A novel fluidised bed bioreactor for fermentation of glucose to ethanol using alginate immobilised yeast

Summary A novel fluidised bed bioreactor has been developed which avoids the problems of particle flotation and gas logging seen in such bioreactors by arranging for the bed to be simultaneously fluidised from the top and bottom with fluid removal from a side port. With alginate immobilised yeast, the bed converted glucose to ethanol at a 27% higher rate than a similar conventional fluidised bed bioreactor using similar conditions.     

A fluidised bed vessel for the culture of immobilised plant cells and its application for the continuous production of fine cell suspensions

With the expansion of immobilised plant cell technology the need has arisen for a suitable vessel in which systems can be efficiently manipulated.Described is a vessel which incorporates many of the features of a fluidised bed, together with some of those from airlift technology to enable immobilised plant cells to be cultured at high biomass concentrations while maintaining a high mass transfer and controlled aeration under continuous flow conditions. In the case described, the vessel has been used for the continuous production of fine plant cell suspensions, although it is easily adaptable for use in cell mediated biotransformation or de novo synthesis studies.Abbreviations 2,4D 2.4 dichlorophenoxyacetic acid     

Design and assembly of solid-phases for the effective recovery of nanoparticulate bioproducts in fluidised bed contactors

Practical recovery of nanoparticulate bioproducts from suspension feedstocks has been studied in batch, fixed bed and fluidised bed adsorptive contactors. The performance of five discrete configurations of adsorbent solid phase has been critically evaluated in the anion exchange recovery of mg quantities of BSA nanoparticles. These have served as surrogate size mimics of less easily sourced viral and plasmid gene therapy vectors, characterised by high value and a shortage of supply in quantities sufficient for research and development. Performance parameters of binding capacity, efficacy of washing, desorption efficiency and total cycle time were strongly influenced by the external and internal topographies of solid phases, together with the localised concentrations of interacting chemical ligands which modulate adsorption. In respect of a full operational recovery cycle, porous adsorbents developed for refined chromatographic fractionation of macromolecules, appear less suited overall than solid, non-porous particles, or solid particles coated with a shallow pellicle of active adsorbent material. Such findings have been confirmed in a detailed demonstration of the recovery of plasmid DNA (7.8 Kb) from chemical lysates of Escherichia coli.     

Anaerobic fluidised bed for the purification of effluents from chemical and mechanical pulping

Anaerobic treatment has seldom been used for wastewaters from the pulp and paper industry and other branches of the chemical industry. Escape of volatile pollutants to the atmosphere, which always occurs during aerobic treatment, is avoided, and much less sludge is being produced than in an aerobic process. The greatest obstacle for using anaerobic treatment in the pulp and paper industry is the large wastewater volume, which necessitates short hydraulic detention times, because the treatment is to occur in an enclosed space. We used solid carrier particles to prevent wash-out of biomass from the reactor at high hydraulic loading, and an up-flow system in order to be able to use very small carrier particles, maximizing the surface area for biomass attachment. In this paper we describe and discuss the results obtained with this type of anaerobic reactor (fluidised bed) at bench and semitechnical scale for wastewaters from pressurized ground wood pulping and paper manufacture, sulphite pulp evaporator condensate and bleach waste. Earlier work with Kraft pulp bleaching effluent and thermomechanical pulping wastewater and evaporator condensates using anaerobic reactors is also discussed. The results obtained thus far show that there are several wastewater streams from the pulping industry, where 60 to 90% of the dissolved organic pollutants (measured as COD_Cr or TOC) was biodegraded within 4 to 24 h. The high strength waste streams (COD_Cr 2000 mg O₂ 1⁻¹) allowed organic space load of 4 to 10 kg COD_Cr m⁻³ reactor volume d⁻¹. With low strength wastes the hydraulic loading was the limiting factor.     

Emissions modeling of fluidised bed co-combustion of poultry litter and peat

Gaseous emissions from the fluidised bed co-combustion of 50% w/w chicken litter and peat were monitored and recorded. Emission data were used to create a dispersion model for a proposed site on a poultry farm in Ireland. Variables within the combustion unit influenced both combustion and emission levels of pollutants such as SO(2) and NO(x), CO. Concentrations of atmospheric pollutants decreased with use of the correct ratio between fluidising and secondary air. Dispersion modelling of combustion at a proposed poultry unit predicted that ground level concentrations for the set of emissions data would be below the limits and guidelines set by air quality standards.     

Pilot-scale culture ofCoffea arabica in a novel loop fluidised bed reactor

A novel bubble free loop fluidized bed reactor for plant cell cultures was developed and tested usingCoffea arabica as a model cell line. The effects of main operational parameters like morphology and size of inoculum, oxygen supply as well as recirculation of sparingly soluble gases on cell growth and alkaloid production rates in this reactor were studied and the results were compared with standard shake flask experiments. By on-line monitoring of biomass and oxygen uptake rates the main kinetic parameters for cell growth and alkaloid production were evaluated. It was demonstrated that the novel reactor is easy to run and is particularly adequate for measuring kinetic parameters necessary for scale up.     

The influence of yeast metabolism on dispersion in a fluidised yeast bed

Summary The effect of yeast metabolism on the dispersion characteristics of a fluidised bed fermentor containing flocs of the yeast Saccharomyces carlsbergensis was investigated. Dispersion in the metabolizing fluidised yeast floc system was compared with the dispersion in an inert yeast floc system and in a glass bead system. Breakdown in plug-flow was found to occur in the metabolically active yeast bed when the flow rate was increased over a relatively narrow operating range (up to a dilution rate of 0.08 h^-1). The superficial liquid velocity at which perfect mixing was approximated was some 18 times greater in the inert yeast floc system than in the metabolizing yeast floc system.Abbreviations C Tracer concentration - C ₀ Concentration of tracer at time t=0 - V Mixing chamber volume - v Volumetric flow rate - l Time - t Mean residence time - N Number of tanks in series - D/l Dispersion number - ² variance     

Mechanical stability of some immobilised preparations for glucose isomerisation in fixed and agitated bed columns

Immobilised preparations mechanical stability was studied by an analysis of particle disintegration in fixed and agitated beds of laboratory reactors. The analysis was carried out by comparing the particle size distributions obtained for three different states within the overall time-course of operation: The initial state, an intermediate state and the final one. It was found that mechanical stability by both fixed bed operation and paddle mixing was high. Agitation by flat discs was shown to cause intensive particle attrition. The relationships observed are discussed with regard to the flow pattern of the various types of reactors.     

The use of macroporous gelatin carriers for the cultivation of mammalian cells in fluidised bed reactors

A fluidised bed system for the cultivation of mammalian cells on a new type of macroporous gelatin microcarrier is described. The volumetric cell densities achieved under controlled conditions for two standard cell lines (VERO, CHO) were one order of magnitude higher compared to conventional techniques using spherical microcarriers. The system can be potentially used for both anchorage dependent and independent cells.     

Denitrification kinetics in a fluidised bed using methanol as the carbon source

Summary Denitrification in a fluidised bed was found to follow the Michaelis-Menten rate equation. The kinetic constants for the process were determined using a shallow bed in the laboratory for the prediction of deep bed performance on a pilot plant.     

Use of a micro-expanded bed containing immobilised lysozyme for cell disruption in flow injection analysis

A method for cell disruption in Flow Injection Analysis (FIA) systems has been developed. The principle involves on-line cell disruption by means of immobilised lysozyme followed by an ultrasonic treatment. In order to avoid flow problems in the analytical system, the lysozyme was immobilised to Streamline^reg that was used in an expanded bed in the flow system. Samples of suspensions of Micrococcus lysodeikticus were treated and the success of the treatment was evaluated in terms of released protein and as a decrease in the optical density at 450 nm. The new technology offers a powerful tool in flow injection analyses for quantification of intracellular compounds. The concept of integration, i.e. combining cell disruption with handling of cell debris and assay procedure in one continuous flow process facilitates its use and increases the probability of reaching reproducible and reliable results.     

Bed expansion and pressure drop studies in a liquid-solid inverse fluidised bed reactor

Hydrodynamic characteristics (bed expansion and pressure drop) of a different type of two phase fluidised bed reactor termed as liquid-solid inverse fluidised bed reactor in which low density particles are fluidised with downward flow of liquid are studied in the present investigation. Experiments were conducted using 6?mm diameter spherical particles of low density polyethylene (LDPE) and polypropylene (PP) with water and aqueous solutions of Carboxy methyl cellulose (CMC). It was found that the minimum fluidisation velocity, U _mf decreased with increase in CMC concentration and solid density. A dimensionless correlation was proposed for the prediction of bed height at fully fluidised conditions.     

Experimental behavior of a whole cell immobilized dextransucrase biocatalyst in batch and packed bed reactors

Dextransucrases from Leuconostoc mesenteroides have been used to produce a diversity of controlled structure oligosaccharides with potential industrial applications. This is the case of !(1̄) branched glucooligosaccharides produced by L. mesenteroides NRRL B-1299 dextransucrase. In order to establish an industrial scale process with the immobilized enzyme, a biocatalyst was produced by whole cell entrapment in alginate beads. The main physical and physicochemical properties of the biocatalyst were determined and the hydrodynamic behavior in a packed bed reactor studied. It was possible to produce spherical beads of 0.2 cm diameter containing the insoluble part of L. mesenteroides culture (cells and insoluble polymer) with an activity of 4 IU/g. Immobilization yield reached 93% with an effectiveness factor of 0.995 for particles of d_p < 0.2 cm. Due to the complexity of dextransucrase mechanism and kinetics, data obtained from initial rate measurements failed to describe the results obtained from the batch and continuous reactors. Therefore, apparent K_M and V_max data were used for the reactor modeling. It was found that under the conditions studied, the reaction rate was controlled by external mass transfer limitations.     

Fabrication and characterisation of a novel pellicular adsorbent customised for the effective fluidised bed adsorption of protein products

A dense pellicular solid matrix has been fabricated by coating 4% agarose gel on to dense zirconia-silica (ZS) spheres by water-in-oil emulsification. The agarose evenly laminated the ZS bead to a depth of 30 μm, and the resulting pellicular assembly was characterised by densities up to 2.39 g/mL and a mean particle diameter of 136 μm. In comparative fluidisation tests, the pellicular solid phase exhibited a two-fold greater flow velocity than commercial benchmark adsorbents necessary to achieve common values of bed expansion. Furthermore, the pellicular particles were characterised by improved qualities of chromatographic behaviour, particularly with respect to a three-fold increase in the apparent effective diffusivity of lysozyme within a pellicular assembly modified with Cibacron Blue 3GA. The properties of rapid protein adsorption/desorption were attributed to the physical design and pellicular deployment of the reactive surfaces in the solid phase. When combined with enhanced feedstock throughput, such practical advantages recommend the pellicular assembly as a base matrix for the selective recovery of protein products from complex, particulate feedstocks (whole fermentation broths, cell disruptates and biological extracts).     

The influence of variable organic loading on the start-up of an anaerobic fluidised bed reactor

Summary A stepped-loading start-up regime utilising variable organic influent concentrations in the range 1650–11600 mgCOD1^–1 was applied to an anaerobic fluidised bed bioreactor at 37°C. The reactor was sensitive to variable influent COD concentrations, but the stepped-loading aided rapid recovery from transient organic loading shocks. Variable effluent COD levels were produced but a COD removal efficiency of 76% was obtained at a final HRT of 0.5 d and an organic loading rate of 5.3 kg COD m^–3 d^–1.