The paper presents modeling and simulation results of the operation of a three-phase fluidized bed bioreactorwith partial recirculation of biomass. The proposed quantitative description of the bioreactor takes into account biomass growth on inert carriers, microorganisms decay and interphase biomass transfer. Stationary characteristics of the bioreactor and local stability of steady-stateswere determined. The influence of microbiological growth kinetics on the multiplicity of steady-states was discussed. The relationship between biofilm growth and boundaries of fluidized bed existence was shown.
The quantitative description of an airlift bioreactor, in which aerobic biodegradation limited by carbonaceous substrate and oxygen dissolved in a liquid takes place, is presented. This process is described by the double-substrate kinetics. Mathematical models based on the assumption of plug flow and dispersion flow of liquid through the riser and the downcomer in the reactor were proposed. Calculations were performed for two representative hydrodynamic regimes of reactor operation, i.e. with the presence of gas bubbles only within the riser and for complete gas circulation. The analysis aimed at how the choice of a mathematical model of the process would enable detecting the theoretical occurrence of oxygen deficiency in the airlift reactor. It was demonstrated that the simplification of numerical calculations by assuming the “plug flow” model instead of dispersion with high Péclet numbers posed a risk of improper evaluation of the presence of oxygen deficiency zones. Conclusions related to apparatusmodelling and process design were drawn on the basis of the results obtained. The paper is a continuation of an earlier publication (Grzywacz, 2012a) where an analysis of single-substrate models of the airlift reactor was presented.
A kinetic model to describe lovastatin biosynthesis by Aspergillus terreus ATCC 20542 in a batch culture with the simultaneous use of lactose and glycerol as carbon sources was developed. In order to do this the kinetics of the process was first studied. Then, the model consisting of five ordinary differential equations to balance lactose, glycerol, organic nitrogen, lovastatin and biomass was proposed. A set of batch experiments with a varying lactose to glycerol ratio was used to finally establish the form of this model and find its parameters. The parameters were either directly determined from the experimental data (maximum biomass specific growth rate, yield coefficients) or identified with the use of the optimisation software. In the next step the model was verified with the use of the independent sets of data obtained from the bioreactor cultivations. In the end the parameters of the model were thoroughly discussed with regard to their biological sense. The fit of the model to the experimental data proved to be satisfactory and gave a new insight to develop various strategies of cultivation of A. terreus with the use of two substrates.
The paper presents microbiological characteristic of sewage sludge composted in controlled conditions together with biowastes (wheat, maize and rapeseed straw, sawdust and refined glycerol). An experiment was carried out in which the material was mixed at appropriate weight proportions and then placed in bioreactor chambers of constant air flow (4 lźmin-1). The performed composting process aimed at determining the developmental dynamics of heterotrophic bacteria, molds, actinomycetes as well as bacteria from Salmonella genus and Enterobacteriaceae family. Microbiological analyses were performed on selective substrates using Koch's plate method. Moreover, using the floatation method, the presence of live eggs of ATT (Ascaris spp., Trichuris spp., Toxocara spp.) intestinal parasites was assessed and levels of dehydrogenase activity were determined using 1% triphenylotetrazole chloride as a substrate. It was concluded, on the basis of the obtained research results, that the composting process reduced bacterial counts of heterotrophic bacteria, molds and the activity of dehydrogenases activity in all experimental treatments. On the other hand, no reduction was observed in quantities of actinomycetes in the composted materials whose changes in numbers were found to correlate positively most strongly with levels of dehydrogenases activity. In addition, it was found that changes in numbers of the analysed groups of microorganisms depended, primarily, on the pH value and concentrations of ammonia released from the composted materials. Furthermore, the obtained research results also revealed that the sewage sludge used in the experiment did not contain Salmonella spp. bacteria and live eggs of ATT intestinal parasites, and that the composting process reduced completely numbers of bacteria from the Enterobacteriaceae family in all compost treatments. The obtained composts fulfilled all sanitary standards complying with the requirements issued by the Minister of Agriculture and Rural Development (2008) as well as with the EU regulation (EC) No. 185/2007 from February 2007 changing EEC regulations No. 809/2003 and No. 810/2003 connected with the extension of the period of transitional requirements for composting and biogas plants as provided by the EU regulation No. 1774/2002 of the European Parliament and Council.
Typically applied static (i.e. non-agitated) cultures do not provide sufficient conditions for efficient propagation of suspended non-adherent cells, in general. Feasibility of small-scale wave-type agitated single-use bioreactors for gentle agitation underlies applicability of such systems for scaling-up of fragile biomass of animal cells. The basic aim of the study was to compare the results of non-adherent HL-60 cell propagation performed referentially as the batch culture in typical static (i.e. non-agitated) disposable culture flasks (50 cm3 of culture medium) and in ReadyToProcess WAVETM25 bioreactor system (GE Healthcare) equipped with disposable culture bag (300 cm3 of culture medium) subjected to continuous wave-type agitation. The density and viability of HL-60 cells were significantly higher for the bioprocess subjected to wave-type agitation, than in the reference static culture. The values of the specific rate of glucose consumption per cell (rglc=cell) exhibited by HL-60 cells maintained in the system with continuous wave-type agitation was significantly lower (i.e. up to more than 42%) than the values noted for the static culture, for exactly the same time-points of two compared cultures. The results of the studies undoubtedly and comprehensively confirmed the applicability of the studied disposable bioreactor with wave-induced agitation as the right platform for proceeding the propagation of non- adherent HL-60 cells and for providing the culture conditions required by HL-60 cells for sustainable metabolism.
A simple analytical method for determination of basic hydrodynamic characteristics of hybrid fluidized-bed air-lift devices was presented. These devices consist of two parts: a two-phase air-lift part and a two-phase liquid-solid fluidized-bed part. Forced circulation of fluid in the air-lift part is used for fluidization of solid particles in the fluidized-bed part. According to the opinion given in the literature, if such apparatus is used for aerobic microbiological processes, its advantage is lower shear forces acting on the biofilm immobilized on fine-grained material compared with shear forces in three-phase fluidized-bed bioreactors. Another advantage is higher biomass concentration due to its immobilization on fine particles, compared with two-phase airlift bioreactors. A method of calculating gas hold-up in the air-lift part, and gas and liquid velocities in all zones of the analyzed apparatus is presented.
In this study, β-galactosidase enzyme from Kluyveromyces fragilis was immobilised on a commercial polyethersulfone membrane surface, 10 kDa cut-off. An integrated process, concerning the simultaneous hydrolysis-ultrafiltration of whey lactose was studied and working conditions have been fixed at 55°C and pH 6.9, the same conditions that are used for the industrial process of protein concentration. For the immobilisation, best results were obtained using 5% (v/v) of glutaraldehyde solution and 0.03 M galactose; the total activity recovery coefficient (TARC) was 44.2%. The amount of immobilised enzyme was 12.49 mg with a total activity of 86.3 LAU at 37°C, using 5% (w/v) lactose solution in phosphate buffer (100 mM pH 6.9). The stability of the immobilised enzyme was approximately 585 fold higher in comparison with the stability of free enzyme. Multipoint covalent immobilisation improves the stability of the enzyme, thereby enhancing the decision to use the membrane as a filtering element and support for the enzyme immobilisation.
A mathematical model for a two-phase fluidised bed bioreactor with liquid recirculation and an external aerator was proposed. A stationary nonlinear analysis of such a bioreactor for an aerobic process with double-substrate kinetics was carried out. The influences of a volumetric fraction of solid carriers in the liquid phase, the rate of active biomass transfer from the biofilm to the liquid, the concentration of carbonaceous substrate, the mean residence time of the liquid and the efficiency of the external aerator on the steady state characteristics of the bioreactor were described. A method for determination of the minimal recirculation ratio related to oxygen demand and fluidised bed conditions was presented. On the basis of the obtained results, it is possible to choose reasonable operating conditions of such plants and to determine constraints, while considering acceptable concentrations of a toxic substrate being degraded.
Biological regeneration of water and organic sorbents used in the absorption of hydrophilic and hydrophobic pollutants, respectively, was studied. In both cases biodegradation takes place in a membrane bioreactor. In the case of organic sorbents regeneration of the biodegradation process is integrated with the extraction of a given pollutant to water phase. In experiments carried out in this work, the proposed systems were tested using a strain of Pseudomonas fluorescens. For hydrophilic compounds experiments were performed using alcohols (1-butanol and 2-propanol) as model substrates. Applying the mathematical model of a membrane bioreactor elaborated previously, the values of pollutant concentration were determined and positively verified in the experiments. This system of water sorbent regeneration is fully satisfying. The process of biodegradation integrated with extraction was analysed on the basis of model compounds such as benzene and toluene. The study confirmed a possibility of organic sorbent (silicone oil) regeneration. However, due to a very high partition coefficient of benzene or toluene between the organic and aqueous phases, the process could be considered only for the case of their high concentrations in the gas directed to absorption.
The paper presents the dynamic characteristics of a continuous tank bioreactor for microbiological process, with a developed predator-prey food chain. The presence of the predator microorganism considerably influences the position and stability character of steady-states. There appears to exist a wide range of unstable steady-states and high-amplitude oscillations of state variables. Without automatic control, the system can operate only in unsteady conditions. From technological point of view, this circumstance is unfavorable. It was shown that oscillations can be removed by employing automatic control with continuous P or PI controllers. Moreover, the use of a controller with integrating element causes removal of the predator from the bioreactor. The paper discusses an application of this phenomenon for practical purposes.
Two types of submerged membrane bioreactors (MBR): hollow fiber (HF) and hollow sheet (HS), have been studied and compared in terms of energy consumption and average shear stress over the membrane wall. The analysis of energy consumption was made using the correlation to determine the blower power and the blower power demand per unit of permeate volume. Results showed that for the system geometries considered, in terms the of the blower power, the HF MBR requires less power compared to HS MBR. However, in terms of blower power per unit of permeate volume, the HS MBR requires less energy. The analysis of shear stress over the membrane surface was made using computational fluid dynamics (CFD) modelling. Experimental measurements for the HF MBR were compared with the CFD model and an error less that 8% was obtained. For the HS MBR, experimental measurements of velocity profiles were made and an error of 11% was found. This work uses an empirical relationship to determine the shear stress based on the ratio of aeration blower power to tank volume. This relationship is used in bubble column reactors and it is extrapolate to determine shear stress on MBR systems. This relationship proved to be overestimated by 28% compared to experimental measurements and CFD results. Therefore, a corrective factor is included in the relationship in order to account for the membrane placed inside the bioreactor.
One of the actual challenges in tissue engineering applications is to efficiently produce as high of number of cells as it is only possible, in the shortest time. In static cultures, the production of animal cell biomass in integrated forms (i.e. aggregates, inoculated scaffolds) is limited due to inefficient diffusion of culture medium components observed in such non-mixed culture systems, especially in the case of cell-inoculated fiber-based dense 3D scaffolds, inside which the intensification of mass transfer is particularly important. The applicability of a prototyped, small-scale, continuously wave-induced agitated system for intensification of anchorage-dependent CP5 chondrocytes proliferation outside and inside three-dimensional poly(lactic acid) (PLA) scaffolds has been discussed. Fibrous PLA-based constructs have been inoculated with CP5 cells and then maintained in two independent incubation systems: (i) non-agitated conditions and (ii) culture with wave-induced agitation. Significantly higher values of the volumetric glucose consumption rate have been noted for the system with the wave-induced agitation. The advantage of the presented wave-induced agitation culture system has been confirmed by lower activity of lactate dehydrogenase (LDH) released from the cells in the samples of culture medium harvested from the agitated cultures, in contrast to rather high values of LDH activity measured for static conditions. Results of the proceeded experiments and their analysis clearly exhibited the feasibility of the culture system supported with continuously wave-induced agitation for robust proliferation of the CP5 chondrocytes on PLA-based structures. Aside from the practicability of the prototyped system, we believe that it could also be applied as a standard method offering advantages for all types of the daily routine laboratory-scale animal cell cultures utilizing various fiber-based biomaterials, with the use of only regular laboratory devices.
The results of numerical computations concerning momentum transfer processes in an air – biophase – liquid system agitated in a bioreactor equipped with baffles and a Smith turbine (CD 6 impeller) are presented in this paper. The effect of sucrose concentration on the distributions of the velocity of the continuous phase, gas hold-up and the size of gas bubbles in the system was analysed. Simulation results were presented in the form of the contours of the analysed magnitudes. The effect of sucrose concentration on the averaged values (i.e. determined on the basis of local values) of gas hold-up and gas bubbles size was evaluated. The results of the numerical computations of gas hold-up were compared with our own experimental data.
Biosynthesis of lovastatin (a polyketide metabolite of Aspergillus terreus) in bioreactors of different working volume was studied to indicate how the change of scale of the process influences the formation of this metabolite. The experiments conducted in shake flasks of 150 ml working volume allowed to obtain lovastatin titres at the level of 87.5 mg LOV l-1, when two carbon sources, namely lactose and glycerol were used. The application of the same components in a large stirred-tank bioreactor of 5.3-litre working volume caused a decrease of lovastatin production by 87% compared to the shake flask culture. The deficiency of nitrogen in this bioreactor did not favour the formation of lovastatin, in contrast to the small bioreactor of 1.95-litre working volume, in which lovastatin titres comparable to those in the shake flasks could be achieved, when organic nitrogen concentration was two-fold decreased. When the control of pH and/or pO2 was used simultaneously, an increase in lovastatin production was observed in the bioreactors. However, these results were still slightly lower than lovastatin titres obtained in the shake flasks.