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.

Resistance of bioenergy sorghum to Diatraea saccharalis (Lepidoptera: Crambidae) / Resistência de sorgo bioenergia à diatraea saccharalis (lepidoptera: crambidae)

This study evaluated the effects of the sugarcane borer Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) on cultivars of sweet and biomass sorghum for the selection of resistant cultivars. The present work consisted of two trials, with natural pest infestation. In the first one, 10 sweet sorghum cultivars were analyzed for the following variables: plant height, number of healthy and damaged internodes, gallery position and size, stem infestation level and soluble solids content (°Brix). In the second trial, it was analyzed 16 genotypes of high biomass sorghum, with the same variables above mentioned, in addition to the lignin, cellulose and hemicellulose contents. Among sweet sorghum genotypes evaluated, the genotype CMSXS647 stood out due to the traits: plant height, infestation level, gallery size and soluble solids content. Among the sorghum genotypes evaluated, CMSXS7030, CMSXS7012 and CMSXS7028 presented ideal characteristics for infestation level, plant height and number of lignocellulosic compounds. Such information, in addition to supporting the bioenergy sorghum breeding program, will assist in integrated pest management for sorghum cultivation.

Foram estudados os efeitos causados pela broca-do-colmo Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae), em cultivares de sorgo sacarino e biomassa visando seleção de cultivares resistentes à praga. O presente trabalho foi constituído de dois ensaios, com infestação natural da praga. No primeiro, 10 cultivares de sorgo sacarino foram analisadas quanto às seguintes variáveis: altura das plantas, quantidade de internódios sadios e com injúrias, posição e tamanho da galeria, intensidade de infestação de colmos e teor de sólidos solúveis (°Brix). No segundo ensaio, foram analisados 16 genótipos de sorgo biomassa, com as mesmas variáveis supracitadas, além dos teores de lignina, celulose e hemicelulose. Entre os genótipos de sorgo sacarino avaliados, o genótipo CMSXS647 foi o que se destacou em função das características: altura de plantas, intensidade de infestação, tamanho de galerias e teor de sólidos solúveis. Entre os genótipos de sorgo biomassa avaliados: CMSXS7030, CMSXS7012 e CMSXS7028 apresentaram características ideais para intensidade de infestação, altura de plantas e quantidade de compostos lignocelulósico. Tais informações, além de prover o programa de melhoramento de sorgo energia podem ajudar o programa de MIP para a cultura do sorgo, uma vez que o produtor conhece a suscetibilidade dos materiais escolhidos.

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.

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.

Effect of synthetic and natural media on lipid production from Fusarium oxysporum

Background: Dependence on fossil resources, for the production of fuels and energy, has resulted in environmental and financial problems, which require our immediate action in order to reverse the situation. Use of renewable sources for the production of fuels and energy is an important alternative with biodiesel remains as one of the promising options. Aim of this work is to evaluate the fungus Fusarium oxysporum for its potentials to accumulate microbial lipids when grown on synthetic media and saccharified sweet sorghum stalks. Results: The effect of different carbon sources, nitrogen sources and C/N ratio on the lipid production was initially examined, which resulted in a lipid concentration of 4.4 g/L, with lipid content of 42.6% w/w. Sweet sorghum stalks were able to support growth and lipid production of the fungus, both as carbon source and as nitrogen source. It was also shown that saccharification of the dried stalks is an important step to increase lipid production. Removal of the remaining stalk solids enabled the lipid production during cultivation in increased initial solids of up to 16 w/w. This resulted in a lipid production of 3.81 g/L. Conclusions: It was demonstrated that F. oxysporum can be used as an efficient oleaginous microorganism, with sweet sorghum serving as an excellent raw material for the cultivation of the fungus. The lipids obtained during this work were also found to have a fatty acid profile with good potentials to be used for biodiesel production.

High-temperature ethanol production using thermotolerant yeast newly isolated from Greater Mekong Subregion

Abstract The application of high-potential thermotolerant yeasts is a key factor for successful ethanol production at high temperatures. Two hundred and thirty-four yeast isolates from Greater Mekong Subregion (GMS) countries, i.e., Thailand, The Lao People's Democratic Republic (Lao PDR) and Vietnam were obtained. Five thermotolerant yeasts, designated Saccharomyces cerevisiae KKU-VN8, KKU-VN20, and KKU-VN27, Pichia kudriavzevii KKU-TH33 and P. kudriavzevii KKU-TH43, demonstrated high temperature and ethanol tolerance levels up to 45 °C and 13% (v/v), respectively. All five strains produced higher ethanol concentrations and exhibited greater productivities and yields than the industrial strain S. cerevisiae TISTR5606 during high-temperature fermentation at 40 °C and 43 °C. S. cerevisiae KKU-VN8 demonstrated the best performance for ethanol production from glucose at 37 °C with an ethanol concentration of 72.69 g/L, a productivity of 1.59 g/L/h and a theoretical ethanol yield of 86.27%. The optimal conditions for ethanol production of S. cerevisiae KKU-VN8 from sweet sorghum juice (SSJ) at 40 °C were achieved using the Box-Behnken experimental design (BBD). The maximal ethanol concentration obtained during fermentation was 89.32 g/L, with a productivity of 2.48 g/L/h and a theoretical ethanol yield of 96.32%. Thus, the newly isolated thermotolerant S. cerevisiae KKU-VN8 exhibits a great potential for commercial-scale ethanol production in the future.

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.

Effects of drought stress on biomass and carbohydrate contents of two sweet sorghum cultivars.

Sweet sorghum is adapted to the hot and dry climatic conditions. It can be used for different products such as food, feed, fiber and fuel. This study was carried out to evaluate the effects of four irrigation intervals and three harvesting dates on stem height, stem diameter, biomass, juice volume, brix, sucrose and invert sugar of two sweet sorghum cultivars. The results showed that delay in irrigation intervals from 7 to 21 day significantly decreased all the above characteristics expect for invert sugar which was increased. The highest biomass (56.50 t ha-1) and sucrose content (11.35 %) were obtained at 7 to 10 day irrigation interval. Both biomass and sucrose contents decreased significantly as harvesting date delayed from physiological maturity. Since both sweet sorghum cultivars, Keller and Sofra had similar biomass, sucrose content, brix and juice volume across irrigation intervals and harvesting dates, therefore it is suggested to plant either Keller or Sofra irrigated on 7 to 10 day interval and harvested at physiological maturity.

The use of dried spent yeast as a low-cost nitrogen supplement in ethanol fermentation from sweet sorghum juice under very high gravity conditions

Dried spent yeast (DSY) was used as a low-cost nitrogen supplement for ethanol fermentation from sweet sorghum juice under very high gravity (VHG) conditions by Saccharomyces cerevisiae NP 01. The fermentation was carried out at 30ºC in a 5-litre bioreactor. The results showed that DSY promoted ethanol production efficiencies. The ethanol concentration (P), productivity (Qp) and yield (Yp/s) of the sterile juice (total sugar of 280 g l-1) supplemented with 8 g l-1 of DSY were not different from those supplemented with yeast extract and/or peptone at the same amount. The initial yeast cell concentration of 5 x 10(7) cells ml-1 was found to be optimal for scale-up ethanol production. In addition, an increase in sugar concentration in inoculum preparation medium (from 10 to 100 g l-1) improved the ability of the inoculum to produce ethanol under the VHG conditions. When S. cerevisiae NP 01 grown in the juice containing 100 g l-1 of total sugar was used as the inoculum for ethanol fermentation, the P, Qp and Yp/s obtained were 108.98 +/- 1.16 g l-1, 2.27 +/- 0.06 g l-1 h-1 and 0.47 +/- 0.01 g g-1, respectively. Similar results were also observed when the ethanol fermentation was scaled up to a 50-litre bioreactor under the same conditions. The cost of the sweet sorghum for ethanol production was US$ 0.63 per litre of ethanol. These results clearly indicate the high potential of using sweet sorghum juice supplemented with DSY under VHG fermentation for ethanol production in industrial applications.

Ethanol production from sweet sorghum juice under very high gravity conditions: batch, repeated-batch and scale up fermentation

Batch ethanol fermentations from sweet sorghum juice by Saccharomyces cerevisiae NP 01 were carried out in a 500 ml air-locked Erlenmeyer flask under very high gravity (VHG) and static conditions. The maximum ethanol production efficiency was obtained when 9 g l-1 of yeast extract was supplemented to the juice. The ethanol concentration (P), productivity (Qp) and yield (Yp/s) were 120.24 +/- 1.35 g l-1, 3.01 +/- 0.08 g l-1 h-1 and 0.49 +/- 0.01, respectively. Scale up ethanol fermentation in a 5-litre bioreactor at an agitation rate of 100 rev min-1 revealed that P, Qp and Yp/s were 139.51 +/- 0.11 g l-1, 3.49 +/- 0.00 g l-1 h-1 and 0.49 +/- 0.01, respectively, whereas lower P (119.53 +/- 0.20 g l-1) and Qp (2.13 +/- 0.01 g l-1 h-1) were obtained in a 50-litre bioreactor. In the repeated-batch fermentation in the 5-litre bioreactor with fill and drain volume of 50 percent of the working volume, lower P and Qp were observed in the subsequent batches. P in batch 2 to 8 ranged from 103.37 +/- 0.28 to 109.53 +/- 1.06 g l-1.

Effects of nitrogen treatments and harvesting stages on the aconitic acid, invert sugar and fiber in sweet sorghum cultivars.

Sweet sorghum is adapted to the hot and dry climatic conditions and its tolerance to slat is moderately. It can be used for different products such as food, feed, fiber and fuel. This study was carried out to evaluate the effects, three nitrogen treatments, and three harvesting stages on the aconitic acid, fiber and invert sugar of three sweet sorghum cultivars in the experimental station and the results showed that the effects were significant. Among nitrogen treatments, application of 100 kg ha-1 urea at planting and 200 kg ha-1 urea at 4 leaf stage had the highest aconitic acid (0.26%) and invert sugar (3.44%). Among sweet sorghum cultivars, IS2325 and Vespa had the highest aconitic acid (0.26%) and invert sugar (3.86%), respectively. Plant harvested at 4 leaf stage had the highest aconitic acid (0.26%) and the highest invert sugar (3.85%). Rio had higher fiber content than Vespa and IS2325 and all cultivars had the highest fiber content before chilling harvesting stage. In general, since high invert sugar and high aconitic acid interfere crystallization of sugar so, it is suggested that to plant Vespa, apply urea 100 kg ha-1 urea at planting, 100 kg ha-1 urea at 4 leaf stage and 100 kg ha-1 urea at booting and harvested before chilling that had lowest aconitic acid and invert sugar. Thereby, it is recommended to plant Vespa, apply urea 100 kg ha-1 urea at planting, 100 kg ha-1 urea at 4 leaf stage and 100 kg ha-1 urea at booting and harvested at 4 leaf stage that had the highest aconitic acid.