Quantification of brewers' yeast flocculation in a stirred tank: effect of physical parameters on flocculation.

Quantification of yeast flocculation under defined conditions will help to understand the physical mechanisms of the flocculation process used in beer fermentation. Flocculation was quantified by measuring the size of yeast flocs and the number of single cells. For this purpose, a method to measure floc size and number of single cells in situ was developed.

Effect of agitation intensities on fungal morphology of submerged fermentation.

Both parallel fermentations with Aspergillus awamori (CBS 115.52) and a literature study on several fungi have been carried out to determine a relation between fungal morphology and agitation intensity. The studied parameters include hyphal length, pellet size, surface structure or so-called hairy length of pellets, and dry mass per-wet-pellet volume at different specific energy dissipation rates.

Scale-up of stirring as foam disruption (SAFD) to industrial scale.

Foam disruption by agitation-the stirring as foam disruption (SAFD) technique-was scaled up to pilot and production scale using Rushton turbines and an up-pumping hydrofoil impeller, the Scaba 3SHP1. The dominating mechanism behind SAFD-foam entrainment-was also demonstrated at production scale. The mechanistic model for SAFD defines a fictitious liquid velocity generated by the (upper) impeller near the dispersion surface, which is correlated with complete foam disruption.

Physiological responses to mixing in large scale bioreactors.

Escherichia coli fed-batch cultivations at 22 m3 scale were compared to corresponding laboratory scale processes and cultivations using a scale-down reactor furnished with a high-glucose concentration zone to mimic the conditions in a feed zone of the large bioreactor. Formate accumulated in the large reactor, indicating the existence of oxygen limitation zones. It is suggested that the reduced biomass yield at large scale partly is due to repeated production/re-assimilation of acetate from overflow metabolism and mixed acid fermentation products due to local moving zones with oxygen limitation.

Partitioning behavior of amino acids in aqueous two-phase systems with recyclable volatile salts.

As part of an ongoing research effort on aqueous two-phase systems (ATPSs) with volatile salts, this work describes the partitioning behavior of a series of amino acids, namely L-serine, glycine, L-alanine, L-valine, L-methionine, L-isoleucine, and L-phenylalanine, in these systems. The results show that amino acids partition in a similar way in polymer-volatile salt ATPSs and in traditional polymer-salt ATPSs. Increasing amino acid hydrophobicities lead to increasing partition coefficients.

Feasible boundaries of aqueous two-phase systems with NH(3) and CO(2) as recyclable volatile salts.

Aqueous two-phase systems (ATPSs) have great potential for use in the downstream processing of fermentation products. A major drawback of these systems, limiting application in industrial practice up till now, is the consumption of large amounts of auxiliary materials such as polymers and salts. Making use of alternative auxiliaries can diminish this relatively large discharge.

An efficient model development strategy for bioprocesses based on neural networks in macroscopic balances: Part II.

There is a need for efficient modeling strategies which quickly lead to reliable mathematical models that can be applied for design and optimization of (bio)-chemical processes. The serial gray box modeling strategy is potentially very efficient because no detailed knowledge is needed to construct the white box part of the model and because covenient black box modeling techniques like neural networks can be used for the black box part of the model. This paper shows for a typical biochemical conversion how the serial gray box modeling strategy can be applied efficiently to obtain a model with good frequency extrapolation properties.

Modeling and measurements of fungal growth and morphology in submerged fermentations.

Generalizing results from fungal fermentations is difficult due to their high sensitivity toward slight variation in starting conditions, poor reproducibility, and difference in strains. In this study a mathematical model is presented in which oxygen transfer, agitation intensity, dissolved oxygen tension, pellet size, formation of mycelia, the fraction of mycelia in the total biomass, carbohydrate source consumption, and biomass growth are taken into account. Two parameters were estimated from simulation, whereas all others are based on measurements or were taken from literature.

Effects of dissolved oxygen tension and mechanical forces on fungal morphology in submerged fermentation.

The effects of dissolved oxygen tension and mechanical forces on fungal morphology were both studied in the submerged fermentation of Aspergillus awamori. Pellet size, the hairy length of pellets, and the free filamentous mycelial fraction in the total biomass were found to be a function of the mechanical force intensity and to be independent of the dissolved oxygen tension provided that the dissolved oxygen tension was neither too low (5%) nor too high (330%). When the dissolved oxygen concentration was close to the saturation concentration corresponding to pure oxygen gas, A.

Modeling brewers' yeast flocculation.

Flocculation of yeast cells occurs during the fermentation of beer. Partway through the fermentation the cells become flocculent and start to form flocs. If the environmental conditions, such as medium composition and fluid velocities in the tank, are optimal, the flocs will grow in size large enough to settle.

Centrifugal partition chromatographic reaction for the production of chiral amino acids.

The use of a centrifugal partition chromatographic reactor is investigated for the production of chiral amino acids from racemic mixtures. Chirally selective enzymatic hydrolysis of N-acetyl-L-methionine into acetic acid and L-methionine was carried out in the chromatographic reactor to demonstrate the concept of integrated reaction and separation in centrifugal partition chromatography (CPC). The products L-methionine and acetic acid, as well as the unconverted substrate, N-acetyl-D-methionine are obtained separately.

Poly(ethylene glycol)-salt aqueous two-phase systems with easily recyclable volatile salts.

Aqueous two-phase systems (ATPSs) have great potential in the downstream processing of fermentation products. However, the consumption of large amounts of auxiliary materials limits application in industrial practice. Promising alternatives to the salts used so far are volatile salts such as ammonium bicarbonate and ammonium carbamate, which can be recycled to the extraction system as gaseous carbon dioxide and ammonia.

An efficient model development strategy for bioprocesses based on neural networks in macroscopic balances.

In the serial gray box modeling strategy, generally available knowledge, represented in the macroscopic balance, is combined naturally with neural networks, which are powerful and convenient tools to model the inaccurately known terms in the macroscopic balance. This article shows, for a typical biochemical conversion, that in the serial gray box modeling strategy the identification data only have to cover the input-output space of the inaccurately known term in the macroscopic balances and that the accurately known terms can be used to achieve reliable extrapolation. The strategy is demonstrated successfully on the modeling of the enzymatic (repeated) batch conversion of penicillin G, for which real-time results are presented.

Effects of glutamine supply on growth and metabolism of mammalian cells in chemostat culture.

Glutamine is a major source of energy, carbon, and nitrogen for mammalian cells. The amount of glutamine present in commercial mammalian cell media is, however, not necessarily balanced with cell requirements. Therefore, the effects of glutamine limitation on the physiology of two mammalian cell lines were studied in steady-state chemostat cultures fed with IMDM medium with 5% serum.

Classification, error detection, and reconciliation of process information in complex biochemical systems.

Bioprocess identification starts with collection of process information. Usually there is a variety of information available, consisting of actual measurement data, historical data, empirical kinetic and yield correlations, and general knowledge available from literature. A central problem is to find out how the various pieces of information should be integrated.

Variation of the pH of the background electrolyte due to electrode reactions in capillary electrophoresis: theoretical approach and in situ measurement.

Electrode reactions during the electrophoretic process may change the pH of the buffer and subsequently the migration behavior of solutes with resultant loss of reproducibility. A theoretical treatment of pH variations due to electrolytic processes is presented. The choice of buffer appears to have a dramatic influence on the pH variations observed, even if substantial buffer action is expected at the pH chosen.

Linear constrain relations in biochemical reaction systems III. Sequential application of data reconciliation for sensitive detection of systematic errors.

This article presents a method to test the presence of relatively small systematic measurement errors; e.g., those caused by inaccurate calibration or sensor drift.

Separation of selenium analogues of sulphur-containing amino acids by high-performance liquid chromatography and high-resolution gas chromatography.

The historically conditioned adaptation of living organisms to chemically corresponding elements is influenced in nature by anthropogenic activities in many regions, the selenium-sulphur pair being one example of such a case. The separation of selenomethionine, selenoethionine and selenocystine was studied by HPLC and high-resolution GC. Ion-exchange chromatography followed by temperature-programmed GC gives the possibility of the analytical separation of trace amounts of selenomethionine in a complex mixture of common amino acids.

Linear constraint relations in biochemical reaction systems: II. Diagnosis and estimation of gross errors.

Conservation equations derived from elemental balances, heat balances, and metabolic stoichiometry, can be used to constrain the values of conversion rates of relevant components. In the present work, their use will be discussed for detection and localization of significant errors of the following types: 1.At least one of the primary measurements has a significant error (gross measurement error).

Linear constraint relations in biochemical reaction systems: I. Classification of the calculability and the balanceability of conversion rates.

Measurements provide the basis for process monitoring and control as well as for model development and validation. Systematic approaches to increase the accuracy and credibility of the empirical data set are therefore of great value. In (bio)chemical conversions, linear conservation relations such as the balance equations for charge, enthalpy, and/or chemical elements, can be employed to relate conversion rates.

Influence of oxygen adsorption on the dynamic K(L)a measurement in three-phase slurry reactors.

To check for possible mass transfer limitations of oxygen and/or carbon dioxide in kinetic experiments on microbial desulphurization of coal, it is important to properly measure the volumetric mass transfer coefficient (k(L)a) especially at high slurry densities. Volumetric mass transfer coefficients of oxygen, at different solid hold-up values (epsilon(s) = 0 to 0.28) of coal slurries (d(par) < 100 * 10(-6) m), were measured in a lab scale fermentor and in a lab scale pachuca tank, using the dynamic gas-liquid absorption method.

Comparison of two experimental methods for the determination of Michaelis-Menten kinetics of an immobilized enzyme.

For the application of immobilized enzymes, the influence of immobilization on the activity of the enzyme should be Known. This influence can be obtained by determining the intrinsic kinetic parameters of the immobilized enzyme, and by comparing them with the kinetic parameters of the suspended enzyme. This article deals with the determination of the intrinsic kinetic parameters of an agarose-gel bead immobilized oxygen-consuming enzyme: L-lactate 2-monooxygenase.

Modeling and experimental validation of carbon dioxide evolution in alkalophilic cultures.

The chemical reactions involving carbon dioxide in mineral culture media are considered. A mathematic model is set up, based on published data, which is valid at pH values below 9, and in which the nonideality of the solution is taken into account. The crucial parameter is the constant expressing the equilibrium between carbon dioxide and bicarbonate, K(1).

Integrated product formation and recovery in fermentation.

Fermentation processes are hampered by a variety of problems originating from the accumulation of products in the fermenter. Integration of fermentation and a primary product separation step can accelerate the product formation, improve the product yield, and facilitate downstream processing. The advantages of integrated bioprocesses, however, are counteracted by the incompatibility of the subprocesses.

Determination of growth and coupled nitrification/denitrification by immobilized Thiosphaera pantotropha using measurement and modeling of oxygen profiles.

An oxygen microsensor in combination with mathematical modeling was used to determine the behavior of immobilized Thiosphaera pantotropha. This organism can convert ammonia completely to nitrogen gas under aerobic conditions (coupled nitrification/denitrification) and denitrifies nitrate at highest rates under anaerobic conditions. Immobilization of T.