Product inhibition in packed-bed immobilized enzyme reactors for complex processes: Modelling of two-substrate processes

An analytical model was developed for describing the performance of packed-bed enzymic reactors operating with two cosubstrates, and when one of the reaction products is inhibitory to the enzyme. To this aim, the compartmental analysis technique was used. The relevant equations obtained were solved numerically, and the effect of the main operational parameters on the reactor characteristics were studied.Notation C _infa,i ^sup* local concentration of products in the pores of stage i - C _j,i concentration of substrate j in the pores of stage i - D _infa ^sup* internal (pore) diffusion coefficient for the reaction product a - D _j internal (pore) diffusion coefficient of substrate j - J _infa,i ^sup* net flux of product a, taking place from the pores of stage i into the corresponding bulk phase - J _j,i net flux of substrate j, taking place from the bulk phase of stage i into the corresponding pores - K _b inhibition constant - K _m,1, K _m,2 Michaelis constants for substrate 1 and 2, respectively - K _q inhibition constant - n total number of elementary stages in the reactor - Q volumetric flow rate throughout the reactor - R _j,i, R _infa,i ^sup* local reaction rates in pores of stage i, in terms of concentration of substrate j and product a respectively - S _infa,i ^sup* , S _infa,i-1 ^sup* bulk concentration of the reaction product a, in the stages i and i — 1, respectively - S _j,0 concentration of substrate j in the reactor feed - S _j,i-1, S _j,i concentration of substrate j in the bulk phase leaving stages i — 1 and i, respectively - V total volume of the reactor - V _m maximal reaction rate in terms of volumetric units - y axial coordinate of the pores - y ₀ depth of the pores - ^* dimensionless parameter, defined in Equation (22) - ₁ dimensionless parameter, defined in Equation (6) - ₂ dimensionless parameter, defined in Equation (6) - ₁ dimensionless parameter, defined in Equation (6) - ₂ dimensionless parameter, defined in Equation (6) - ^* dimensionless parameter, defined in Equation (22) - ₁ dimensionless parameter, defined in Equation (6) - ₂ dimensionless parameter, defined in Equation (6) - ^* dimensionless parameter, defined in Equation (22) - ^* dimensionless parameter, defined in Equation (22) - volumetric packing density of catalytic particles (dimensionless) - porosity of the catalytic particles (dimensionless) - V _infi ^sup* dimensionless concentration of reaction product in pores of stage i, defined in Equation (17) - j,i dimensionless concentration of substrate j in pores of stage i; defined in Equation (6) - _j,i-1, _j.i dimensionless concentration of substrate j in the bulk phase of stage i; defined in Equation (6) - dimensionless position along the pore; defined in Equation (6)     

Analysis of a chemostat model with variable yield coefficient: Tessier kinetics

We investigate a chemostat model in which the growth rate is given by a Tessier expression with a variable yield coefficient. We combine analytical results with path-following methods. The washout conditions are found. When washout does not occur we establish the conditions under which the reactor performance and reactor productivity are maximised. We also determine the parameter region in which oscillations may be generated in the reactor. We briefly discuss the implications of our results for comparing the performance of a single bioreactor against a cascade of two bioreactors.     

Influence of immobilized biomolecules on magnetic bead plug formation and retention in capillary electrophoresis

Significant changes in the formation and retention of magnetic bead plugs in a capillary during electrophoresis were studied, and it was demonstrated that these effects were due to the type of biological molecule immobilized on the surface of these beads. Three biological molecules, an antibody, an oligonucleotide, and alkaline phosphatase (AP), were attached to otherwise identical streptavidin-coated magnetic beads through biotin-avidin binding in order to isolate differences in bead immobilization in a magnetic field resulting from the type of biological molecule immobilized on the bead surface. AP was also attached to the magnetic beads using epoxy groups on the bead surfaces (instead of avidin-biotin binding) to study the impact of immobilization chemistry. The formation and retention of magnetic bead plugs were studied quantitatively using light scattering detection of magnetic particles eluting from the bead plugs and qualitatively using microscopy. Both the types of biomolecule immobilized on the magnetic bead surface and the chemistry used to link the biomolecule to the magnetic bead impacted the formation and retention of the bead plugs.     

Spectrophotometric cell comprising parallel rotating and stationary bioreactors: Application to the determination of glucose in serum samples

A spectrophotometric cell comprising parallel bioreactors facing each other and containing immobilized enzyme preparations is described. The lower reactor rotates to minimize diffusional constraints, and the upper reactor is fixed to provide an integrated design for the realization of coupled enzyme-catalyzed reactions. The operating characteristics of the cell are illustrated with the determination of glucose using glucose oxidase [EC 1.1.3.4] and horseradish peroxidase [EC 1.11.1.7] as immobilized enzymes (horseradish peroxidase on the rotating reactor and glucose oxidase on the stationary one). The H₂O₂ produced in the dissolved-oxygen oxidation of β- -glucose enters into oxidative coupling in a reaction with N,N-dimethylaniline and 4-aminophenazone which is catalyzed by horseradish peroxidase; the absorbance of the colored complex formed provides the basis for monitoring. The cell was incorporated into a continuous-flow/stopped-flow/continuous-flow operation, and the determination was based on the rate of response under stopped-flow conditions. The overall approach was applied to the determination of glucose in standards of human serum and samples of bovine blood serum.     

A Comparison of Simple Rheological Parameters and Simulation Data for Zymomonas mobilis Fermentation Broths with High Substrate Loading in a 3-L Bioreactor

Traditionally, as much as 80% or more of an ethanol fermentation broth is water that must be removed. This mixture is not only costly to separate but also produces a large aqueous stream that must then be disposed of or recycled. Integrative approaches to water reduction include increasing the biomass concentration during fermentation. In this paper, experimental results are presented for the rheological behavior of high-solids enzymatic cellulose hydrolysis and ethanol fermentation for biomass conversion using Solka Floc as the model feedstock. The experimental determination of the viscosity, shear stress, and shear rate relationships of the 10 to 20% slurry concentrations with constant enzyme concentrations are performed with a variable speed rotational viscometer (2.0 to 200 rpm) at 40 °C. The viscosities of enzymatic suspension observed were in range of 0.0418 to 0.0144, 0.233 to 0.0348, and 0.292 to 0.0447 Pa s for shear rates up to 100 reciprocal seconds at 10, 15, and 20% initial solids (w/v), respectively. Computational fluid dynamics analysis of bioreactor mixing demonstrates the change in bioreactor mixing with increasing biomass concentration. The portion-loading method is shown to be effective for processing high-solids slurries.     

Bioethanol Production Optimization: A Thermodynamic Analysis

In this work, the phase equilibrium of binary mixtures for bioethanol production by continuous extractive process was studied. The process is composed of four interlinked units: fermentor, centrifuge, cell treatment unit, and flash vessel (ethanol-congener separation unit). A proposal for modeling the vapor–liquid equilibrium in binary mixtures found in the flash vessel has been considered. This approach uses the Predictive Soave–Redlich–Kwong equation of state, with original and modified molecular parameters. The congeners considered were acetic acid, acetaldehyde, furfural, methanol, and 1-pentanol. The results show that the introduction of new molecular parameters r and q in the UNIFAC model gives more accurate predictions for the concentration of the congener in the gas phase for binary and ternary systems.     

Simultaneous quantification of poly-dispersed anionic, amphoteric and nonionic surfactants in simulated wastewater samples using C18 high-performance liquid chromatography-quadrupole ion-trap mass spectrometry

This paper describes the development of a guantitative method for direct and simultaneous determination of three frequently encountered surfactants, amphoteric (cocoamphoacetate, CAA), anionic (sodium laureth sulfate, SLES), and nonionic (alcohol ethoxylate, AE) using a reversed-phase C18 HPLC coupled with an ESI ion-trap mass spectrometer (MS). Chemical composition, ionization characteristics and fragmentation pathways of the surfactants are presented. Positive ESI was effective for all three surfactants in agueous methanol buffered with ammonium acetate. The method enables rapid determinations in small sample volumes containing inorganic salts (up to 3.5 g L(-1)) and multiple classes of surfactants with high specificity by applying surfactant specific tandem mass spectrometric strategies. It has dynamic linear ranges of 2-60, 1.5-40, 0.8-56 mg L(-1) with R2 egual or greater than 0.999, 0.98 and 0.999 (10 microL injection) for CAA, SLES, and AE, respectively.