A comparison between the reaction rates in biofilm reactors and free suspended cells bioreactors

A theoretical equation for comparison of the bioreaction rates in a biofilm reactor and in a bioreactor with free suspended cells at different input substrate concentrations and substrate conversions is proposed. The main assumption is that there is no kinetic change after fixation of the cells. Some previously published experimental data are used for verification of this equation. It is shown that difference between the experimental and calculated data is within 19%.     

Gaseous concentration gradients in tray type solid state fermentors — Effect on yields and productivities

In aerobic solid state fermentation systems, interaction of mass transfer effects with bioreaction plays an important role on the yields and productivities of the bioreactors. Experimental observations on the oxygen and carbon dioxide concentration gradients in a tray type solid state fermentation system are reported in this paper. Steep gradients are experienced in deep beds making large portions of the bioreactor ineffective. The results are useful in the design of the bioreactor in terms of efficient mass transfer as well as critical thickness of the substrate bed to be used.     

The solution of hollow fiber bioreactor design equations.

A methodology for simplifying the solution procedure for hollow fiber bioreactor design equations has been described. Such a procedure facilitates decoupling of membrane and spongy matrix equations from the tube side equations. The equivalence between the reduced equations and the hemodialyzer problem has been explicitly obtained.     

A microscale model of bacterial and biofilm dynamics in porous media

A microscale model for the transport and coupled reaction of microbes and chemicals in an idealized two-dimensional porous media has been developed. This model includes the flow, transport, and bioreaction of nutrients, electron acceptors, and microbial cells in a saturated granular porous media. The fluid and chemicals are represented as a continuum, but the bacterial cells and solid granular particles are represented discretely. Bacterial cells can attach to the particle surfaces or be advected in the bulk fluid. The bacterial cells can also be motile and move preferentially via a run and tumble mechanism toward a chemoattractant. The bacteria consume oxygen and nutrients and alter the profiles of these chemicals. Attachment of bacterial cells to the soil matrix and growth of bacteria can change the local permeability. The coupling of mass transport and bioreaction can produce spatial gradients of nutrients and electron acceptor concentrations. We describe a numerical method for the microscale model, show results of a convergence study, and present example simulations of the model system.     

Fully automated single-use stirred-tank bioreactors for parallel microbial cultivations

Single-use stirred tank bioreactors on a 10-mL scale operated in a magnetic-inductive bioreaction block for 48 bioreactors were equipped with individual stirrer-speed tracing, as well as individual DO- and pH-monitoring and control. A Hall-effect sensor system was integrated into the bioreaction block to measure individually the changes in magnetic field density caused by the rotating permanent magnets. A restart of the magnetic inductive drive was initiated automatically each time a Hall-effect sensor indicates one non-rotating gas-inducing stirrer. Individual DO and pH were monitored online by measuring the fluorescence decay time of two chemical sensors immobilized at the bottom of each single-use bioreactor. Parallel DO measurements were shown to be very reliable and independently from the fermentation media applied in this study for the cultivation of Escherichia coli and Saccharomyces cerevisiae. The standard deviation of parallel pH measurements was pH 0.1 at pH 7.0 at the minimum and increased to a standard deviation of pH 0.2 at pH 6.0 or at pH 8.5 with the complex medium applied for fermentations with S. cerevisiae. Parallel pH-control was thus shown to be meaningful with a tolerance band around the pH set-point of ± pH 0.2 if the set-point is pH 6.0 or lower.     

An analysis of a trickle-bed bioreactor: carbon disulfide removal

An analysis of the local processes occurring in a trickle-bed bioreactor (TBB) with a first-order bioreaction shows that the identification of the TBB operating regime requires knowledge of the substrate concentration in the liquid phase. If the substrate liquid concentration is close to 0, the rate-controlling step is mass transfer at the gas-liquid interface; when it is close to the value in equilibrium with the gas phase, the controlling step is the phenomena occurring in the biofilm. CS2 removal rate data obtained in a TBB with a Thiobacilii consortia biofilm are analyzed to obtain the mass transfer and kinetic parameters, and to show that the bioreactor operates in a regime mainly controlled by mass transfer. A TBB model with two experimentally determined parameters is developed and used to show how the bioreactor size depends on the rate-limiting step, the absorption factor, the substrate fractional conversion, and on the gas and liquid contact pattern. Under certain conditions, the TBB size is independent of the flowing phases' contact pattern. The model effectively describes substrate gas and liquid concentration data for mass transfer and biodegradation rate controlled processes.     

Modeling of continuous L(+)-lactic acid production with immobilized R. oryzae in an airlift bioreactor

Continuous L(+)-lactic acid production was carried out in an airlift bioreactor with immobilized R. oryzae in polyurethane foam cubes. In a pseudo-steady state, the productivity of lactic acid increased with increasing dilution rate or feeding glucose concentration. A double-layer reaction-diffusion model for the pseudo-steady state process was developed to describe the bioreaction system. Using independently determined model parameters, the model prediction agreed well with the experimental results. Therefore, the model can be employed to understand the fermentation behavior, and for the process design and optimization.     

Perfluorocarbon facilitated O2 transport in a hepatic hollow fiber bioreactor

A mathematical model describing O₂ transport in a hepatic hollow fiber (HF) bioreactor supplemented with perfluorocarbons (PFCs) in the circulating cell culture media was developed to explore the potential of PFCs in properly oxygenating a bioartificial liver assist device (BLAD). The 2‐dimensional model is based on the geometry of a commercial HF bioreactor operated under steady‐state conditions. The O₂ transport model considers fluid motion of a homogeneous mixture of cell culture media and PFCs, and mass transport of dissolved O₂ in a single HF. Each HF consists of three distinct regions: (1) the lumen (conducts the homogeneous mixture of cell culture media and PFCs), (2) the membrane (physically separates the lumen from the extracapillary space (ECS), and (3) the ECS (hepatic cells reside in this compartment). In a single HF, dissolved O₂ is predominantly transported in the lumen via convection in the axial direction and via diffusion in the radial direction through the membrane and ECS. The resulting transport equations are solved using the finite element method. The calculated O₂ transfer flux showed that supplementation of the cell culture media with PFCs can significantly enhance O₂ transport to the ECS of the HF when compared with a control with no PFC supplementation. Moreover, the O₂ distribution and subsequent analysis of ECS zonation demonstrate that limited in vivo‐like O₂ gradients can be recapitulated with proper selection of the operational settings of the HF bioreactor. Taken together, this model can also be used to optimize the operating conditions for future BLAD development that aim to fully recapitulate the liver's varied functions. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

Selective fengycin production in a modified rotating discs bioreactor

Production of lipopeptides fengycin and surfactin in rotating discs bioreactor was studied. The effects of rotation velocity and the addition of agitators between the discs on volumetric oxygen transfer coefficient k _L a were firstly studied in model media. Then the production of lipopeptides was also studied at different agitation conditions in the modified bioreactor (with agitators). The effect of agitation on dissolved oxygen, on submerged and immobilized biomass, on lipopeptide concentrations and yields and on the selectivity of the bioreaction was elucidated and discussed. The proposed modified rotating discs bioreactor allowed to obtain high fengycin concentrations (up to 787 mg L^?1), but also better selectivity of the bioreaction towards fengycin (up to 88 %) and better yields of fengycin per glucose (up to 62.9 mg g^?1), lipopeptides per glucose (up to 71.5 mg g^?1), fengycin per biomass (up to 309 mg g^?1) and lipopeptides per biomass (up to 396 mg g^?1) than those reported in the literature. Highest fengycin production and selectivity were obtained at agitation velocity of 30 min^?1. The proposed non-foaming fermentation process could contribute to the scale-up of lipopeptide fermentors and promote the industrial production of fengycin. The proposed bioreactor and bioprocess could be very useful also for the production of other molecules using bioprocesses requiring bubbleless oxygen supply.     

The flux-force relations across complex membranes with active transport

Equations are derived for the total material flux, and the total electric current flux, across a complex membrane system with active transport. The equations describe the fluxes as linear functions of forces across the system, and specifically of electrical potential, hydrostatic pressure, chemical potentials, and active transport rates. The equations can be simplified for experimental studies by making one or more of the forces equal to zero. The osmotic pressure difference across a membrane system is shown to be a function of the electrical potential and chemical potential differences and of the active transport rates. The transmembrane potential is shown to be the sum of a diffusion potential and an active transport potential. A simple equation is derived describing the current across a membrane as a linear function of the electrical potential and the active transport rate. Specific examples of the application of the equations to nerve membrane potentials are considered.     

A novel membrane bioreactor for detoxifying industrial wastewater: II. Biodegradation of 3-chloronitrobenzene in an industrially produced wastewater

A novel membrane bioreactor has been used for the treatment of an industrially produced wastewater arising in the manufacture of 3-chloronitrobenzene. This wastewater is not amenable to direct biological treatment without some form pretreatment or dilution, due to the inorganic composition (pH <1, salt concentration 4% w/w) of the wastewater. In the membrane bioreactor, the organic pollutants are first separated from the wastewater by selective membrane permeation, and then biodegraded in the biological growth compartment of the bioreactor. At a wastewater flow rate of 64 mL h(-1) (corresponding to a contact time of approximately 1.7 hours) over 99% of the 3-chloronitrobenzene and over 99% of the nitrobenzene in the wastewater were degraded. Degradation of 3-chloronitrobenzene was accompanied by evolution of chloride ions in a stoichiometric ratio. Both 3-chloronitrobenzene and nitrobenzene degradation were accompanied by the evolution of carbon dioxide; approximately 80% of the carbon entering the system was oxidized to CO(2) carbon. Analysis of mass transport across the membrane revealed that 3-chloronitrobenzene and nitrobenzene are transported more rapidly than phenol. This is explained in terms of a resistances-in-series model, which shows phenol transfer to be rate limited by the membrane diffusion step, whereas the chloronitrobenzene and nitrobenzene transfer are rate limited by the liquid film mass transfer. (c) 1993 Wiley & Sons, Inc.     

Effect of unstirred layers on binding and reaction kinetics at a membrane surface

The effects of solution unstirred layers on the time course of chemical reactions and transport processes at a membrane surface are determined. A set of equations which describes non-steady-state diffusion through an unstirred layer coupled with chemical reaction at a membrane surface or transport through a membrane is developed. A numerical solution to the equations is obtained by uncoupling diffusive and chemical processes in an iterative manner. The diffusive process is solved by the Crank-Nicolson method; the chemical process is solved by integrating the differential equations describing the kinetics. Diffusive processes in one dimension, in three dimensions, and in the presence of an arbitrary potential near the membrane surface are solved. General characteristics of the calculated reaction time course are discussed using surface binding and membrane transport examples. Small, neglected, unstirred layers are shown to sometimes yield erroneous values of rate parameters for a surface reaction and to simulate competitive reaction kinetics. Experimental approaches for measuring unstirred layer thickness are reviewed.     

Oxygen transport and consumption by suspended cells in microgravity: a multiphase analysis

A rotating bioreactor for the cell/tissue culture should be operated to obtain sufficient nutrient transfer and avoid damage to the culture materials. Thus, the objective of the present study is to determine the appropriate suspension conditions for the bead/cell distribution and evaluate oxygen transport in the rotating wall vessel (RWV) bioreactor. A numerical analysis of the RWV bioreactor is conducted by incorporating the Eulerian-Eulerian multiphase and oxygen transport equations. The bead size and rotating speed are the control variables in the calculations. The present results show that the rotating speed for appropriate suspensions needs to be increased as the size of the bead/cell increases: 10 rpm for 200 microm; 12 rpm for 300 microm; 14 rpm for 400 microm; 18 rpm for 600 microm. As the rotating speed and the bead size increase from 10 rpm/200 microm to 18 rpm/600 microm, the mean oxygen concentration in the 80% midzone of the vessel is increased by approximately 85% after 1-h rotation due to the high convective flow for 18 rpm/600 microm case as compared to 10 rpm/200 microm case. The present results may serve as criteria to set the operating parameters for a RWV bioreactor, such as the size of beads and the rotating speed, according to the growth of cell aggregates. In addition, it might provide a design parameter for an advanced suspension bioreactor for 3-D engineered cell and tissue cultures.     

Dual-chambered membrane bioreactor for coculture of stratified cell populations

We have developed a dual-chambered bioreactor (DCB) that incorporates a membrane to study stratified 3D cell populations for skin tissue engineering. The DCB provides adjacent flow lines within a common chamber; the inclusion of the membrane regulates flow layering or mixing, which can be exploited to produce layers or gradients of cell populations in the scaffolds. Computational modeling and experimental assays were used to study the transport phenomena within the bioreactor. Molecular transport across the membrane was defined by a balance of convection and diffusion; the symmetry of the system was proven by its bulk convection stability, while the movement of molecules from one flow line to the other is governed by coupled convection-diffusion. This balance allowed the perfusion of two different fluids, with the membrane defining the mixing degree between the two. The bioreactor sustained two adjacent cell populations for 28 days, and was used to induce indirect adipogenic differentiation of mesenchymal stem cells due to molecular cross-talk between the populations. We successfully developed a platform that can study the dermis–hypodermis complex to address limitations in skin tissue engineering. Furthermore, the DCB can be used for other multilayered tissues or the study of communication pathways between cell populations.     

膜生物反应器的研究进展

膜生物反应器是近年来发展的废水处理新技术,具有活性污泥浓度高、污泥龄长、占地面积小、投资省的特点。利用膜生物反应器进行污水处理不仅可以大大节约水资源,还可以大大节约能源,节省设备和运行费用,已成为二十一世纪研究热点。膜生物反应器是通过高效膜分离技术与活性污泥相结合,增大污泥中的特效菌来加快生化反应速率,提高废水处理效果。目前处理对象已从生活污水扩展到高浓度的有机废水和难降解的工业废水。本文综述了膜生物反应器在废水中的应用研究情况,并分析比较了各种膜材质的特点、适用范围以及膜的污染因素和清洗方法,展望了膜生物反应器的应用前景及进一步研究方向。     

Enhancing Cell Viability with Pulsating Flow in a Hollow Fiber Bioartificial Liver

A pulsating flow of medium was used to alleviate diffusion and transport limitations in a hollow fiber bioreactor containing a human hepatoblastoma cell line. The strategy is easy to implement but effective. The pulsating flow is introduced by a solenoid pinch valve at the outlet of the bioreactor and regulated by a timing circuit. In a permeability test, the system with pulsating flow had much less membrane fouling as compared to the control, a conventional hollow fiber unit. In hepatocyte culture test runs, the pulsating-flow bioreactor demonstrated the ability to maintain a higher cell viability. Histological sections indicated significantly smaller necrotic regions in the pulsating-flow bioreactor as compared to the conventional unit.     

Multivariate statistical modelling of mass transfer in a membrane-supported biofilm reactor

The main objective of this work is to develop an overall mass transfer model applicable to a particular case of membrane supported biofilm, the ion-exchange membrane bioreactor (IEMB). A multivariate projection to latent structures (PLS) model of the anionic membrane transport in an IEMB was developed and analyzed to establish the mass transfer limiting variables for the removal of anionic pollutants (nitrate and perchlorate) from drinking water. The proposed PLS model accounts for the biological contribution to the mass transfer and predicts the anionic fluxes across the ion-exchange membrane with a prediction improvement of at least 50% when compared with a simplified mechanistic Donnan dialysis-based transport model. The PLS model allowed for predicting the transport of target anions using only operational physicochemical data, therefore, the use of several assumptions as in mechanistic model building was avoided as well as the need for biofilm characterization. To decrease the model complexity, several techniques which select the most informative predictors were also successfully used. The analyses of important predictors to each anionic transport model show that transport driving force related variables were the most important. Moreover, at least 30% of the model information is related with biocompartment bulk variables.     

Design and performance study of a novel immobilized hollow fiber membrane bioreactor

A dual-layer coaxial hollow fiber (DLHF) bioreactor for cell immobilization developed to overcome nutrients transport limitation is presented. Cells were contained in the annular space between two coaxial hollow fibers, and nutrients were supplied by a forced convective transport from the shell side through the annular space to the lumen side. With judicious selection of the membrane materials, a low operating transmembrane pressure of 50 kPa, and using E. coli as the model organism, a high cell density of 10(11) cells/mL annular space volume and a high cell viability of (up to 80%) were obtained.     

Hemoglobin‐based oxygen carrier and convection enhanced oxygen transport in a hollow fiber bioreactor

A mathematical model was developed to study O₂ transport in a convection enhanced hepatic hollow fiber (HF) bioreactor, with hemoglobin‐based O₂ carriers (HBOCs) present in the flowing cell culture media stream of the HF lumen. In this study, four HBOCs were evaluated: PEG‐conjugated human hemoglobin (MP4), human hemoglobin (hHb), bovine hemoglobin (BvHb) and polymerized bovine hemoglobin (PolyBvHb). In addition, two types of convective flow in the HF extra capillary space (ECS) were considered in this study. Starling flow naturally occurs when both of the ECS ports are closed. If one of the ECS ports is open, forced convective flow through the ECS will occur due to the imposed pressure difference between the lumen and ECS. This type of flow is referred to as cross‐flow in this work, since some of the fluid entering the HF lumen will pass across the HF membrane and exit via the open ECS port. In this work, we can predict the dissolved O₂ concentration profile as well as the O₂ transport flux in an individual HF of the bioreactor by solving the coupled momentum and mass transport equations. Our results show that supplementation of the cell culture media with HBOCs can dramatically enhance O₂ transport to the ECS (containing hepatocytes) and lead to the formation of an in vivo‐like O₂ spectrum for the optimal culture of hepatocytes. However, both Starling flow and cross‐flow have a very limited effect on O₂ transport in the ECS. Taken together, this work represents a novel predictive tool that can be used to design or analyze HF bioreactors that expose cultured cells to defined overall concentrations and gradients of O₂. Biotechnol. Bioeng. 2009;102: 1603–1612. © 2008 Wiley Periodicals, Inc.     

A comparative study of gel entrapped and membrane attached microbial reactors for biodegrading phenol

A comparative study between two reactors, one using microorganisms entrapped in calcium alginate gel, and the other using microorganisms attached on the surface of a membrane (polymeric microporous sheeting, MPS^TM) to biodegrade phenol is performed. Results indicate that the alginate bead bioreactor is efficient at higher phenol concentrations while the membrane bioreactor shows better performance at lower phenol concentrations. This unique response is primarily attributed to the different techniques by which the microorganisms are immobilized in the two reactors.In batch mode, below a starting concentration of 100 ppm phenol, biodegradation rates in the membrane bioreactor are (7.58 to 12.02 mg phenol/h · g dry biomass) atleast 10 times the rates in alginate bead bioreactor (0.74 to 1.32 mg phenol/h · g dry biomass). Biodegradation rates for the two reactors match at a starting concentration of 250 ppm phenol. Above 500 ppm phenol, the rates in the alginate bead bioreactor are (7.3 to 8.1 mg phenol/h · g dry biomass) on an average 5.5 times the corresponding rates in the membrane bioreactor (2.18 to 1.03 mg phenol/h · g dry biomass).In continuous feed mode the steady state degradation rates in the membrane bioreactor are one to two orders of magnitude higher than the alginate bead bioreactor below 150 ppm inlet phenol concentration. At an inlet concentration around 250 ppm phenol the rates are comparable. Above 500 ppm of phenol the rates in the alginate bioreactor are an order of magnitude high than the membrane bioreactor.Due to substrate inhibition, and its inability to sustain a high biomass concentration, the membrane bioreactor shows poor efficiencies at phenol concentrations above 250 ppm. At low phenol concentrations the apparent reaction rates in the alginate bead bioreactor decrease due to the diffusional resistance of the gel matrix, while biodegradation rates in the membrane bioreactor remain high due to essentially no external diffusional resistance.Results indicate that a combined reactor system can be more effective for bioremediation than either separate or attached microbial reactors.