Scale-down/scale-up studies leading to improved commercial beer fermentation

Scale-up/scale-down techniques are vital for successful and safe commercial-scale bioprocess design and operation. An example is given in this review of recent studies related to beer production. Work at the bench scale shows that brewing yeast is not compromised by mechanical agitation up to 4.5 W/kg; and that compared with fermentations mixed by CO(2) evolution, agitation ≥ 0.04 W/kg is able to reduce fermentation time by about 20%. Work at the commercial scale in cylindroconical fermenters shows that, without mechanical agitation, most of the yeast sediments into the cone for about 50% of the fermentation time, leading to poor temperature control. Stirrer mixing overcomes these problems and leads to a similar reduction in batch time as the bench-scale tests and greatly reduces its variability, but is difficult to install in extant fermenters. The mixing characteristics of a new jet mixer, a rotary jet mixer, which overcomes these difficulties, are reported, based on pilot-scale studies. This change enables the advantages of stirring to be achieved at the commercial scale without the problems. In addition, more of the fermentable sugars are converted into ethanol. This review shows the effectiveness of scale-up/scale-down studies for improving commercial operations. Suggestions for further studies are made: one concerning the impact of homogenization on the removal of vicinal diketones and the other on the location of bubble formation at the commercial scale.     

Characterization of plant polysaccharide- and mucin-fermenting anaerobic bacteria from human feces

Organisms able to grow on arabinogalactan, pectin, xylan, wheat bran, guar, apple cell walls, and mucin were isolated by enrichment from human feces. The number of polysaccharide fermenters and the properties of the predominant bacteria varied between subjects. The ability to use one polysaccharide was not related to the ability to use others. Some organisms (e.g., Bacteroides spp.) isolated on other substrates also utilized mucin, but were not isolated in the mucin enrichment. The mucin fermenters isolated by enrichment had a very restricted ability to utilize complex polysaccharides and their constituent monosaccharides, suggesting that the presence of plant polysaccharides in the human colon is unlikely to prevent the use of colonic mucin as an energy source by bacteria. Characterization with a range of biochemical tests showed that many of the isolates, but especially the mucin fermenters, did not resemble organisms described previously.     

Characterization of plant polysaccharide- and mucin-fermenting anaerobic bacteria from human feces.

Organisms able to grow on arabinogalactan, pectin, xylan, wheat bran, guar, apple cell walls, and mucin were isolated by enrichment from human feces. The number of polysaccharide fermenters and the properties of the predominant bacteria varied between subjects. The ability to use one polysaccharide was not related to the ability to use others. Some organisms (e.g., Bacteroides spp.) isolated on other substrates also utilized mucin, but were not isolated in the mucin enrichment. The mucin fermenters isolated by enrichment had a very restricted ability to utilize complex polysaccharides and their constituent monosaccharides, suggesting that the presence of plant polysaccharides in the human colon is unlikely to prevent the use of colonic mucin as an energy source by bacteria. Characterization with a range of biochemical tests showed that many of the isolates, but especially the mucin fermenters, did not resemble organisms described previously.     

Modelling of concentration profiles in tower fermenter. The influence of the axial dispersion coefficient

The production of CO₂ in tower fermenters which can cause back mixing, and thereby reducing the efficiency of such fermenters has been modelled with respect to fermentation. It is confirmed that the effect is significant. The effect of variations in the kinetic parameters, the maximum specific growth rate and the biomass yield coefficient was found to be insignificant. A good correlation between experimental and calculated data was obtained.     

A kLa identification technique for a dynamic model of the coupled system: Air-lift fermenter — oxygen probes

The time delay of oxygen probe response to the signal from a fermenter makes identification of the volumetric oxygen transfer coefficient k_La by the dynamic method more complicated. A coupled model involving the transient-state oxygen balance of the fermenter together with the dynamic model of the oxygen probe must be then formulated, solved and identified. In this paper two simple models of air-lift loop fermenters have been proposed and a coupled mathematical model of the fermenter – oxygen probe system has been developed. The identification procedure was used to estimate k_La values in the fermenter with internal circulation flow on the basis of experimental measurements. A comparison of evaluated and experimental indications of the probes placed at various heights of the column proves that the model presented gives a possibility of the first-step approximation of k_La in loop fermenters.     

Production of Escherichia coli Heat-labile Enterotoxin in Fermenter Dialysis Culture

Production of Escherichia coli heat-labile enterotoxin was investigated with one porcine and one human strain in three different media under different cultivation conditions. Cultivation in aerated fermenters at pH 7·0 yielded 10–20 times more enterotoxin/ml of culture fluid than cultivation in shake flasks. A trypton-yeast extract medium was optimal in fermenter cultures. Comparatively good yields of enterotoxin in fermenters were also obtained in a glucose-salts medium. Continuous feeding of glucose and salts during fermenter cultivation resulted in a lower production of enterotoxin/mg of bacterial cells. Since this decrease in specific yield could be reversed by using dialysis culture, it was concluded that inhibition of toxin formation was due to the accumulation of extracellular low molecular weight metabolites. The highest yield of enterotoxin in dialysis culture was 80 ED₅₀ ml⁻¹ (rabbit jejunal loop test) which is at least eight times more toxin than in ordinary fermenter culture and 80 times more toxin than in shake flask cultures.     

Influence of reactor geometry on mixing characteristics in a down flow jet loop bioreactor: newtonian and non-newtonian fluids

Mixing characteristics in the downcomer and the riser of a continuous down-flow jet loop bioreactor was studied with Newtonian and non-Newtonian fluids. The mixing parameters were determined through the curve fitting of the experimental impulse response data with the solution of one dimensional axial dispersion model. It was found that circulation number and axial dispersion coefficient increased with an increase in liquid flow rate and draft tube to column diameter ratio and the axial dispersion coefficient was comparatively higher in the riser. The circulation number increased with decrease in nozzle diameter. The model predicted the experimental data well within 8% deviation for both the systems (water and CMC). Correlations were obtained to predict axial dispersion coefficients in the riser and downcomer of the reactor.     

Jet aeration as alternative to overcome mass transfer limitation of stirred bioreactors

In industrial biotechnology increasing reactor volumes have the potential to reduce production costs. Whenever the achievable space time yield is determined by the mass transfer performance of the reactor, energy efficiency plays an important role to meet the requirements regarding low investment and operating costs. Based on theoretical calculations, compared to bubble column, airlift reactor, and aerated stirred tank, the jet loop reactor shows the potential for an enhanced energetic efficiency at high mass transfer rates. Interestingly, its technical application in standard biotechnological production processes has not yet been realized. Compared to a stirred tank reactor powered by Rushton turbines, maximum oxygen transfer rates about 200% higher were achieved in a jet loop reactor at identical power input in a fed batch fermentation process. Moreover, a model‐based analysis of yield coefficients and growth kinetics showed that E. coli can be cultivated in jet loop reactors without significant differences in biomass growth. Based on an aerobic fermentation process, the assessment of energetic oxygen transfer efficiency [kgO₂ kW^?1 h^?1] for a jet loop reactor yielded an improvement of almost 100%. The jet loop reactor could be operated at mass transfer rates 67% higher compared to a stirred tank. Thus, an increase of 40% in maximum space time yield [kg m^?3 h^?1] could be observed.     

Modelling of multi-phase flow in jet loop reactors

A model for the steady state flow in jet loop reactors with two-phase flow has been developed. The model can be used to predict the critical effect in a reversed flow jet loop fermenter. The calculated critical effect according to the model corresponds well to experimental data.     

Fibre digestibility in large herbivores as related to digestion type and body mass — An in vitro approach

The coexistence of different ungulate species in a given ecosystem has been the focus of many studies. Differences between ruminant foregut fermenters and hindgut fermenters were remarkable for example in the way they ingest and digest high fibre diets. Digestion trials based on total collections are difficult to conduct or are sometimes even not possible for wild animals in the field or in zoos. To gain information on the fibre digestion achieved by these animals and the influence of body mass (BM) thereon, a method using spot sampling is desirable. In this study, in vitro fermentation of faecal neutral detergent fibre (NDF) was used as a measure of fibre digestion in large ungulates. Food and faecal samples of 10 ruminant foregut fermenting and 7 hindgut fermenting species/breeds were collected. All animals received 100% grass hay with ad libitum access. The NDF of food and faeces was fermented in vitro in a Hohenheim gas test (HGT) for 96 h. The digestion type generally had an effect on the gas production (GP) of faecal NDF in the HGT with hindgut fermenters showing higher values than ruminant foregut fermenters. At any time interval of incubation, BM had no influence on GP. The results are in accordance with both findings that ruminant foregut fermenters have longer mean retention times and more comprehensive particle reduction and findings of a lack of influence of BM on digesta mean retention time. It can be stated that the HGT (96 h) is a useful and quick method to show also small differences within groups in fibre digestion.     

Design and operation of pulsed anaerobic digesters

The development of a pulse-driven loop reactor (PDLR), a pulsed anaerobic filter (PAF) and a pulsed anaerobic baffled reactor (PABR) is described. In an anaerobic PDLR internal circulation is achieved by a specially designed pulse-nozzle. In a PAF and PABR an oscillation is superimposed onto the liquid content of the fermenters by means of a pulse pump without any moving devices in the reactors. Pulsed digesters faciliate degassing, avoid reactor clogging as well as short-circuiting and allow a variety of packed-bed to fluidized-bed operations. Anaerobic fermentation of acetic acid and distillery slops in pulsed digesters on a laboratory scale shows that hydrodynamic stress caused by pulsation is well compatible to degrading bacteria.List of Symbols PAF Pulsed Anaerobic Filter - PDLR Pulse-Driven Loop Reactor - PABR Pulsed Anaerobic Baffled Reactor     

Influence of oxygen and substrate concentrations on the ideal film thickness and the maximum overall substrate uptake rate in microbial film fermenters

The criterion for the oxygen limitation of substrate uptake in microbial film fermenters is expressed in terms of diffusion coefficients, utilization coefficients, and the free solution concentrations of substrate and oxygen. It is proposed that the ideal film thickness in such fermenters is equal to the penetration depth of the limiting substrate. The ideal film thickness is calculated, in terms of the parameters contained in the criterion for oxygen limitation, for three separate kinetic rate expressions. It is found that for the air–glucose–microbe system a simplified kinetic rate expression can be used and the region of dependence on two substrates is shown to be very limited. This is not true for other systems. Maximum uptake rates are calculated for a range of concentrations. Finally, it is shown that the procedure used can be generalized to determine the limiting substrate in a multisubstrate system and to calculate ideal film thickness and uptake rates for any pair of substrates where the kinetics of substrate uptake are known for the individual microorganism.     

Quantification of damage to suspended insect cells as a result of bubble rupture

It is proposed that when cells are either attached to, or very near, a rupturing bubble, the hydrodynamic forces associated with the rupture are sufficient to kill the cells. Four types of experiments were conducted to quantify the number and location of these killed cells. We determined: (1) the number of cells killed as a result of a single, 3.5-mm bubble rupture; (2) the number and viability of cells in the upward jet that results when a bubble ruptures; (3) the number of cells on the bubble film; and (4) the fate of cells attached to the bubble film after film rupture. All experiments were conducted with Spodoptera frugiperda (SF-9) insect cells, in TNM-FH and SFML medium, with and without Pluronic F-68. Experiments indicate that approximately 1050 cells are killed per single, 3.5-mm bubble rupture in TNM-FH medium and approximately the same number of dead cells are present in the upward jet. It was also observed that the concentration of cells in this upward jet is higher than the cell suspension in TNM-FH medium without Pluronic F-68 by a factor of two. It is believed that this higher concentration is the result of cells adhering to the bubble interface. These cells are swept up into the upward jet during the bubble rupture process. Finally, it is suggested that a thin layer around the bubble containing these absorbed cells is the "hypothetical killing volume" presented by other researchers. (c) 1994 John Wiley & Sons, Inc.     

A Comparison of pH-controlled and Dissolved Oxygen-controlled Nutrient Addition for the Maintenance of Steady-state in a Mixed Continuous Culture

Controlled substrate addition was used to maintain mixed microbial cultures in fermenters at either pH 7.0 or 70% dissolved oxygen saturation. Control of pH permitted a greater volume of substrate to be processed. Ammonium nitrogen concentrations were similar for both fermenters but concentrations of oxidized nitrogen varied. Nitrification/denitrification sequences appeared to be initiated by unscheduled changes in dissolved oxygen concentration. It was possible to maintain a steady state with respect to a controlled parameter and end-product quality but other parameters fluctuated.     

Local overall volumetric gas-liquid mass transfer coefficients in gas-liquid-solid reversed flow jet loop bioreactor with a non-Newtonian fluid

The local overall volumetric gas-liquid mass transfer coefficients at the specified point in a gas-liquid-solid three-phase reversed flow jet loop bioreactor (JLB) with a non-Newtonian fluid was experimentally investigated by a transient gassing-in method. The effects of liquid jet flow rate, gas jet flow rate, particle density, particle diameter, solids loading, nozzle diameter and CMC concentration on the local overall volumetric gas-liquid mass transfer coefficient (K(L)a) profiles were discussed. It was observed that local overall K(L)a profiles in the three-phase reversed flow JLB with non-Newtonian fluid increased with the increase of gas jet flow rate, liquid jet flow rate, particle density and particle diameter, but decreased with the increase of the nozzle diameter and CMC concentration. The presence of solids at a low concentration increased the local overall K(L)a profiles, and the optimum of solids loading for a maximum profile of the local overall K(L)a was found to be 0.18x10(-3)m(3) corresponding to a solids volume fraction, varepsilon(S)=2.8%.     

Discussion on entrained droplets in fermenters used for the cultivation of single-celled mircoorganisms

The validity of regarding the liquid phase in vigorously agitated sparged fermenters during the cultivation of single-celled aerobic microorganisms as essentially homogeneous is examined. Droplet formation from bursting bubbles and physical effects of the spray environment on single-celled microorganisms are discussed. The implications of droplet removal from the head space of fermenters by collision with and drainage down the walls are considered, particularly factors concerned with wall growth above the liquid level in fermenters.     

Nickel as an accelerator of biogas production in water hyacinth (Eichornia crassipes solms.)

The effect of nickel ions on biogas production has been investigated in 3-litre fermenters. It was observed that nickel is stimulatory up to 5 ppm, with an optimum at 2·5 ppm, in a water hyacinth-bovine excreta substrate. A similar effect with nickel was also observed at 5 ppm in bovine excreta.     

Comparative nutrient extraction from forages by grazing bovids and equids: a test of the nutritional model of equid/bovid competition and coexistence

Summary Ruminants are unevenly distributed across the range of body sizes observed in herbivorous mammals; among extant East African species they predominate, in numbers and species richness, in the medium body sizes (10–600 kg). The small and the large species are all hind-gut fermenters. Some medium-sized hind-gut fermenters, equid perissodactyls, coexist with the grazing ruminants, principally bovid artiodactyls, in grassland ecosystems. These patterns have been explained by two complementary models based on differences between the digestive physiology of ruminants and hind-gut fermenters. The Demment and Van Soest (1985) model accounts for the absence of ruminants among the small and large species, while the Bell/Janis/Foose model accounts both for the predominance of ruminants, and their co-existence with equids among the medium-sized species (Bell 1971; Janis 1976; Foose 1982). The latter model assumes that the rumen is competitively superior to the hind-gut system on medium quality forages, and that hind-gut fermenters persist because of their ability to eat more, and thus to extract more nutrients per day from high fibre, low quality forages. Data presented here demonstrate that compared to similarly sized grazing ruminants (bovids), hind-gut fermenters (equids) have higher rates of food intake which more than compensate for their lesser ability to digest plant material. As a consequence equids extract more nutrients per day than bovids not only from low quality foods, but from the whole range of forages eaten by animals of this size. Neither of the current nutritional models, nor refinements of them satisfactorily explain the preponderance of the bovids among medium-sized ungulates; alternative hypotheses are presented.     

Hydrodynamics and mixing in down flow jet loop bioreactor with a non-Newtonian fluid

Gas holdup and liquid circulation time were measured in a down flow jet loop bioreactor with a non-Newtonian fluid. It was observed that the circulation time decreases with increase in nozzle diameter, draft tube to column diameter ratio and shear thinning of the media. The gas holdup increases with increase in gas and liquid velocities. The optimum draft tube to column diameter ratio was found to be 0.438. Correlations for gas holdup and circulation time involving operational and geometrical variables were presented.     

The energetics of coprophagy: a theoretical analysis

Lagomorphs, many rodents and some other small mammals eat their faeces during the part of the day when they are not foraging for fresh food. One of the possible benefits of this habit of coprophagy is that it may enable them to extract more energy from their food. A computer model is used to assess the likely benefits and explore their relationships to food, feeding rate and gut morphology. The predicted benefits are much larger for hindgut fermenters than for foregut fermenters, and especially large for hindgut fermenters with relatively small fermentation chambers. They are larger for poor foods (with lower proportions of cell contents) than for richer ones. At low feeding rates the energetic advantage of coprophagy may disappear if the faeces from food eaten during one feeding period emerge largely during the next, but this can be avoided by adjusting the rate of passage of gut contents during the intervening rest period.