Modelling mixing parameters in concentric-tube airlift bioreactors

The mixing behaviour of the liquid phase in concentric-tube airlift bioreactors of different scale (RIMP: V_L=0.070 m³; RIS-1: V_L=2.50 m³; RIS-2: V_L=5.20 m³) in terms of mixing time was investigated. This mixing parameter was determined from the output curves to an initial Dirac pulse, using the classical tracer response technique, and analyzed in relation to process and geometrical parameters, such as: gas superficial velocity, x_SGR; top clearance, h_S; bottom clearance, h_B, and ratio of the resistances at downcomer entrance, A_d/A_R. A correlation between the mixing time and the specified operating and geometrical parameters was developed, which was particularized for two flow regimes: bubbly and transition (x_SGRА.08 m/s) and churn turbulent flow (x_SGR> 0.08 m/s) respectively. The correlation was applied in bioreactors of different scale with a maximum error of ᆲ%.     

Concentric-tube airlift bioreactors

Mass transfer coefficients were measured in three concentric-tube airlift reactors of different scales (RIMP, V _L =0.07 m³;RIS?1,V _L =2.50 m³;RIS?2, V _L =5.20 m³). The effects of top and bottom clearance and flow resistances at downcorner entrance were studied in water-air system. Experimental results show that h _s ,h _B and A _d /A _R ratio affect K _L a values as a result of their influence on gas holdup and liquid velocity. The gas-liquid mass-transfer coefficients for all the geometric variables were successfully correlated as Sherwood number with Froude and Galilei numbers, the bottom spatial ratio (B=h _B /D _R ), the top spatial ratio , the gas separation ratio and the downcomer flow resistance ratio (R=A _d /A _R ). The proposed empirical model satisfactorily fitted the experimental data obtained in large airlift reactors and some data presented in literature.     

Concentric-tube airlift bioreactors

Gas holdup investigations were performed in three concentric-tube airlift reactors of different scales of operation (RIMP: 0.070 m³; RIS-1: 2.5 m³; RIS-2: 5.2 m³; nominal volumes). The influences of the top and bottom clearances and the flow resistances at the downcomer entrance were studied using tap water as liquid phase and air as gaseous phase, at atmospheric pressure. It was found that the gas holdup in the individual zone of the reactor: riser, downcomer and gas-separator, as well as that in the overall reactor is affected by the analyzed geometrical parameters in different ways, depending on their effects on liquid circulation velocity. Gas holdup was satisfactorily correlated with Fr, Ga, bottom spatial ratio (B), top spatial ratio (T), gas separation ratio (Y) and downcomer flow resistance ratio (A _d /A _R ). Correlations are presented for gas holdup in riser, downcomer, gas separator and for the total gas holdup in the reactor. All the above stressed the importance of the geometry in dynamic behaviour of airlift reactors.     

Concentric-tube airlift bioreactors

Liquid circulation velocity was investigated in three concentric-tube airlift reactors of different scales (RIMP, V _L =0.07 m³; RIS-1, V _L =2.5 m³; RIS-2, V _L =5.20 m³). The effects of top and bottom clearance and resistance in flow pathway at downcomer entrance on the riser liquid superficial velocity, the circulation time, the friction coefficient and flow radial profiles of the gas holdup and the liquid superficial velocity in riser, using water-air as a biphasic system, were studied. It was found that the riser liquid superficial velocity is affected by the analyzed geometrical parameters in different ways, depending on their effects on the pressure loss. The riser liquid superficial velocity, the friction coefficient and the parameters of the drift-flux model were satisfactorily correlated with the bottom spatial ratio (B), gas separation ratio (Y) and downcomer flow resistance ratio (A _d /A _D ), resulting empirical models, with correlation coefficients greater than 0.85.     

Liquid circulation in external-loop airlift bioreactors

A simple model for prediction of liquid velocity in external-loop airlift bioreactors has been developed. Theoretical correlations for friction factor of gas-non-Newtonian two-phase flows and for liquid velocity in the riser were derived using the concept of an eddy diffusivity. The predictions of the proposed model were compared with the available experimental data for the friction factor and the liquid velocity in the riser of external-loop airlift contactors. Satisfactory agreement was obtained.     

Gas hold-up in external-loop airlift bioreactors

A new model of gas hold-up is proposed for external-loop airlift bioreactors. It is based on the similarity between the liquid circulation due to the local variation of gas hold-up in airlift bioreactors and the natural convection due to temperature difference. The model is developed to include the case of non-Newtonian fermentation media which are involved in many industrially bioprocesses. The capability of the model is examined using a wide range of experimental results including the present data. Reasonable agreement is obtained between the proposed model and the experimental data both for Newtonian and non-Newtonian media.     

Mathematical models for mixing in deep-jet bioreactors: Calculation of parameters

The macroscopic mathematical model based on compartments with ideal mixing zones and tanks-in series was evaluated. Based on the experimental data obtained in a 300 dm³ pilot reactor and the dependence of mixing time on the volume of liquid phase, we have found mathematical relations between the ratio of vessel diameter to liquid level, adjustable parameters of model and the mixing time.List of Symbols V dm³ total volume of bioreactor - V _g dm³ total volume of liquid - V ₁ dm³ volume of ideally mixed zone in the vessel - V ₂ dm³ volume of macromixer in inner circulation flows - V ₃ dm³ volume of liquid phase in the pump - V ₄ dm³ volume of liquid phase in the pipe between the vessel and the pump - V ₅ dm³ volume of liquid phase in the pipe between the pump and air input system included falling jet - V _LT dm³ volume of liquid in the tank - V _LC dm³ volume of liquid in the circulation system - F _E dm³/s inner volumetric circulation flow rate across the macromixers - F _cir dm³/s external volumetric circulation flow rate, pumping capacity - t _A s time interval of the pulse application - t _AA s time point of the pulse application related to the free choosen starting point of the experiment - t _m s mixing time - t _c s circulation time - t _end s end time of simulation - C _*,* kg/m³ concentration of tracer in the indicated compartment - C ₀ kg/m³ concentration of the tracer before the injection - C _t kg/m³ concentration of the tracer at the indicated time - C kg/m³ theoretical concentration of the full mixed tracer - C _sim kg/m³ calculated concentration of tracer during numerical integration method - i index of an arbitrary tank - D _T m diameter of bioreactor - D 1/s dilution rate - H _L m level of liquid in the unaerated vessel - vector of inhomogenities     

The specific interfacial area in external-loop airlift bioreactors

The specific interfacial areas in two external-loop airlift bioreactors of laboratory and pilot scale were determined, mainly by the chemical reaction method (sulphite oxidation). The parameter studied in water/salt and starch/salt solutions was greately affected by gas superficial velocity, A _D /A _R ratio, by H _R ?H _D /H _D ratio and η _ap , respectively. Correlations for the specific interfacial area in the two systems, considering the effects of the above-mentioned parameters, were proposed.     

Hydrodynamics of non-Newtonian liquids in external-loop airlift bioreactors

Gas holdup investigation was performed in two external-loop airlift bioreactors of laboratory (V _L =1.189·10^?3? 1.880·10^?3 m³; H _R =1.16 ? 1.56 m; H _D = 1.10 m; A _D /A _R = 0.111 ? 1.000) and pilot scale (V _L =0.157?0.170 m³; H _R =4.3?4.7 m; H _D =4.0?4.4 m;A _D /A _R =0.04?0.1225), respectively, using as liquid phase non-Newtonian starch solutions of different concentration with K=0.061?3.518 Pa sⁿ and n=0.86?0.39 and fermentation broths of P. chrysogenum, S. griseus, S. erythreus, B. licheniformis and C. acremonium at different hours since inoculation and from different batches. The influence of bioreactor geometry, liquid properties and the amount of introduced compressed air was investigated. The effect of sparger design on gas holdup was found to be negligible. It was found that gas holdup depends on the flow media index, ?_GR decreasing with the increase of liquid pseudoplasticity, A _D /A _R ratio and H _R /H _D ratio. The experimental data are in agreement with those presented in literature by Popovic and Robinson, which take into account liquid properties, geometric parameters and gas superficial velocity, with a maximum error of ±30%. It was obtained a correlation for gas holdup estimation taking into account the non-Newtonian behaviour of the fermentation broths and the dry weight of the solid phase, as well. The concordance between the experimental data and those calculated with the proposed correlation was good, with a maximum error of ±17%. Also, a dimensionless correlation for gas holdup involving superficial velocities of gas and liquid, cross sectional areas ratio, dispersion height to riser diameter ratio, as well as Froude and Morton numbers, was obtained.     

Hydrodynamics in external-loop airlift bioreactors with static mixers

Liquid circulation superficial velocity and gas holdup behaviours were investigated in an external-loop airlift bioreactor of 0.170?m³ liquid volume in gas-induced and forced-circulation-loop operation modes, in the presence of static mixers made of corrugated stainless steel pieces, resulting in packets with the height-to-diameter ratio equal to unity and using non-Newtonian starch solutions as liquid phase. The static mixers were disposed in the riser in three blocks, each with three mixing packets, successively turned 90° to the adjacent mixing element. It was found that in the presence of static mixers and forced-loop operation mode, liquid circulation superficial velocity in the riser section was significantly diminished, while gas holdup increased in a great measure. It was considered that static mixers split the fluid into individual streams and break up the bubbles, resulting in small bubble sizes with a relative homogeneous bubble distribution over riser cross section. They act as supplementary resistances in liquid flow, reducing riser cross sectional area, equivalent with A _D /A _R area ratio diminishing.     

Hydrodynamics of non-Newtonian liquids in external-loop airlift bioreactors

In order to obtain further information on the behaviour and optimal design of external-circulation-loop airlift bioreactors, the liquid circulating velocity was studied using highly viscous pseudoplastic solutions of starch and antibiotic biosynthesis liquids of Penicillium chrysogenum, Streptomyces griseus, Streptomyces erythreus, Bacillus licheniformis and Cephalosporium acremonium. Measurements of liquid circulation velocity were made in laboratory and pilot plant external-loop airlift bioreactors, under various conditions concerning gas flow rate, riser liquid height at constant downcomer height, A _D /A _R ratio, using the impulse-response technique. It has been found that these parameters had a significant effect on liquid circulation velocity together with the apparent viscosity and dry weight of the solid phase in the biosynthesis liquids. For the tested liquids, the superficial liquid velocity in the riser section of an external-loop airlift bioreactor may be described by the following equation: where the exponents and the constant c take different values depending on the liquid phase properties and flow regime.     

Stochastic modelling of axial dispersion in external-loop airlift bioreactors

The paper presents a model of the motion of a particle subjected to several transport processes in connection with mixing in two phase flow. A residence time distribution technique coupled with a one-dimensional dispersion model was used to obtain the axial dispersion coefficient in the liquid phase, D_ax. The proposed model of D_ax for an external-loop airlift bioreactor is based on the stochastic analysis of the two-phase flow in a cocurrent bubble column and modified for the specific flow in the airlift reactor. The model takes into account the riser gas superficial velocity, the riser liquid superficial velocity, the Sauter bubble diameter, the riser gas hold-up, the downcomer-to-riser cross sectional area ratio. The proposed model can be applied with an average error of ᆨ.     

Dynamic modeling and simulation of continuous airlift bioreactors

For dynamic behaviors of continuous airlift bioreactors, a mathematical model based on a tanks-in-series model with backflow has been developed. The equations describing the dynamics of airlift bioreactors are material balances for micro-organism, substrate, dissolved oxygen and oxygen in gas-phase and heat balances. Non-ideal mixing of liquid and gas phases is taken into account using a tanks-in-series model with backflow. The batch operation, startup operation and the consequence of plant failure were simulated and the effects of design and operating parameters for an airlift bioreactor on its dynamic behaviors were discussed. The concentration profiles of micro-organism, substrate, dissolved oxygen and oxygen in gas-phase and the temperature profile in an airlift bioreactors and their dynamics were obtained. The computational results indicate that the transients of a chemostat in the case of bubble column bioreactor are slower compared with those in the case of airlift bioreactor. The proposed simulator is more precise as compared with models published previously in the literature and therefore provides more reliable and rational examination of continuous airlift bioreactor performance.     

Study of the liquid circulation velocity in external-loop airlift bioreactors

Liquid circulation velocity was studied in externalloop air-lift bioreactors of laboratory and pilot scale, respectively for different gas input rates, downcomer-to-riser cross-sectional area ratio, A _D/A_R and liquid phase apparent viscosities.It was found that, up to a gas superficial velocity in the riser v _SGR 0.04 m/s the dependency of v _SLR on v _SGR is in the following form: v _SLR = a v _SGR ^b , with the exponent b being 0.40. Over this value of v _SGR, only a small increase in liquid superficial velocity, v _SLR is produced by an increase in v _SGR. A _D/A_R ratio affects the liquid superficial velocity due to the resistance in flow and overall friction.For non-Newtonian viscous liquids, the circulation liquid velocity in the riser section of the pilot external-loop airlift bioreactor is shown to be dependent mainly on the downcomer-to-riser cross-sectional area ratio, A _D/A_R, the effective (apparent) liquid viscosity, _eff and the superficial gas velocity, v _SGR.The equation proposed by Popovic and Robinson [11] was fitted well, with an error of ± 20%.List of Symbols A _D m² downcomer cross-sectional area - A _Rm² riser cross-sectional area - a = coefficient in Eq. (7) - b = exponent in Eq. (7) - c _s m^–1 Coefficient in Eq. (3) - D _D m downcomer diameter - D _R m riser diameter - g m²/s gravitational acceleration - H _D m dispersion height - H _L m ungassed liquid height - K Pa s ⁿ consistency index - K _B = friction factor at the bioreactor bottom - K _F = friction factor - K _T = friction factor at the bioreactor top - V _L m³ liquid volume in the bioreactor - V _D m³ liquid volume in downcomer - V _R m³ liquid volume in riser - v _LDm/s downcomer linear liquid velocity - v _LR m/s riser linear liquid velocity - v _SGR m/s riser superficial liquid velocity - v _SLR m/s riser superficial liquid velocity - s^–1 shear rate - _GD = downcomer gas holdup - _GR = riser gas holdup - _eff Pa s effective (apparent) viscosity - Pa shear stress The authors wish to thank Mrs. Rodica Roman for the help in experimental data collection and to Dr. Stefanluca for the financial support.     

Microbial biomass production from rice straw hydrolysate in airlift bioreactors

Rice straw is a by-product of rice production, and a great bioresource as raw biomass material for manufacturing value-adding protein for animal feedstock, which has been paid more and more attention. In the present work, utilizing rice straw hydrolysate as a substrate for microbial biomass production in 11.5L external-loop airlift bioreactors was investigated. Rice straw hydrolysate obtained through acid-hydrolyzing rice straw was used for the culture of yeast Candida arborea AS1.257. The influences of gas flow rate, initial liquid volume, hole diameter of gas sparger and numbers of sieve plates on microbial biomass production were examined. The best results in the external-loop airlift bioreactor were obtained under 9.0 L initial liquid volume, 1.1 (v/v)/min gas flow rate during culture time of 0-24 h and 1.4 (v/v)/min gas flow rate of 24-48 h at 29+/-1 degrees C. The addition of the sieve plates in the riser of the external-loop airlift bioreactor increased productivity. After 48 h, under optimized operation conditions, crude protein productivity with one sieve and two sieves were 13.6 mg/mL and 13.7 mg/mL, respectively, comparing 12.7 mg/mL without sieves in the airlift bioreactor and 11.7 mg/mL in the in the 10-L mechanically stirred tank bioreactor. It is feasible to operate the external-loop airlift bioreactors and possible to reduce the production cost for microbial biomass production from the rice straw hydrolysate.     

Engineering approach to mixing quantification in bioreactors

Homogeneity-time is defined and introduced as the criterion for mixing quality in bioreactors. The criterion could replace the mixing time, in the case, when more than one measuring point (sensors) is included in the measuring system. Results based on the homogeneity-time and the temperature pulse method, achieved in stirred tank reactors under aerated conditions as well as in a jet-mixed tank, are presented.List of Symbols C _p,p kJ/kg K Heat capacity of the pulse medium - C _p,s kJ/kg K Heat capacity of the reactor-medium - F m³/s Flow rate of the pulse-input - i Inhomogeneity - I _N Inhomogeneity-number - M (t) °C Ideal response curve - m deNumber of combinations for certain number of sensors acc. to Table 1 - n Number of sensor - _p kg/m³ Density of the pulse medium - kg/m³ Density of the tank medium - s ₁ °C Mean absolute deviation of the sensor temperatures related on the ideal response curve s₂ s Mean absolute deviation of the homogeneity-times related on the time achieved with 6 sensors - t s Time - t (i) s Homogeneity-time - t _ps s Starting time of tracer injection - t _PE s End time of tracer injection - T _E °C Mean medium temperature at the end of experiment - T _k °C Temperature at k-th sensor position - T _p °C Pulse temperature - T _s °C Mean medium temperature before the tracer injection - V _s m³ Tank volume before pulse input     

Hydrodynamic behaviour of animal cell microcarrier suspensions in split-cylinder airlift bioreactors

Hydrodynamic characteristics of suspensions of microcarriers used for culturing anchorage dependent animal cells are reported in split-cylinder internal-loop airlift bioreactors. Cell culture media are simulated using salt solutions that duplicate the ionic strengths of typical media. Effects of solids loading (0–30 kg·m^–3), microcarrier particle size (150–300×10^–6 m diameter) and density (1030–1050 kg·m^–3) on gas induced circulation of the slurry, mixing time, gas holdup and gas velocity requirements to attain complete suspension of solids are discussed for two reactors with aspect ratios of 7.6 and 14.5, but equal riser-to-downcomer cross-sectional area ratios of 1.0, aerated at low air flow rates (0–8×10^–6 m³·s^–1) through a sintered glass sparger with 110×10^–6 m diameter pores. The study covers the ranges of solids concentrations, types, densities, particle sizes and aeration rates that are of relevance in animal cell culture applications.Airlift bioreactors displayed suitable hydrodynamic characteristics for potentially supporting anchorage dependent cell cultures on microcarriers at carrier loadings similar to those that are currently used in stirred tank bioreactors. The minimum gas flow rates and the induced liquid circulation rates necessary to achieve and maintain suspension of the heaviest and the largest microcarriers were well within practicable limits, limits which have been shown to be withstood by animal cells in non-anchorage dependent suspension culture in airlift bioreactors. No floatation problems were encountered with the carriers, nor was sedimentation a problem so long as the identified minimum suspension criteria were met. Chisti's liquid circulation equation, originally intended for two-phase flow, applied to the three-phase gas-liquid-microcarrier systems.     

Amelioration of ligninolytic enzyme production by Phanerochaete chrysosporium in airlift bioreactors

Maximum activities of manganese-dependent peroxidase (MnP) and lignin peroxidase (LiP) in free cultures of Phanerochaete chrysosporium (ATCC 24725) were 258 U l^–1 and 103 U l^–1, respectively, in an airlift bioreactor. Immobilisation of the fungus on an inert carrier as well as several design modifications of the bioreactor employed gave MnP activities around 500–600 U l^–1 during 9 days' operation. The continuous operation of the latter led to MnP and LiP activities about 140 U l^–1 and 100 U l^–1, respectively, for two months, without operational problems. Furthermore, the extracellular liquid secreted decolourised the polymeric dye Poly R-478 about 56%.     

Large-scale production of monoclonal antibodies in defined serum-free media in airlift bioreactors

Forty- and ninety-liter airlift bioreactors have been used successfully to grow hybridoma cell lines in chemically defined serum-free media. In the airlift bioreactor, hybridoma cell growth and monoclonal antibody productivity are comparable to that obtained by conventional cell culture. At sparging rates of 0.60-1.20 vvh (volume of sparged gas per bioreactor volume per hour), the airlift bioreactor achieves rapid mixing and adequate oxygen mass transfer. Foaming is minimal and inconsequential for serum-free media and media supplemented with 5%-10% fetal bovine serum. The use of serum-free medium facilitates monoclonal antibody purification and enhances the purity of the final MAb product.     

Performance of airlift bioreactors in the cultivation of some antibiotic producing microorganisms

The specific aspects of airlift reactors emphasizing their function relevance to particular application as bioreactors are presented. The two main groups of airlift reactors – external-loop and concentric-tube reactors – were investigated on a pilot-plant scale with regard to their performance during the cultivation of unicellular and filamentous microorganisms which produce Bacitracin, Cephalosporin C and Nystatin. Some results were compared to those obtained in conventional stirred tank bioreactors. The comparison was carried out based on physical properties (oxygen transfer rate (OTR), volumetric mass transfer coefficient (k_La) and efficiency of oxygen transfer (E)), cell mass, productivity and substrate consumption, secondary metabolite production, and efficiency of the product formation with regard to the specific power input. It was shown that B. licheniformis, C. acremonium and S. noursei fermentations occurred similarly to those performed in stirred vessels, proving that the capacity of the airlift bioreactors surpassed the problems which arise from the morphology and rheology of the broths. From the chemical engineering point of view, it was obvious that the primary tasks of a bioreactor (uniform distribution of microorganisms and nutrients over the entire fermenter volume, appropriate supply of biomass with nutrients and oxygen) were fulfilled by the airlift bioreactors tested. In addition, the efficiency of oxygen transfer (OTR referred to power input) in the airlift fermenters proved to be about 38% higher than in the stirred tank bioreactors (expressed as average values), while the sorption efficiency (OTR referred to antibiotic production) was found to be 22% greater in the airlift system than in an STR. Therefore, the biosyntheses were performed with about a 30–40% increase in energy efficiency and energy savings compared to the conventional system. Moreover, the lack of mechanical devices in the airlift system provides greater safety and a gentler environment for the cultivation of microorganisms.