated the influence of the concentration of dry matter on the enzymatic hydrolysis process of starch from sweet potato for ethanol production. Through the sweet potato was produced a flour using a low-cost method and easy operation equipments. The sweet potato flour was characterized physical and chemically and from these results was prepared the treatments for enzymatic hydrolysis. The experimental design considered as independent variable the dry matter concentration of the sweet potato flour in 3 levels; 10, 15 and 20% in the formulation of suspensions. The other variables were keeping constant being: temperature in the 1° hydrolysis step of 90°C and time of 2 hours; temperature in the 2° saccharification step of 60°C and time of 17 hours. The hydrolysates obtained at the three assays were transferred to six liter enlerynmeyer and inoculated with a biologic catalyst, Saccharomyces, dehydrated yeasts of Saccharomyces cerevisiae CAT 1, at a rate of 5% in weight. The flasks were placed in a shaker type orbital with controlled temperature of 30°C during a time of 15 hours. The initial reducer sugars concentration and respective ethanol concentrations in wine were: 11.2% glucose and 2.16% ethanol in the suspension with 10% of dry matter; 13.5% glucose and 4.39% ethanol with 15% and 17.5% glucose and 6.03% ethanol in suspension with 20% of dry matter. ix The results showed that the higher percentage of dry matter carried out to higher sugar yield in hydrolyzed. It was possible observed that products quality improved with a higher concentration of dry matter

Aplicações de coquetéis de celulases para a sacarificação enzimática do bagaço de cana-de-açúcar

Pós-graduação em Química - IBILCE

Produção de enzimas celulolíticas pelos fungos Trichoderma reesei e Myceliophthora thermophila e aplicação na sacarificação do bagaço de cana-de-açucar

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação térmica e estrutural do bagaço de cana de açúcar pré-tratado com ozônio, ultrassom e micro-ondas para produção de etanol celulósico por hidrólise enzimática

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Produção de ácidos orgânicos por cepas leveduras assimiladoras de xilose

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Evaluation of microwave-assisted pretreatment of lignocellulosic biomass immersed in alkaline glycerol for fermentable sugars production

A pretreatment with microwave irradiation was applied to enhance enzyme hydrolysis of corn straw and rice husk immersed in water, aqueous glycerol or alkaline glycerol. Native and pretreated solids underwent enzyme hydrolysis using the extract obtained from the fermentation of Myceliophthora heterothallica, comparing its efficiency with that of the commercial cellulose cocktail Celluclast (R). The highest saccharification yields, for both corn straw and rice husk, were attained when biomass was pretreated in alkaline glycerol, method that has not been previously reported in literature. Moreover, FTIR, TG and SEM analysis revealed a more significant modification in the structure of corn straw subjected to this pretreatment.Highest global yields were attained with the crude enzyme extract, which might be the result of its content in a great variety of hydrolytic enzymes, as revealed zymogram analysis. Moreover, its hydrolysis efficiency can be improved by its supplementation with commercial beta-glucosidase.

Studies on the application of Myceliophthora thermophila JCP1-4 cellulases cocktail on sugarcane bagasse pretreated by different methods

The hydrolysis step for sugar production in biorefineries is crucial for the sequential processes involved and cellulases cocktails behave differently according to the pretreatment employed. In this study, the application of the cellulases cocktail produced by the fungus Myceliophthora thermophila JCP1-4 was studied on the saccharification of sugarcane bagasse pretreated by ozonolysis and thermic ferric nitrate (TFN), and the results were compared with commercial enzymes (Novozymes Celluclast 1.5L, Novozym 188). The fungal cellulases cocktail hold an activity of FPU:β-glucosidase of 1:4(U/mL); time, temperature, FPU by g of cellulose load and percentage of dry matter (DM) were studied. The analysis of central composite design of TFN pretreated showed that fungal cellulases works better in DM values of 3–3.5% (4.5% for commercial), temperatures higher than 50 °C (<45 °C for commercial) and 15FPU for both; commercial enzymes yielded 7.78 g/L of reducing sugars and the fungal enzymes 5.42 g/L. With the ozone pretreated, the fungal enzymes presented a higher thermostability with faster kinects, being able to produce 5.56 g/L of reducing sugars (60 °C, 8 h), against 5.20 g/L for commercial enzymes (50 °C, 24 h), (10FPU, 3%DM for both). The FPU derivate analysis revels better yields with 7.5FPU, and the increase of DM to 7.5% resulted 13.28 g/L of reducing sugars.

Desenvolvimento de um bioprocesso utilizando-se resíduos para produção de amilases por Rhizopus oligosporus e etanol por Saccharomyces cerevisiae

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeito de parâmetros variáveis de fermentação sobre o crescimento celular e composição química do fermentado de batata-doce

This study aimed to evaluate the use of sweet potato as a substrate for the production of spirits. In order to promote an alternative technologically feasible, an experimental design was performed to minimize the operations of preparation, defining the most adequate conditions for the fermentation process. From sweet potato flour obtained by milling and dehydration process of the roots was carried out an enzymatic process of hydrolysis-saccharification of suspension of sweet potato flour with 18% dry matter. The hydrolyzate was used in the fermentation process which followed the 23 full factorial experimental design with central and axial points, and the independent variables were the concentration of reducing sugars, concentration of viable yeast and fermentation temperature. The dependent variables were viable cells, residual sugar, ethanol, glycerol and methanol. The dependent variables were quantified by liquid chromatography. The data analysis indicate that the best fermentation conditions among the tested conditions were: concentration of yeast 5 x 107-1 x 108 in number of viable cells, total reducing sugars from 12.5 to 13.5% and temperature between 33 -34ºC.

Purificação parcial e caracterização bioquímica de uma isoforma de β-glicosidase do fungo termofílico Myceliophthora thermophila M.7.7

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Production of alcoholic beverage from ginger: study of fermentation process and final product quality

Introduction: In Brazil part of the production of ginger is of inadequate quality for export. The production of spirit from felt-over rhizomes is an alternative of great interest to producers of these rhizomes. Aim: Aiming to increase the value of felt-over rhizomes, this work aimed to study the use of ginger as a raw material for alcoholic beverage production. It was evaluated the effect of fermentation conditions on the components of fermented alcoholic, as well as, the quality of alcoholic distilled beverage of ginger. Methods: Dehydrated ginger passed by enzymatic hydrolysis-saccharification processes. The hydrolysate obtained was analyzed for sugar profile in HPLC. The alcoholic fermentation process followed the central composite rotational design for three factors: fermentation temperature (23 to 37ºC), time of fermentation (17 to 33 h) and concentration of inoculum (0.22 to 3.00%). The fermented alcoholic obtained was analyzed in HPLC for the contents of ethanol, methanol, glycerol and residual sugars. The distillated alcoholic beverage of ginger was analyzed for ethanol, methanol, acetaldehyde, ethyl acetate and higher alcohols in the gas chromatography (GC). In addition, copper content and acidity were analyzed Results: Sugar profile of the ginger hydrolysate revealed the presence of 77.8% of glucose. Data analysis of fermentation process showed influence of temperature on ethanol and methanol content of the fermented alcoholic of ginger. Time of fermentation had effect on glycerol content. All parameters of process had influence on residual sugars contents. The HPLC analysis has shown presence of methanol, ethyl acetate, aldehyde, acids, higher alcohols and esters in distilled alcoholic beverage of ginger. Conclusion: Fermented alcoholic of ginger with higher levels of ethanol can be obtained under the conditions of 1.5% w/w of inoculum, 30°C of temperature and 24 hours of fermentation time. In this condition of fermentation process the beverage of ginger had good quality.

Hydrothermal pretreatment of sugarcane bagasse using response surface methodology improves digestibility and ethanol production by SSF

Sugarcane bagasse was characterized as a feedstock for the production of ethanol using hydrothermal pretreatment. Reaction temperature and time were varied between 160 and 200A degrees C and 5-20 min, respectively, using a response surface experimental design. The liquid fraction was analyzed for soluble carbohydrates and furan aldehydes. The solid fraction was analyzed for structural carbohydrates and Klason lignin. Pretreatment conditions were evaluated based on enzymatic extraction of glucose and xylose and conversion to ethanol using a simultaneous saccharification and fermentation scheme. SSF experiments were conducted with the washed pretreated biomass. The severity of the pretreatment should be sufficient to drive enzymatic digestion and ethanol yields, however, sugars losses and especially sugar conversion into furans needs to be minimized. As expected, furfural production increased with pretreatment severity and specifically xylose release. However, provided that the severity was kept below a general severity factor of 4.0, production of furfural was below an inhibitory concentration and carbohydrate contents were preserved in the pretreated whole hydrolysate. There were significant interactions between time and temperature for all the responses except cellulose digestion. The models were highly predictive for cellulose digestibility (R (2) = 0.8861) and for ethanol production (R (2) = 0.9581), but less so for xylose extraction. Both cellulose digestion and ethanol production increased with severity, however, high levels of furfural generated under more severe pretreatment conditions favor lower severity pretreatments. The optimal pretreatment condition that gave the highest conversion yield of ethanol, while minimizing furfural production, was judged to be 190A degrees C and 17.2 min. The whole hydrolysate was also converted to ethanol using SSF. To reduce the concentration of inhibitors, the liquid fraction was conditioned prior to fermentation by removing inhibitory chemicals using the fungus Coniochaeta ligniaria.

Adding value to the Brazilian sisal: acid hydrolysis of its pulp seeking production of sugars and materials

The present work is inserted into the broad context of the upgrading of lignocellulosic fibers. Sisal was chosen in the present study because more than 50% of the world's sisal is cultivated in Brazil, it has a short life cycle and its fiber has a high cellulose content. Specifically, in the present study, the subject addressed was the hydrolysis of the sisal pulp, using sulfuric acid as the catalyst. To assess the influence of parameters such as the concentration of the sulfuric acid and the temperature during this process, the pulp was hydrolyzed with various concentrations of sulfuric acid (30-50%) at 70 A degrees C and with 30% acid (v/v) at various temperatures (60-100 A degrees C). During hydrolysis, aliquots were withdrawn from the reaction media, and the solid (non-hydrolyzed pulp) was separated from the liquid (liquor) by filtering each aliquot. The sugar composition of the liquor was analyzed by HPLC, and the non-hydrolyzed pulps were characterized by viscometry (average molar mass), and X-ray diffraction (crystallinity). The results support the following conclusions: acid hydrolysis using 30% H2SO4 at 100 A degrees C can produce sisal microcrystalline cellulose and the conditions that led to the largest glucose yield and lowest decomposition rate were 50% H2SO4 at 70 A degrees C. In summary, the study of sisal pulp hydrolysis using concentrated acid showed that certain conditions are suitable for high recovery of xylose and good yield of glucose. Moreover, the unreacted cellulose can be targeted for different applications in bio-based materials. A kinetic study based on the glucose yield was performed for all reaction conditions using the kinetic model proposed by Saeman. The results showed that the model adjusted to all 30-35% H2SO4 reactions but not to greater concentrations of sulfuric acid. The present study is part of an ongoing research program, and the results reported here will be used as a comparison against the results obtained when using treated sisal pulp as the starting material.

Alkaline sulfite/anthraquinone pretreatment followed by disk refining of Pinus radiata and Pinus caribaea wood chips for biochemical ethanol production

BACKGROUND: Alkaline sulfite/anthraquinone (ASA) cooking of Pinus radiata and Pinus caribaea wood chips followed by disk refining was used as a pretreatment for the production of low lignified and high fibrillated pulps. The pulps produced with different delignification degrees and refined at different energy inputs (250, 750 and 1600 Wh) were saccharified with cellulases and fermented to ethanol with Saccharomyces cerevisiae using separated hydrolysis and fermentation (SHF) or semi-simultaneous saccharification and fermentation (SSSF) processes. RESULTS: Delignification of ASA pulps was between 25% and 50%, with low glucans losses. Pulp yield was from 70 to 78% for pulps of P. radiata and 60% for the pulp of P. caribaea. Pulps obtained after refining were evaluated in assays of enzymatic hydrolysis. Glucans-to-glucose conversion varied from 20 to 70%, depending on the degree of delignification and fibrillation of the pulps. The best ASA pulp of P. radiata was used in SHF and SSSF experiments of ethanol production. Such experiments produced maximum ethanol concentration of 20 g L-1, which represented roughly90% of glucose conversion and an estimated amount of 260 L ethanol ton(-1) wood. P. caribaea pulp also presented good performance in the enzymatic hydrolysis and fermentation but, due to the low amount of cellulose present, only 140 L ethanol would be obtained from each ton of wood. CONCLUSION: ASA cooking followed by disk refining was shown to be an efficient pretreatment process, which generated a low lignified and high-fibrillated substrate that allowed the production of ethanol from the softwoods with high conversion yields. (C) 2012 Society of Chemical Industry

Unraveling the structure of sugarcane bagasse after soaking in concentrated aqueous ammonia (SCAA) and ethanol production by Scheffersomyces (Pichia) stipitis

Abstract Background Fuel ethanol production from sustainable and largely abundant agro-residues such as sugarcane bagasse (SB) provides long term, geopolitical and strategic benefits. Pretreatment of SB is an inevitable process for improved saccharification of cell wall carbohydrates. Recently, ammonium hydroxide-based pretreatment technologies have gained significance as an effective and economical pretreatment strategy. We hypothesized that soaking in concentrated aqueous ammonia-mediated thermochemical pretreatment (SCAA) would overcome the native recalcitrance of SB by enhancing cellulase accessibility of the embedded holocellulosic microfibrils. Results In this study, we designed an experiment considering response surface methodology (Taguchi method, L8 orthogonal array) to optimize sugar recovery from ammonia pretreated sugarcane bagasse (SB) by using the method of soaking in concentrated aqueous ammonia (SCAA-SB). Three independent variables: ammonia concentration, temperature and time, were selected at two levels with center point. The ammonia pretreated bagasse (SCAA-SB) was enzymatically hydrolysed by commercial enzymes (Celluclast 1.5 L and Novozym 188) using 15 FPU/g dry biomass and 17.5 Units of β-glucosidase/g dry biomass at 50°C, 150 rpm for 96 h. A maximum of 28.43 g/l reducing sugars corresponding to 0.57 g sugars/g pretreated bagasse was obtained from the SCAA-SB derived using a 20% v/v ammonia solution, at 70°C for 24 h after enzymatic hydrolysis. Among the tested parameters, pretreatment time showed the maximum influence (p value, 0.053282) while ammonia concentration showed the least influence (p value, 0.612552) on sugar recovery. The changes in the ultra-structure and crystallinity of native SCAA-SB and enzymatically hydrolysed SB were observed by scanning electron microscopy (SEM), x-ray diffraction (XRD) and solid-state 13C nuclear magnetic resonance (NMR) spectroscopy. The enzymatic hydrolysates and solid SCAA-SB were subjected to ethanol fermentation under separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) by Scheffersomyces (Pichia) stipitis NRRL Y-7124 respectively. Higher ethanol production (10.31 g/l and yield, 0.387 g/g) was obtained through SSF than SHF (3.83 g/l and yield, 0.289 g/g). Conclusions SCAA treatment showed marked lignin removal from SB thus improving the accessibility of cellulases towards holocellulose substrate as evidenced by efficient sugar release. The ultrastructure of SB after SCAA and enzymatic hydrolysis of holocellulose provided insights of the degradation process at the molecular level.