Ethanol production in an integrated process of fermentation and ethanol recovery by pervaporation

In ethanol fermentations inhibition of the microorganism by ethanol limits the amount of substrate in the feed that can be converted. In a process high feed concentrations are desirable to minimize the flows. Such high feed concentrations can be realized in integrated processes in which ethanol is recovered from the fermentation broth as it is formed. In this study ethanol recovery by pervaporation was coupled to glucose fermentations by baker's yeast. Pervaporation was carried out with commercial silicone based hollow-fibre membrane modules with relatively high fluxes. Three different types of process configurations with pervaporation were investigated. Two of these configurations also included cell retention by microfiltration, in order to optimize the productivity. In the systems with pervaporation a feed containing 360 kg/m³ glucose could be converted almost completely. This feed concentration is a factor three higher than in a process without ethanol recovery. The productivity was 14 kg/m³ h in a system with pervaporation only, and could be increased to 43 kg/m³ h in the system with all recycle by microfiltration. The kinetic data suggest that accumulation of inhibitory compounds occurs in the integrated system. The integrated process was relatively easy in operation.     

Ethanol from hydrolyzed whey permeate using Saccharomyces cerevisiae in a membrane recycle bioreactor

A diauxic fermentation was observed during batch fermentation of enzyme-hydrolyzed whey permeate to ethanol by Saccharomyces cerevisiae. Glucose was consumed before and much faster than galactose. In the continuous membrane recycle bioreactor (MRB), sugar utilization was a function of dilution rate and concentration of sugars. At a cell concentration of 160 kg/m³, optimum productivity was 31 kg/(m³ · h) at ethanol concentration of 65 kg/m³. Low levels of acetate (0.05–0.1 M) reduced cell growth during continuous fermentation, but also reduced galactose utilization.     

Effects of ethanol concentration on immobilized Zymomonas mobilis for continuous production of ethanol

The effects of ethanol concentration on the ethanol productivity and activity of immobilized Zymomonas mobilis cells during continuous fermentation of glucose has been studied at various ethanol concentrations. On changing the inlet ethanol concentration, Po, from 0.0 kg/m³ to any other level, 8 h were required to fully experience the effects of a change in Po, whereas 8 h to 2 days, depending on Po, were required to reach the steady state on switching back to the ethanol free medium. The volumetric ethanol productivity decreased from 92.5 to 0.0 kg/m³·h as the ethanol concentration in the bioreactor was changed from 46.3 to 126 kg/m³. The activity of the immobilized cells recovered up to 63% in 2 days even after exposing the cells to 126 kg/m³ of ethanol.     

Propionic acid fermentation: improvement of performances by coupling continuous fermentation and ultrafiltration

The present paper presents a study of propionic acid production from whey by using Propionibacterium acidipropionici in batch and continuous fermentation with cell recycle. The experimental investigation is carried through with a biomass concentration (DW) of 112kg/m³. The highest propionic acid productivity is 2.14 kg/(m³ h). Biomass concentration is 9 times as high, propionic acid productivity 6 times as high as compared to batch results.     

Production of high ethanol concentrations from glucose using a vertical rotating immobilized cell reactor of the bacterium zymomonas mobilis

Long-term continuous ethanol production of up to 80 g.l¹ with a volumetric ethanol productivity of 63 g. l^?1. h^?1 was maintained for more than 72 days using a Vertical Rotating Immobilized Cell Reactor of the bacterium Z. mobilis. Continuous production of higher ethanol concentration was unsuccessful due to an inhibition of cell growth by long exposure to high ethanol concentrations. However, ethanol concentration as high as 120g. l^?1 and volumetric ethanol productivity of 13g. l^?1. h^?1 were achieved in a repeated-batch fermentation system using the same bioreactor. By a simple washing operation at the end of each run, immobilized biomass could be effectively regenerated and used to carry out more than 10 successive fermentation cycles.     

The influence of cyclic glucose feeding on a continuous bakers' yeast culture

Conclusions Except for the pronounced adaptation-hysteresis effect, the pulse experiments exhibited the expected trend: deviation from the steady feed reference curve was greatest at lowest dilution rates. Under conditions of lowest glucose level the effect of pulsing would be expected to cause the largest fluctuations in glucose, causing a certain fraction of the cells to ferment. Generally over the entire dilution rate range the biomass production was decreased and the ethanol was increased by pulsing the feed stream. There is, however, some evidence that pulse feeding can trigger quite unexpected results. Point (6) at D=0.3 h^–1 in Fig. 1 exhibit a biomass productivity which was about 20% greater than the continuous feeding reference value (DX=3.6 kg m^–3 h^–1 as compared with 3.0 kg m^–3 h^–1). Such performance would be of significant commercial value, but the poor reproducibility due to adaptation, as seen here, certainly would preclude its application.The results obtained should also be applicable to fed batch operation at the corresponding glucose level. Further experiments including the variation of the glucose feeding period would be necessary to obtain a conclusive picture. The observed phenomena are likely to occur in other fermentations and could eventually explain some of the problems existing with scale up of fermentation processes.Symbols D dilution rate h^–1 - P product (ethanol) concentration kg m^–3 - Q_O2 specific oxygen uptake rate mol kg^–1 s^–1 - Q_CO2 specific CO₂ production rate mol kg^–1 s^–1 - S substrate (glucose) concentration kg m^–3 - X biomass concentration kg m^–3 - Y_P/S yield of ethanol from glucose kg kg^–1 - Y_X/S yield of biomass from glucose kg kg^–1     

Solid-state fermentation of carob pods for ethanol production

The production of ethanol from carob pods by Saccharomyces cerevisiae in solid-state fermentation was investigated. The maximal ethanol concentration (160±3 g/kg dry pods), ethanol productivity (6.7 ± 0.2 g/kg per hour), ethanol yield (40 ± 1.8%), biomass concentration (7.5 ± 0.4 x 10⁸ cells/g carob pulp) and fermentation efficiency (80 ± 2%) were obtained at an inoculum amount of 3%, a particle size of 0.5 mm, a moisture level of 70%, a pH of 4.5 and a temperature of 30°C. Under the same fermentation conditions both sterilized and non-sterilized carob pods pulp gave the same maximum ethanol concentration.     

Effect of substrate concentration on ethanol production by Zymomonas mobilis on cellulose hydrolysate

Summary A cellulose hydrolysate from Aspen wood, containing mainly glucose, was fermented into ethanol by a thermotolerant strain MSN77 of Zymomonas mobilis. The effect of the hydrolysate concentration on fermentation parameters was investigated. Growth parameters (specific growth rate and biomass yield) were inhibited at high hydrolysate concentrations. Catabolic parameters (specific glucose uptake rate, specific ethanol productivity and ethanol yield) were not affected. These effects could be explained by the increase in medium osmolality. The results are similar to those described for molasses based media. Strain MSN77 could efficiently ferment glucose from Aspen wood up to a concentration of 60 g/l. At higher concentration, growth was inhibited.Nomenclature S glucose concentration (g/l) - X biomass concentration (g/l) - P ethanol concentration (g/l) - C conversion of glucose (%) - t fermentation time (h) - q_S specific glucose uptake rate (g/g.h) - q_p specific ethanol productivity (g/g.h) - YINX/S biomass yield (g/g) - Y_p/S ethanol yield (g/g) - specific growth rate (h^-1)     

Oxalic acid production by Aspergillus niger

Aspergillus niger is able to produce a quite high concentration of oxalic acid using sucrose as carbon and energy source. Operating at pH higher than 6 and an enriched N and P medium is necessary in order to conduct the fermentation towards oxalic acid production. A pH?shift technique, operating at acid pH?in the first two days and then setting pH?to 6, allowed the productivity to slightly increase in shaking flasks cultures up to 3.0?kg/m³?·?d, with a final oxalic acid concentration of 29?kg/m³. When operating at more controlled conditions, in a stirred tank, both productivity and oxalic acid concentration were improved (4.1?kg/m³?·?d and 33.8?kg/m³, respectively). However the main drawback of this fermentation is the low yield attained (about 0.3?kg oxalic acid/kg sucrose) because most of glucose, resulting from the hydrolysis of sucrose by the extracellular enzymes secreted at the beginning of the fermentation, is very quickly oxidised to gluconic acid, a process which is favoured at a pH?close to 6. Milk whey was proved to be a very good substrate as it allows oxalic acid to be produced with a similar productivity (2.5?kg/m³?·?d in shaking flasks) giving excellent yields of almost 0.6?kg oxalic acid/kg lactose.     

In-situ recovery of butanol during fermentation

End-product inhibition in the acetone-butanol fermentation was reduced by using extractive fermentation to continuously remove acetone and butanol from the fermentation broth. In situ removal of inhibitory products from Clostridium acetobutylicum resulted in increased reactor productivity; volumetric butanol productivity increased from 0.58 kg/(m³h) in batch fermentation to 1.5 kg/(m³h) in fed-batch extractive fermentation using oleyl alcohol as the extraction solvent. The use of fed-batch operation allowed glucose solutions of up to 500 kg/m³ to be fermented, resulting in a 3.5- to 5-fold decrease in waste water volume. Butanol reached a concentration of 30–35 kg/m³ in the oleyl alcohol extractant at the end of fermentation, a concentration that is 2–3 times higher than is possible in regular batch or fed-batch fermentation. Butanol productivities and glucose conversions in fed-batch extractive fermentation compare favorable with continuous fermentation and in situ product removal fermentations.List of Symbols C _g kg/m³ concentration of glucose in the feed - C _w dm³/m³ concentration of water in the feed - F(t) cm³/h flowrate of feed to the fermentor at time t - V(t) dm³ broth volume at time t - V _i dm³ initial broth volume - V _si dm³ volume of the i-th aqueous phase sample - effective fraction of water in the feed Part 1. Bioprocess Engineering 2 (1987) 1–12     

New cell recycle ethanol fermentation with periodic cleaning of filter with gas

High ethanol productivities were obtained by cell recycle cultures of yeast and bacterial strains at a dry cell concentration of 200 kg cells m^–3 using a new membrane bioreactor system. The filtration rates of the cultures were stabilized by removing the microbial cake on the filter with periodic back flows of the fermentation gas through the filter. For instance, the filtration flux of 0.023 m³m^–2h^–1 was maintained for 30 h with the periodic cleaning of the filter, whereas it decreased at a half time of 2 h without the cleaning. Ethanol productivity, ethanol concentration and filtration flux attained were: 68.7 kg/(m³ · h), 62.7 kg/m³ and 0.029 m³m^–2h^–1 for Saccharomyces carlsbergensis, LAM1068, the respective values for Zymomonas mobilis, ZM4, were: 93.7, 33 5 and 0.074.     

Simulation of the dynamics in the Baker's yeast process

Simulation of the dynamics in a fed batch process for production of Baker's yeast is discussed and applied. Experimental evidences are presented for a model of the energy metabolism. The model involves the concept of a maximum respiratory capacity of the cell. If the sugar concentration is increased above a critical value, corresponding to a critical rate of glycolysis and a maximum rate of respiration, then all additional sugar consumed at higher sugar concentrations is converted into ethanol.In a fed batch process with constant sugar feed the sugar concentration declines slowly. If ethanol is present when the sugar concentration declines below the critical value of 110 mg/dm³ fructose +glucose the metabolism switches rapidly into combined oxidation of sugar and ethanol. Thus, no diauxic growth is involved under process conditions. The rate of ethanol consumption is determined by the free capacity of respiration under these conditions. The invertase activity of the cells was found to be so high that mainly fructose and glucose were present in the medium, typically in the concentration range around 100 mg/dm³. These components are consumed at the same rate but with fructose at a higher concentration, indicating a higher K _s for fructose consumption.The model was used in simulation experiments to demonstrate the dynamics of the Baker's yeast process and the influence of different process conditions.List of Symbols DOT % air sat dissolved oxygen tension - F dm³/h rate of inlet medium flow - H kg/(dm³ % air sat.) oxygen solubility - K kg/m³ saturation constant specified by index - K _L a 1/h volumetric oxygen transfer coefficient - m g/(g · h) maintenance coefficient specified by index - P kg/(m³ · h) mean productivity of biomass in the process - q g/(g · h) specific consumption or production rate - S kg/m³ concentration of sugar in reactor - S ₀ kg/m³ concentration of inlet medium sugar medium t h process time - V dm³ medium volume - X kg/m³ concentration of biomass - Y g/g yield coefficient specified by index - 1/h specific growth rate Index aa anaerobic condition - c critical value - e ethanol - ec ethanol consumption - ep ethanol production - max maximum value - o oxygen - oe oxygen for growth on ethanol - os oxygen for growth on sugar - s sugar - x biomass     

Effects of glycerol on alcohol fermentation. Inhibition mechanism and diffusion limitations

Batch alcohol fermentations have been carried out varying the starting level of glycerol in the broth and keeping constant all the other fermentation parameters, in order to study the effect of the accumulation of this metabolite on the fermentation kinetics. A linear slow decrease of the maximum specific ethanol productivity with increasing glycerol level has been followed by a sharp fall of this parameter over a glycerol concentration threshold. A kinetic study through unstructured integrated models demonstrates that, at low concentrations, glycerol behaves as a non competitive inhibitor of fermentation, while, over a concentration threshold (105 kg/m³), an additional effect takes place, likely ascribable to diffusion limitations provoked by excess viscosity of the broth.     

Optimization of the microbial synthesis of dihydroxyacetone from glycerol with <Emphasis Type="Italic">Gluconobacter oxydans</Emphasis>

An optimized repeated-fed-batch fermentation process for the synthesis of dihydroxyacetone (DHA) from glycerol utilizing Gluconobacter oxydans is presented. Cleaning, sterilization, and inoculation procedures could be reduced significantly compared to the conventional fed-batch process. A stringent requirement was that the product concentration was kept below a critical threshold level at all times in order to avoid irreversible product inhibition of the cells. On the basis of experimentally validated model calculations, a threshold value of about 60 kg m^-3 DHA was obtained. The innovative bioreactor system consisted of a stirred tank reactor combined with a packed trickle-bed column. In the packed column, active cells could be retained by in situ immobilization on a hydrophilized Ralu-ring carrier material. Within 17 days, the productivity of the process could be increased by 75% to about 2.8 kg m^-3 h^-1. However, it was observed that the maximum achievable productivity had not been reached yet.Abbreviations K _O Monod half saturation constant of dissolved oxygen (kg m^-3) - K _S Monod half saturation constant of substrate glycerol (kg m^-3) - O Dissolved oxygen concentration (kg m^-3) - P Product concentration (kg m^-3) - P _crit Critical product concentration constant (kg m^-3) - S Substrate concentration (kg m^-3) - t Time (s) - X Biomass concentration (dry weight) (kg m^-3) - Y _P/S Yield coefficient of product from substrate - Y _X/S Yield coefficient of biomass from substrate - Growth dependent specific production rate constant (kg m^-3) - Growth independent specific production rate constant (s^-1) - Specific growth rate (s^-1) - _max Maximum specific growth rate constant (s^-1)     

Acclimation Strategy of a Biohydrogen Producing Population in a Continuous‐Flow Reactor with Carbohydrate Fermentation

Poor startup of biological hydrogen production systems can cause an ineffective hydrogen production rate and poor biomass growth at a high hydraulic retention time (HRT), or cause a prolonged period of acclimation. In this paper a new startup strategy was developed in order to improve the enrichment of the hydrogen‐producing population and the efficiency of hydrogen production. A continuously‐stirred tank reactor (CSTR) and molasses were used to evaluate the hydrogen productivity of the sewage sludge microflora at a temperature of 35 °C. The experimental results indicated that the feed to microorganism ratio (F/M ratio) was a key parameter for the enrichment of hydrogen producing sludge in a continuous‐flow reactor. When the initial biomass was inoculated with 6.24 g of volatile suspended solids (VSS)/L, an HRT of 6 h, an initial organic loading rate (OLR) of 7.0 kg chemical oxygen demand (COD)/(m³ × d) and an feed to microorganism ratio (F/M) ratio of about 2–3 g COD/(g of volatile suspended solids (VSS) per day) were maintained during startup. Under these conditions, a hydrogen producing population at an equilibrium state could be established within 30 days. The main liquid fermentation products were acetate and ethanol. Biogas was composed of H₂ and CO₂. The hydrogen content in the biogas amounted to 47.5 %. The average hydrogen yield was 2.01 mol/mol hexose consumed. It was also observed that a special hydrogen producing population was formed when this startup strategy was used. It is supposed that the population may have had some special metabolic pathways to produce hydrogen along with ethanol as the main fermentation products.     

High yeast concentration in continuous fermentation with cell recycle obtained by tangential microfiltration

Summary Continuous fermentation fed by 150 kg/m³ of glucose with total cell recycling by tangential microfiltration enabled yeasts concentration of 300 kg/m³ (dry weight) to be reached with a dilution rate of 0,5h^–1 and a cell viability greater than 75%. The stability of this system was tested for 50 residence times of the permeate. The method can be used both for the production of cell concentrates and for high rates of metabolite production.Nomenclature D. W. dry weight - X_T (kg/m³) total cell concentration D.W. - X_V (kg/m³) viable cell concentration D.W. - V viability of cell culture in per cent of total cell concentration - S (kg/m³) glucose concentration - P (kg/m³) ethanol concentration - D (h) dilution rate - R (kg/kg) fermentation yield - (h) specific growth rate - vp(kg/kg/h) specific alcohol production rate - (m) yeast size - (kg/kg) kg of intracellular water per kg of dry cells     

Model aided design of repeated fed-batch penicillin fermentation

Structured models of antibiotic fermentation that quantify maturation and aging of product forming biomass are fitted to experimental data. Conditions of superiority of repeated fed batch cultivation are characterized on the basis of a performance criterion that includes penicillin productivity and costs of operation. Emphasis is placed on the relevance of such research to the model aided design of optimal cyclic operation.List of Symbols c IU/mg cost factor - D s^–1 dilution rate - J IU · cm^–3 · h^–1 net productivity - k _p IU · mg^–11 · h^–1 specific product formation rate - k _pm IU · mg^–1 · h^–1 maximum specific product formation rate - p IU/cm³ concentration of penicillin - T s final time of fermentation - t s fermentation time - X kg/m³ concentration of biomass dry weight - X ₁kg/m³ concentration of young, immature biomass - X ₂ kg/m³ concentration of mature product forming biomass - X _c kg/m³ biomass concentration of the end of growth phase - X _mkg/m³ maximum biomass concentration Greek Letters s^–1 specific maturation rate - s^–1 specific aging rate - s^–1 specific growth rate - _m s^–1 maximum specific growth rate - _p s^–1 specific growth rate during the product formation phase - s cycle time - % volume fraction of draw-off Abbreviations CC chemostat culture - RFBC repeated fed batch culture - RBC repeated batch culture     

Process and technoeconomics of ethanol production by immobilized cells

Summary A two-stage fermentation process has been developed for continuous ethanol production by immobilized cells of Zymomonas mobilis. About 90–92 kg/m³ ethanol was produced after 4 h of residence time. Entrapped cells of Zymomonas mobilis have a capability to convert glucose to ethanol at 93% of the theoretical yield. The immobilized cell system has functioned for several weeks, and experience indicates that the carrageenan gel apparently facilitates easy diffusion of glucose and ethanol.The simplicity and the high productivity of the plug-flow reactor employing immobilized cells makes it economically attrative. An evaluation of process economics of an immobilized cell system indicates that at least 4 c/l of ethanol can be saved using the immobilized cell system rather than the conventional batch system. The high productivity achieved in the immobilized cell reactor results in the requirement for only small reactor vessels indicating low capital cost. Consequently, by switching from batch to immobilized processing, the fixed capital investment is substantially reduced, thus increasing the profitability of ethanol production by fermentation.     

Fed-batch cultivation of <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> on lignocellulosic hydrolyzate

Saccharomyces cerevisiae grows very poorly in dilute acid lignocellulosic hydrolyzate during the anaerobic fermentation for fuel ethanol production. However, yeast cells grown aerobically on the hydrolyzate have increased tolerance for the hydrolyzate. Cultivation of yeast on part of the hydrolyzate has therefore the potential of enabling increased ethanol productivity in the fermentation of the hydrolyzate. To evaluate the ability of the yeast to grow in the hydrolyzate, fed-batch cultivations were run using the ethanol concentration as input variable to control the feed-rate. The yeast then grew in an undetoxified hydrolyzate with a specific growth rate of 0.19 h⁻¹ by controlling the ethanol concentration at a low level during the cultivation. However, the biomass yield was lower for the cultivation on hydrolyzate compared to synthetic media: with an ethanol set-point of 0.25 g/l the yield was 0.46 g/g on the hydrolyzate, compared to 0.52 g/g for synthetic media. The main reason for the difference was not the ethanol production per se, but a significant production of glycerol at a high specific growth rate. The glycerol production may be attributed to an insufficient respiratory capacity.     

Enhanced ethanol production by continuous fermentation in a two-tank system with cell recycling

An experimental method for producing ethanol continuously was designed and tested with a cell-recycling two-tank system, which was composed of two fermentors, each of which was individually equipped with a settler for recycling flocculent yeast. This system was effective for the continuous fermentation of ethanol from sucrose at high cell-recycling (r = 0.8–0.9) and dilution (up to 0.48 h^?1) rates. The system has several advantages; the high cell concentration in the fermentors and relief of substrate and product inhibition. Thus, the enhanced productivity using this continuous fermentation with the two-tank cell-recycling system was significantly higher compared with that of the batch fermentation. The results indicate that increased recycling ratios caused an increase in biomass concentration and subsequently, product concentration in the tank. The ethanol productivity increased with the dilution rate, but higher dilution rates could render increasing amounts of sugar unconverted. Continuous fermentation with the sugar feed concentration of 160 g/l at r = 0.9 and dilution rate of 0.2 h^?1 achieved the highest productivity with less than 2% of the unconverted sugar in the product steam. Under the same cell recycling ratios a productivity range of 6.9–7.5 g/l h^?1 could be achieved with feeding concentrations of 80–200 g/l, while batch fermentation at these sugar concentrations led to productivities of 3.85–4.48 g/l h^?1.