Enhancement of ethanol production efficiency in repeated-batch fermentation from sweet sorghum stem juice: effect of initial sugar, nitrogen and aeration

BACKGROUND: Ethanol concentration (PE), ethanol productivity (QP) and sugar consumption (SC) are important values in industrial ethanol production. In this study, initial sugar and nitrogen (urea) concentrations in sweet sorghum stem juice (SSJ) were optimized for high PE (≥10%, v/v), QP, (≥2.5 g/L·h) and SC (≥90%) by Saccharomyces cerevisiae SSJKKU01. Then, repeated-batch fermentations under normal gravity (NG) and high gravity (HG) conditions were studied. RESULTS: The initial sugar at 208 g/L and urea at 2.75 g/L were the optimum values to meet the criteria. At the initial yeast cell concentration of ~1 × 108 cells/mL, the PE, QP and SC were 97.06 g/L, 3.24 g/L·h and 95.43%, respectively. Repeated-batch fermentations showed that the ethanol production efficiency of eight successive cycles with and without aeration were not significantly different when the initial sugar of cycles 2 to 8 was under NG conditions (~140 g/L). Positive effects of aeration were observed when the initial sugar from cycle 2 was under HG conditions (180­200 g/L). The PE and QP under no aeration were consecutively lower from cycle 1 to cycle 6. Additionally, aeration affected ergosterol formation in yeast cell membrane at high ethanol concentrations, whereas trehalose content under all conditions was not different. CONCLUSION: Initial sugar, sufficient nitrogen and appropriated aeration are necessary for promoting yeast growth and ethanol fermentation. The SSJ was successfully used as an ethanol production medium for a high level of ethanol production. Aeration was not essential for repeated-batch fermentation under NG conditions, but it was beneficial under HG conditions.

Greek yogurt with added sorghum flours: antioxidant potential and sensory acceptance / Yogurt griego adicionado de harinas integrales de sorgo: potencial antioxidante y evaluación sensorial

ABSTRACT The aim of this study was to develop and evaluate the antioxidant characteristics and sensory acceptance of Greek yogurt with added whole sorghum flours (WSF) from genotypes BRS305 (tannin) and BR501 (tannin-free). Five formulations were elaborated: 1) Control (without WSF); 2) 2% BR501; 3) 4% BR501; 4) 2% BRS305 and 5) 4% BRS305 and evaluated for antioxidant capacity, total anthocyanins, total phenols, condensed tannins, sensory acceptance and purchase intention. The 4% BRS305 formulation presented higher antioxidant properties, but lower sensory acceptance. No significant difference was observed for: color, flavor and overall acceptability between the control and 2% BR501; nor for texture and overall acceptability between the control and 4% BR501. Principal component analysis explained that all variables associated with antioxidant properties were positively correlated with the first major component (PC1: 82.7%). The 2% and 4% BR501 formulations were more highly correlated with PC1 (92.7%), as well as the control in relation to sensorial characteristics. The addition of WSF BRS305 improved the antioxidant properties of yogurts and the addition of WSF BR501 did not interfere with the sensory acceptance of the formulations. The similarity of the sensory acceptance of the yogurt containing WSF from the BR 501 genotype to the control, opens perspectives for the insertion of sorghum in human food, using dairy products as suitable matrices, adding potential functionality to this type of product.

RESUMEN El trabajo tuvo como propósito desarrollar y evaluar las características antioxidantes y la aceptación sensorial de yogurt griego con adición de harinas integrales de sorgo (HIS) de los genotipos BRS305 (con taninos) y BR501 (sin taninos). Cinco formulaciones fueron elaboradas: 1) Control (sin HIS); 2) 2% BR501; 3) 4% BR501; 4) 2% BRS305 y 5) 4% BRS305. Las formulaciones fueron evaluadas en función de su actividad antioxidante, antocianinas totales, fenólicos totales, taninos condensados, aceptación sensorial e intención de compra. La formulación 4% BRS305 presentó más propiedades antioxidantes, pero menor aceptación sensorial que las otras formulaciones. No se observó diferencia significativa entre el control y el 2% BR501 para el color, sabor e impresión global, ni tampoco para textura e impresión global en relación al 4% BR501. El análisis de componente principal ha explicado que todas las variables asociadas a las propiedades antioxidantes han sido positivamente correlacionadas con el primer componente principal (PC1: 82,7%). Las formulaciones 2% y 4% BR501 se han correlacionado más con el PC1 (92,7%), así como el control con las características sensoriales. La adición de HIS BRS305 ha mejorado las propiedades antioxidantes de los yogurts y la adición de HIS BR501 no ha interferido en la aceptación sensorial de las formulaciones. La similitud de la aceptación sensorial del yogurt que contiene el HIS del genotipo BR501 con el control, abre perspectivas para la inserción del sorgo en la alimentación humana, utilizando productos lácteos como matrices adecuadas, agregando funcionalidad potencial a este tipo de producto.

Partial purification and biochemical characterization of an alkaline esterase from Sorghumbicolor

Esterases are enzymes that present good potential for industrial applications since they catalyze the formation or cleavage of ester bonds in water-soluble substrates, and sorghumseeds can represent an alternative source of this enzyme. The extraction of esterase from sorghumseeds is an economical alternative to obtain an enzyme of great interest. Esterases may improve the quality or accelerate the maturation of cheeses, cured bacon and fermented sausages and may also resolve racemic mixtures. Recently, seed esterases have been the focus of much attention as biocatalysts. In some cases, these enzymes present advantages over animal and microbial lipases due to some quite interesting features such as specificity and low cost, being a great alternative for their commercial exploitation as industrial enzymes The esterase studied here was extracted from sorghumseeds and some of its biochemical properties determined using synthetic substrates (p-nitrophenyl butyrate, caprylate, laurate and palmitate). The enzyme presented optimum activity at pH 8.0 and was stable in all the pH ranges studied. The optimum temperature for its activity was 40ºC but it showed low stability at this temperature (40% relative activity). The values derived for Km and Vmax were 0.67mM and 125 U.mg-1, respectively, obtained using p-nitrophenyl butyrate as the substrate. The enzyme showed an increase in activity when K2HPO4was added to the reaction medium, but the ions Mn2+, CO+, Hg+and Fe2+strongly inhibited the enzyme activity. This enzyme showed a preference for the hydrolysis of short chain fatty acids. The characteristics of sorghumesterase are very similar to those of the microbial esterases used in detergent processing.

Co-production of ethanol and biodiesel from sweet sorghum juice in two consecutive fermentation steps

Background: Sugars from sweet sorghum stalks can be used to produce ethanol and also to grow oleaginous yeasts. Instead of two separate processes, in this paper we propose a different route producing ethanol and microbial oil in two consecutive fermentation steps. Results: Three yeasts were compared in the first ethanol producing step. In the second step four different oleaginous yeasts were tested. Sweet sorghum juice was first clarified and concentrated. High gravity ethanol fermentation was carried out with concentrated juice with 23.7 g/100 mL of total sugars and without added nutrients. Total sugars were 2.5 times more than the original clarified juice. One yeast gave the best overall response over the two other tested; relative high ethanol productivity, 1.44 g ethanol/L•h−1 , and 90% of sugar consumption. Aeration by flask agitation produced superior results than static flasks for all yeasts. Microbial oil production was done employing the residual liquid left after ethanol separation. The pooled residual liquid from the ethanol distillation contained 7.08 g/mL of total carbohydrates, rich in reducing sugars. Trichosporon oleaginosus and Lipomyces starkeyi produced higher dry biomass, total sugar consumption and oil productivity than the other two oleaginous yeasts tested; with values around 25 g/L, 80%, and 0.55 g oil/L•h−1 respectively. However, the biomass oil content in all yeasts was relatively low in the range of 14 to 16%. Conclusion: The two step process is viable and could be considered an integral part of a consolidated biorefinery from sweet sorghum.

Multi-enzyme complex of white rot fungi in saccharification of lignocellulosic material

ABSTRACT The multi-enzyme complex (crude extract) of white rot fungi Pleurotus ostreatus, Pleurotus eryngii, Trametes versicolor, Pycnosporus sanguineus and Phanerochaete chrysosporium were characterized, evaluated in the hydrolysis of pretreated pulps of sorghum straw and compared efficiency with commercial enzyme. Most fungi complexes had better hydrolysis rates compared with purified commercial enzyme.

Ethanol production from sweet sorghum by Saccharomyces cerevisiae DBKKUY-53 immobilized on alginate-loofah matrices

Abstract Ethanol production from sweet sorghum juice (SSJ) using the thermotolerant Saccharomyces cerevisiae strain DBKKUY-53 immobilized in an alginate-loofah matrix (ALM) was successfully developed. As found in this study, an ALM with dimensions of 20 × 20 × 5 mm3 is effective for cell immobilization due to its compact structure and long-term stability. The ALM-immobilized cell system exhibited greater ethanol production efficiency than the freely suspended cell system. By using a central composite design (CCD), the optimum conditions for ethanol production from SSJ by ALM-immobilized cells were determined. The maximum ethanol concentration and volumetric ethanol productivity obtained using ALM-immobilized cells under the optimal conditions were 97.54 g/L and 1.36 g/L h, respectively. The use of the ALM-immobilized cells was successful for at least six consecutive batches (360 h) without any loss of ethanol production efficiency, suggesting their potential application in industrial ethanol production.

Improvement of ethanol production from sweet sorghum juice under batch and fed-batch fermentations: effects of sugar levels, nitrogen supplementation, and feeding regimes

Background: Fermentation process development has been very important for efficient ethanol production. Improvement of ethanol production efficiency from sweet sorghum juice (SSJ) under normal gravity (NG, 160 g/L of sugar), high gravity (HG, 200 and 240 g/L of sugar) and very high gravity (VHG, 280 and 320 g/L of sugar) conditions by nutrient supplementation and alternative feeding regimes (batch and fed-batch systems) was investigated using a highly ethanol-tolerant strain, Saccharomyces cerevisiae NP01. Results: In the batch fermentations without yeast extract, HG fermentation at 200 g/L of sugar showed the highest ethanol concentration (PE, 90.0 g/L) and ethanol productivity (QE, 1.25 g/L·h). With yeast extract supplementation (9 g/L), the ethanol production efficiency increased at all sugar concentrations. The highest PE (112.5 g/L) and QE (1.56 g/L·h) were observed with the VHG fermentation at 280 g/L of sugar. In the fed-batch fermentations, two feeding regimes, i.e., stepwise and continuous feedings, were studied at sugar concentrations of 280 g/L. Continuous feeding gave better results with the highest PE and QE of 112.9 g/L and 2.35 g/L·h, respectively, at a feeding time of 9 h and feeding rate of 40 g sugar/h. Conclusions: In the batch fermentation, nitrogen supplementation resulted in 4 to 32 g/L increases in ethanol production, depending on the initial sugar level in the SSJ. Under the VHG condition, with sufficient nitrogen, the fed-batch fermentation with continuous feeding resulted in a similar PE and increased QP by 51% compared to those in the batch fermentation.

Lipophilic metabolite profiling of maize and sorghum seeds and seedlings, and their pest spotted stem borer larvae: A standardized GC-MS based approach.

In order to better understand the biochemical interactions and to identify new biomarkers for plant resistance against insects, we proposed a suitable lipophilic profiling method for insects and their host plants. The critical components of GC-MS based analysis are: sample amount, extraction, derivatization, temperature gradient, run time, and identification of peaks. For lipophilic metabolite profiling of maize and sorghum, and their insect pest, spotted stem borer larvae, we recommend 100 mg sample weight for seeds and insect samples (whole insect body), and 200 mg for seedlings. Maize and sorghum seeds required less time for fat extraction in comparison to their seedlings and the pest fed on these seedlings. GC-MS was standardized for better separation and intensity of peaks using different temperature gradients in the range of 180-300 C. A total of 48 lipophilic compounds encompassing various classes based on their functional groups such as fatty acids, fatty alcohols, hydrocarbons, sterols and terpenoids, vitamin derivative, etc. were separated in the seedlings (30), seeds (14), and the pest (26) in the retention time range of 3.22 to 29.41 min. This method could be useful to study nutritional aspects of different field crops in relation to various stresses apart from the analysis of lipophilic compounds for better understanding of insect-plant interactions.

Dilute acid pretreatment and enzymatic hydrolysis of sorghum biomass for sugar recovery—A statistical approach.

Sorghum is one of the commercially feasible lignocellulosic biomass and has a great potential of being sustainable feedstock for renewable energy. As with any lignocellulosic biomass, sorghum also requires pretreatment which increases its susceptibility to hydrolysis by enzymes for generating sugars which can be further fermented to alcohol. In the present study, sorghum biomass was evaluated for deriving maximum fermentable sugars by optimizing various pretreatment parameters using statistical optimization methods. Pretreatment studies were done with H2SO4, followed by enzymatic saccharification. The efficiency of the process was evaluated on the basis of production of the total reducing sugars released during the process. Compositional analysis was done for native as well as pretreated biomass and compared. The biomass pretreated with the optimized conditions could yield 0.408 g of reducing sugars /g of pretreated biomass upon enzymatic hydrolysis. The cellulose content in the solid portion obtained after pretreatment using optimised conditions was found to be increased by 43.37% with lesser production of inhibitors in acid pretreated liquor.

Avaliação das silagens do sorgo BRS-610 em sete estádios de maturação pela técnica in vitro semiautomática de produção de gases / Evaluation of the silages of sorghum BRS-610 at seven stages of maturation by semi-automated in vitro gas production technique

Avaliou-se a qualidade da silagem do sorgo BRS-610 produzida em sete estádios de maturação dos grãos, pela técnica in vitro semiautomática de produção de gases. O delineamento foi inteiramente ao acaso, com quatro repetições por tratamento, e as médias foram comparadas pelo teste Scott-knott. Os parâmetros potencial máximo de produção de gases, tempo de colonização, taxa de produção de gases e degradabilidade efetiva da matéria seca para as taxas de passagem de 2, 5 e 8 por cento/h foram obtidos pelo modelo de France et al. O potencial máximo de produção de gases não diferiu entre as silagens e indicou média de 170,5mL/g de matéria seca. Entre leitoso/pastoso e pastoso houve menor tempo de colonização. As maiores taxas de fermentação foram observadas nas silagens produzida entre os estádios pastoso/farináceo a seco. As degradabilidades efetivas foram semelhantes entres as silagens, com média de 46,2 por cento. O sorgo BRS-610 produziu silagem de qualidade satisfatória, com destaque para a silagem produzida com a planta no estádio pastoso/farináceo, por associar melhor taxa de fermentação e menor tempo de colonização.

The quality of the silages of sorghum BRS-610 produced at seven stages of grain maturation was evaluated by the semi-automatic gas production technique. It was used a complete randomized design, with four repetitions per treatment, and the means were compared by the Scott-knott test (P<0.05). The potential of production of gases, lag phase, rates of gas production, and effective dry matter degradability for 2, 5, and 8 percent/h rates of passage were obtained by the model of France et al. The potential of gas production did not differ among the silages, showing mean of 170.5mL/dry matter. The stages between milky/soft dough to soft dough showed shorter lag phase. The highest rate of gas production was observed in the silages produced between soft dough/floury to dry stages. The effective degradabilities were similar among the silages, averaging 46.2 percent. The sorghum BRS-610 produced satisfactory quality silage, mainly for the silage produced with the plant at soft dough/floury stage, for associating better rate of gas production and shorter lag phase.

Composição bromatológica e parâmetros físicos de grãos de sorgo com diferentes texturas do endosperma / Chemical composition and physical parameters of sorghum with differents endosperm textures

Chemical composition and physical parameters of 33 sorghum grain genotypes with different endosperm textures were obtained. Dry matter mean of genotypes was 88.77 percent; crude protein varied from 9.85 percent to 18.28 percent; starch varied from 62.07 percent to 78.74 percent; ether extract from 1.76 percent to 3.68 percent; crude fibra from 0.35 percent to 6.60 percent. Tannins were observed only for BR 305, SHS 600 and A 9904. The 1,000 grains weight varied from 12.58g for CMSXS 214 to 27,18g for A 9904. Density values varied from 1.40g/cm3 for texiota 5 to 1.50g/cm3 for SC 283. The vitreosity values varied from 1.10 to SC 283 to 4.20 for Early Hegari. Great variability for chemical and physical parameters of sorghum genotypes was observed

Effects of planting date and time of nitrogen application on yield and sugar content of sweet sorghum.

This experiment was conducted at Azad University experiment station to evaluate the effects of planting date and time of nitrogen application on yield and sugar content of sweet sorghum in 2003. Four planting dates (May 4, March 19, June 3 and June 18) and three stages of nitrogen application (3-5/8 leaf stage, boot stage and soft dough stage) were assigned to the main and subplots, respectively. Plant height and diameter, fresh stalk yield, total dry weight, brix value (soluble solids), sugar content and grain yield were affected by planting date. For all the characteristics, the maximum value was obtained by the first planting date. Only stem height, diameter and total plant dry weights were affected by time of nitrogen application. In these cases, nitrogen application at (3-5/8) leaf stage was superior to others. According to the results, sweet sorghum should be planted on the beginning of May and nitrogen be applied at (3-5/8) leaf stage.