Continuous Ethanol Production Research Articles

Continuous ethanol production using immobilized yeast cells entrapped in loofa-reinforced alginate carriers

A culture of Saccharomyces cerevisiae M30 entrapped in loofa-reinforced alginate was used for continuous ethanol fermentation in a packed-bed reactor with initial sugar concentrations of 200-248 g/L. Maximum ethanol productivity of 11.5 g/(L·h) was obtained at an ethanol concentration of 57.4 g/L, an initial sugar concentration of 220 g/L and a dilution rate (D) of 0.2 h-1. However, a maximum ethanol concentration of 82.1 g/L (productivity of 9.0 g/(L·h)) was obtained at a D of 0.11 h-1. Ethanol productivity in the continuous culture was 6-8-fold higher than that in the batch culture. Due to the developed carrier's high biocompatibility, high porosity, and good mechanical strength, advantages such as cell regeneration, reusability, altered mechanical strength, and high capacity to trap active cells in the reactor were achieved in this study. The immobilized cell reactor was successfully operated for 30 days without any loss in ethanol productivity. The average conversion yield was 0.43-0.45 throughout the entire operation, with an immobilization yield of 47.5%. The final total cell concentration in the reactor was 37.3 g/L (17.7 g/L immobilized cells and 19.6 g/L suspended cells). The concentration of suspended cells in the effluent was 0.8 g/L.

Continuous ethanol production by the synchronous saccharification and fermentation using food wastes

The synchronous saccharification and fermentation (SSF) by continuous fill and draw method was investigated in order to develop a continuous ethanol fermentation process using the food wastes (FW) available among Korea’s organic wastes. The activity of the hydrolytic enzymes was maintained constantly in the continuous culture by their intermittent addition together with medium exchange. The concentrations of reducing sugar in the culture were maintained at a steady state by regulating supplemented enzyme concentration and exchange rate of medium, reflecting on the consumption rate of reducing sugar caused by the fermentation. When the temperature of the SSF was maintained at the fixation of 35 °C, which enabled us to perform both enzymatic hydrolysis and enzyme fermentation simultaneously, the rate of reducing sugar consumption was 3.61 g/L-hr. For the enzymatic saccharification of FW, when 0.01 BGU as Viscozyme/g-FW and 0.05 AGU as Spirizyme Plus/g-FW were used, the production rate of reducing sugar was 3.93 g/L-hr, indicating a little higher rate of production than that of consumption. A decompression device with ethanol condensing ability was used to continuously pull out ethanol from the culture broth at −600 mmHg, where the ethanol evaporation ability would be maximized and the water evaporation minimized during the process. As a result of the continuous SSF performance, the reducing sugar concentration was maintained at around 30 g/L. The amylase activity was maintained at 8.93±2.17 U/mL. During a 352 hour culture, the whole ethanol productivity was 2.24 g/L-hr, indicating a considerable productivity compared with the other result reported in the continuous SSF.

Fuel ethanol production from concentrated food waste hydrolysates in immobilized cell reactors by Saccharomyces cerevisiae H058

BACKGROUND: Continuous ethanol fermentation of concentrated food waste hydrolysates has been studied. The process was carried out in an immobilized cell reactor with beads of calcium-alginate containing immobilized Saccharomyces cerevisiae H058 at temperature 30 °C and pH 5.0. RESULTS: The total residual sugar decreased with increase of hydraulic retention time (HRT) under various reducing sugar concentrations. Ethanol production by immobilized cells increased with increase in HRT, regardless of the substrate concentrations employed. The highest ethanol concentration of 89.28 g L−1 was achieved at an HRT of 5.87 h and reducing sugar concentration of 200 g L−1. At an HRT of 1.47 h, the maximum volumetric ethanol productivity of 49.88 g L−1 h−1 and the highest ethanol yield of 0.48 g g−1 were achieved at reducing sugar concentration of 160 and 200 g L−1, respectively. The difference between the fresh and the 30-day Ca–alginate immobilized cell was also shown by scanning electronic micrographs of beads taken from their outer and inner surfaces. CONCLUSIONS: Continuous ethanol production from concentrated food waste hydrolysates using immobilized yeast cells is promising in view of the high ethanol productivity obtained at relatively high conversion and excellent reactor stability. Copyright © 2011 Society of Chemical Industry

Continuous ethanol production from wheat straw hydrolysate by recombinant ethanologenic Escherichia coli strain FBR5

Continuous production of ethanol from alkaline peroxide pretreated and enzymatically saccharified wheat straw hydrolysate by ethanologenic recombinant Escherichia coli strain FBR5 was investigated under various conditions at controlled pH 6.5 and 35 °C. The strain FBR5 was chosen because of its ability to ferment both hexose and pentose sugars under semi-anaerobic conditions without using antibiotics. The average ethanol produced from the available sugars (21.9-47.8 g/L) ranged from 8.8 to 17.3 g/L (0.28-0.45 g/g available sugars, 0.31-0.48 g/g sugar consumed) with ethanol productivity of 0.27-0.78 g l(-1)h(-1) in a set of 14 continuous culture (CC) runs (16-105 days). During these CC runs, no loss of ethanol productivity was observed. This is the first report on the continuous production of ethanol by the recombinant bacterium from a lignocellulosic hydrolysate.

Continuous ethanol production using immobilized yeast cells entrapped in loofa-reinforced alginate carriers.

A culture of Saccharomyces cerevisiae M30 entrapped in loofa-reinforced alginate was used for continuous ethanol fermentation in a packed-bed reactor with initial sugar concentrations of 200-248 g/L. Maximum ethanol productivity of 11.5 g/(L·h) was obtained at an ethanol concentration of 57.4 g/L, an initial sugar concentration of 220 g/L and a dilution rate (D) of 0.2 h-1. However, a maximum ethanol concentration of 82.1 g/L (productivity of 9.0 g/(L·h)) was obtained at a D of 0.11 h-1. Ethanol productivity in the continuous culture was 6-8-fold higher than that in the batch culture. Due to the developed carrier’s high biocompatibility, high porosity, and good mechanical strength, advantages such as cell regeneration, reusability, altered mechanical strength, and high capacity to trap active cells in the reactor were achieved in this study. The immobilized cell reactor was successfully operated for 30 days without any loss in ethanol productivity. The average conversion yield was 0.43-0.45 throughout the entire operation, with an immobilization yield of 47.5%. The final total cell concentration in the reactor was 37.3 g/L (17.7 g/L immobilized cells and 19.6 g/L suspended cells). The concentration of suspended cells in the effluent was 0.8 g/L.

Observed kinetics of the continuous growth and ethanol production of the genetically engineered yeast BHL01

Observed kinetics of the continuous growth and ethanol production of the genetically engineered yeast BHL01

Improvement continuous ethanol production from sweet sorghum juice using a multistage bioreactor

Improvement continuous ethanol production from sweet sorghum juice using a multistage bioreactor

Cane molasses fermentation for continuous ethanol production in an immobilized cells reactor by Saccharomyces cerevisiae

Sodium-alginate immobilized yeast was employed to produce ethanol continuously using cane molasses as a carbon source in an immobilized cell reactor (ICR). The immobilization of Saccharomyces cerevisiae was performed by entrapment of the cell cultured media harvested at exponential growth phase (16 h) with 3% sodium alginate. During the initial stage of operation, the ICR was loaded with fresh beads of mean diameter of 5.01 mm. The ethanol production was affected by the concentration of the cane molasses (50, 100 and 150 g/l), dilution rates (0.064, 0.096, 0.144 and 0.192 h −1) and hydraulic retention time (5.21, 6.94, 10.42 and 15.63 h) of the media. The pH of the feed medium was set at 4.5 and the fermentation was carried out at an ambient temperature. The maximum ethanol production, theoretical yield ( Y E / S ), volumetric ethanol productivity ( Q P ) and total sugar consumption was 19.15 g/l, 46.23%, 2.39 g l −1 h −1 and 96%, respectively.

Continuous production of ethanol from hexoses and pentoses using immobilized mixed cultures of Escherichia coli strains

We have developed highly efficient ethanologenic Escherichia coli strains that selectively consume pentoses and/or hexoses. Mixed cultures of these strains can be used to selectively adjust the sugar utilization kinetics in ethanol fermentations. Based on the kinetics of sugar utilization, we have designed and implemented an immobilized cell system for the optimized continuous conversion of sugars into ethanol. The results confirm that immobilized mixed cultures support a simultaneous conversion of hexoses and pentoses into ethanol at high yield and at a faster rate than immobilized homogenous cells. Continuous ethanol production has been maintained for several weeks at high productivity with near complete sugar utilization. The control of sugar utilization using immobilized mixed cultures can be adapted to any composition of hexoses and pentoses by adjusting the strain distribution of immobilized cells. The approach, therefore, holds promise for ethanol fermentation from lignocellulosic hydrolysates where the feedstock varies in sugar composition.

Ethanol Production from Different Substrates by a Flocculent Saccharomyces cerevisiae Strain

During the last years there has been an increasing interest in using ethanol as a substitute for fossil fuels. The bioethanol used today is mainly produced from sugar cane and cereals, but reducing the production costs of ethanol is still crucial for a viable economic process. Cellulose from vegetable biomass will be the next cheap raw material for second generation fuel ethanol production and agricultural by-products with a low commercial value, as corn stover, corn fiber and cane bagasses would become an attractive feedstock for bioethanol production.In this study, different strains of Saccharomyces cerevisiae have been screened for the ability of bioethanol production. Yeasts were grown in a synthetic liquid medium containing sucrose in batch regime and the growth rates, ethanol and biomass productions were determined as well as their growth ability in cane molasses.The results indicate that a flocculent yeast, isolated in our lab and designated by strain F, was the most promising yeast strain among those tested for continuous ethanol production. This strain was isolated from corn hydrolysates, obtained from a Portuguese distillery facility (DVT, Torres Novas, Portugal) showing highest growth rate (0.49h-1), highest ethanol yield (0.35g/g) and high flocculation capacity.The study on ethanol production in continuous reactor process with the selected yeast strain (strain F) was made on sucrose and cane molasses at different dilution rates (0.05-0.42 h-1). A steady flocculating yeast fluidized bed reactor system was established allowing the functioning of the reactor for 1000 h. Data shows that when the dilution rate rose to 0.42h-1, the highest productivity (20g/Lh) was obtained attaining an ethanol concentration in the reactor of 47g/L for sucrose and molasses media.

Hybrid cybernetic model-based simulation of continuous production of lignocellulosic ethanol: Rejecting abruptly changing feed conditions

Fermenting various sugars derived from lignocellulosic biomass promises to be attractive for producing ethanol, an important alternative fuel. Diversity of lignocellulosic biomass sources and pre-processing variations mean entering sugars are expected to experience large, though infrequent, changes. Recent developments in hybrid cybernetic modeling allow efficient in silico studies. This enables studying sequential linearization-based model predictive control for ensuring high productivity and conversion, for a chemostat seeded with yeast capable of co-fermentation. An appropriate controlled variable (conversion) and control formulation are ascertained. Also, a recently proposed hidden-Markov disturbance model, capable of describing the aforesaid changes, results in closed-loop performance superior to the typical integrated white-noise assumption.

Continuous Ethanol Production from Cassava Through Simultaneous Saccharification and Fermentation by Self-Flocculating Yeast Saccharomyces Cerevisiae CHFY0321

In this study, a fermentor consisting of four linked stirred towers that can be used for simultaneous saccharification and fermentation (SSF) and for the accumulation of cell mass was applied to the continuous production of ethanol using cassava as the starchy material. For the continuous process with SSF, the pretreated cassava liquor and saccharification enzyme at total sugar concentrations of 175 g/L and 195 g/L were continuously fed to the fermentor with dilution rates of 0.014, 0.021, 0.031, 0.042, and 0.05 h(-1). Considering the maximum saccharification time, the highest volumetric productivity and ethanol yield were observed at a dilution rate of 0.042 h(-1). At dilution rates in the range of 0.014 h(-1) to 0.042 h(-1), high production rates were observed, and the yeast in the first to fourth fermentor showed long-term stability for 2 months with good performance. Under the optimal culture conditions with a feed sugar concentration of 195 g/L and dilution rate of 0.042 h(-1), the ethanol volumetric productivity and ethanol yield were 3.58 g/L x h and 86.2%, respectively. The cell concentrations in the first to fourth stirred tower fermentors were 74.3, 71.5, 71.2, and 70.1 g dry cell/L, respectively. The self-flocculating yeast, Saccharomyces cerevisiae CHFY0321, developed by our group showed excellent fermentation results under continuous ethanol production.

Continuous ethanol production from sweet sorghum stem juice using stirred tank and tubular bioreactors

Continuous ethanol production from sweet sorghum stem juice using stirred tank and tubular bioreactors

Continuous Production of Ethanol from Starch Using Glucoamylase and Yeast Co-Immobilized in Pectin Gel

This work presents a continuous simultaneous saccharification and fermentation (SSF) process to produce ethanol from starch using glucoamylase and Saccharomyces cerevisiae co-immobilized in pectin gel. The enzyme was immobilized on macroporous silica, after silanization and activation of the support with glutaraldehyde. The silica-enzyme derivative was co-immobilized with yeast in pectin gel. This biocatalyst was used to produce ethanol from liquefied manioc root flour syrup, in three fixed bed reactors. The initial reactor yeast load was 0.05 g wet yeast/ml of reactor (0.1 g wet yeast/g gel), used in all SSF experiments. The enzyme concentration in the reactor was defined by running SSF batch assays, using different amount of silica-enzyme derivative, co-immobilized with yeast in pectin gel. The chosen reactor enzyme concentration, 3.77 U/ml, allowed fermentation to be the rate-limiting step in the batch experiment. In this condition, using initial substrate concentration of 166.0 g/l of total reducing sugars (TRS), 1 ml gel/1 ml of medium, ethanol productivity of 8.3 g/l/h was achieved, for total conversion of starch to ethanol and 91% of the theoretical yield. In the continuous runs, feeding 163.0 g/l of TRS and using the same enzyme and yeast concentrations used in the batch run, ethanol productivity was 5.9 g ethanol/l/h, with 97% of substrate conversion and 81% of the ethanol theoretical yield. Diffusion effects in the extra-biocatalyst film seemed to be reduced when operating at superficial velocities above 3.7 x 10(-4) cm/s.

Mucor indicus as a biofilter and fermenting organism in continuous ethanol production from lignocellulosic hydrolyzate

The dimorphic filamentous fungus Mucor indicus was used as a biofilter and fermenting microorganism in a bioreactor for continuous cultivation of dilute-acid lignocellulosic hydrolyzates up to dilution rate ( D) 0.3 h −1. The hydrolyzate was so toxic that the fungus could not grow in traditional stirred-tank bioreactors even at D 0.1 h −1. On the other hand, the fungus could be manipulated to grow as a mixture of short dispersed filaments and yeast-like cells. This morphology was suitable to create a biological filter, when attached to a specific screen located across the outlet stream. This filter was fully permeable to the liquid, but not the biomass inside the bioreactor. While the biomass concentration in the outflow was between 0.51 and 3.43 g/l at dilution rates 0.3–0.1 h −1, the biomass concentration inside the bioreactor was held at 8.34–9.91 g/l. The cells consumed 95.7%, 94.3%, and 75.3% of hexoses and 64.5%, 25.9%, and 7.6% of xylose present in the hydrolyzate at D 0.1, 0.2, and 0.3 h −1, respectively. The ethanol yields at these dilution rates were 0.37, 0.45, and 0.46 g/g consumed sugars, respectively. The only important by-products were glycerol and xylitol, which at D 0.2 h −1 yielded 0.036 and 0.022 g/g consumed sugars, respectively.

The performance of serial bioreactors in rapid continuous production of ethanol from dilute-acid hydrolyzates using immobilized cells

The performance of single, and series of, continuous stirred-tank (CSTBR) and fluidized-bed bioreactor (FBBR) in anaerobic continuous cultivation of glucose in defined media and dilute-acid hydrolyzates at dilution rates 0.22, 0.43, 0.65 and 0.86 h −1 using immobilized Saccharomyces cerevisiae CBS 8066, was investigated. While the single CSTBR and FBBR could not take up more than 77% and 92% of glucose in a defined medium at dilution rate 0.86 h −1, addition of the second bioreactor decreased the residual glucose to less than 1.1% of the incoming sugar. A similar trend was obtained in cultivation of dilute-acid hydrolyzates. A CSTBR could take up 75% and 54% of the initial fermentable sugars at dilution rates 0.43 and 0.86 h −1, while the addition of the FBBR improved the assimilation of the sugars to 100% and 86%, respectively. The ethanol yields from the hydrolyzate were between 0.41 and 0.48 g/g in all the experiments. The specific and volumetric ethanol productivities were 1.13 g/g h and 5.98 g/L h for the single bioreactor and 0.98 g/g h and 5.49 g/L h for the serial bioreactor at the highest dilution rate, respectively. Glycerol was the only important by-product in terms of concentration, and yielded 0.05–0.07 g/g from the hydrolyzate. From the initial 3.98 g/L acetic acid present in the hydrolyzate, 0.1–0.8 g/L was assimilated by the cells. The yeast cells were accumulated close to the surface of the beads. While the cells had a dry-weight concentration of 129 g/L close to the surface of the beads, the concentration in the core was only 13 g/L.

Optimal biocatalyst loading in a fixed bed

The optimal distribution of biocatalyst in a fixed bed operating at steady state was determined to minimize the length of the bed for a fixed conversion of 95%. The distribution in terms of the biocatalyst loading on an inert support depends upon the axial distance from the bed entrance (continuous solution) as well as a set of dimensionless parameters that reflect the bed geometry, the bulk flow, reaction kinetics and diffusional characteristics. A mathematical model of the system with the following features was used to obtain the results: (1) convective mass transfer and dispersion in the bulk phase; (2) mass transfer from the bulk phase to the surface of the catalyst particle; and (3) simultaneous diffusion and chemical reaction in the catalyst particle with Michaelis-Menton kinetics and a reliable diffusion model (Zhao and DeLancey in Biotechnol Bioeng 64:434-441, 1999, 2000). The solution to the mathematical model was obtained with Mathematica utilizing the Runge Kutta 4-5 method. The dimensionless length resulting from the continuous solution was compared with the optimal length restricted to a uniform or constant cell loading across the entire bed. The maximum difference in the dimensionless length between the continuous and uniform solutions was determined to be 6.5%. The model was applied to published conversion data for the continuous production of ethanol that included cell loading (Taylor et al. in Biotechnol Prog 15:740-751, 2002). The data indicated a minimum production cost at a catalyst loading within 10% of the optimum predicted by the mathematical model. The production rate versus cell loading in most cases displayed a sufficiently broad optimum that the same (non-optimal) rate could be obtained at a significantly smaller loading such as a rate at 80% loading being equal to the rate at 20% loading. These results are particularly important because of the renewed interest in ethanol production (Novozymes and BBI International, Fuel ethanol: a technological evolution, 2004).

Efficiency of a fixed-bed and a gas-lift three-column reactor for continuous production of ethanol by pectate-and alginate-immobilized Saccharomyces cerevisiae cells

AbstractEthanol-tolerant and thermo-tolerant yeast strain Saccharomyces cerevisiae C11-3 cells immobilized in calcium pectate and calcium alginate gels were used for ethanol fermentation in a three-reactor system with a gradient temperature control. The fermentation process has been tested in a fixed-bed and a gas-lift arrangement. The gas-lift system was more efficient due to a better mass transport between the phases. Abrasion was more evident in calcium alginate particles, while calcium pectate beads were not significantly damaged. Two different concentrations of alginate were tested and calcium pectate gel was demonstrated to be more suitable as an immobilization material in comparison with calcium alginate due to its mechanical resistance and favourable diffusion parameters, providing an ethanol production of more than 7.5 g dm−3 h−1 over a period of 630 h.

High efficiency ethanol fermentation by entrapment of Zymomonas mobilis into LentiKatsR

To examine the potential of Zymomonas mobilis entrapped into polyvinylalcohol (PVA) lens-shaped immobilizates in batch and continuous ethanol production. Cells, free or immobilized in PVA hydrogel-based lens-shaped immobilizates - LentiKats, were cultivated on glucose medium in a 1 l bioreactor. In comparison with free cell cultivation, volumetric productivity of immobilized batch culture was nine times higher (43.6 g l(-1) h(-1)). The continuously operated system did not improve the efficiency (volumetric productivity of the immobilized cells 30.7 g l(-1) h(-1)). We demonstrated Z. mobilis capability, entrapped into LentiKats, in the cost-efficient batch system of ethanol production. The results reported here emphasize the potential of bacteria in combination with suitable fermentation technology in industrial scale. The innovation compared with traditional systems is characterized by excellent long-term stability, high volumetric productivity and other technological advantages.

Continuous ethanol production using self-flocculating yeast in a cascade of fermentors

Continuous ethanol fermentation using a self-flocculating yeast strain SPSC01 was examined in a four-stage tanks in series fermentation system with a total working volume of 4000 ml. The first tank was designated for seed cultivation and the others for ethanol fermentation. Two-stage enzymatic hydrolysate of corn powder containing total sugar of 120 g l −1, supplemented with 2.0 g l −1 (NH 4) 2HPO 4 and K 2HPO 4, respectively, was used as seed culture substrate and fed into the seed fermentor at the dilution rate of 0.017 h −1. Meanwhile, the hydrolysate containing total sugar of 220 g l −1, supplemented with 0.5 g l −1 (NH 4) 2HPO 4 and 0.15 g l −1 K 2HPO 4 was used as ethanol fermentation substrate and fed into the second tank at the dilution rates of 0.017, 0.025, 0.033, 0.040 and 0.050 h −1, respectively. Steady states were observed when the fermentation system was operated at the dilution rates of 0.017, 0.025, 0.033 and 0.050 h −1. However, when the fermentation system was operated at the dilution rate of 0.040 h −1, unsteady states and oscillations that characterized by big fluctuations of residual sugar, ethanol and biomass were detected. Compared with freely suspended yeast cell ethanol fermentation system, this self-flocculating yeast strain could partly self-immobilize within the tanks. The effluent containing 95.6 g l −1 ethanol, 1.5 g l −1 reducing sugar and 2.5 g l −1 total sugar was steadily produced when the fermentation system was operated at the dilution rate of 0.033 h −1. The ethanol productivity of 3.44 g l −1 h −1 was achieved, which almost doubled that of freely suspended yeast cell system when the same levels of ethanol and residual sugars were reached. The conversion yield of ethanol to total sugar in the media was calculated to be 0.465, equivalent to 91.1% of its theoretical value of 0.511.