A continuous, farm-scale, solid-phase fermentation process for fuel ethanol and protein feed production from fodder beets

Fuel ethanol (95%) was produced from fodder beets in two farm-scale processes. In the first process, involving conventional submerged fermentation of the fodder beets in a mash, ethanol and a feed (PF) rich in protein, fat, and fiber were produced. Ethanol yields of 70 L/metric ton (7 gal/ton) were obtained; however, resulting beers had low ethanol concentrations [3-5% (v/v)]. The high viscosity of medium and low sugar, beet mashes caused mixing problems which prevented any further increase of beet sugar in the mash. The severely limited the maximum attainable ethanol concentration during fermentation, thereby making the beer costly to distill into fuel ethanol and the process energy inefficient. In order to achieve distillably worthwhile ethanol concentrations of 8-10% (v/v), we developed and tested a solid-phase fermentation process (continuous). In preliminary trials, this system produced fermented pulp with over 8% (v/v) ethanol corresponding to an ethanol yield of 87 L/metric ton (21 gal/ton). Production costs with this novel process are $0.47/L ($1.77/gal) and the energy balance is 2.11. These preliminary cost estimates indicate that fodder beets are potentially competitive with corn as an ethanol feedstock. Additional research, however, is warranted to more precisely refine individual costs, energy balances and the actual value of the PF.     

Semicontinuous diffusion fermentation of fodder beets for fuel ethanol and cubed protein feed production

A novel, semicontinuous diffusion fermentation system was used to produce fuel ethanol and a cubed protein feed (CPF) from fodder beets at an intermediate scale. In the process, fodder beet cubes were augered diagonally upward against a flow of 0.26N H(2)SO(4) and yeast in a tubular fermentor. Exiting one end of the fermentor was CPF, while fermented beer [6-9% (v/v) ethanol] exited the other end. Retention times for beer and CPF were 264 and 72 h, respectively. Contamination was controlled by maintaining the fermentation pH between 2.1 and 2.6 using H(2)SO(4). Production costs for a greatly scaled-up (times 1400) conceptual version of this system (using a continuous rather than a semicontinuous processing mode) were projected by calculation to be $0.529/L for 95% ethanol (net of a $0.112/L credit for CPF). The calculated energy balance (energy output-energy input ratio) was estimated to be 3.04. In calculating the energy balance, the output energy of the CPF and input energy for growing the fodder beets were not included. A design for the scaled-up plant is provided.     

Ethanol production from fodder beets

Various yeasts such as two strains of Saccharomyces cerevisiae, Saccharomyces diastaticus, and Kluyveromyces marxianus were investigated for their ability to ferment fodder beet juice to alcohol. Juice extracted from fodder beet roots without any additives was used as a fermentation substrate. The fermentation kinetic parameters were determined and compared for each species of yeast tested. The best species for fodder beet juice fermentation was chosen and products obtained by fermentation of one hectare of fodder beet plants are given.     

Effect of pulp pH on solid phase fermentation of fodder beets for fuel ethanol production

Summary The pH of fodder beet pulp was varied to see how this affected solid phase fermentation by yeast. The process is for fuel ethanol production. When pulp was adjusted to a pre-inoculation pH of 3.0–3.5, ethanol yields (78–85% of theoretical, averaging 8.9% v/v) and fermentation efficiencies (97–99%) were greatest, the fermentation time was the shortest (30–39 h) and no bacterial contamination occurred.     

Effect of fodder beet cube size on ethanol production via diffusion fermentation

Summary The size of fodder beet cubes used to produce fuel ethanol via diffusion fermentation was varied to see how this affected various fermentation parameters. The highest yeast populations, shortest fermentation times, and highest ethanol yields and fermentation efficiencies were observed when 2.54 cm square cubes (or smaller) were utilized. Ethanol concentrations here averaged 4.21% (v/v) while the highest concentration reached was 4.83%. To minimize energy for slicing beets and still optimize yields, cubes of 1.91–2.54 cm should be used.     

Solid phase fermentation of foder beets for ethanol production: Effect of grind size

Solid phase fermentation of pulped fodder beets was studied to see what effect beet particle size had on various fermentation parameters. All trials were run in 4-l stainless stell containers and hammermilled pulp was initially adjusted to pH 3.0 to control bacterial contaminants. The maximum yeast population that built up in the pulp was independent of the hammermill screen size (0.476–1.905 cm) and averaged 2.0−2.3 × 10⁸ cells/ml. Pulp from finer screens (0.476–0.953 cm) took 19–22 h to reach a peak yeast population while pulp from coarser screens (1.270–1.905 cm) took a slightly longer 24–28 h. The time to reach maxium ethanol concentration was not affected by screeen size and averaged 28–30 h. Ethanol yields dropped slightly form 85–87% of theoretical with the finest screens to 83–84% with the coarset screens. The maximum ethanol concentration observed was 7.96% (v/v) and the average of all runs was 7.63% (v/v). Fermentation efficiency averaged 98–99% thoughout. The lack of a response to grinding fodder beets with different screens was due to their wet fibrous nature which hindered free flow of pulp though the screens. Pulp was, instead, extruded though the screens, forming particles of generally similar size. Our results indicate that the primary consideration for grind size is energy consumption for grinding. Therefore, if a hammermill is used, a large screen (1.270–1.905 cm) which requires less energy should be employed so as to minimize energy consumption. This strategy does not result in longer fermentation times or reduced ethanol yields.     

Optimization of fermentation conditions for ethanol production from whey

Summary Optimal conditions for ethanol production in 7% whey solutions by the yeast Candida pseudotropicalis ATCC 8619 included initial pH of 4.57 and 30°C. Complete fermentation of the available lactose took place without supplementary nutrients; additions of nitrogen or phosphorus salts, yeast extract or corn steep liquor resulted in increased yeast production and lower ethanol yields. A positive correlation was observed between increases in yeast inocula and lactose utilization and ethanol production rates; 8.35 g/l of ethanol was obtained within 22 h by using yeast inoculum of 13.9 g/l. No differences in fermentation rates or ethanol yields were observed when whole or deproteinized whey solutions were used. Concentrated whey permeates, obtained after removal of the valuable proteins from whey, can be effectively fermented for ethanol production.     

海带发酵制取生物乙醇的影响因素与优化条件

针对海带的碳水化合物不易被单一菌株发酵转化为乙醇的难题,通过酸化、匀浆和消化等预处理和正交试验,利用多酶系多菌种微生物复合发酵剂的酿酒曲,研究海带发酵制取生物乙醇的影响因素与优化条件。结果表明:在预处理试验中,加入一定量的Na2CO3,可以提高海带液中还原性糖和总糖的含量;消化温度对总糖影响相对较大,而对还原性糖的影响较小;过滤不利于得到较高浓度的乙醇;在优化条件中,发酵液的初始酸碱度是最重要的,其次是发酵温度和基质浓度,发酵液体积的影响程度相对较小。在基质(海带)质量浓度为0.15 g/L、温度34℃、起始pH 6.5和发酵液体积200 mL时,可以获得最大的乙醇产量4.09 g(以100 g海带计)。     

Modelling ethanol production from cellulose: separate hydrolysis and fermentation versus simultaneous saccharification and fermentation

In ethanol production from cellulose, enzymatic hydrolysis, and fermentative conversion may be performed sequentially (separate hydrolysis and fermentation, SHF) or in a single reaction vessel (simultaneous saccharification and fermentation, SSF). Opting for either is essentially a trade-off between optimal temperatures and inhibitory glucose concentrations on the one hand (SHF) vs. sub-optimal temperatures and ethanol-inhibited cellulolysis on the other (SSF). Although the impact of ethanol on cellobiose hydrolysis was found to be negligible, formation of glucose and cellobiose from cellulose were found to be significantly inhibited by ethanol. A previous model for the kinetics of enzymatic cellulose hydrolysis was, therefore, extended with enzyme inhibition by ethanol, thus allowing a rational evaluation of SSF and SHF. The model predicted SSF processing to be superior. The superiority of SSF over SHF (separate hydrolysis and fermentation) was confirmed experimentally, both with respect to ethanol yield on glucose (0.41 g g^?1 for SSF vs. 0.35 g g^?1 for SHF) and ethanol production rate, being 30% higher for an SSF type process. High conversion rates were found to be difficult to achieve since at a conversion rate of 52% in a SSF process the reaction rate dropped to 5% of its initial value. The model, extended with the impact of ethanol on the cellulase complex proved to predict reaction progress accurately.     

Aeration alleviates ethanol inhibition and glycerol production during fed-batch ethanol fermentation

In this study, we investigated the effects of aeration on ethanol inhibition and glycerol production during fed-batch ethanol fermentation. When aeration was conducted at 0.13, 0.33, and 0.8 vvm, the ethanol productivity, specific ethanol production rate, and ethanol yield in the presence of greater than 100 g/L of ethanol were higher than when aeration was not conducted. In addition, estimation of the parameters (α and β) in a model equation of ethanol inhibition kinetics indicated that aeration alleviated ethanol inhibition against the specific growth rate and the specific ethanol production rate. Specifically, when aeration was conducted, the glycerol yield and specific glycerol production rate decreased approximately 50 and 70%, respectively. Finally, the results of this study indicated that aeration during fed-batch ethanol fermentation may improve the ethanol concentration in the final culture broth, as well as the ethanol productivity.     

Technology and economics of fermentation alcohol — An update

Fermentation alcohol is being widely studied as an alternative fuel, and production is increasing, especially in Brazil, where the goal is more than 10 billion litres per year by 1985.Fuel markets are hundreds of times greater than the traditional ethanol markets which the existing industry supplies. To make a material contribution to fuel supply, fermentation ethanol must be treated as a major chemical and produced in large-volume, highly efficient plants. Such plants must be assured of a continuous supply of low-cost raw materials for which suitable processes have been developed and commercially proven. Sugar cane in the tropics and grains in some temperate countries meet these requirements; cellulosics do not qualify at present, nor will they in the foreseeable future, without major breakthroughs.Using techniques borrowed from the starch sweetener industry, starchy materials may be economically hydrolysed to fermentable sugars; rapid acid hydrolysis may prove superior to enzymatic processes. Major projects are under way to replace traditional batch or cascade fermentations with rapid, single-vessel continuous units, but these have not yet been fully proven. Where suitable, yeast recycle is being used as a means of increasing alcohol yields, and energy-efficient distillation methods of the petrochemical industry are being adopted. The consequent large reduction in steam consumption greatly reduces the appeal of other methods which have been proposed to remove water.Opportunities for process improvements abound, especially in developing (1) the means to provide cellulosic raw materials in large quantities at acceptable costs, (2) economically effective methods of pretreating and hydrolysing cellulosics, (3) practical organisms for converting five-carbon sugars to ethanol and (4) higher fermentation yields and efficiencies using bacteria or immobilized yeast.     

Enhanced ethanol production via fermentation of rice straw with hydrolysate-adapted Candida tropicalis ATCC 13803

Neutralized hydrolysate and pretreated rice straw obtained from a 2% (w/v) sulfuric acid pretreatment were mixed at 10% (w/v) and subjected to simultaneous saccharification and co-fermentation (SSCF), with cellulase, β-glucosidase, and Candida tropicalis cells at 15 FPU/g-ds, 15 IU/g-ds and 1 × 10⁹ cells/ml, respectively. A 36-h SSCF with adapted cells resulted in YP/S and ethanol volumetric productivity of 0.36 g/g and 0.57 g/l/h, respectively. In addition to ethanol, insignificant amounts of glycerol and xylitol were also produced. Adapted C. tropicalis cells produced nearly 1.6 times more ethanol than non-adapted cells. Ethanol yield (Yp/s), ethanol volumetric productivity and a xylitol concentration of 0.48 g/g, 0.33 g/l/h and 0.89 g/l, respectively, were produced from fermentation of remaining hydrolysate with adapted C. tropicalis cells. The 0.20 g/g ethanol yield and 77% production efficiency from SSCF of pretreated rice straw indicate scale-up potential for the process. This study demonstrated that C. tropicalis produced ethanol and xylitol from a mixed-sugar stream, although cell adaptation affected ethanol and xylitol yields. Scanning electron microscopy indicated agglomeration of cellulose microfibrils and globular deposition of lignin in acid-pretreated rice straw.     

Anaerobic bio-hydrogen production from ethanol fermentation: the role of pH

Hydrogen was produced by an ethanol-acetate fermentation at pH of 5.0 +/- 0.2 and HRT of 3 days. The yield of hydrogen was 100-200 ml g Glu(-1) with a hydrogen content of 25-40%. This fluctuation in the hydrogen yield was attributed to the formation of propionate and the activity of hydrogen utilizing methanogens. The change in the operational pH for the inhibition of this methanogenic activity induced a change in the main fermentation pathway. In this study, the main products were butyrate, ethanol and propionate, in the pH ranges 4.0-4.5, 4.5-5.0 and 5.0-6.0, respectively. However, the activity of all the microorganisms was inhibited below pH 4.0. Therefore, pH 4.0 was regarded as the operational limit for the anaerobic bio-hydrogen production process. These results indicate that the pH plays an important role in determining the type of anaerobic fermentation pathway in anaerobic bio-hydrogen processes.     

Development of a low-cost fermentation medium for ethanol production from biomass

Nutrient cost is an important aspect in the fermentation of biomass to ethanol. With a goal of developing a cost-effective fermentation medium, several industrially available nutrient sources were evaluated for their effectiveness in the simultaneous saccharification and fermentation of pretreated poplar with Saccharomyces cerevisiae D₅A. These studies showed that a low-cost medium containing 0.3% corn steep liquor and 2.5 mM MgSO₄ · 7H₂O was similar in performance to a nutrient-rich medium. Besides its low cost, this alternative medium consists of components that are available on a commercial scale, thereby making it industrially relevant. Received: 14 August 1996 / Received revision: 7 January 1997 / Accepted: 24 January 1997     

Continuous tower fermentation for power ethanol production

Summary The capability of the continuous tower fermenter to accumulate and retain high cell densities (70–90 g dry wt/1) when using naturally flocculant yeasts is demonstrated with semi-defined glucose feed at concentrations of 120–200 g/1 and high hydraulic loadings. Conversion and ethanol productivity data are given as a function of throughput and feed glucose concentration.     

Processing and fermentation of Jerusalem artichoke for ethanol production

Summary Processing and fermentation trials on Jerusalem artichoke (Helianthus tuberosus L.) tubers, and on pure inulin media were carried out. Acid and thermal treatments, pure and mixed cultures of yeast, and enzyme preparations were investigated. Best ethanol yields on either substrate were obtained with pH 2 thermal treatments, resulting in 131.6 liters ethanol per metric ton fresh tuber.     

Valorization of olive stones for xylitol and ethanol production from dilute acid pretreatment via enzymatic hydrolysis and fermentation by Pachysolen tannophilus

Olive stones are an agro-industrial by-product abundant in the Mediterranean area that is regarded as a potential lignocellulosic feedstock for sugar production. Statistical modeling of dilute-sulphuric acid hydrolysis of olive stones has been performed using a response surface methodology, with treatment temperature and process time as factors, to optimize the hydrolysis conditions aiming to attain maximum d-xylose extraction from hemicelluloses. Thus, solid yield and composition of solid and liquid phases were assessed by empirical modeling. The highest yield of d-xylose was found at a temperature of 195 °C for 5 min. Under these conditions, 89.7% of the total d-xylose was recovered from raw material. The resulting solids from optimal conditions were assayed as substrate for enzymatic hydrolysis, while fermentability of hemicellulosic hydrolysates was tested using the d-xylose-fermenting yeast Pachysolen tannophilus. Both bioprocesses were considerably influenced by enzyme loading and inoculum size. In the enzymatic hydrolysis step, about 56% of cellulose was converted into d-glucose by using an enzyme/solid ratio of 40 FPU g⁻¹, while in the fermentation carried out with a cell concentration of 2 g L⁻¹ a yield of 0.44 g xylitol/g d-xylose and a global volumetric productivity of 0.11 g L⁻¹ h⁻¹ were achieved.