Studies on mixing in horizontal rotating tubular bioreactor

A horizontal rotating tubular bioreactor (HRTB) is a combination of a “thin-layer bioreactor” and a “biodisc” reactor. Its interior is divided by O-ring shaped partition walls. Mixing properties of this new type of the bioreactor were investigated by using a temperature step method. The mixing simulations were done by Runge-Kutta-Fehlberg numerical integration. Adjustable parameters of the “spiral flow” model were optimised by Monte-Carlo method. In this investigation, the structured “spiral flow” model (containing four adjustable parameters) was tested in a wide range of experimental conditions. The results show that the structured “spiral flow” model is capable to describe the mixing in HRTB in the whole range of both bioreactor operational parameters (n and D).     

Studies on mixing in horizontal rotating tubular bioreactor

In previous investigations on mixing in a horizontal rotating tubular bioreactor (HRTB) the structured “spiral flow” model was developed which contained four adjustable parameters [1, 2]. In order to incorporate the mixing model in a semifundamental scale-up procedure it was necessary to make a relation between the adjustable model parameters and process parameters of the bioreactor expressed as dimensionless numbers. Mathematical equations which relate adjustable model parameters with dimensionless numbers were developed by non-linear and surface regression methods. These equations were applied to develop the prediction systems for adjustable model parameters. In total, nine systems of equations for the prediction of the adjustable model parameters were established and examined by simulation. Three of them (SC-2, SC-6 and SC-9) were selected as adequate to describe the mixing performance of HRTB in a wide range of process conditions.     

Estimation of axial dispersion in horizontal rotating tubular bioreactor by means of a structured model

A horizontal rotating tubular bioreactor (HRTB) is designed as the combination of a "thin layer bioreactor" and a "biodisc" reactor. The investigation of mixing in HRTB was done by the temperature step method in a wide range of process conditions [residence time (t_z=360036000 s) and bioreactor rotation speed (n=0.0830.917 s^у)]. In all experiments heat losses were detected. A mathematical model based on "tank in series" concept was developed to describe the mixing in HRTB - a "spiral flow" model (SFM) which has incorporated heat losses. However, the simulations of SFM could be used for calculation of temperature response curves for the case when there is no heat losses. These corrected curves were used then to estimate Bodenstein number as a parameter of standard dispersion model (SDM). The obtained Bodenstein numbers were in the range 10-17. The simulations showed that SFM was more capable to describe the mixing in HRTB giving better fitting with experimental measurements than SDM, indicating that mixing pattern in HRTB is too complex to be described with this relatively simple, one-parameter model.     

Heterotrophic cultivation of Paracoccus denitrificans in a horizontal rotating tubular bioreactor

In this work, the heterotrophic cultivation of bacterium Paracoccus denitrificans has been studied in a horizontal rotating tubular bioreactor (HRTB). After development of a microbial biofilm on the inner surface of the HRTB, conditions for one-step removal of acetate and ammonium ion were created. The effect of bioreactor process parameters [medium inflow rate (F) and bioreactor rotation speed (n)] on the bioprocess dynamics in the HRTB was studied. Nitrite and nitrogen oxides (NO and N₂O) were detected as intermediates of ammonium ion degradation. The biofilm thickness and the nitrite concentration were gradually reduced with increase of bioreactor rotation speed when the medium inflow rate was in the range of 0.5–1.5 l h⁻¹. Further increase of inflow rate (2.0–2.5 l h⁻¹) did not have a significant effect on the biofilm thickness and nitrite concentration along the HRTB. Complete acetate consumption was observed when the inflow rate was in the range of 0.5–1.5 l h⁻¹ at all bioreactor rotation speeds. Significant pH gradient (cca 1 pH unit) along the HRTB was only observed at the highest inflow rate (2.5 l h⁻¹). The results have clearly shown that acetate and ammonium ion removal by P. denitificans can be successfully conducted in a HRTB as a one-step process.     

Mixing time in a down-flow jet loop bioreactor

Mixing time was determined in a down-flow jet loop bioreactor with Newtonian and non-Newtonian fluids. It was observed that the mixing time decreased with an increase in linear liquid velocity, superficial gas velocity, draft tube to column diameter ratio, nozzle diameter and shear thinning of media. The optimum draft tube to column diameter ratio was found to be about 0.44. Correlations were presented for prediction of mixing time.List of Symbols A m² cross sectional area of the column - C kmol/m³ local tracer concentration - A _D m² flow area,A _D =/4 (D _Z ² -D _TO ² ) - D m column diameter - D _E m draft tube diameter - D _TO m outside diameter of the air tube - D _TFL m equivalent flow diameter,D _TFL =(D _Z ² -D _TO ² )^0.5 - D _z m nozzle diameter - g m/s² gravitational acceleration - h % inhomogeneity - H m height of the column - H _B m distance between the lower edge of the draft tube and the impact plate - H _T m distance between the upper edge of the draft tube and the liquid nozzle - K Pa.sⁿ consistency index in power-law model - L _E m length of the draft tube - n flow index in the power-law model - Re _j jet Reynolds number,Re _j =(D _TFL×w₁×_L)/ _eff - t _M s mixing time - t _sg m/s superficial gas velocity based onA - W _l m/s linear liquid velocity based onD _D Greek Letters N/m² shear stress - s shear rate - kg/m³ density of liquid - N/m surface tension of the liquid - Pa.sⁿ viscosity of liquid Indices X concentration at infinite time maximum value of tracer concentration - eff effective - L Liquid - obs observed - pred Predicted     

Fermentative bioconversion in a horizontal rotating tubular bioreactor

The performance of a horizontal rotating tubular bioreactor (HRTB) was investigated with a biological system under non-sterile conditions. A spontaneously developed microbial culture was cultivated in a simple glucose/yeast extract medium. A fermentative bioconversion was examined by different combinations of process parameters (bioreactor rotation speed 5–30 min⁻¹ and medium inflow rate 1–10 l h⁻¹). Bioconversion dynamics in HRTB was monitored by withdrawing the samples from five positions along the bioreactor. Investigation in HRTB showed a rapid and an efficient glucose conversion into different products of metabolism. Glucose consumption rate along the HRTB depended on medium inflow rate, while bioreactor rotation speed did not have a significant influence. Complete glucose conversion in HRTB was observed at inflow rates of up to 6.5 l h⁻¹. The pH gradient along the HRTB was detected at higher medium inflow rates (6.5 and 10 l h⁻¹), but did not significantly influence substrate conversion efficiency. A discussion of its potential use and a comparison of HRTB with other bioreactors are also presented.     

Modelling mixing parameters in concentric-tube airlift bioreactors

Axial dispersion of the liquid phase was investigated in a concentric-tube airlift bioreactor (RIMP: V _L=0.70?m³) as a whole and in the separate zones (riser, downcomer, gas-separator) using the axial dispersion model. The axial dispersion number Bo and the axial dispersion coefficient, D _ax were determined from the output curves to an initial Dirac pulse, using the tracer response technique. They were analyzed in relation to process and geometrical parameters, such as: gas superficial velocity, ν_SGR; top clearance, h _S; bottom clearance, h _B, and resistances at downcomer entrance expressed as A _d/A _R ratio. Correlations between Bodenstein numbers in the overall bioreactor and riser and downcomer sections (Bo_T,Bo_R,Bo_D) and the geometrical and process parameters were developed, which can allow to assess the complex influence of these parameters on liquid axial dispersion.     

Application of a Novel Oscillatory Flow Micro-bioreactor to the Production of γ-decalactone in a Two Immiscible Liquid Phase Medium

A novel micro-bioreactor based on the oscillatory flow technology was applied to the scale-down of the biotechnological production of γ-decalactone. A decrease up to 50% of the time required to obtain the maximum concentration of the compound was observed, when compared with other scaled-down platforms (stirred tank bioreactor or shake flask). A three-fold increase in γ-decalactone productivity was obtained by increasing oscillatory mixing intensity from Re_o ~482 to Re_o ~1447. This was presumably related to the effective contribution of the reactor geometry to enhanced mass transfer rates between the two immiscible liquid phases involved in the process by increasing the interfacial area. Revisions requested 11 October 2005; Revisions received 4 January 2006     

Effect of dilution rate on growth characteristics of Candida utilis in multistage culture systems with different interstage mixing

The effect of dilution rate on important process parameters of biomass production in two multistage culture systems with different interstage mixing has been examined. Experiments were performed in a multistage tower fermenter and in a cascade of fermenters. Measurements were made at steady-state of continuous culture under constant and identical values of ethanol concentration of 50 gl^?1 in the feed, temperature, OTR and pH in both culture systems used. The microorganism used was Candida utilis. Ethanol inhibition influenced cell growth rate due to the complete dissimilation of the restricted quantity of acetate to H₂O and CO₂, leading to insufficient energy generation. The value of ethanol concentration at which ethanol started to inhibit cell growth was a combined function of OTR, S_R and D. The presence of the interstage mixing resulted in more efficient ethanol conversion to biomass in the whole range of dilution rates and significantly lowered the risk of washing-out at high values of both S_R and D.     

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.     

Mathematical modeling of mixing in a horizontal rotating tubular bioreactor: “Spiral flow” model

A horizontal rotating tubular bioreactor (HRTB) was designed as a combination of a thin-layer bioreactor and a biodisc reactor whose interior was divided by O-ring shaped partition walls. For the investigation of mixing in HRTB the temperature step method was applied. Temperature changes in the bioreactor were monitored by six Pt-100 sensors (t ₉₀ response time 0.08 s and resolution 0.002 °C) which were connected with an interface unit and a personal computer. In this work a modified tank in series concept was used to establish a mathematical model. The heat balance of the model compartments was established according to the physical model and the spiral flow pattern. Numerical integration was done by the Runge-Kutta-Fehlberg method. The mathematical mixing model called spiral flow model contained four adjustable parameters (N1, Ni, F _cr and F _p) and five parameters which characterized the plant and experimental conditions. The spiral flow model was capable to describe the mixing in HRTB properly, and its applicability was much better than with the simple flow model, presented earlier.List of Symbols A _ui m² ithpart of inner surface of bioreactor's wall - A _vi m² ith part of outlet surface of bioreactor's wall - C _p kJ kg^–1 K^–1 heat capacity of liquid - c _pr kJ kg^–1 K^–1 heat capacity of bioreactor's wall - D h^–1 dilution rate - E °C°C^–1h^–1 error of mathematical model - F _cr dm³ s^–1 circulation flow in the model - F _p dm³s^–1 back flow in the model - F _t dm³ s^–1 inlet flow in bioreactor - I °C intensity of temperature step, the difference in temperature between the temperature of the inlet liquid flow and the temperature of liquid in bioreactor before temperature step - K1 Wm^–2 K ^–1 heat transfer coefficient between the liquid and bioreactor's wall - K2 Wm^–2 K ^–1 heat transfer coefficient between the bioreactor's wall and air - L m length of bioreactor - m _s kg mass of bioreactor's wall - n min^–1 rotational speed of bioreactor - n _s number of temperature sensors - N1 number of cascades - Ni number of compartments inside cascade - r _u m inner diameter of bioreactor - r _v m outside diameter of bioreactor - s(t) step function - t s time - T °C temperature - T _c °C calculated temperature - T _m °C measured temperature - T _N 1,Ni°C temperature of liquid in defined compartments inside the cascade - T _N 1,S°C temperature of defined part of bioreactor's wall - T _N i,z°C temperature of surrounding air - V _t dm³ volume of liquid in the bioreactor     

Mathematical modeling of Fe(II), Cu(II), Ni(II) and Zn(II) removal in a horizontal rotating tubular bioreactor

Industrial wastewaters polluted with toxic heavy metals are serious ecological and environmental problem. Therefore, in this study multi-heavy metals (Fe²⁺, Cu²⁺, Ni²⁺ and Zn²⁺) removal process with mixed microbial culture was examined in the horizontal rotating tubular bioreactor (HRTB) by different combinations of process parameters. Hydrodynamic conditions and biomass sorption capacity have main impact on the removal efficiency of heavy metals: Fe²⁺ 95.5–79.0%, Ni²⁺ 92.7–54.8%, Cu²⁺ 87.7–54.9% and Zn²⁺ 81.8–38.1%, respectively. On the basis of experimental results, integral mathematical model of removal heavy metals in the HRTB was established. It combines hydrodynamics (mixing), mass transfer and kinetics to define bioprocess conduction in the HRTB. Mixing in the HRTB was described by structured cascade model and metal ion removal by two combined diffusion–adsorption models, respectively. For Langmuir model, average variances between experimental and simulated concentrations of metal ions were in the range of 1.22–10.99 × 10⁻³ and for the Freundlich model 0.12–3.98 × 10⁻³, respectively. On the basis of previous facts, it is clear that developed integral bioprocess model with Freundlich model is more efficient in the prediction of concentration of metal ions in the HRTB. Furthermore, the results obtained also pointed out that the established model is at the same time accurate and robust and therefore it has great potential for use in the scale-up procedure.     

Induced reversal of order of cell division and DNA replication in Tetrahymena.

The sequence of macronuclear DNA replication (S) and cell division (D) in two normal or synchronized cell cycles in (amicronucleate) Tetrahymena can be symbolized S₁, D₁, S₂, D₂. Using heat shock synchronized cells, the final heat shock (33.8 °C) has been extended from 20 min to 5 h. This leads to an altered sequence: S₁, S₂, D₁, D₂, characterized by reversal of D₁ and S₂. S₂ occurs during the extended shock, D₁ comes in standard time after the shock has been discontinued. Thus prolonged stay at elevated temperature can dissociate two cell cycles into a subcycle of DNA replication followed by a subcycle of cell division. S₁ and S₂ charge the cells with four times the amount of DNA in newly divided cells, and D₁ and D₂ partition this DNA to four cells. Synthesis of DNA between D₁ and D₂ is not required, and it occurs in only few cells.     

Effects of geometrical design on hydrodynamic and mass transfer characteristics of a rectangular-column airlift bioreactor

Gas holdup and gas–liquid mass transfer coefficients were measured in a 21-L rectangular-column airlift bioreactor with aspect ratio of 10 and working volumes ranging from 10 to 16 L. The effect of the bottom and top clearances was investigated using water and mineralized CMC solutions and covering a range of effective viscosity from 0.02 to 0.5 Pa s and surface tension from 0.065 to 0.085 N m⁻¹. The gas holdup and mass transfer results were successfully correlated using expressions derived via dimensional analysis. The separator gas holdup was found to be similar to the total gas holdup in the airlift bioreactor. The downcomer gas holdup (ɛ_d) increased two-fold when the bottom clearance (h_b) was increased from 0.014 to 0.094 m while the top clearance (h_t) had no effect. Increasing h_b decreased the mass transfer by 50% compared to 31% when the top clearance (h_t/D_hr) was increased. It was found that the gas–liquid separator diameter ratio (D_hs/D_hc) exerted the maximal influence of over 65% on mass transfer as compared to both clearances.     

Kinetics of ion exchange process for separation of glutamic acid

Kinetics of the separation of L-glutamic acid (GLU) by ion exchange has been studied with strongly acidic H⁺-type cation exchange resin Amberlite IR-122. Since glutamic acid is a trivalent ampholyte and dissociates according to three equilibrium reactions, separation of G⁺ ions by a cation exchange process is accompanied by the dominant reversible reaction, i.e. G⁺+H⁺ ? G⁰. Accompanying reversible reaction has an effect on the ion exchange rate, and decreases the performance of the process comparing with the ideal case that the exchanging ions retain their identity. The analysis was performed first with the ion exchange column, DIC (L/D=0.52); and then with the ion exchange column, IC (L/D=10.9). The data were collected with model glutamic acid solutions for both DIC and IC columns/reactors. IC experimental results were then compared with that of DIC and the effect of scale up on ion exchange process was investigated. The experimental results have provided an adequate basis for the design calculations, and the design parameters were determined. Rate coefficients for the liquid phase mass transfer controlled cation exchange process were calculated and interrelated with a plot of j _Mfactor versus Reynolds number.     

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.     

Biodegradation of Methyl Orange by alginate-immobilized Aeromonas sp. in a packed bed reactor: external mass transfer modeling

Azo dyes are recalcitrant and xenobiotic nature makes these compounds a challenging task for continuous biodegradation up to satisfactorily levels in large-scale. In the present report, the biodegradation efficiency of alginate immobilized indigenous Aeromonas sp. MNK1 on Methyl Orange (MO) in a packed bed reactor was explored. The experimental results were used to determine the external mass transfer model. Complete MO degradation and COD removal were observed at 0.20 cm bead size and 120 ml/h flow rate at 300 mg/l of initial dye concentration. The degradation of MO decreased with increasing bead sizes and flow rates, which may be attributed to the decrease in surface of the beads and higher flux of MO, respectively. The experimental rate constants (k _ps) for various beads sizes and flow rates were calculated and compared with theoretically obtained rate constants using external film diffusion models. From the experimental data, the external mass transfer effect was correlated with a model J _D = K Re ^?(1 ? n). The model was tested with K value (5.7) and the Colburn factor correlation model for 0.20, 0.40 and 0.60 bead sizes were J _D = 5.7 Re ^?0.15, J _D = 5.7 Re ^?0.36 and J _D = 5.7 Re ^?0.48, respectively. Based on the results, the Colburn factor correlation models were found to predict the experimental data accurately. The proposed model was constructive to design and direct industrial applications in packed bed reactors within acceptable limits.     

Effects of cations on the plasma membrane of Blastocladiella emersonii zoospores. An ESR study

The physical properties of the plasma membrane of the aquatic phycomycete Blastocladiella emersonii were investigated, in particular the effects of cations on membrane structure. Intact zoospores and lipid extracts were labelled with the spin-labels 5-nitroxystearate (5-NS), 12-nitroxystearate (12-NS), and 2,2,6,6-tetramethylpiperidine-1-oxyl (Tempo). Electron spin resonance spectroscopy indicated a total of three breaks in plots of the hyperfine splitting parameter, 2T_|, order parameter, S, and the partition coefficient, f, vs. temperature. The first and third break points (T_L and T_H) were found to be independent of the external K⁺, Ca²⁺, or Mg²⁺ concentrations. They were similar to the break points found in aqueous dispersions of lipid extracts and correlate well with the temperature limits for zoospore viability. In contrast, the middle break point (T_M) was markedly influenced by the external Ca²⁺ concentration. Ca²⁺ increased T_M from 12°C (no Ca²⁺ added) to 22°C (10 mM Ca²⁺), i.e., growth temperature. K⁺ reversed this Ca²⁺ effect, downshifting T_M from 22°C to 10°C. A comparison of the physico-chemical effects of these ions on the membrane, as revealed by the cation-induced shift in T_M, is closely correlated with the temperature dependence and physiological effects of cations on zoospore differentiation. This suggests that cations may modify the physical state of the plasma membrane and be involved in regulating the initial changes during zoospore encystment.     

Genetic diversity of nine faba bean (Vicia faba L.) populations revealed by isozyme markers

Seven isozyme systems (Sod, 6-Pgd, Me, Est, Skdh, Fdh and Gdh) representing nine loci were used to study the genetic diversity of nine faba bean populations. Seven loci revealed polymorphic bands and showed the same quaternary structure as that found in several species. They revealed a high number of phenotypes. Indeed, from 3 to 9 phenotypes per locus were investigated in this study. The percentage of polymorphic loci (P = 59.3 %) was higher than that mentioned in the autogamous species (P = 20.3 %) and less than the optimum (P=96 %) indicated for allogamous plants. Total genetic diversity (H _T) and within population genetic diversity (H _S) were estimated with the isozyme markers. The contribution of among population genetic diversity (D _ST) to total genetic diversity was 22%. Enzyme markers pointed out an average inbreeding level for whole population (F _IT) and within population (F _IS). Within population genetic diversity represents 78% of total diversity. Intra-population genetic diversity (H _S = 0.206) was ranged with the respect of allogamous species and was clearly higher than that of among population genetic diversity (D _ST = 0.057) indicating an out-crossing predominance in the studied populations. The expected heterozygosity was higher than that observed heterozygosity at the allogamous species was confirmed in this study. Although, the mean estimated gene flow was less than 1(Nm=0.814), the dendrogram based on Nei’s genetic distance of the 9 populations using UPGMA method showed some genetic drift between populations.