Scale-down studies on the hydrodynamics of two-liquid phase biocatalytic reactors

The maintenance of constant interfacial area per unit volume is a key parameter for the successful scale-up of two-liquid phase bioconversion processes. To date, however, there is little published information on the hydrodynamics of such systems and a suitable basis for scale-up has yet to be defined and verified. Here we report power input and hydrodynamic data for a whole-cell bioconversion process using resting cells of Rhodococcus R312 to catalyse the hydration of a poorly water-soluble substrate 1,3-dicyanobenzene (1,3-DCB). Experiments were performed in geometrically similar 3-L and 75-L reactors, each fitted with a three-stage Rushton turbine impeller system. The two-phase system used comprised of 20% v/v toluene dispersed in 0.1 M aqueous phosphate buffer containing up to 10 g(ww) x L(-1) of resuspended biocatalyst and 20 g x L(-1) 1,3-DCB. The power input to the 3-L reactor was first determined using an air-bearing technique for both single-phase and two-phase mixing. In both cases, the power number attained a constant value of 11 at Re>10,000, while the measured power inputs were in the range 0.15-3.25 kW x m(-3). Drop size distributions and Sauter mean drop diameters (d(32)) were subsequently measured on-line in both reactors, using an in-situ light-backscattering technique, for scale-up on the basis of either constant power input per unit volume or constant tip speed. At both scales d(32) decreased with increasing agitation rate, while the drop size distributions obtained were log-normal. All the measured d(32) values were in the range of 30-50 microm, with the lowest values being obtained in systems with biocatalyst present. In all cases, constant power input per unit volume was found to be the most suitable basis for scale-up. This gave virtually identical d(32) values and drop size distributions at both scales. A number of correlations were also identified that would allow reasonable prediction of d(32) values for various agitation rates at each scale. While the results obtained are for a particular phase system, the scale-down methodology presented here would allow the rapid evaluation of other bioconversion processes in the 3-L reactor with a 25-fold reduction in scale. In this way, potential problems that might be encountered at the larger scale, such as the carryover of antifoam from the fermentation stage, could be quickly and efficiently identified.     

Multiple-impeller systems with a special emphasis on bioreactors: a critical review

The multiple-impeller agitated systems are compared with single-impeller agitated systems with a special focus on its applications for bioreactors. Correlations reported in the literature for gas phase hold-up, mass transfer coefficient and power consumption under gassed and ungassed conditions are compared and recommendations have been made regarding their suitability for design and scale-up of bioreactors. The multiple-impeller systems are found to be superior as compared to single-impeller systems in all the above mentioned aspects, except liquid mixing. For all kinds of reactors where the sole purpose is mass transfer, multiple-impeller systems are advantageous and there would be large savings on an industrial scale, especially for the bioreactors where the reaction periods are long and the power consumption cost could be a significant component to the overall production costs.     

A methodology for the design of scale-down bioreactors by the use of mixing and circulation stochastic models

Scale-up is traduced in practice by an increase of the dimensions of the bioreactors, leading to a modification of the time scale and thus of the process dynamics. In the present work, a methodology to study the effect of scale-up on bioreactors hydrodynamics and to put in place scale-down reactors representative of the flow properties encountered in real scales bioreactors is detailed.In order to simplify the analysis, we have proposed the use of a stochastic model which is directly affected by the time scale. Indeed, to run simulations with such models, we have to specify the time taken to achieve a transition Δt. Stochastic models are thus reliable to study scale-up effect on stirred reactors hydrodynamics. In addition, these models permit to have an insight on the internal dynamic of the process.In the case of the circulation process, qualitative aspects have to be taken into account and induce a modification of the flow regions arrangement of the model. The stochastic analysis of large-scale bioreactors permits to propose a translating methodology into a scale-down context. Optimised scale-down reactors can be used further to carry out fermentation tests with the hydrodynamic conditions of the industrial scale. In a general rule, the performances of stochastic model allow to facilitate greatly the analysis of the scale-up effect and the hydrodynamic characteristics of both large-scale and scale-down reactors.     

Trickle-bed root culture bioreactor design and scale-up: growth, fluid-dynamics, and oxygen mass transfer

Trickle-bed root culture reactors are shown to achieve tissue concentrations as high as 36 g DW/L (752 g FW/L) at a scale of 14 L. Root growth rate in a 1.6-L reactor configuration with improved operational conditions is shown to be indistinguishable from the laboratory-scale benchmark, the shaker flask (mu=0.33 day(-1)). These results demonstrate that trickle-bed reactor systems can sustain tissue concentrations, growth rates and volumetric biomass productivities substantially higher than other reported bioreactor configurations. Mass transfer and fluid dynamics are characterized in trickle-bed root reactors to identify appropriate operating conditions and scale-up criteria. Root tissue respiration goes through a minimum with increasing liquid flow, which is qualitatively consistent with traditional trickle-bed performance. However, liquid hold-up is much higher than traditional trickle-beds and alternative correlations based on liquid hold-up per unit tissue mass are required to account for large changes in biomass volume fraction. Bioreactor characterization is sufficient to carry out preliminary design calculations that indicate scale-up feasibility to at least 10,000 liters.     

Essential prerequisites for successful bioprocess development of biological CH4 production from CO2 and H2

The production and storage of energy from renewable resources steadily increases in importance. One opportunity is to utilize carbon dioxide (CO₂)-type hydrogenotrophic methanogens, which are an intriguing group of microorganisms from the domain Archaea, for conversion of hydrogen and CO₂ to methane (CH₄). This review summarizes the current state of the art of bioprocess development for biological CH₄ production (BMP) from pure cultures with pure gasses. The prerequisites for successful quantification of BMP by using closed batch, as well as fed-batch and chemostat culture cultivation, are presented. This review shows that BMP is currently a much underexplored field of bioprocess development, which mainly focuses on the application of continuously stirred tank reactors. However, some promising alternatives, such as membrane reactors have already been adapted for BMP. Moreover, industrial-based scale-up of BMP to pilot scale and larger has not been conducted. Most crucial parameters have been found to be those, which influence gas-limitation fundamentals, or parameters that contribute to the complex effects that arise during medium development for scale-up of BMP bioprocesses, highly stressing the importance of holistic BMP quantification by the application of well-defined physiological parameters. The much underexplored number of different genera, which is mainly limited to Methanothermobacter spp., offers the possibility of additional scientific and bioprocess development endeavors for the investigation of BMP. This indicates the large potential for future bioprocess development considering the possible application of bioprocessing technological aspects for renewable energy storage and power generation.     

Solid state fermentation reactors: from lab scale to pilot plant

Relatively many workers in the world are studying different aspects in SSF processes but few are working on reactor design and scale-up. From about 10 years, we are developing reactors from lab scale to pilot plant, based on the same technology, reactor design and flowsheet to allow fermentation with a deep layer (up to 1 m in the pilot plant). These reactors have all a forced aeration and the possibility or not to agitate. Regulations of temperature and water content of the culture are monitored by a special device.     

Wastewater treatment with particulate biofilm reactors

The review presented in this paper focuses on applications of particulate biofilm reactors (e.g. Upflow Sludge Blanket, Biofilm Fluidized Bed, Expanded Granular Sludge Blanket, Biofilm Airlift Suspension, Internal Circulation reactors). Several full-scale applications for municipal and industrial wastewater treatment are presented and illustrated, and their most important design and operation aspects (e.g. biofilm formation, hydrodynamics, mass transfer, mixing) are analysed and discussed. It is clear from the review that this technology can be considered a grown up technology for which good design and scale-up guidelines are available.     

Characterization of agitation environments in 250 ml spinner vessel, 3 L,and 20 L reactor vessels used for animal cell microcarrier culture

Three dimensional particle tracking velocimetry (3-D PTV) was used to characterize the flow fields in the impeller region of three microcarrier reactor vessels. Three typical cell culture bioreactors were chosen: 250 ml small-scale spinner vessels, 3 L bench-scale reactor, and 20 L medium-scale reactor. Conditions studied correspond to the actual operating conditions in industrial setting and were determined based on the current scale-up paradigm: the Kolmogorov eddy length criterion. In this paper we present characterization of hydrodynamics on the basis of flow structures produced because of agitation. Flow structures were determined from 3-D mean velocity results obtained using 3-D PTV. Although the impellers used in 3 L and 20 L reactors were almost identical, the flow structures produced in the two reactors differed considerably. Results indicate that near geometric scale up does not necessarily amount to scale-up of flow patterns and indicates that intensity as well as distribution of energy may vary considerably during such a scale-up.     

Why use bubble-column bioreactors?

Among the plethora of bioreactors available for aerobic culture, bubble columns, which are composed of a cylindrical vessel fitted with a gas sparger, are gaining in use. The simple construction of bubble-column reactors makes them easy to maintain. In addition, it is possible to control the degree of shear, uniformly within the reactor, which is critical to the growth of plant and animal cells in particular. This article reviews in detail the hydrodynamic, heat and mass-transfer characteristics of bubble-column bioreactors - parameters that are important for industrial scale-up.     

Design of a reactor bioleach process for refractory gold treatment

Abstract: CRA has been developing bioleaching for the treatment of low-grade refractory gold resources. In lhe course of dcveloping a biolcach process for a pyrite concentrate at Bougainville Copper Limited (BCL), CRA has confronted the myriad of problems associated with proving a concept at a small scale, to the design of a conceptual flowsheet. A phased programme was initiated to develop the project. Laboratory scale batch studies indicated that the pyrite concentrate was amenable to bacterial leaching and subsequent cyanidatkm gold recovery. Large scale continuous leaching was then performed to delincate the major operating wtriables. In conjunction with this programme, CRA has also been addressing the problem of reactor scale-up. The success of the bioleach process is dependent on the design of large, energy-efficient reactors, with reactor sizes of the order of 1000 m³ contemplated. Results from these scale-up studies are presented in this paper.     

Scale-down of microalgae cultivations in tubular photo-bioreactors--a conceptual approach

Rational design of large-scale bioreactors is still suffering from inadequate scale-up of technical parameters from lab to large scale and from missing kinetic information concerning the physiological reactions of the specific strain under cultivation. Therefore, simulations of processes expected in large-scale have to be carried out as far as possible and experiments have to be performed in small-scale reactors mimicking the situation in large scale. This procedure is referred to as scale-down. In this paper a concept to accomplish this task is proposed. Firstly, interactions between light transfer, fluid dynamics, and microbial metabolism are described. Secondly, a procedure is given to decompose the interactions by simulation on the one hand and by finding physiological parameters in model reactors on the other. Light transfer can be calculated by Monte Carlo methods, while fluid dynamics is handled by CFD. Ideally illuminated model photo-bioreactors and pilot reactors with enforced flow field are proposed to measure physiological parameters especially induced by light/dark cycles generated by interaction of turbulences and light attenuation.     

Scale-up criteria of square tank surface aerator

Oxygen transfer rate and the corresponding power requirement to operate the rotor are vital for design and scale-up of surface aerators. Present study develops simulation or scale-up criterion correlating the oxygen transfer coefficient and power number along with a parameter governing theoretical power per unit volume (X, which is defined as equal to F(4/3)R(1/3), where F and R are impellers' Froude and Reynolds number, respectively). Based on such scale-up criteria, design considerations are developed to save energy requirements while designing square tank surface aerators. It has been demonstrated that energy can be saved substantially if the aeration tanks are run at relatively higher input powers. It is also demonstrated that smaller sized tanks are more energy conservative and economical when compared to big sized tanks, while aerating the same volume of water, and at the same time by maintaining a constant input power in all the tanks irrespective of their size. An example illustrating how energy can be reduced while designing different sized aerators is given. The results presented have a wide application in biotechnology and bioengineering areas with a particular emphasis on the design of appropriate surface aeration systems.     

Scale-up analysis of geometrically dissimilar single-use bioreactors

Cell culture scale-up is a challenging task due to the simultaneous change of multiple hydrodynamic process characteristics and their different dependencies on the bioreactor size as well as variation in the requirements of individual cell lines. Conventionally, the volumetric power input is the most common parameter to select the impeller speed for scale-up, however, it is well reported that this approach fails when there are huge differences in bioreactor scales. In this study, different scale-up criteria are evaluated. At first, different hydrodynamic characteristics are assessed using computational fluid dynamics data for four single-use bioreactors, the Mobius® CellReady 3 L, the Xcellerex™ XDR-10, the Xcellerex™ XDR-200, and the Xcellerex™ XDR-2000. On the basis of this numerical data, several potential scale-up criteria such as volumetric power input, impeller tip speed, mixing time, maximum hydrodynamic stress, and average strain rate in the impeller zone are evaluated. Out of all these criteria, the latter is found to be most appropriate, and the successful scale-up from 3 to 10 L bioreactor and to 200 L bioreactor is confirmed with cell culture experiments using Chinese Hamster Ovary cell cultivation.     

The organism as bioreactor. Interpretation of the reduction law of metabolism in terms of heterogeneous catalysis and fractal structure

Organisms and bioreactors are open, dissipative systems in steady state. They are functionally equivalent with respect to turnover and kinetics, and structurally analogous with respect to fractal organization and self-similar scaling. As heterogeneous catalytic systems both are governed by interaction of mass transport and reaction. The structural equivalent to turbulence in the reactor, yielding high efficiency, is the fractal folding and branching of the transport systems of the organism. Dimensionally and in terms of fractals, organisms and reactors are therefore area-volume hybrids. The physiological consequence of this is the reduction law of metabolism. Introducing limits into allometric functions describing scale-up of similar organisms yields probability density distributions of their realization.     

High-performance recombinant protein production with <Emphasis Type="Italic">Escherichia coli</Emphasis> in continuously operated cascades of stirred-tank reactors

The microbial expression of intracellular, recombinant proteins in continuous bioprocesses suffers from low product concentrations. Hence, a process for the intracellular production of photoactivatable mCherry with Escherichia coli in a continuously operated cascade of two stirred-tank reactors was established to separate biomass formation (first reactor) and protein expression (second reactor) spatially. Cascades of miniaturized stirred-tank reactors were implemented, which enable the 24-fold parallel characterization of cascade processes and the direct scale-up of results to the liter scale. With PAmCherry concentrations of 1.15 g L^?1 cascades of stirred-tank reactors improved the process performance significantly compared to production processes in chemostats. In addition, an optimized fed-batch process was outperformed regarding space–time yield (149 mg L^?1 h^?1). This study implicates continuous cascade processes to be a promising alternative to fed-batch processes for microbial protein production and demonstrates that miniaturized stirred-tank reactors can reduce the timeline and costs for cascade process characterization.     

Scale up aspects of sparged insect-cell bioreactors

Conclusion In this chapter we have attempted to evaluate the most important parameters which can be useful for the pur-pose of design and scale up. Insect cells and animal cells in general can be grown well in large vessels. However, none of the theories and parameters discussed in this chapter have been validated on a larger scale than laboratory and small pilot reactors. Selection of the most suitable design and scale-up method there-fore needs in particular studies in larger vessels. The Kolmogorov theory and the killing-volume model are in this respect the most promising approaches for the optimal design of large-scale animal-cell bioreactors.     

Anchoring high-throughput screening methods to scale-up bioproduction of siderophores

The use of high throughput strategies is of acknowledged relevance since the rational use of small-scale reactors, coupled with suitable analytic tools, is contributing to the acceleration of process development in several areas of biotechnology. These small-scale reactors are available in different working volumes and configurations, being useful in a wide array of applications, from cell screening to process optimization.The present work was focused on the development of a high-throughput strategy, combining microtiter plates and analytic methodologies, to screen an in-house library of environmental bacteria in order to identify good siderophore producers. From a library of roughly 500 marine microorganisms, it was possible to ultimately obtain 11 bacterial strains with high production capabilities. Two of them had not been previously identified as siderophore producers. The bioprocess was scaled-up from microtiter plates to a 5 L stirred tank reactor, while maintaining the overall volumetric productivity, using the k_La similarity as scale-up criterion.This novel approach is a suitable alternative to traditional screening tools.     

Aerobic Biodegradation of Mononitrotoluenes in Batch and Continuous Reactor Systems

Degradations of nitrotoluenes, individually and in a mixture, were carried out in batch and continuous aerobic reactors by a defined mixed microbial culture. The degradation rates and efficiencies of the isomers were evaluated in batch and continuous reactors. The results demonstrated that all the three-mononitrotoluene isomers were degraded simultaneously and completely in presence of excess oxygen. The nitro group position on the benzene ring influenced the degradation rates of the individual MNT isomers in batch systems. In the continuous biofilm reactor with a sufficient biocatalyst quantity, quality and excessive oxygen supply rate, the degradation rates of the mononitrotoluenes were almost identical as long as the compounds were present individually and their loading did not exceed the capacity of the catabolic master reaction. The microbial composition of the biofilm changed qualitatively and quantitatively during long-term continuous operation under aerobic non-aseptic conditions. This complex investigation resulted in data that can be applied for the scale-up procedure for field experiments.     

Comparison of the performances of stirred tank and airlift tower loop reactors.

Following a consideration of the prerequisites for reactor comparison and the fundamental differences between stirred tank and airlift tower loop reactors, their performances are compared for the production of secondary metabolites: penicillin V by Penicillium chrysogenum, cephalosporin C by Cephalosporium acremonium, and tetracycline by Streptomyces aureofaciens. In stirred tank reactors, cell mass concentrations, volumetric productivities, and specific power inputs are higher than in airlift tower loop reactors. In the latter, efficiencies of oxygen transfer are higher, and specific productivities with regard to power input, substrate and oxygen consumptions, and yield coefficients of product formation with regard to substrate and oxygen consumptions are considerably higher than in stirred tank reactors. The prerequisites for improved performance are discussed.     

Nuclear power: levels of safety

The rise and fall of the nuclear power industry in the United States is a well-documented story with enough socio-technological conflict to fill dozens of scholarly, and not so scholarly, books. Whatever the reasons for the situation we are now in, and no matter how we apportion the blame, the ultimate choice of whether to use nuclear power in this country is made by the utilities and by the public. Their choices are, finally, based on some form of risk-benefit analysis. Such analysis is done in well-documented and apparently logical form by the utilities and in a rather more inchoate but not necessarily less accurate form by the public. Nuclear power has failed in the United States because both the real and perceived risks outweigh the potential benefits. The national decision not to rely upon nuclear power in its present form is not an irrational one. A wide ranging public balancing of risk and benefit requires a classification of risk which is clear and believable for the public to be able to assess the risks associated with given technological structures. The qualitative four-level safety ladder provides such a framework. Nuclear reactors have been designed which fit clearly and demonstrably into each of the possible qualitative safety levels. Surprisingly, it appears that safer may also mean cheaper. The intellectual and technical prerequisites are in hand for an important national decision. Deployment of a qualitatively different second generation of nuclear reactors can have important benefits for the United States. Surprisingly, it may well be the "nuclear establishment" itself, with enormous investments of money and pride in the existing nuclear systems, that rejects second generation reactors. It may be that we will not have a second generation of reactors until the first generation of nuclear engineers and nuclear power advocates has retired.