Cyclohexane Removal in a Dual-Tube Membrane Bioreactor

A dual-tube dense-phase silicone rubber membrane bioreactor was investigated for control of cyclohexane-contaminated air as part of a jet propulsion (JP-8) fuel remediation investigation strategy. The reactor was seeded with a mixed bacterial consortium isolated from the water/fuel interface of a JP-8 jet fuel sample and activated sludge, capable of aromatic and cyclic compound biodegradation. Cyclohexane removal ranged from 1.1 to 28.6 mg L^?1, with removal percentages ranging from 4.6% to 37.6%. Removal in the bioreactor ranged from 29.4 to 596.6 mg min^?1 m^?2 and measured elimination capacities ranged from 46.7 to 947.9 g m^?3 h^?1. Removal rates and elimination capacity increased with increasing biofilm growth and with increasing loading rates of cyclohexane. Loading rates ranged from 395.9 to 2189.5 mg min^?1 m^?2. Results of this study showed effective removal of cyclohexane using the membrane bioreactor, suggesting that this technology may have applicability for treating vapors contaminated with cyclic hydrocarbons.     

Oxygen and CO2 transfer of a polypropylene dimpled membrane lung with variable secondary flows

Gas transfer performance is presented for one form of the Oxford membrane lung in which vortex mixing is induced in blood flow across a dimpled polypropylene membrane. Dimensional analysis has been used to define the parameters characterizing mass transfer in the device, and of three fluid mechanical parameters: Reynolds number based on peak pulsation velocity, Strouhal number, and ratio of mean to oscillatory flows, only the first has been found to affect mass transfer significantly in the ranges studied. For oxygenation, rated flows in excess of 5 l min⁻¹ m⁻² are measured. A new definition is presented for a rated flow for extracorporeal CO₂ removal and values in excess of 1.2 l min⁻¹ m⁻² are obtained.     

Bioproduction of phenylethanoid glycosides by plant cell culture of Scrophularia striata Boiss.: from shake-flasks to bioreactor

Phenylethanoid glycosides (PeG) are a class of polyphenols found in some plants that have pharmaceutical effects as anti-inflammatories and anti-oxidants. The presence of PeG (acteoside) in the aerial parts of Scrophularia striata Boiss. has been demonstrated. Considerable progress has been made using plant cell cultures to stimulate formation and accumulation of secondary metabolites. The present study optimized phenylethanoid production from shake flasks to bioreactor using a cell culture of S. striata. The optimal conditions for production of cell biomass by scale-up to a bioreactor were determined to be a pH of 4.8, air flow rate of 0.5–1.5 l min⁻¹, and mixing speed of 110–170 rpm at 25 ± 1 °C in darkness. Growth parameters and PeG production were measured and compared with the results from the shake flasks. The results showed that cell biomass was high in the bioreactor (15.64 g l⁻¹ DW) and in the shake flasks (14.16 g l⁻¹ DW). The acteoside content in the bioreactor was 1404.20 μg g⁻¹ DW, which is threefold higher than in the shake flasks (459.71 μg g⁻¹ DW). The echinacoside concentration in the bioreactor was 1449.39 μg g⁻¹, 1.36-fold lower than in the shake flasks (1973.03 μg g⁻¹ DW). This study established an efficient way for production of acteoside, the major PeG, in a bioreactor.

     

A horizontal bioreactor for ethanol production by immobilized cells

The one-parameter-tanks-in-series model was found to be an adequate model for the characterization of flow dynamics in a horizontal immobilized cell reactor, when blue dextran was used as tracer. Isobutanol proved to be inadequate, because it diffused inside the beads and thus caused tailing in RTD. The CO₂ evolution rate displayed the most pronounced effect on axial liquid dispersion. At high CO₂ production rates and low dilution rates each stage of the reactor behaved like a well-mixed reactor. At lower CO₂ evolution rates the number of tanks (N) related to the reactor increased up to 10. The medium flow rate affects axial dispersion to a minor degree. An increase of the dilution rate from 0.328 to 1.34 h^?1 resulted in a slight rise of N from 3.5 to 5 at high CO₂ production and from 4 to 7 at medium CO₂ production rates. Variation in the bead hold up showed the same characteristic axial mixing behavior as reflected by changing the medium flow rate. The quantitative correlation between axial mixing and the most significant fermentation parameters (dilution rate, CO₂ evolution rate and bead hold up) allow to develop an overall model, which besides kinetic expressions also contains terms related to the flow dynamics of the reactor. In the third part of this communication such a model will be presented and compared with actual fermentation data.     

Continuous production of ellagic acid in a packed-bed reactor

Ellagic acid is a high-value bioactive compound that is used in the food, cosmetic and pharmaceutical industries. The aim of this work was to develop a continuous system for ellagic acid production. Ellagitannase produced by solid-state fermentation and attached to polyurethane foam particles was used as a biocatalyst in a continuous bioreactor for the hydrolysis of ellagitannins from pomegranate by-product. A packed-bed reactor containing the biocatalyst (22.22 Units per gram of dry solid, U gds⁻¹) was fed with a pomegranate ellagitannins solution (0.1%, w/v) at a flow rate of 0.27 mL min⁻¹ at 60 °C. The bioreactor completed several biotransformations while maintaining the hydrolysis rate (60%) with a half-life of 10 continuous cycles of ellagic acid production. Volumetric productivity and ellagic acid yield were 1.09 g L⁻¹ h⁻¹ and 235.89 mg g⁻¹ of pomegranate ellagitannins during the first 70 min of hydrolysis, respectively. The developed biocatalyst showed good operational and mechanical stability and may be successfully used for ellagitannin hydrolysis in a continuous system. This is the first report of high-yield continuous production of ellagic acid using an auto-immobilized enzyme.     

A noninvasive technique for the measurement of the energetic state of free‐suspension mammalian cells

A perfusion small‐scale bioreactor allowing on‐line monitoring of the cell energetic state was developed for free‐suspension mammalian cells. The bioreactor was designed to perform in vivo nuclear magnetic resonance (NMR) spectroscopy, which is a noninvasive and nondestructive method that permits the monitoring of intracellular nutrient concentrations, metabolic precursors and intermediates, as well as metabolites and energy shuttles, such as ATP, ADP, and NADPH. The bioreactor was made of a 10‐mm NMR tube following a fluidized bed design. Perfusion flow rate allowing for adequate oxygen supply was found to be above 0.79 mL min^?1 for high‐density cell suspensions (10⁸ cells). Chinese hamster ovary (CHO) cells were studied here as model system. Hydrodynamic studies using coloration/decoloration and residence time distribution measurements were realized to perfect bioreactor design as well as to determine operating conditions bestowing adequate homogeneous mixing and cell retention in the NMR reading zone. In vivo ³¹P NMR was performed and demonstrated the small‐scale bioreactor platform ability to monitor the cell physiological behavior for 30‐min experiments. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

Gas–liquid mass transfer in an up-flow cocurrent packed-bed biofilm reactor

Gas–liquid mass transfer was investigated in an up-flow cocurrent packed-bed biofilm reactor. In aerobic processes gas–liquid mass transfer can be considered as a key operational parameter as well as in reactor scale-up. The present paper investigates the influence of the liquid phase mixing in the determination of the volumetric gas–liquid mass transfer coefficient (k_La) coefficient. Residence time distribution (RTD) experiments were performed in the reactor to determine the flow pattern of the liquid phase and to model mathematically the liquid phase mixing. The mathematical model derived from RTD experiments was used to evaluate the influence of the liquid mixing on the experimental estimation of the (k_La) in this reactor type. The methods used to estimate the k_La coefficient were: (i) dynamic gassing-out, (ii) sulphite method, and (iii) in-process estimation through biological conversion obtained in the reactor. The use of standard chemical engineering correlations to determine the k_La in this type of bioreactors is assessed. Experimental and modelling results show how relevant can be to take into consideration the liquid phase mixing in the calculations of the most-used methods for the estimation of k_La coefficient. k_La coefficient was found to be strongly heterogeneous along the reactor vertical axis. The value of the k_La coefficient for the packed-bed section ranged 0.01–0.12 s⁻¹. A preliminary correlation was established for up-flow cocurrent packed-bed biofilm reactors as a function of gas superficial velocity.     

Epoxidation of 1,7‐octadiene by Pseudomonas oleovorans in a membrane bioreactor

A growing cell culture of Pseudomonas oleovorans was used to biotransform 1,7‐octadiene to 1,2‐epoxy‐7,8‐octene in a continuous‐flow bioreactor with an external membrane module. A dense silicone rubber membrane was used to contact an organic phase, containing both the reactant (1,7‐octadiene) and the growth substrate (heptane), with an aqueous biomedium phase containing the biocatalyst. Heptane and octadiene delivery to the aqueous phase, and epoxide extraction into the solvent, occurred by diffusion across the dense membrane under a concentration‐driving force. In addition, a liquid feed of heptane and octadiene was pumped directly into the bioreactor to increase the rate of delivery of these compounds to the aqueous phase. In this system 1,2‐epoxy‐7,8‐octene accumulated in a pure solvent phase, thus, product recovery problems associated with emulsion formation were avoided. Furthermore, no phase breakthrough of either liquid across the membrane was observed. In this system, the highest volumetric productivity obtained was 30 U.L⁻¹, and this was achieved at a dilution rate of 0.07 h⁻¹, 70 m².m⁻³ of membrane area, and a steady‐state biomass concentration of 2.5 g.L⁻¹. The system was stable for over 1250 h. Decreasing the dilution rate led to an increased biomass concentration, however, the specific activity was significantly reduced, and therefore, an optimal dilution rate was determined at 0.055 h⁻¹. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 63: 601–611, 1999.     

Impact of nozzle operation on mass transfer in jet aerated loop reactors. Characterization and comparison to an aerated stirred tank reactor

The impact of mass transfer on productivity can become a crucial aspect in the fermentative production of bulk chemicals. For highly aerobic bioprocesses the oxygen transfer rate (OTR) and productivity are coupled. The achievable space time yields can often be correlated to the mass transfer performance of the respective bioreactor. The oxygen mass transfer capability of a jet aerated loop reactor is discussed in terms of the volumetric oxygen mass transfer coefficient k_La [h^?1] and the energetic oxygen transfer efficiency E [kgO₂ kW^?1 h^?1]. The jet aerated loop reactor (JLR) is compared to the frequently deployed aerated stirred tank reactor. In jet aerated reactors high local power densities in the mixing zone allow higher mass transfer rates, compared to aerated stirred tank reactors. When both reactors are operated at identical volumetric power input and aeration rates, local k_La values up to 1.5 times higher are possible with the JLR. High dispersion efficiencies in the JLR can be maintained even if the nozzle is supplied with pressurized gas. For increased oxygen demands (above 120 mmol L^?1 h^?1) improved energetic oxygen transfer efficiencies of up to 100 % were found for a JLR compared to an aerated stirred tank reactor operating with Rushton turbines.     

Open-flow respirometry under field conditions: How does the airflow through the nest influence our results?

Open-flow respirometry is a common method to measure oxygen-uptake as a proxy of energy expenditure of organisms in real-time. Although most often used in the laboratory it has seen increasing application under field conditions. Air is drawn or pushed through a metabolic chamber or the nest with the animal, and the O₂ depletion and/or CO₂ accumulation in the air is analysed to calculate metabolic rate and energy expenditure. Under field conditions, animals are often measured within the microclimate of their nest and in contrast to laboratory work, the temperature of the air entering the nest cannot be controlled. Thus, the aim of our study was to determine the explanatory power of respirometry in a set-up mimicking field conditions. We measured O₂ consumption of 14 laboratory mice (Mus musculus) using three different flow rates [50 L*h⁻¹ (834 mL*min⁻¹), 60 L*h⁻¹ (1000 mL*min⁻¹) and 70 L*h⁻¹ (1167 mL*min⁻¹)] and two different temperatures of the inflowing air; either the same as the temperature inside the metabolic chamber (no temperature differential; 20 °C), or cooler (temperature differential of 10 °C). Our results show that the energy expenditure of the mice did not change significantly in relation to a cooler airflow, nor was it affected by different flow rates, despite a slight, but significant decrease of about 1.5 °C in chamber temperature with the cooler airflow. Our study emphasises the validity of the results obtained by open-flow respirometry when investigating energy budgets and physiological responses of animals to ambient conditions. Nevertheless, subtle changes in chamber temperature in response to changes in the temperature and flow rate of the air pulled or pushed through the system were detectable. Thus, constant airflow during open-flow respirometry and consequent changes in nest/chamber temperature should be measured.     

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.     

Continuous Solvent/Detergent Virus Inactivation Using a Packed‐Bed Reactor

Continuous virus inactivation (VI) remains one of the missing pieces while the biopharma industry moves toward continuous manufacturing. The challenges of adapting VI to the continuous operation are two‐fold: 1) achieving fluid homogeneity and 2) a narrow residence time distribution (RTD) for fluid incubation. To address these challenges, a dynamic active in‐line mixer and a packed‐bed continuous virus inactivation reactor (CVIR) are implemented, which act as a narrow RTD incubation chamber. The developed concept is applied using solvent/detergent (S/D) treatment for inactivation of two commonly used model viruses. The in‐line mixer is characterized and enables mixing of the viscous S/D chemicals to ±1.0% of the target concentration in a small dead volume. The reactor's RTD is characterized and additional control experiments confirm that the VI is due to the S/D action and not induced by system components. The CVIR setup achieves steady state rapidly before two reactor volumes and the logarithmic reduction values of the continuous inactivation process are identical to those obtained by the traditional batch operation. The packed‐bed reactor for continuous VI unites fully continuous processing with very low‐pressure drop and scalability.     

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.     

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

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

The investigation of dairy industry wastewater treatment in a biological high performance membrane system

The dairy industry is generally considered to be the largest source of food processing wastewater in many countries. The highly variable nature of dairy wastewaters in terms of volumes and flowrates and in terms of high organic materials contents such as COD 921–9004 mg L⁻¹, BOD 483–6080 mg L⁻¹, TN of 8–230 mg L⁻¹ and SS of 134–804 mg L⁻¹ makes the choice of an effective wastewater treatment regime difficult. A high performance bioreactor, an aerobic jet loop reactor, combined with a ceramic membrane filtration unit, was used to investigate its suitability for the treatment of the dairy processing wastewater. The oxygen transfer rates of the bioreactor were found to be very high (100–285 h⁻¹) on the operating conditions. A loading rate of 53 kg COD m⁻³ d⁻¹ resulted in 97–98% COD removal efficiencies under 3 h hydraulic retention time. The high MLSS concentrations could be retained in the system (up to 38,000 mg L⁻¹) with the contribution of UF (ultrafiltration) unit. During the filtration of activated sludge, the fluxes decreased with increasing MLSS. Cake formation fouling was determined as dominant fouling mechanisms. The results demonstrate that jet loop membrane bioreactor system was a suitable and effective treatment choice for treating dairy industry wastewater.     

Citric acid production and morphology of Aspergillus niger as functions of the mixing intensity in a stirred tank and a tubular loop bioreactor

The relationship between Aspergillus niger morphology and citric acid production was investigated in two reactor systems with different configurations, a tubular loop and a stirred tank bioreactor, with operating volumes of 6 and 8 dm³, respectively. Morphology was quantified by image analysis. In each system, morphology, characterized by the parameter P (mean convex perimeter of clumps), and citric acid production, were agitation-dependent and closely linked. Increased agitation caused a reduction of clump sizes and results when both reactors demonstrate that the parameter P should not exceed a threshold value in order to achieve increased productivities. The results obtained from the two reactors were in agreement, both qualitatively and quantitatively. Reducing the fundamentally different mixing conditions of the two bioreactors to the order of the dimensionless mixing parameter relative mixing time (τ_m), results showed that the loop simulated the stirred tank. Also, relationships valid for one system accurately described the results obtained from the other system, demonstrating the validity of the relationship between morphology and productivity for the particular fermentation, regardless of the reactor type. Previous attempts to evaluate the use of loop configurations as scale-up tools and their performance as bioreactors, neglected the morphology of the producer micro-organisms. This study demonstrated the close link between morphology and productivity for citric acid production by A. niger, and identified a morphology parameter that was used successfully to characterize the process performance.     

CFD of mixing of multi‐phase flow in a bioreactor using population balance model

Mixing in bioreactors is known to be crucial for achieving efficient mass and heat transfer, both of which thereby impact not only growth of cells but also product quality. In a typical bioreactor, the rate of transport of oxygen from air is the limiting factor. While higher impeller speeds can enhance mixing, they can also cause severe cell damage. Hence, it is crucial to understand the hydrodynamics in a bioreactor to achieve optimal performance. This article presents a novel approach involving use of computational fluid dynamics (CFD) to model the hydrodynamics of an aerated stirred bioreactor for production of a monoclonal antibody therapeutic via mammalian cell culture. This is achieved by estimating the volume averaged mass transfer coefficient (k_La) under varying conditions of the process parameters. The process parameters that have been examined include the impeller rotational speed and the flow rate of the incoming gas through the sparger inlet. To undermine the two‐phase flow and turbulence, an Eulerian‐Eulerian multiphase model and k‐ε turbulence model have been used, respectively. These have further been coupled with population balance model to incorporate the various interphase interactions that lead to coalescence and breakage of bubbles. We have successfully demonstrated the utility of CFD as a tool to predict size distribution of bubbles as a function of process parameters and an efficient approach for obtaining optimized mixing conditions in the reactor. The proposed approach is significantly time and resource efficient when compared to the hit and trial, all experimental approach that is presently used. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:613–628, 2016     

Removal of benzene in a hybrid bioreactor

A novel hybrid bioreactor was designed to remove volatile organic compounds from wastewater and its performance was investigated. The bioreactor was composed of a biofilter section and a bubble column bioreactor section. Benzene was used as a model compound and the influent benzene was removed by immobilized cells in a bubble column bioreactor. Gas phase benzene stripped by air injection was removed in a biofilter. When the superficial air flow rate was 21.1 m h⁻¹ (0.76 min of residence time in a biofilter), up to 2.2 ppm of benzene in gas phase was removed completely in a biofilter and the maximum removal rate was 4.71 mg day⁻¹ cm⁻³. The concentration profile of benzene along the biofilter column was dependent on the superficial air flow rate and the degree of microbial adaptation. Air flow rate and residence time were found to be the most important operation parameters for the hybrid bioreactor. By manipulating these operational parameters, the removal efficiency and capacity of the hybrid bioreactor could be enhanced. The organic load on the hybrid bioreactor could be shared by the biofilter and bubble column bioreactors and the fluctuation of load on the hybrid bioreactor could be absorbed by changing the distribution of benzene between biofilter and bubble column bioreactors. The maximum removal capacity of the hybrid bioreactor in the experimental range was obtained when the biofilter took 50.3% of influent benzene while 100% of removal efficiency was achieved when the biofilter took 72.3% of influent benzene.     

Regime analysis of a Baeyer–Villiger bioconversion in a three-phase (air–water–ionic liquid) stirred tank bioreactor

The aim of this work was to conduct a regime analysis on a three-phase (air–water–ionic liquid) stirred tank bioreactor of the Baeyer–Villiger bioconversion process, using [MeBuPyrr][BTA] ionic liquid as the dispersed phase. The regime analysis based on characteristic times of the different mechanisms involved (mixing, mass transfer, reaction) can yield a quantitative estimate of bioreactor performance. The characteristic time obtained for oxygen uptake rate (54 s⁻¹) was among the characteristic times determined for oxygen transfer (13–129 s⁻¹) under different operating conditions, suggesting that the oxygen transfer rate under certain operating conditions could be a limiting step in the bioconversion process. Further enhancement of oxygen transfer rates requires proper selection of the bioreactor operational conditions, and improved design of the ionic liquid used as oxygen transfer vector.     

Performance of a filter loop reactor using Zymomonas mobilis and validation of the ethanol production model — II

A special loop recycle reactor with capillary crossflow filters was designed to enhance ethanol productivity. The new set-up did not comprise a classical reaction vessel with a stirrer but the working volume consisted only of the void volumes of the filters and the peripheral equipment, however, it behaved as a well mixed CSTR. A circulating pump provided for both mixing and recirculation of the culture fluid. Degassing was accomplished with a cyclone type device. Even if the circulating pump destroyed cells and automated backflushing of the filters could not prevent membrane fouling, maximum biomass concentration reached 98 g l⁻¹ at a dilution rate of 4 h⁻¹ and the maximum ethanol productivity achieved was 224 g l⁻¹ h⁻¹. Using the loop recycle reactor, the ethanol production model proposed in Part I of this study (Nipkow et al. (1986) J. Biotechnol. 3, 35–47) extended with a variable rate accounting for mechanical destruction of cells was verified. It was demonstrated that the predicted productivity of 350 g l⁻¹ h⁻¹ — exploiting the biological potential of Zymomonas mobilis — should be attainable if improved mechanical equipment in a filter recycle system is employed.