Alcoholic Fermentation with Self‐Flocculating Yeast in a Tower Upflow Reactor

Ethanol production using self‐flocculating yeast in a batch tower upflow reactor system operating with a recirculation loop was examined. Ethanol productivity, yield, and residual sucrose concentration were evaluated experimentally according to a central composite design with initial cell and sucrose concentrations and recirculation flow rate as independent variables. Yeast cell concentration strongly influenced the reactor performance. Alcoholic fermentation was conducted using this strain and reactor configuration which allowed for high productivity and high sucrose conversion. The ethanol yield was comparable with industrial yields. A kinetic study of the fermentation process under optimized conditions was performed using the experimental data and considering inhibition by sucrose and ethanol.     

Effect of biological ageing of wine on its nitrogen composition for producing high quality vinegar

There is increasing interest in producing novel high quality vinegars. In this work, we examined the effect of biological ageing of white wine on its nitrogen composition (viz. amino acids, urea and ammonium ion, which constitute the main sources of nitrogen for acetic bacteria during the acetification process) with a view to confirming the suitability of aged wine for producing quality vinegar.Available nitrogen contents in biologically aged wine were lower than in young (unaged) wine; this resulted in a slightly lower acetification rate and production with the former. The nitrogen composition of the two vinegars was very similar, with l-proline and l-cysteine as the major amino acids. By exception, only the vinegar from the young wine contained ammonium ion. The ageing process by flor yeast produces urea which is eliminated by the bacteria in the next stage.Available nitrogen for use by acetic bacteria in biologically aged wine is seemingly no limiting factor for acetification. The problem posed by the formation of urea during wine ageing was suppressed by the subsequent acetification process. The study could be an example of how different microorganisms use the available substrate in serial biotransformations.     

The effects of dilution rate and glucose concentration on continuous acetone–butanol–ethanol fermentation by Clostridium acetobutylicum immobilized on bricks

BACKGROUND: Owing to the rapid depletion of petroleum fuel, the production of butanol through biological routes has attracted increasing attention. However, low butanol productivity severely impedes its potential industrial production. It is known that the immobilization of whole cells can enhance productivity in the acetone‐butanol‐ethanol (ABE) continuous fermentation process. Therefore, the objective of this study was to develop a low‐cost continuous operation for butanol production. RESULTS: Bricks were chosen as cell support because of their low cost and ease of use for immobilization. The solvent productivity for the bricks with immobilized cells was 0.7 g L^?1 h^?1, 1.89 times that of free cells (0.37 g L^?1 h^?1) at a dilution rate of 0.054 h^?1. The productivity improvement can contribute to greater retention of biomass inside the reactor due to immobilization. The increase in glucose feed concentration raised total solvent production. However, it resulted in a decrease in yield (grams of solvents produced per gram of glucose introduced). Continuous operation with immobilized cells at a dilution rate of 0.107 h^?1 resulted in a solvent productivity of 1.21 g L^?1 h^?1, 2.1 times that of the operation at 0.027 h^?1. However, the yield (butanol produced per glucose consumed) was decreased to 0.19 from 0.29 under the same glucose feeding condition of 60 g L^?1. CONCLUSION: The increase in dilution rate and feed glucose concentration enhanced productivity, but decreased the utilization of substrates and the final solvent concentration. Therefore, a balance between productivity and glucose utilization is required to ensure continuous process operation. Copyright © 2011 Society of Chemical Industry     

Dynamic modelling,simulation and optimization of an extractive continuous alcoholic fermentation process

The conventional alcoholic fermentation is a typical inhibitory process, leading to low productivity and yield. The ethanol produced inhibits yeast cells, causing a reduction in the alcohol production rate and cell growth rate. In this work, modelling and simulation have shown that continuous extractive fermentation, coupling a fermenter with an extractive vacuum flash chamber, is technically possible. In this case, the ethanol is partially removed, increasing drastically the productivity. Additionally, temperature control can be performed without using heat exchangers. The optimization was carried out using the method of factorial design and response surface analysis, leading to the determination of the most relevant variables, which were: 1.2 h residence time, 0.4 flash recycle rate, 180 g dm⁻³ sugar concentration and 0.35 cell recycle rate. The results, using optimized variables, were 98% conversion and 23 g dm⁻³ h⁻¹ productivity, which represent a three times higher productivity than in a conventional continuous process. © 1999 Society of Chemical Industry     

Wood blocks as a carrier for Saccharomyces Cerevisiae used in the production of ethanol and fructose

Saccharomyces cerevisiae ATCC 39859 was immobilized onto small cubes of wood in order to produce very enriched fructose syrup from synthetic glucose-fructose mixtures, through the selective fermentation of glucose. The kinetics of growth and ethanol production rates were studied. Several tests to assess the influence of substrate and product concentration on the production rates were carried out and appropriate rate equations were proposed as a design basis for continuous immobilized reactors. The ethanol production rate and cell growth rate were found to be inhibited linearly by both substrate and product concentrations. A maximum ethanol productivity of 21.9 g 1⁻¹ h⁻¹ was attained from a feed containing 10% (by weight) glucose and 10% (by weight) fructose. The ethanol concentration was 29.6 g 1⁻¹, the glucose conversion was 78% and a fructose yield of 99% was obtained. This resulted in a final fructose:glucose ratio of 2.7. At lower ethanol productivity levels the fructose:glucose ratio increased, as did the ethanol concentration in the effluent. The ethanol productivities obtained in this study were 33%–132% higher than those obtained in a previous study using the same system, under similar conditions, with the cells immobilized in alginate beads.     

Different process configurations for bioethanol production from pretreated olive pruning biomass

BACKGROUND: In Mediterranean countries, olive tree pruning provides a widely available renewable agricultural residue with, currently, no industrial application. This residue could provide feedstock for the bioethanol industry. In the present study, olive tree pruning biomass pretreated with both ‘liquid hot water’ and ‘dilute‐sulfuric acid’ was tested as a substrate for ethanol production. Three different process configurations, separate hydrolysis and fermentation (SHF), simultaneous saccharification, fermentation and prehydrolysis (PSSF), and simultaneous saccharification and fermentation (SSF), were compared at different water‐insoluble solids concentrations. RESULTS: High ethanol concentration of about 3.7% (v/v) was obtained by separate hydrolysis and fermentation or prehydrolysis and simultaneous saccharification and fermentation of liquid hot water pretreated at 23% (w/w) substrate loading. CONCLUSION: The nature of the pretreated residue allows high substrate concentration (≥17% w/w) to be used in the enzymatic hydrolysis step. Substrate loading of 17% DM has been shown to provide a compromise between hydrolysis efficiency and glucose concentrations for the same enzyme/substrate ratio. Prehydrolysis prior to simultaneous saccharification and fermentation facilitated SSF performance at high substrate loading on liquid hot water pretreated olive pruning residue. This effect was not observed with dilute‐acid pretreated substrate. Copyright © 2011 Society of Chemical Industry     

ANALYSIS OF A TWO-STAGE FERMENTOR WITH RECYCLE FOR CONTINUOUS ETHANOL PRODUCTION

The performance of a recycle two-stage fermentor with cell separators after each stage is analyzed numerically for the continuous production of ethanol. In this system, the bleed withdrawn from the first stage is introduced to the second tank in order to reuse the cells in the bleed. Kinetic expressions and parametric values are taken from the literature. The effects of operating parameters on the concentrations in the stages are examined. Also investigated are the critical dilution rate for washout, the ethanol productivity and the substrate conversion. In addition, the influences of the saturation constant and the equilibrium partition coefficient of ethanol are examined. With respect to the overall productivity and conversion, it is found that the present two-stage system is more efficient than either a recycle chemostat or a recycle two-stage fermentor with a single separator after the final stage.     

不同类型载气对乙醇气提发酵的影响

研究了在乙醇气提发酵过程中,分别使用空气、二氧化碳、氮气和高纯氮作为气提载气对细胞生理及乙醇发酵的影响. 结果表明,空气能维持细胞生长和活力,但发酵效果最差,二氧化碳使细胞发酵能力波动不稳定且抑制副产物甘油合成;而以浓度为99.5%～99.8%的氮气为气提载气取得了较好的效果,乙醇和甘油的生产速率比不气提批式发酵分别提高2倍和1.9倍. 对以高纯氮(99.999%)为气提载气的研究发现,发酵进行到47 h开始出现菌种大量死亡现象,此时适当通氧和补充生长所需营养物质,细胞数量回升,活性恢复,最终乙醇生产速率比不气提时提高1.3倍.     

Optimisation of a continuous flash fermentation for butanol production using the response surface methodology

Factorial design and response surface techniques were used in combination with mathematical modelling and computational simulation to optimise an innovative industrial bioprocess, the production of biobutanol employing the flash fermentation technology. A parametric analysis performed by means of a full factorial design at two levels determined the influence of operating variables on butanol yield and productivity. A second set of simulations were carried out based on the central composite rotatable design. This procedure generated simplified statistical models that describe butanol yield and productivity as functions of the significant operating variables. From these models, response surfaces were obtained and used to optimise the process. For a range of substrate concentration from 130 to 180 g/l, the optimum operating ranges ensure butanol productivity between 7.0 and 8.0 g/l h, butanol yield between 19 and 22%, substrate conversion above 90% and final butanol concentration around 25 g/l.     

Kinetic study on ethanol production using Saccharomyces cerevisiae ITV‐01 yeast isolated from sugar cane molasses

BACKGROUND: Bio‐ethanol production from renewable sources, such as sugar cane, makes it a biofuel that is both renewable and environmentally friendly. One of the strategies to reduce production costs and to make ethanol fuel economically competitive with fossil fuels could be the use of wild yeast with osmotolerance, ethanol resistance and low nutritional requirements. The aim of this work was to investigate the kinetics of ethanol fermentation using Saccharomyces cerevisiae ITV‐01 yeast strain in a batch system at different glucose and ethanol concentrations, pH values and temperature in order to determine the optimum fermentation conditions. RESULTS: This strain showed osmotolerance (its specific growth rate (µ_max) remained unchanged at glucose concentrations between 100 and 200 g L^?1) as well as ethanol resistance (it was able to grow at 10% v/v ethanol). Activation energy (Ea) and Q₁₀ values calculated at temperatures between 27 and 39 °C, pH 3.5, was 15.6 kcal mol^?1 (with a pre‐exponential factor of 3.8 × 10¹² h^?1 (R² = 0.94)) and 3.93 respectively, indicating that this system is biologically limited. CONCLUSIONS: The optimal conditions for ethanol production were pH 3.5, 30 °C and initial glucose concentration 150 g L^?1. In this case, a maximum ethanol concentration of 58.4 g L^?1, ethanol productivity of 1.8 g L^?1 h^?1 and ethanol yield of 0.41 g g^?1 were obtained. Copyright © 2010 Society of Chemical Industry     

Bioethanol production from corn meal by simultaneous enzymatic saccharification and fermentation with immobilized cells of Saccharomyces cerevisiae var. ellipsoideus

The simultaneous enzymatic saccharification and fermentation (SSF) of corn meal using immobilized cells of Saccharomycescerevisiae var. ellipsoideus yeast in a batch system was studied. The yeast cells were immobilized in Ca-alginate by electrostatic droplet generation method. The process kinetics was assessed and determined and the effect of addition of various yeast activators (mineral salts: ZnSO₄ · 7H₂O and MgSO₄ · 7H₂O, and vitamins: Ca-pantothenate, biotin and myo-inositol) separately or mixed, was investigated. Taking into account high values of process parameters (such as ethanol concentration, ethanol yield, percentage of the theoretical ethanol yield, volumetric productivity and utilized glucose) and significant energy savings the SSF process was found to be superior compared to the SHF process. Further improvement in ethanol production was accomplished with the addition of mineral salts as yeast activators which contributed to the highest increase in ethanol production. In this case, the ethanol concentration of 10.23% (w/w), percentage of the theoretical ethanol yield of 98.08%, the ethanol yield of 0.55 g/g and the volumetric productivity of 2.13 g/l·h were obtained.     

Response Surface Optimization of Ultrasonic-Assisted Oxyresveratrol Extraction from the Bark of Cultured Black Mulberry (Morus nigra L.)

Ultrasonic-assisted extraction was applied to extract oxyresveratrol from cultured black mulberry bark (OCBMB). The process was optimized by response surface methodology. According to the results of the signal factor test, the Box–Behnken design was employed to investigate the effects of four chosen independent variables (ultrasonic power, extraction time, ethanol concentration, and acetic acid concentration) on the oxyresveratrol extraction, and the response surface plots showed that the parameters exhibited interactive effects. The optimal conditions were as follows: extraction temperature, 50°C; ratio of solution to solid (v/m), 20:1; ultrasonic power, 496.23 W; extraction time, 113 min; ethanol concentration, 70.98%; acetic acid concentration, 0.77%. Under these optimal conditions, experimental values well matched with the predicted values, which indicated the model used had extremely good fitting degree and the response surface methodology for optimizing Ultrasonic-assisted extraction of OCBMB was successful.     

Second-generation ethanol production from steam exploded barley straw by Kluyveromyces marxianus CECT 10875

Barley straw is nowadays being considered a potential lignocellulosic raw material for fuel-ethanol production as an alternative to starch- or sugar-containing feedstock. In this work, several configuration strategies for ethanol production from steam-exploded barley straw by Kluyveromyces marxianus CECT 10875 have been studied with the aim of obtaining higher ethanol concentrations.Different substrate loading (2-15%, w/v) were studied during enzymatic hydrolysis. The xylanase contribution on glucose production and glucan conversion at different substrate loading was also investigated. In addition, three different process configurations, separate hydrolysis and fermentation, simultaneous saccharification and fermentation and presaccharification and simultaneous saccharification, were compared at different water insoluble solids concentration (5%, 10% and 15%). The influence of xylanase addition on the ethanol yield was studied as well.Results show that endo-xylanases improved glucan conversion and ethanol yield compared with a standard enzymatic mixture, markedly at low substrate concentration. The positive effect of added xylanase was most evident at early stages of enzymatic hydrolysis. Regarding process configurations for the period of 72 h, SSF with endo-xylanases provided the best ethanol yield, nearly 70%, for 10% WIS. Nonetheless, the higher ethanol concentration, 29.4 g/l, was obtained at 15% WIS.     

An approach to optimization of enzymatic hydrolysis from sugarcane bagasse based on organosolv pretreatment

BACKGROUND: The organosolv pretreatment followed by enzymatic hydrolysis of the pretreated material and subsequent fermentation of the hydrolysate produced, was the strategy used for ethanol production from sugarcane bagasse. The effect of different operational variables affecting the pretreatment (the catalyst type and its concentration, and the pretreatment time) and enzymatic hydrolysis stage (substrate concentration, cellulase loading, addition of xylanase and Tween 20, and the cellulase/β‐glucosidase ratio), were investigated. RESULTS: The best values of glucose concentration (28.8 g L^?1) and yield (25.1 g per 100 g dry matter) were obtained when the material was pretreated with 1.25% (w/w) H₂SO₄ for 60 min, and subsequently hydrolyzed using 10% (w/v) substrate concentration in a reaction medium supplemented with xylanase (300 UI g^?1) and Tween 20 (2.5% w/w). Fermentation of the broth obtained under these optimum conditions by Saccharomyces cerevisiae resulted in an ethanol yield of 92.8% based on the theoretical yield, after 24 h. CONCLUSION: Organosolv pretreatment of sugarcane bagasse under soft conditions, and subsequent enzymatic hydrolysis of the pretreated material with a cellulolytic system supplemented with xylanase and Tween 20, is a suitable procedure to obtain a glucose rich hydrolysate efficiently fermentable to ethanol by Sacharomyces cerevisiae yeasts. Copyright © 2010 Society of Chemical Industry     

同步闪蒸汽提发酵制乙醇

为提高乙醇生产发酵强度,提出了同步汽提闪蒸乙醇发酵新过程。该过程集合了闪蒸发酵和汽提发酵的优点,在发酵的同时能更有效地在位分离乙醇,从而提高发酵强度。通过与普通发酵、闪蒸发酵、汽提发酵等过程进行间歇实验比较,同步闪蒸汽提发酵过程具有发酵强度高的优势;而且,采用耐高温酵母高温发酵、提高通气量、闪蒸罐的进料流速可进一步提高其发酵强度。该过程简单、高效,具有工业应用的潜力。     

On the Degradation and Stabilization of Poly(Methyl Methacrylate) in a Continuous Process

Thermally unstable polymers such as poly(methyl methacrylate) are degraded considerably during industrial processing. This degradation and its reduction to a minimum have been investigated in both lab and continuous pilot‐scale experiments. A three‐step degradation mechanism, starting at 180 °C, was proved by Thermogravimetrical Analysis (TGA) and a kinetic approach to describe it was derived. The knowledge of this degradation behavior was then applied to a pilot‐scale process with a production rate of 10 kg/h and the process yield loss during the devolatilization step was investigated. Using heat stabilizers, the overall process yield could be improved by 10 %, whereas the residual organic volatiles concentration (VOC) was drastically reduced to values below 1000 ppm. In order to preserve the molecular weight of the final product these stabilizers were added into the process, separately, at the end of the polymerization reaction but before the devolatilization step.     

Production of bioethanol under high pressure of CO2: The effect of process conditions

The aim of this work is to investigate the possibility of producing ethanol by glucose fermentation under high pressure of carbon dioxide up to 48 bar, in order to exploit both ethanol and denser CO₂ as a by-product of the process. The fermentation is carried out using Ethanol Red™-Lesaffre Saccharomyces cerevisiae yeast strain, which is commonly applied in industrial bioethanol production.The experiments were performed in six small reactors (2 mL of volume each) connected in parallel, to investigate the effect of the process variables at the same conditions of temperature and pressure, and in one pilot reactor (1 L of volume) to confirm the results obtained at the lower scale.The influence of operative variables, such as carbon dioxide pressure (0–48 bar), temperature (32 and 36 °C), glucose (150–250 g/L), inositol (0–400 mg/L) and biomass concentration (OD 2 and 3.5), was measured in terms of ethanol concentration (by gas chromatography) and ethanol productivity (expressed as grams of ethanol per CFU of yeast). Both of these parameters were found to be strongly dependent on glucose concentration and CO₂ pressure, which negatively affects the fermentation. Nevertheless, also at 50 bar it is possible to produce appreciable amounts of ethanol.     

Production of pure ethanol from azeotropic solution by pressure swing adsorption

Pressure swing adsorption (PSA) is attractive for final separation in the process of water removal especially for fuel ethanol production. Despite many researches on simulation and experimental works on adsorption of water on 3A zeolite in a fixed bed, none have studied a process with the actual PSA system. The purpose of this research was to study the PSA process with two adsorbers and effects of several parameters. The research also included analysis of kinetic and thermodynamic data of ethanol-water adsorption on commercial 3A zeolites in a single fixed bed. A two-level factorial design experiment was used in this research work to preliminarily screen the influence and interaction among the factors. Effects of important parameters such as initial temperature, feed concentration and feed rate were investigated. It was proven that the Langmuir isotherm could best predict the experimental results. In the PSA pilot test, the principal factors, which had effects on the performance, were feed rate, feed concentration, adsorption pressure and the cycle time. Prediction of the process efficiency in terms of ethanol recovery and enrichment was proposed in the form of regression models. The results of the study in a fixed bed adsorber could help designing a pilot-scale PSA unit. The experiments proved to be successful in terms of producing high concentration ethanol with high percentage of ethanol recovery. With further simulation work the process could be scaled up for an industrial use.     

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.     

Comparison of bioethanol production of simultaneous saccharification & fermentation and separation hydrolysis & fermentation from cellulose-rich barley straw

Cellulose rich barley straw, which has a glucan content of 62.5%, followed by dilute acid pretreatment, was converted to bioethanol by simultaneous saccharification and fermentation (SSF). The optimum fractionation conditions for barley straw were an acid concentration of 1% (w/v), a reaction temperature of 158 °C and a reaction time of 15 min. The maximum saccharification of glucan in the fractionated barley straw was 70.8% in 72 h at 60 FPU/gglucan, while the maximum digestibility of the untreated straw was only 18.9%. With 6% content WIS (water insoluble solid) for the fractionated barley straw, 70.5 and 83.2% of the saccharification yield were in SHF and SSF (representing with glucose equivalent), respectively, and a final ethanol concentration of 18.46 g/L was obtained under the optimized SSF conditions: 34 °C with 15 FPU/g-glucan enzyme loading and 1 g dry yeast cells/L. The results demonstrate that the SSF process is more effective than SHF for bioethanol production by around 18%.