ENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 3 2009
Bhagyalakshmi Neelwarne
Abstract To arrive at an appropriate bioreactor design and in situ recovery of the products, red beet hairy roots were used as a model system where the levels of betalain pigments (betacyanins and betaxanthins) were followed as secondary metabolite and the peroxidase enzyme as primary metabolite. Medium volume and other kinetic parameters were found to play significant roles by way of directly affecting the biomass yield rather than a specific metabolite. The hydrodynamic stress created on the roots by large culture volume could be minimized by pulse-feeding of medium in shake-flasks; and by separating the biomass chamber from the aerated medium reservoir in circulatory fed-batch bioreactor. Accordingly the bioreactor was modified to provide anchorage and air-enrichment chamber which resulted in higher formation of both the metabolites than in shake-flasks. Various down-stream processing aspects such as in situ release of pigments by non-destructive methods, followed by adsorption through a column and recovery by desorption were optimized for betalains. A strategy for simultaneous recovery of pigment and peroxidase was worked out using aqueous two phase extraction (ATPE). [source]

Study of the Biosynthesis of 1-Octen-3-ol Using a Crude Homogenate of Agaricus bisporus in a Bioreactor

JOURNAL OF FOOD SCIENCE, Issue 3 2008
R.O. Morawicki
ABSTRACT:, 1-Octen-3-ol and 10-oxo- trans -8-decenoic acid are metabolites of the breakdown of linoleic acid (LA) by mushroom enzymes. These compounds can be produced in a bioreactor using a crude mushroom homogenate and the exogenous addition of LA and oxygen. The factors' duration of blending, mushroom,buffer ratio, effect of a surfactant, whole against partially clarified reaction broths, purity of LA, and utilization of stumps instead of whole mushrooms were studied for their effect on reaction yield using a 1-L bioreactor. The results showed the feasibility of using the more inexpensive 60%-pure LA instead of the 99%-pure LA even when a yield loss was involved. Waste stumps could be used instead of whole mushrooms with a yield decline of 26%. [source]

Neural Optimization of Fed-batch Streptokinase Fermentation in a Non-ideal Bioreactor

THE CANADIAN JOURNAL OF CHEMICAL ENGINEERING, Issue 5 2002
Pratap R. PatnaikArticle first published online: 19 MAY 200
Abstract Microbial fermentations involving two or more kinds of competing cells and operating under realistic conditions are difficult to monitor, model and optimize by model-based methods. They deviate from ideal behavior in two significant aspects: incomplete dispersion in the broth and the influx of disturbances. The approach here has been to optimize the filtered noise and dispersion on-line through neural networks. This method has been applied to the fed-batch production of streptokinase (SK). The culture has two kinds of cells , active (or productive) and inactive , and their growth is inhibited by the substrate and the primary metabolite (lactic acid). Using simulated data, the fermentation was optimized by a system of three neural networks, updated continually during successive time intervals. Such sequential optimization with dynamic filtering of inflow noise generated better cell growth and SK activity than static optimization and even an ideal fermentation. Les fermentations microbiennes faisant intervenir deux ou plusieurs sortes de cellules en compétition et se déroulant dans des conditions réelles, sont difficiles à surveiller, à modéliser et à optimiser par des méthodes basées sur des modèles. De telles fermentations s'écartent du comportement idéal dans deux voies importantes : la dispersion incomplète dans le bouillon et la venue de perturbations. Notre approche consiste ici à optimiser le bruit filtré et la dispersion en continu par des réseaux neuronaux. Cette méthode a été appliquée à la production à alimentation discontinue de streptokinase (SK). La culture comporte deux sortes de cellules , actives (ou productives) ou inactives , et leur croissance est inhibée par le substrat et la métabolite primaire (acide lactique). À l'aide de données simulées, la fermentation a été optimisée par un système de trois réseaux neuronaux, qui ont été mis à jour continuellement à des intervalles de temps successifs. Une telle optimisation séquentielle avec filtrage dynamique du bruit génère une meilleure croissance des cellules et activité du SK que l'optimisation statique et même la fermentation idéale. [source]

Modified volume expansion method for measuring gas holdup

THE CANADIAN JOURNAL OF CHEMICAL ENGINEERING, Issue 2 2002
Annie X. Meng
Abstract A simple, modified volume expansion method, or inclined tube method, is compared to the pressure difference method for determining gas holdup in an airlift bioreactor. The modified volume expansion method could be used for all pneumatic bioreactors where fluid fluctuation is vigorous and visual observation of the continuous phase rise is difficult. The overall gas holdup data measured using the inclined tube method are shown to be very close to overall gas holdup determined using a gamma ray density monitor system. However, the overall gas holdup measured by the pressure difference method is found to be significantly different. This difference is due to energy dissipation in the External Loop Airlift Bioreactor (ELAB) used in this study, which causes the pressure difference method to be incorrect. On compare une méthode d'expansion de volume modifiée simple, ou méthode à tubes inclinés, à une méthode de différence de pression pour déterminer la rétention des gaz dans un bioréacteur à air ascendant. La méthode d'expansion de volume modifiée pourrait être utilisée pour tous les bioréacteurs pneumatiques oú la fluctuation du fluide est vigoureuse et l'observation visuelle de l'ascension de la phase continue difficile. Les données globales de rétention de gaz mesurées à l'aide de la méthode à tubes inclinés s'avèrent très proches de la rétention de gaz globale déterminéd à l'aide d'un moniteur de densité à rayons gamma. Toutefois, on a trouvé que la rétention de gaz globale mesurée par la méthode de différence de pression était significativement différente. Cette différence est due à la dissipation d'énergie dans le bioréacteur à air ascendant à boucle externe (ELAB) utilisé dans I'étude, qui rend la méthode de différence de pression incorrecte. [source]

New Pulsatile Hydrostatic Pressure Bioreactor for Vascular Tissue-engineered Constructs

ARTIFICIAL ORGANS, Issue 2 2010
Faisal M. Shaikh
Abstract Mechanical conditioning represents a potential means to enhance the biochemical and biomechanical properties of tissue-engineered cell constructs. Bioreactors that can simulate physiologic conditions can play an important role in the preparation of tissue-engineered constructs. Although various forms of bioreactor systems are currently available, these have certain limitations, particularly when these are used for the creation of vascular constructs. The aim of the present report is to describe and validate a novel pressure bioreactor system for the creation of vascular tissue. Here, we present and discuss the design concepts, criteria, as well as the development of a novel pressure bioreactor. The system is compact and easily housed in an incubator to maintain sterility of the construct. Moreover, the proposed bioreactor, in addition to mimicking in vivo pressure conditions, is flexible, allowing different types of constructs to be exposed to various physiologic pressure conditions. The core bioreactor elements can be easily sterilized and have good ergonomic assembly characteristics. This system is a fundamental tool, which may enable us to make further advances in bioreactor technology and tissue engineering. The novel system allows for the application of pressure that may facilitate the growth and development of constructs needed to produce a tissue-engineered vascular graft. [source]

In Vitro and In Vivo Evaluation of Albumin Synthesis Rate of Porcine Hepatocytes in a Flat-Plate Bioreactor

ARTIFICIAL ORGANS, Issue 7 2001
Masaya Shito
Abstract: Several configurations of extracorporeal bioartificial liver devices have been developed for the potential treatment of fulminant hepatic failure or as a bridge to liver transplantation. Recently, we developed a microchannel flat-plate bioreactor with an internal membrane oxygenator in which porcine hepatocytes are cultured as a monolayer on the bottom glass surface. In the present study, we investigated synthetic function of porcine hepatocytes in the bioreactor in both in vitro and in vivo flow circuit models. In vitro, albumin synthesis was stable in the bioreactor for up to 4 days of perfusion. In vivo, with the extracorporeal connection of the bioreactor to rat vasculature, porcine albumin was detectable for 24 h in the rat plasma. We also developed a simple mathematical model to predict the in vivo porcine albumin concentration in rat plasma. These results indicate that this configuration of a microchannel flat-plate bioreactor has potential as a liver support device and warrants further investigation. [source]

Performance of Dual-Media Expanded Bed Bioreactor

ASIA-PACIFIC JOURNAL OF CHEMICAL ENGINEERING, Issue 5-6 2005
R. Abdul-Rahman
Abstract Adsorption and biological treatment are two possible approaches to remove chloro-organic and organic compounds. Granular activated carbon (GAC) biofilm reactors combine these two features, the adsorptive capacity and irregular shape of GAC particles providing niches for bacterial colonisation protected from high fluid forces, while the variety of functional groups on the surface enhance the attachment of microorganisms. The biofilm process is compact and offers reactions in both aerobic and anoxic states. Studies on removal of nitrogen constituents by a biofilm process were carried out using a dual-media expanded bed bioreactor, with GAC and plastic media as support media. The plastic media also acts as a filter for the effluent. Experiments were carried out at F:M of about 0.45 and hydraulic residence times (HRT) of 48, 24 and 12 hours. Bed expansion was maintained at 20,30% by recirculation flow. Aerobic condition was maintained at dissolved oxygen (DO) of about 2 mg/l throughout the bed. Chemical oxygen in demand (COD) in feed was 1000 mg/L while the total-N was 100 mg/L. Analysis showed that the process is able to maintain very stable conditions, achieving substantial COD removal of about 85% and total-N removal of about 80%. Biofilm biomass measurements showed an increase from 400 mg/l at HRT of 48 hours to 10,100 mg/l at HRT 12 hours, showing that much higher biomass concentrations may be contained in a biofilm process as compared to a conventional suspended biomass process. Bioreactors contain their own ecosystems, the nature of the community and the state of microorganisms define the kinetics and determine reactor performance. Growth kinetic parameters obtained are YH = 0.3421 mg/mg, m,H = 0.2252 day,1, KH = 319.364 mg/l and bH = 0.046 day,1. The denitrification kinetic parameters obtained are YHD = 0.9409 mg/mg, m,HD = 0.1612 day,1, KHD = 24.6253 mg/l and bHD = 0.0248 day,1. These parameters enable prediction of required reactor sizes and operational parameters. The plastic media has greatly improved effluent clarification by 98% as compared to single-media (GAC) only reactor. [source]

Bioproduction of the aroma compound 2-Phenylethanol in a solid,liquid two-phase partitioning bioreactor system by Kluyveromyces marxianus

BIOTECHNOLOGY & BIOENGINEERING, Issue 2 2009
Fang Gao
Abstract The rose-like aroma compound 2-phenylethanol (2-PE) is an important fragrance and flavor ingredient. Several yeast strains are able to convert l -phenylalanine (l -phe) to 2-PE among which Kluyveromyces marxianus has shown promising results. The limitation of this process is the low product concentration and productivity primarily due to end product inhibition. This study explored the possibility and benefits of using a solid,liquid Two-Phase Partition Bioreactor (TPPB) system as an in situ product removal technique. The system applies polymer beads as the sequestering immiscible phase to partition 2-PE and reduce the aqueous 2-PE concentration to non-inhibitory levels. Among six polymers screened for extracting 2-PE, Hytrel® 8206 performed best with a partition coefficient of 79. The desired product stored in the polymer was ultimately extracted using methanol. A 3,L working volume solid,liquid batch mode TPPB using 500,g Hytrel® as the sequestering phase generated a final overall 2-PE concentration of 13.7,g/L, the highest reported in the current literature. This was based on a polymer phase concentration of 88.74,g/L and aqueous phase concentration of 1.2,g/L. Even better results were achieved via contact with more polymers (approximately 900,g) with the aqueous phase applying a semi-continuous reactor configuration. In this system, a final 2-PE concentration (overall) of 20.4,g/L was achieved with 1.4,g/L in the aqueous and 97,g/L in the polymer phase. The overall productivities of these two reactor systems were 0.38 and 0.43,g/L,h, respectively. This is the first report in the literature of the use of a polymer sequestering phase to enhance the bioproduction of 2-PE, and exceeds the performance of two-liquid phase systems in terms of productivity as well as ease of operation (no emulsions) and ultimate product recovery. Biotechnol. Bioeng. 2009; 104: 332,339 © 2009 Wiley Periodicals, Inc. [source]

Design and Characterization of a Rotating Bed System Bioreactor for Tissue Engineering Applications

BIOTECHNOLOGY PROGRESS, Issue 1 2008
Fabienne Anton
The main challenge in the development of bioreactors for tissue engineering is the delivery of a sufficient nutrient and oxygen supply for cell growth in a 3D environment. Thus, a new rotating bed system bioreactor for tissue engineering applications was developed. The system consists of a culture vessel as well as an integrated rotating bed of special porous ceramic discs and a process control unit connected with the reactor to ensure optimal culturing conditions. The aim of the project was the design and construction of a fully equipped rotating bed reactor, and in particular, the characterization and optimization of the system with regard to technical parameters such as mixing time and pH-control to guarantee optimal conditions for cell growth and differentiation. Furthermore, the applicability of the developed system was demonstrated by cultivation of osteoblast precursor cells. The porous structure of the ceramic discs and the external medium circulation loop provide an optimal environment for tissue generation in long-term cultivations. Mass transfer limitations were minimized by the slow rotation, which also provides the cells with sufficient nutrients and oxygen through alternate contact to air and medium. An osteoblast precursor cell line was successfully cultivated in this bioreactor for 28 days. [source]

Bioreactor Coupled with Electromagnetic Field Generator: Effects of Extremely Low Frequency Electromagnetic Fields on Ethanol Production by Saccharomycescerevisiae

BIOTECHNOLOGY PROGRESS, Issue 5 2007
Victor H. Perez
The effect of extremely low frequency (ELF) magnetic fields on ethanol production by Saccharomyces cerevisiae using sugar cane molasses was studied during batch fermentation. The cellular suspension from the fermentor was externally recycled through a stainless steel tube inserted in two magnetic field generators, and consequently, the ethanol production was intensified. Two magnetic field generators were coupled to the bioreactor, which were operated conveniently in simple or combined ways. Therefore, the recycle velocity and intensity of the magnetic field varied in a range of 0.6,1.4 m s,1 and 5,20 mT, respectively. However, under the best conditions with the magnetic field treatment (0.9,1.2 m s,1 and 20 mT plus solenoid), the overall volumetric ethanol productivity was approximately 17% higher than in the control experiment. These results made it possible to verify the effectiveness of the dynamic magnetic treatment since the fermentations with magnetic treatment reached their final stage in less time, i.e., approximately 2 h earlier, when compared with the control experiment. [source]

Optimization of Rosmarinic Acid Production by Lavandula vera MM Plant Cell Suspension in a Laboratory Bioreactor

BIOTECHNOLOGY PROGRESS, Issue 2 2005
Atanas I. Pavlov
The all-round effect of dissolved oxygen concentration, agitation speed, and temperature on the rosmarinic acid production by Lavandula veraMM cell suspension was studied in a 3-L laboratory bioreactor by means of the modified Simplex method. Polynomial regression models were elaborated for description of the process of rosmarinic acid production (Y) in the bioreactor as a consequence of the variation of the dissolved oxygen (X1) concentration between 10% and 50%; agitation (X2) between 100 and 400 rpm; and temperature (X3) between 22 and 30 °C. The optimization made it possible to establish the optimal conditions for the biosynthesis of rosmarinic acid by L. veraMM: dissolved oxygen (X1*), 50% of air saturation; agitation (X2*), 400 rpm; and temperature (X3*), 29.9 °C, where maximal yield (Ymax) of 3489.4 mg/L of rosmarinic acid was achieved (2 times higher compared with the shake-flasks cultivation). [source]

Production of a Secreted Glycoprotein from an Inducible Promoter System in a Perfusion Bioreactor

BIOTECHNOLOGY PROGRESS, Issue 5 2004
Matthew L. Lipscomb
The primary advantage of an inducible promoter expression system is that production of the recombinant protein can be biochemically controlled, allowing for the separation of unique growth and production phases of the culture. During the growth phase, the culture is rapidly grown to high cell density prior to induction without the extra metabolic burden of exogenous protein production, thus minimizing the nonproductive period of the culture. Induction of the culture at high cell density ensures that the volumetric production will be maximized. In this work, we have demonstrated the feasibility of overexpressing a reporter glycoprotein from the inducible MMTV promoter in recombinant Chinese hamster ovary (CHO) cells cultured in a high cell density perfusion bioreactor system. Retention of suspension-adapted CHO cells was achieved by inclined sedimentation. To maximize volumetric production of the culture, we have demonstrated that high cell density must be achieved prior to induction. This operating scheme resulted in a 10-fold increase in volumetric titer over the low density induction culture, corresponding directly to a 10-fold increase in viable cell density during the highly productive period of the culture. The amount of glycoprotein produced in this high cell density induction culture during 26 days was 84-fold greater than that produced in a week long batch bioreactor. Long-term perfusion cultures of the recombinant cell line showed a production instability, a phenomenon that is currently being investigated. [source]

Cybernetic Model Predictive Control of a Continuous Bioreactor with Cell Recycle

BIOTECHNOLOGY PROGRESS, Issue 5 2003
Kapil G. Gadkar
The control of poly-,-hydroxybutyrate (PHB) productivity in a continuous bioreactor with cell recycle is studied by simulation. A cybernetic model of PHB synthesis in Alcaligenes eutrophus is developed. Model parameters are identified using experimental data, and simulation results are presented. The model is interfaced to a multirate model predictive control (MPC) algorithm. PHB productivity and concentration are controlled by manipulating dilution rate and recycle ratio. Unmeasured time varying disturbances are imposed to study regulatory control performance, including unreachable setpoints. With proper controller tuning, the nonlinear MPC algorithm can track productivity and concentration setpoints despite a change in the sign of PHB productivity gain with respect to dilution rate. It is shown that the nonlinear MPC algorithm is able to track the maximum achievable productivity for unreachable setpoints under significant process/model mismatch. The impact of model uncertainty upon controller performance is explored. The multirate MPC algorithm is tested using three controllers employing models that vary in complexity of regulation. It is shown that controller performance deteriorates as a function of decreasing biological complexity. [source]

Design of a Tubular Loop Bioreactor for Scale-up and Scale-down of Fermentation Processes

BIOTECHNOLOGY PROGRESS, Issue 5 2003
Maria Papagianni
Microorganisms traveling through circulation loops in large-scale bioreactors experience variations in their environment such as dissolved oxygen concentration and pH gradients. The same changes are not experienced in small bioreactors, and it is suggested that herein lies one of the major reasons for the problems encountered when translating fermentation data from one scale to another. One approach to study this problem is to look at the circulation loop itself. The present work concerns an attempt to simulate the circulation loops inside stirred tank reactors, using a tubular loop reactor specially constructed for the purpose. The reactor carries a number of ports and probes along its length for the determination of concentration gradients within. The broth is circulated around the loop by the use of peristaltic pumps, and the circulation time (tc, s) is used as a measure of simulated reactor size. The reactor system has been evaluated using the citric acid fermentation by Aspergillus niger as a test process. Acid production and fungal morphology, in terms of the mean convex perimeter of mycelial clumps quantified by image analysis, were used as the parameters of evaluation for the two systems in comparison. From comparative experiments carried out in 10 and 200 L stirred tank bioreactors, it appears that the loop reactor simulates the corresponding stirred tank representing a valuable tool in scaling up and scaling down of fermentation process. [source]

Enhanced Production of Podophyllotoxin by Podophyllum hexandrum Using in Situ Cell Retention Bioreactor

BIOTECHNOLOGY PROGRESS, Issue 3 2003
Saurabh Chattopadhyay
The rhizomes of the rare plant Podophyllum hexandrum contain podophyllotoxin, which is a precursor of the anticancer drugs etoposide and teniposide. Batch cultivation of Podophyllumhexandrum was conducted using optimized medium in a 3 L bioreactor, which resulted in biomass and podophyllotoxin concentrations of 21.4 g/L and 13.8 mg/L in 24 and 26 days, respectively. The batch kinetics was used to identify the mathematical model. The model was extrapolated to identify the nutrient feeding rate (150 mL/d) and substrate concentration (105 g/L) in the incoming feed for nonlimiting and noninhibitory glucose concentration in the cell retention bioreactor. An improvement in cell growth to 53 g/L and intracellular podophyllotoxin accumulation of 48.8 mg/L was achieved in 60 days, when the bioreactor was operated in continuous cell retention cultivation mode. [source]

A Feasible Enzymatic Process for d -Tagatose Production by an Immobilized Thermostable l -Arabinose Isomerase in a Packed-Bed Bioreactor

BIOTECHNOLOGY PROGRESS, Issue 2 2003
Hye-Jung Kim
To develop a feasible enzymatic process for d -tagatose production, a thermostable l -arabinose isomerase, Gali152, was immobilized in alginate, and the galactose isomerization reaction conditions were optimized. The pH and temperature for the maximal galactose isomerization reaction were pH 8.0 and 65 °C in the immobilized enzyme system and pH 7.5 and 60 °C in the free enzyme system. The presence of manganese ion enhanced galactose isomerization to tagatose in both the free and immobilized enzyme systems. The immobilized enzyme was more stable than the free enzyme at the same pH and temperature. Under stable conditions of pH 8.0 and 60 °C, the immobilized enzyme produced 58 g/L of tagatose from 100 g/L galactose in 90 h by batch reaction, whereas the free enzyme produced 37 g/L tagatose due to its lower stability. A packed-bed bioreactor with immobilized Gali152 in alginate beads produced 50 g/L tagatose from 100 g/L galactose in 168 h, with a productivity of 13.3 (g of tagatose)/(L-reactor·h) in continuous mode. The bioreactor produced 230 g/L tagatose from 500 g/L galactose in continuous recycling mode, with a productivity of 9.6 g/(L·h) and a conversion yield of 46%. [source]

Fed-Batch Cultivation of Saccharomyces cerevisiae in a Hyperbaric Bioreactor

BIOTECHNOLOGY PROGRESS, Issue 2 2003
I. Belo
Fed-batch is the dominating mode of operation in high-cell-density cultures of Saccharomyces cerevisaein processes such as the production of bakerapos;s yeast and recombinant proteins, where the high oxygen demand of these cultures makes its supply an important and difficult task. The aim of this work was to study the use of hyperbaric air for oxygen mass transfer improvement on S. cerevisiaefed-batch cultivation. The effects of increased air pressure up to 1.5 MPa on cell behavior were investigated. The effects of oxygen and carbon dioxide were dissociated from the effects of total pressure by the use of pure oxygen and gas mixtures enriched with CO2. Fed-batch experiments were performed in a stirred tank reactor with a 600 mL stainless steel vessel. An exponential feeding profile at dilution rates up to 0.1 h,1 was used in order to ensure a subcritical flux of substrate and, consequently, to prevent ethanol formation due to glucose excess. The ethanol production observed at atmospheric pressure was reduced by the bioreactor pressurization up to 1.0 MPa. The maximum biomass yield, 0.5 g g,1 (cell mass produced per mass of glucose consumed) was attained whenever pressure was increased gradually through time. This demonstrates the adaptive behavior of the cells to the hyperbaric conditions. This work proved that hyperbaric air up to 1.0 MPa (0.2 MPa of oxygen partial pressure) could be applied to S. cerevisiaecultivation under low glucose flux. Above that critical oxygen partial pressure value, i.e., for oxygen pressures of 0.32 and 0.5 MPa, a drastic cell growth inhibition and viability loss were observed. The increase of carbon dioxide partial pressure in the gas mixture up to 48 kPa slightly decreased the overall cell mass yield but had negligible effects on cell viability. [source]

Isoprene Formation in Bacillus subtilis: A Barometer of Central Carbon Assimilation in a Bioreactor?

BIOTECHNOLOGY PROGRESS, Issue 5 2002
Megan C. Shirk
Isoprene (2-methyl-1,3-butadiene) is a volatile hydrocarbon of uncertain function in Bacillus subtilis, and we hypothesized that it is an overflow metabolite produced during excess carbon utilization. Here we tested this idea for phase 2 of isoprene release, a phase that occurs during extracellular acetoin accumulation and its reassimilation. Phase 2 isoprene formation could be disrupted in three different ways, all related to acetoin metabolism. Disruption of a gene essential for acetoin biosynthesis (acetolactic acid synthase, alsS) blocked acetoin formation and led to cessation of phase 2 isoprene formation as well as a variety of pleiotropic effects related to loss of pH control. Growth of the alsS mutant with external pH control reversed most of these effects. Disruption of acetoin catabolism (acetoin dehydrogenase, acoA), also eliminated phase 2 isoprene formation and caused cells to transition directly from phase 1 to phase 3; the latter is attributed to amino acid catabolism. A third alteration of acetoin metabolism was detected in the widely used strain 168 ( trpC2) but not in strain MS175, a trpC mutant constructed in the Marburg strain genetic background. Strain 168 exhibited slow acetoin assimilation compared to that of MS175 or the parental strain, with little or no isoprene formation during this growth phase. These findings support the idea that isoprene release occurs primarily when the rate of carbon catabolism exceeds anabolism and that this volatile hydrocarbon is a product of overflow metabolism when precursors are not required for higher isoprenoid biosynthesis. It is suggested that isoprene release might serve as a useful barometer of the rise and fall of central carbon fluxes during the growth of Bacillus strains in industrial bioreactors. [source]

Olive Oil Mill Waste Waters Decoloration and Detoxification in a Bioreactor by the White Rot Fungus Phanerochaeteflavido-alba

BIOTECHNOLOGY PROGRESS, Issue 3 2002
P. Blánquez
Olive oil mill wastewater (OMW) is produced as waste in olive oil extraction. With the purpose of treating this highly polluting waste, a number of experiments were conducted in a laboratory-scale bioreactor with the white rot fungus Phanerochaete flavido-alba ( P.flavido-alba). It is known that this fungus is capable of decolorizing OMW in static or semistatic cultures at Erlenmeyer scale and at 30 °C. The objective of this work was to prove that P. flavido-alba could decolorize OMW in submerged cultures and that it is capable of reducing OMW toxicity at room temperature (25 °C) and in a laboratory-scale bioreactor. In the experiments conducted, manganese peroxidase (MnP) and laccase enzymes were detected; however, unlike other studies, lignin peroxidase was not found to be present. Decoloration obtained after treatment was 70%. The reduction of aromatic compounds obtained was 51%, and the toxicity of the culture medium was reduced by up to 70%. We can therefore state that P.flavido-alba is capable of reducing important environmental parameters of industrial effluents and that prospects are positive for the use of this process at a larger scale, even when working at room temperature. [source]

Design of a Molecular Chaperone-Assisted Protein Folding Bioreactor

BIOTECHNOLOGY PROGRESS, Issue 4 2000
Reto J. Kohler
Escherichia coli molecular chaperone GroEL and co-chaperone GroES are well known to assist the folding/refolding of a diverse range of substrate proteins. Despite this, there have been relatively few reports of the GroEL/GroES molecular chaperone system being used as a biotechnology tool for protein folding/refolding. In this paper, a solution-phase protein folding bioreactor is described that involves the complete GroEL/GroES system. The main features of this bioreactor are the use of a stirred-cell concentrator fitted with a 100 kDa molecular weight cutoff membrane and an attached buffer reservoir. This bioreactor system was used successfully for assisted-batch refolding of guanidinium chloride (Gu-HCl) unfolded mitochondrial malate dehydrogenase (mMDH). We believe that protein folding bioreactor systems of this type could have wide potential utility for the folding/refolding of unfolded protein substrates. [source]

Hairy Root Culture in a Liquid-Dispersed Bioreactor: Characterization of Spatial Heterogeneity

BIOTECHNOLOGY PROGRESS, Issue 3 2000
Gary R. C. Williams
A liquid-dispersed reactor equipped with a vertical mesh cylinder for inoculum support was developed for culture of Atropa belladonna hairy roots. The working volume of the culture vessel was 4.4 L with an aspect ratio of 1.7. Medium was dispersed as a spray onto the top of the root bed, and the roots grew radially outward from the central mesh cylinder to the vessel wall. Significant benefits in terms of liquid drainage and reduced interstitial liquid holdup were obtained using a vertical rather than horizontal support structure for the biomass and by operating the reactor with cocurrent air and liquid flow. With root growth, a pattern of spatial heterogeneity developed in the vessel. Higher local biomass densities, lower volumes of interstitial liquid, lower sugar concentrations, and higher root atropine contents were found in the upper sections of the root bed compared with the lower sections, suggesting a greater level of metabolic activity toward the top of the reactor. Although gas-liquid oxygen transfer to the spray droplets was very rapid, there was evidence of significant oxygen limitations in the reactor. Substantial volumes of non-free-draining interstitial liquid accumulated in the root bed. Roots near the bottom of the vessel trapped up to 3,4 times their own weight in liquid, thus eliminating the advantages of improved contact with the gas phase offered by liquid-dispersed culture systems. Local nutrient and product concentrations in the non-free-draining liquid were significantly different from those in the bulk medium, indicating poor liquid mixing within the root bed. Oxygen enrichment of the gas phase improved neither growth nor atropine production, highlighting the greater importance of liquid-solid compared with gas-liquid oxygen transfer resistance. The absence of mechanical or pneumatic agitation and the tendency of the root bed to accumulate liquid and impede drainage were identified as the major limitations to reactor performance. Improved reactor operating strategies and selection or development of root lines offering minimal resistance to liquid flow and low liquid retention characteristics are possible solutions to these problems. [source]

Experimental Study and Design of a Submerged Membrane Distillation Bioreactor

CHEMICAL ENGINEERING & TECHNOLOGY (CET), Issue 1 2009
J. Phattaranawik
Abstract A hybrid process incorporating membrane distillation in a submerged membrane bioreactor operated at elevated temperature is developed and experimentally demonstrated in this article. Since organic particles are rejected by an ,evaporation' mechanism, the retention time of non-volatile soluble and small organics in the submerged membrane distillation bioreactor (MDBR) is independent of the hydraulic retention time (mainly water and volatiles). A high permeate quality can be obtained in the one-step compact process. The submerged MD modules were designed for both flat-sheet membranes and tubular membrane configurations. The process performance was preliminarily evaluated by the permeate flux stabilities. The module configuration design and air sparging used in the MDBR process were tested. Flux declines were observed for the thin flat-sheet hydrophobic membranes. Tubular membrane modules provided more stable permeate fluxes probably due to the turbulent condition generated from air sparging injected inside the tubular membrane bundles. The experiments with the submerged tubular MD module gave stable fluxes of approximately 5,L/m2 h over 2,weeks at a bioreactor temperature of 56,°C. The total organic carbon in the permeate was consistently lower than 0.7,mg/L for all experiments. [source]

Bioreactor for cultivation of red beet hairy roots and in situ recovery of primary and secondary metabolites

Improved ,-Glucanase Production by a Recombinant Escherichia coli Strain using Zinc-Ion Supplemented Medium

ENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 3 2007
U. Beshay
Abstract In order to investigate the suitability of different metal chelates for affinity chromatography, an expression vector was constructed. It contained a hybrid ,-glucanase as a model protein fused with a His6 -tag and a secretion cassette providing the ability to secrete ,-glucanase into the culture medium. Supplementation of zinc to the medium led to a rapidly increased expression and release of the target protein into the cultivation medium. Results in respect to the supplementation of the commonly used Terrific Broth "TB-medium" with different metal ions are reported with special emphasis on the influence of zinc ions. A concentration of zinc ions in the order of about 0.175 mM led to optimal results. Batch cultivation under well-controlled conditions showed that the growth behavior did not change significantly by adding zinc ions. Growth in a stirred tank bioreactor was much faster in unsupplemented TB-medium compared to shake flask experiments leading to a much higher biomass concentration (15,g/L instead of 3,g/L). The secretion of ,-glucanase under theses conditions started at the transition into the stationary phase and increased to yield an extracellular activity of 1350,U/mL at the end of the fermentation process. An even higher yield of extracellular ,-glucanase (2800,U/mL) was reached when the fermentation was carried out with TB-medium supplemented with 0.175,mM ZnSO4. [source]

Highlights in Biocatalysis , Historical Landmarks and Current Trends

ENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 4 2005
T. Bornscheuer
Abstract Biocatalysis has ancient roots, yet it is developing into a key tool for synthesis in a wide range of applications. Important events in the history of enzyme technology from the 19th century onwards are highlighted. Considering the most relevant progress steps, the production of penicillanic acid and the impact of genetic engineering are traced in more detail. Applied biocatalysis has been defined as the application of a biocatalyst to achieve a desired conversion selectively, under controlled, mild conditions in a bioreactor. Biocatalysts are currently used to produce a wide range of products in the fields of food manufacture (such as bread, cheese, beer), fine chemicals (e.g., amino acids, vitamins), and pharmaceuticals (e.g., derivatives of antibiotics). They not only provide access to innovative products and processes, but also meet criteria of sustainability. In organic synthesis, recombinant technologies and biocatalysts have greatly widened the scope of application. Examples of current applications and processes are given. Recent developments and trends are presented as a survey, covering new methods for accessing biodiversity with new enzymes, directed evolution for improving enzymes, designed cells, and integrated downstream processing. [source]

Post-treatment of anaerobically treated medium-age landfill leachate

ENVIRONMENTAL PROGRESS & SUSTAINABLE ENERGY, Issue 1 2010
Ebru Akkaya
Abstract This study focused on the removal of COD and NH4+ from medium-age leachate. Experiments were performed in a laboratory-scale upflow anaerobic sludge blanket (UASB), a membrane bioreactor (MBR), and using magnesium ammonium phosphate (MAP) precipitation. MBR and MAP were used for the post-treatment steps for anaerobically treated leachate to increase the removal of organics and ammonium. The UASB reactor removed nearly all biodegradable organics and supplied constant effluent COD for all concentration ranges of influent leachate. Ammonium removal efficiency in the UASB reactor was relatively low and the average value was ,7.9%. Integration of MBR to the effluent of UASB reactor increased the average COD removal efficiency from 51.8 to 65.6% and maximum removal efficiency increased to 74.3%. MAP precipitation was applied as a final step to decrease the ammonium concentration in the effluent of UASB+MBR reactors. The effect of pH and the molar ratio of MAP constituents on the removal of ammonium were evaluated. At optimal conditions (pH: 9.0 and Mg/NH4/PO4: 1/1.2/1.2), 96.6% of ammonium was removed and MAP provided additional COD and turbidity treatment. Consequently, the combined system of MBR and MAP precipitation could be used as an appropriate post treatment option for the anaerobically treated medium-age landfill leachate. © 2009 American Institute of Chemical Engineers Environ Prog, 2010 [source]

Industrial wastewater treatment in a membrane bioreactor: A review

ENVIRONMENTAL PROGRESS & SUSTAINABLE ENERGY, Issue 1 2004
B. Marrot
Abstract This paper provides a detailed literature review of wastewater treatment in a membrane bioreactor process (MBR) with special focus on industrial wastewater treatment. MBR systems are compared with conventional wastewater treatment systems. The characteristics of the bioreactor treatment process (biomass concentration and floc size, organic and mass loading rates, etc.) are examined. The membrane separation of microorganisms from the treated wastewater is discussed in detail. Problems of membrane fouling and membrane washing and regeneration, linked to activated sludge characteristics, are examined. © 2004 American Institute of Chemical Engineers Environ Prog, 23: 59,68, 2004 [source]

Potential and Bottlenecks of Bioreactors in 3D Cell Culture and Tissue Manufacturing

ADVANCED MATERIALS, Issue 32-33 2009
David Wendt
Abstract Over the last decade, we have witnessed an increased recognition of the importance of 3D culture models to study various aspects of cell physiology and pathology, as well as to engineer implantable tissues. As compared to well-established 2D cell-culture systems, cell/tissue culture within 3D porous biomaterials has introduced new scientific and technical challenges associated with complex transport phenomena, physical forces, and cell,microenvironment interactions. While bioreactor-based 3D model systems have begun to play a crucial role in addressing fundamental scientific questions, numerous hurdles currently impede the most efficient utilization of these systems. We describe how computational modeling and innovative sensor technologies, in conjunction with well-defined and controlled bioreactor-based 3D culture systems, will be key to gain further insight into cell behavior and the complexity of tissue development. These model systems will lay a solid foundation to further develop, optimize, and effectively streamline the essential bioprocesses to safely and reproducibly produce appropriately scaled tissue grafts for clinical studies. [source]

Treatment of beverage-processing wastewater in a three-phase fluidised bed biological reactor

INTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 6 2008
Samwel Victor Manyele
Summary This paper presents a study on treatment of beverage-processing wastewater (BPWW) in a three-phase fluidised bed bioreactor (TPFBB). Wastewater samples were introduced in the TPFBB and aerated at optimum liquid and gas flow rates while measuring wastewater parameters [pH, chemical oxygen demand (COD), total suspended solids (TSS), total Kjehldahl nitrogen (TKN) and ammonia-nitrogen (NH3 -N)]. Two different initial pH levels were studied, i.e. 9.0 and 11.5. The pH of the wastewater was observed to level off at 9.3 after 1 day. The TSS dropped by 95% after 5 days, for both initial pH levels. The NH3 -N and TKN dropped to similar final concentration independent of initial pH. The COD removal efficiency was observed to depend on the initial pH level. A highest efficiency of 98% and lowest efficiency of 50% were observed at initial pH of 9.0 and 11.5, respectively. The study results show that TPFBB is capable of treating food-processing wastewater under suitable conditions. [source]

Quantifying the heterogeneous heat response of Escherichia coli under dynamic temperatures

JOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2010
E. Van Derlinden
Abstract Aims:, Non-sigmoid growth curves of Escherichia coli obtained at constant temperatures near the maximum growth temperature (Tmax) were previously explained by the coexistence of two subpopulations, i.e. a stress-sensitive and a stress-resistant subpopulation. Mathematical simulations with a heterogeneous model support this hypothesis for static experiments at 45°C. In this article, the behaviour of E. coli, when subjected to a linearly increasing temperature crossing Tmax, is studied. Methods and Results:, Subpopulation dynamics are studied by culturing E. coli K12 MG1655 in brain heart infusion broth in a bioreactor. The slowly increasing temperature (°C h,1) starting from 42°C results in growth up to 60°C, a temperature significantly higher than the known Tmax. Given some additional presumptions, mathematical simulations with the heterogeneous model can describe the dynamic experiments rather well. Conclusions:, This study further confirms the existence of a stress-resistant subpopulation and reveals the unexpected growth of E. coli at temperatures significantly higher than Tmax. Significance and Impact of the Study:, The growth of the small stress-resistant subpopulation at unexpectedly high temperatures asks for a revision of currently applied models in food safety and food quality strategies. [source]