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.

Production of Inulinase by Free and Immobilized Cells of Penicillium funiculosum p.36

The aim of this work was to optimize the growth conditions and continuous production of the enzyme using free and immobilized cells of inulinase by Penicillium funiculosum. The highest yield of enzyme (163.5U/mL) was obtained when the culture was incubated at 27oC and 200 rpm for 96h in a fermentation medium containing both inulin and peptone as sole carbon and nitrogen source, respectively. When the cells of the P. funiculosum were immobilized on different carriers, especially linen fibers, their production ability was successfully maintained for seven successive batches. When the fermentation was carried out using inulin juice prepared from Jerusalem artichoke tubers (in place of pure inulin), inulinase production could be sustained till the second cultivation batch of the P. funiculosum immobilized on linen fibers, yielding 122 U/mL enzyme. Results proved the feasibility of using crude inulin juice as a simple and economic carbon source for the production of inulinase.

Tratamento de efluente sintético de laticínio em reatores em batelada inoculados com Aspergillus niger AN400 / Treatment of synthetic dairy wastewater in batch reactors inoculated with Aspergillus niger AN400

Indústrias de laticínios contêm elevada concentração de matéria orgânica que podem causar poluição. Este trabalho propôs estudar a remoção de matéria orgânica e de nutrientes de um efluente sintético de laticínio, disposto em reatores em bateladas repetidas inoculados com Aspergillus niger AN400. O sistema foi composto por reatores com diferentes configurações e, para cada tipo de montagem, variou-se o material-suporte. O estudo foi realizado em quatro ciclos de cinco dias, com retiradas diárias de alíquotas. A análise dos resultados mostrou eficiência de remoção de demanda química de oxigênio com picos de até 71,7%, concluindo-se que a tecnologia mostrou-se viável na remoção de matéria orgânica. Em relação aos nutrientes, o sistema pôde remover satisfatoriamente nitrato; contudo, para remoção de amônia, não se apresentou promissor.

Dairy industries contain high concentration of organic matter that may cause pollution. This study proposed to study the removal of organic matter and nutrients from a synthetic dairy effluent, arranged in repeated batch reactors inoculated with Aspergillus niger AN400. The system was composed of reactors with different configurations and, for each type of assembly, the support material was varied. The study was conducted in four cycles of five days, with daily withdrawals of portions. The results showed efficiency of removal of chemical oxygen demand with peaks of up to 71.7%. One might conclude that the technology was feasible in the removal of organic matter. For nutrients, the system could successfully remove nitrate; however, for removal of ammonia, it was not promising.

Production of Halophilic α-Amylase by Immobilized Cells of Moderately Halophilic Bacillus sp. Strain TSCVKK.

Aims: To investigate the effect of cell immobilization on amylase production by the moderately halophilic bacterium, Bacillus sp. strain TSCVKK and to compare the properties of the amylase produced under immobilized conditions with the enzyme produced by the free cells. Study Design: Cell immobilization. Place and Duration of Study: Department of Chemistry, Biochemistry Lab, Indian Institute of Technology (IIT Madras), Chennai, Tamil Nadu, between Jan 2009 and March 2009. Methodology: Bacillus sp. strain TSCVKK was immobilized in alginate, agar, polyacrylamide and gelatin. Production of amylase was determined using 3, 5- dinitrosalicylic acid (DNS). Effect of NaCl, pH, temperature on the activity of amylase was determined and compared with the amylase produced by the free cells. Results: Maximum production of 832 mU/ml was achieved with an initial cell load of 1.2% (w/v; wet weight) of 24 h grown cells immobilized in 2% agar of 4 mm3 block size using GSL-2 medium containing 10% NaCl and 1.5% dextrin at pH 8.0 at 30ºC after 36 h of growth. Amylase production was lower when the cells were immobilized in alginate (211 mU/ml) or with the free cells of same biomass concentration as used for immobilization (333 mU/ml). Amylase was not produced when gelatin or polyacrylamide was used as the immobilization matrix. The immobilized cells in 2% agar could be used up to 5 cycles without much reduction in amylase production. Amylase produced through cell immobilization retained all the properties that were shown by amylase produced under submerged fermentation. Conclusion: Agar was the suitable matrix to immobilize Bacillus sp. strain TSCVKK for amylase production. Amylase produced under immobilization conditions retained its temperature, salt and pH requirements. Immobilized cells were used for 5 cycles without much decrease in production.

Study of Sugarcane Pieces as Yeast Supports for Ethanol Production from Sugarcane Juice and Molasses Using Newly Isolated Yeast from Toddy Sap

A repeated batch fermentation system was used to produce ethanol using Saccharomyces cerevisiae strain (NCIM 3640) immobilized on sugarcane (Saccharum officinarum L.) pieces. For comparison free cells were also used to produce ethanol by repeated batch fermentation. Scanning electron microscopy evidently showed that cell immobilization resulted in firm adsorption of the yeast cells within subsurface cavities, capillary flow through the vessels of the vascular bundle structure, and attachment of the yeast to the surface of the sugarcane pieces. Repeated batch fermentations using sugarcane supported biocatalyst were successfully carried out for at least ten times without any significant loss in ethanol production from sugarcane juice and molasses. The number of cells attached to the support increased during the fermentation process, and fewer yeast cells leaked into fermentation broth. Ethanol concentrations (about 72.65~76.28 g/L in an average value) and ethanol productivities (about 2.27~2.36 g/L/hr in an average value) were high and stable, and residual sugar concentrations were low in all fermentations (0.9~3.25 g/L) with conversions ranging from 98.03~99.43%, showing efficiency 91.57~95.43 and operational stability of biocatalyst for ethanol fermentation. The results of the work pertaining to the use of sugarcane as immobilized yeast support could be promising for industrial fermentations.

Efecto de diámetro de esfera y densidad celular en la producción de etanol con levadura inmovilizada en alginato / Effect of pellet diameter and cell density on ethanol production with alginate immobilized yeast

Las células inmovilizadas tienen aplicación potencial en la producción de biocombustibles posibilitando la reutilización de biomasa, el empleo de diversas configuraciones de reactores y sistemas de cultivo, el manejo de altas densidades celulares alcanzando altas productividades volumétricas, y la simplificación de operaciones de procesamiento de salida. El objetivo del presente estudio fue evaluar la influencia del diámetro de las perlas y la densidad celular en la producción de etanol con Saccharomyces uvarum inmovilizada en alginato al 2% (p/v). Para ello se evaluaron tres diámetros de perlas de 2, 2,5 y 3 mm. Las células inmovilizadas fueron cultivadas en medio con 12% (p/v) de glucosa en biorreactores de columna sin agitación a 28 ºC, y se operaron cuatro lotes consecutivos de 48 horas cada uno. En cada lote se cuantificó el consumo de glucosa y se determinó la cantidad de etanol producido. Los rendimientos máximos de etanol para las esferas de 2, 2,5 y 3 mm de diámetro fueron 81, 83 y 97% del rendimiento teórico. La máxima productividad volumétrica de etanol fue 1,2 g/L-1/h-1 con un consumo de glucosa de 99,8% al término del lote, correspondiente a las columnas con perlas de 3 mm y con una producción de 0,017 g de etanol por esfera. La producción de etanol acumulada en cada sistema fue 178, 189 y 200 g/L-1 para 2, 2,5 y 3 mm respectivamente, encontrándose una relación directa con el diámetro de perla e inversa respecto a la densidad celular. Los rendimientos de etanol obtenidos son superiores a los reportados para la misma especie.

Immobilized cells have a potential use in biofuel production. They also allow re-using biomass, using diverse reactor configurations and culture systems, handling high cell densities to obtain high volumetric productivities and to simplify the downstream processing. The purpose of this work was to evaluate the influence of bead diameter and cell density on ethanol production using immobilized Saccharomyces uvarum in 2% (w/v) alginate. For that, three bead diameters (2, 2.5 and 3 mm) were evaluated. Immobilized cells were cultured on a 12% (w/v) glucose medium in column bioreactors without agitation at 28 °C for four 48 h–repeated batches. For each batch, both glucose consumption and ethanol produced were measured. Maximum yields for 2, 2.5 and 3 mm bead diameters were 81, 83 and 97% of theoretical yield. Maximum volumetric productivity of ethanol was 1.2 g/L-1/h-1 with 99.8% glucose consumption at the end of the batch, corresponding to the 3 mm bead diameter and the ethanol production per bead was 0.017 g. Accumulated ethanol production for each system was 178, 189 and 200 g/L-1 for 2, 2.5 y 3 mm bead diameter, respectively, being this directly related to bead diameter and inversely related to cell density. Ethanol yields were higher than those reported for the same species.

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.

Production of Bacillus licheniformis ATCC 21415 alkaline protease in batch, repeated batch and continuous culture

Bacillus licheniformis ATCC 21415 cells were immobilized on different carriers using different methods of immobilization including physical adsorption, covalent binding, ionic binding and entrapment. The immobilized cells were prepared by covalent binding on wool (as a new carrier) through 1% glutaraldehyde had the highest enzyme activity (9.0 U/mL) with the highest specific productivity (6.17 U/g wet cells/h). Alkaline protease production and the stability of biocatalyst were investigated in both free and immobilized cells. The results showed that the immobilized cells were more efficient for enzyme production by repeated batch fermentation (5 cycles, 480 h) with 57% residual activity whereas the free cells retained 35% after 2 cycles. In continuous production the highest enzyme activity (9.9 U/mL) was obtained at a dilution rate of 0.1/h while the highest enzyme yield (763.6 U/h) and the highest reactor productivity (3.32 U/mL/h) were attained at a dilution rate of 0.4/h. Packed-bed bioreactor was a successful method for continuous production of alkaline protease for a long time (168 h) with 53% relative activity. The bioreactor affected the highest specific productivity (118.2 U/g wet cells/h) which was 12-24 times higher than other systems of enzyme production.