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1.
Micromachines (Basel) ; 15(6)2024 Jun 20.
Article in English | MEDLINE | ID: mdl-38930775

ABSTRACT

Acoustic levitation can provide significant benefits for many fundamental research questions. However, it is important to consider that the acoustic field influences the measurement environment. This work focuses on the dissolution of immobilised drops using acoustic levitation in liquid-liquid systems. Previous work demonstrated that the acoustic field of standing waves impacts mass transfer by affecting the spread of dissolved substances in the continuous phase in two distinct ways: (I) solutes may either pass through nodal planes of the standing waves or (II) not pass. The binary systems examined for case (I) are 1-hexanol-water and 1-butanol-water, and for case (II), n-butyl acetate-water and toluene-water. This work quantifies the intensification effect of acoustic levitation on dissolution for the two types of behaviour, by comparing them with reference measurements of mechanically attached dissolving drops. The system was designed to ensure minimal intensification. The minimum intensification of mass transfer for levitating drops in the used setup of case (I) was 25%, and for case (II), it was 65%, both increasing with decreasing surface-equivalent diameter. With this understanding, acoustic levitation can be used more accurately in the field of mass transfer studies.

2.
Magn Reson Chem ; 62(4): 259-268, 2024 Apr.
Article in English | MEDLINE | ID: mdl-37438985

ABSTRACT

The application of compact NMR instruments to hot flowing samples or exothermically reacting mixtures is limited by the temperature sensitivity of permanent magnets. Typically, such temperature effects directly influence the achievable magnetic field homogeneity and hence measurement quality. The internal-temperature control loop of the magnet and instruments is not designed for such temperature compensation. Passive insulation is restricted by the small dimensions within the magnet borehole. Here, we present a design approach for active heat shielding with the aim of variable temperature control of NMR samples for benchtop NMR instruments using a compressed airstream which is variable in flow and temperature. Based on the system identification and surface temperature measurements through thermography, a model predictive control was set up to minimise any disturbance effect on the permanent magnet from the probe or sample temperature. This methodology will facilitate the application of variable-temperature shielding and, therefore, extend the application of compact NMR instruments to flowing sample temperatures that differ from the magnet temperature.

3.
Micromachines (Basel) ; 14(10)2023 Oct 10.
Article in English | MEDLINE | ID: mdl-37893360

ABSTRACT

A low-impact acoustic levitation system has been developed to study immobilised single drops in liquid-liquid systems. The ability to observe liquid drops several millimetres in diameter for days enables fundamental research into a wide range of mechanisms. Non-invasive optical measurements with excellent optical accessibility are possible. This experimental work provides the basis for mass transfer studies, emphasizing the precise volume determination, signal noise, reproducibility, and the impact of the acoustic field on the drop and its surrounding environment. The setup can be effectively controlled and proves beneficial for research objectives provided that all liquid phases are entirely degassed, and there are no compressible voids present within the liquids. In addition to the precise, uniform, and reliable measurement conditions, we observed no acoustic streaming in the proximity of the drop and there was no significant vibration of the drop. Qualitative observations using rainbow schlieren deflectometry indicate that the nodal or anti-nodal planes of the standing waves can act as barriers to the dispersion of inhomogeneous dissolved substances in the continuous phase.

4.
Adv Colloid Interface Sci ; 315: 102907, 2023 May.
Article in English | MEDLINE | ID: mdl-37086624

ABSTRACT

CO2-switchable surfactants have selective surface-activity, which can be activated or deactivated either by adding or removing CO2 from the solution. This feature enables us to use them in the fabrication of responsive colloids, a group of dispersed systems that can be controlled by changing the environmental conditions. In chemical processes, including extraction, reaction, or heterogeneous catalysis, colloids are required in some specific steps of the processes, in which maximum contact area between immiscible phases or reactants is desired. Afterward, the colloids must be broken for the postprocessing of products, solvents, and agents, which can be facilitated by using CO2-switchable surfactants in surfactant-stabilized colloids. These surfactants are mainly cationic and can be activated by the protonation of a nitrogen-containing group upon sparging CO2 gas. Also, CO2-switchable superamphiphiles can be formed by non-covalent bonding between components at least one of which is CO2-switchable. So far, CO2-switchable surfactants have been used in CO2-switchable spherical and wormlike micelles, vesicles, emulsions, foams, and Pickering emulsions. Here, we review the fabrication procedure, chemical structure, switching scheme, stability, environmental conditions, and design philosophy of such responsive colloids. Their fields of application are wide, including emulsion polymerization, catalysis, soil washing, drug delivery, extraction, viscosity control, and oil transportation. We also emphasize their application for the CO2-assisted enhanced oil recovery (EOR) process as a promising approach for carbon capture, utilization, and storage to combat climate change.

5.
Int J Mol Sci ; 24(7)2023 Mar 23.
Article in English | MEDLINE | ID: mdl-37047056

ABSTRACT

Nucleoside analogues are important compounds for the treatment of viral infections or cancers. While (chemo-)enzymatic synthesis is a valuable alternative to traditional chemical methods, the feasibility of such processes is lowered by the high production cost of the biocatalyst. As continuous enzyme membrane reactors (EMR) allow the use of biocatalysts until their full inactivation, they offer a valuable alternative to batch enzymatic reactions with freely dissolved enzymes. In EMRs, the enzymes are retained in the reactor by a suitable membrane. Immobilization on carrier materials, and the associated losses in enzyme activity, can thus be avoided. Therefore, we validated the applicability of EMRs for the synthesis of natural and dihalogenated nucleosides, using one-pot transglycosylation reactions. Over a period of 55 days, 2'-deoxyadenosine was produced continuously, with a product yield >90%. The dihalogenated nucleoside analogues 2,6-dichloropurine-2'-deoxyribonucleoside and 6-chloro-2-fluoro-2'-deoxyribonucleoside were also produced, with high conversion, but for shorter operation times, of 14 and 5.5 days, respectively. The EMR performed with specific productivities comparable to batch reactions. However, in the EMR, 220, 40, and 9 times more product per enzymatic unit was produced, for 2'-deoxyadenosine, 2,6-dichloropurine-2'-deoxyribonucleoside, and 6-chloro-2-fluoro-2'-deoxyribonucleoside, respectively. The application of the EMR using freely dissolved enzymes, facilitates a continuous process with integrated biocatalyst separation, which reduces the overall cost of the biocatalyst and enhances the downstream processing of nucleoside production.


Subject(s)
Nucleosides , Pentosyltransferases , Nucleosides/chemistry , Pentosyltransferases/metabolism , Enzymes, Immobilized/chemistry , Biocatalysis , Deoxyribonucleosides , Purine-Nucleoside Phosphorylase/metabolism
6.
Nanomaterials (Basel) ; 13(4)2023 Feb 11.
Article in English | MEDLINE | ID: mdl-36839066

ABSTRACT

In this study, a first approach to model drop size distributions in agitated nanoparticle-stabilized liquid/liquid systems with population balance equations is presented. Established coalescence efficiency models fail to predict the effect of steric hindrance of nanoparticles at the liquid/liquid interface during the film drainage process. A novel modified coalescence efficiency is developed for the population balance framework based on the film drainage model. The elaborate submodel considers the desorption energy required to detach a particle from the interface, representing an energy barrier against coalescence. With an additional implemented function in the population balance framework, the interface coverage rate by particles is calculated for each time step. The transient change of the coverage degree of the phase interface by particles is thereby considered in the submodel. Validation of the modified submodel was performed with experimental data of agitated water-in-oil (w/o) dispersions, stabilized by well-defined spherical silica nanoparticles. The nanospheres with a size of 28 nm are positively charged and were hydrophobized by silanization with dimethyloctadecyl[3-(trimethoxysilyl)propyl]ammoniumchloride. This modeling approach is a first step toward predicting time-resolved dynamic drop size distributions of nanoparticle-stabilized liquid/liquid systems.

7.
J Environ Manage ; 326(Pt B): 116791, 2023 Jan 15.
Article in English | MEDLINE | ID: mdl-36435135

ABSTRACT

The high water content and low nutrient concentration of digestate complicate its storage, transportation, and utilization. Subsequent digestate processing can effectively remove water and influence nutrient partitioning among digestate fractions and final products. The current study was carried out to evaluate the performance of two typical digestate processing chains, solid and liquid ones, respectively, and to give practical recommendations for optimization. Two fully operating biogas plants with advanced heat utilization were considered as data sources. The digestate mass flow balance of dry matter (DM), water, total N (TN), and ammonium-N mass flows was performed and the efficiency of the examined processing units was calculated. It was found that solid-liquid separation of raw digestate shifted 73-87% of TN and 60-93% of NH4-N to the liquid phase. Subsequent drying of separated solid fraction removed about 6% of the initial water and required 84% less thermal energy per kg N recovered than the processing of separated liquid. The final product, pellets, contained 14% of initial TN, but only 2% of initial NH4-N as a result of microbial conversion of inorganic N during drying. Vacuum evaporation of separated liquid fraction removed 34% of the initial water and left a DM-rich concentrate. At the same time, an ammonium sulfate solution (ASS) containing 21% of initial TN and 34% of initial NH4-N was produced. Both evaluated processing chains showed specific advantages and challenges. Solid products were characterized by a high share of recalcitrant organic compounds and could serve as a soil improver. Liquid processing concentrated plant-available N in ASS, which could be used as valuable inorganic fertilizer.


Subject(s)
Fertilizers , Nitrogen , Anaerobiosis , Biofuels , Fertilizers/analysis , Nitrogen/analysis , Soil , Water
8.
Nanomaterials (Basel) ; 12(15)2022 Aug 01.
Article in English | MEDLINE | ID: mdl-35957079

ABSTRACT

The incorporation of soft hydrophilic particles at the interface of water in non-polar oil emulsion droplets is crucial for several applications. However, the stabilization of water in non-polar oil emulsions with hydrophilic soft material alone is, besides certain exceptions, not possible. In our previous works, we showed that stabilizing the emulsions with well-characterized spherical hydrophobic silica nanospheres (SNs) and soft equally charged microgel particles (MGs) is a robust strategy to stabilize w/o emulsions while still incorporating a large amount of MGs at the interface. In the present study, we address the question of what the maximum amount of MGs at the interface in these kinds of emulsion droplets can be. By using well-characterized mono-disperse SNs, we are able to calculate the fraction of interface covered by the SNs and complementary that of the present MG. We found that it is not possible to decrease the SN coverage below 56% irrespective of MG softness and SN size. The findings elucidate new perspectives to the broader topic of soft/solid stabilized emulsions.

9.
Adv Colloid Interface Sci ; 301: 102601, 2022 Mar.
Article in English | MEDLINE | ID: mdl-35114446

ABSTRACT

Protein surfactant (PS) interactions is an essential topic for many fundamental and technological applications such as life science, nanobiotechnology processes, food industry, biodiesel production and drug delivery systems. Several experimental techniques and data analysis approaches have been developed to characterize PS interactions in bulk and at interfaces. However, to evaluate the mechanisms and the level of interactions quantitatively, e.g., PS ratio in complexes, their stability in bulk, and reversibility of their interfacial adsorption, new experimental techniques and protocols are still needed, especially with relevance for in-situ biological conditions. The available standard techniques can provide us with the basic understanding of interactions mainly under static conditions and far from physiological criteria. However, detailed measurements at complex interfaces can be formidable due to the sophisticated tools required to carefully probe nanometric phenomena at interfaces without disturbing the adsorbed layer. Tensiometry-based techniques such as drop profile analysis tensiometry (PAT) have been among the most powerful methods for characterizing protein's and surfactant's adsorption layers at interfaces via measuring equilibrium and dynamic interfacial tension and dilational rheology analysis. PAT provides us with insightful data such as kinetics and isotherms of adsorption and related surface activity parameters. However, the data analysis and interpretation can be challenging for mixed protein-surfactant solutions via standard PAT experimental protocols. The combination of a coaxial double capillary (micro flow exchange system) with drop profile analysis tensiometry (CDC-PAT) is a promising tool to provide valuable results under different competitive adsorption/desorption conditions via novel experimental protocols. CDC-PAT provides unique experimental protocols to exchange the droplet subphase in a continuous dynamic mode during the in-situ analysis of the corresponding interfacial adsorbed layer. The contribution of diffusion/convection mechanisms on the kinetics of the adsorption/desorption processes can also be investigated using CDC-PAT. Here, firstly, we review the commonly available techniques for characterizing protein-surfactant interactions in the bulk phase and at interfaces. Secondly, we give an overview for applications of the coaxial double capillary PAT setup for investigations of mixed protein-surfactant adsorbed layers and address recently developed protocols and analysis procedures. Exploring the competitive sequential adsorption of proteins and surfactants and the reversibility of pre-adsorbed layers via the subphase exchange are the particular experiments we can perform using CDC-PAT. Also the sequential and simultaneous competitive adsorption/desorption processes of some ionic and nonionic surfactants (SDS, CTAB, DTAB, and Triton) and proteins (bovine serum albumin (BSA), lysozyme, and lipase) using CDC-PAT are discussed. Last but not least, the fabrication of micro-nanocomposite layers and membranes are additional applications of CDC-PAT discussed in this work.


Subject(s)
Serum Albumin, Bovine , Surface-Active Agents , Adsorption , Surface Properties , Surface Tension
10.
Membranes (Basel) ; 12(1)2022 Jan 12.
Article in English | MEDLINE | ID: mdl-35054608

ABSTRACT

Hemicellulose and its derivatives have a high potential to replace fossil-based materials in various high-value-added products. Within this study, two purification cascades for the separation and valorization of hemicellulose and its derived monomeric sugars from organosolv beechwood hydrolyzates (BWHs) were experimentally demonstrated and assessed. Purification cascade 1 included hydrothermal treatment for converting remaining hemicellulose oligomers to xylose and the purification of the xylose by nanofiltration. Purification cascade 2 included the removal of lignin by adsorption, followed by ultrafiltration for the separation and concentration of hemicellulose. Based on the findings of the experimental work, both cascades were simulated on an industrial scale using Aspen Plus®. In purification cascade 1, 63% of the oligomeric hemicellulose was hydrothermally converted to xylose and purified by nanofiltration to 7.8 t/h of a xylose solution with a concentration of 200 g/L. In purification cascade 2, 80% of the lignin was removed by adsorption, and 7.6 t/h of a purified hemicellulose solution with a concentration of 200 g/L was obtained using ultrafiltration. The energy efficiency of the cascades was 59% and 26%, respectively. Furthermore, the estimation of specific production costs showed that xylose can be recovered from BWH at the cost of 73.7 EUR/t and hemicellulose at 135.1 EUR/t.

11.
Langmuir ; 37(44): 12919-12928, 2021 11 09.
Article in English | MEDLINE | ID: mdl-34699224

ABSTRACT

The enzymatic hydrolysis of sunflower oil occurs at the water-oil interface. Therefore, the characterization of dynamic interfacial phenomena is essential for understanding the related mechanisms for process optimizations. Most of the available studies for this purpose deal with averaged interfacial properties determined via reaction kinetics and dynamic surface tension measurements. In addition to the classical approach for dynamic surface tension measurements, here, the evolution of the dilational viscoelasticity of the lipase adsorbed layer at the water-oil interface is characterized using profile analysis tensiometry. It is observed that lipase exhibits nonlinear dilational rheology depending on the concentration and age of the adsorbed layer. For reactive water-oil interfaces, the response of the interfacial tension to the sinusoidal area perturbations becomes more asymmetric with time. Surface-active products of the enzymatic hydrolysis of triglycerides render the interface less elastic during compression compared to the expansion path. The lipolysis products can facilitate desorption upon compression while inhibiting adsorption upon expansion of the interface. Lissajous plots provide an insight into how the hysteresis effect leads to different interfacial tensions along the expansion and compression routes. Also, the droplet shape increasingly deviates from a Laplacian shape, demonstrating an irreversible film formation during aging and ongoing hydrolysis reaction, which supports our findings via interfacial elasticity analysis.


Subject(s)
Lipase , Water , Adsorption , Hydrolysis , Rheology , Triglycerides
12.
Soft Matter ; 17(36): 8258-8268, 2021 Sep 22.
Article in English | MEDLINE | ID: mdl-34550151

ABSTRACT

A general drawback of microgels is that they do not stabilize water-in-oil (w/o) emulsions of non-polar oils. Simultaneous stabilization with solid hydrophobic nanoparticles and soft hydrophilic microgels overcomes this problem. For a fundamental understanding of this synergistic effect the use of well defined particle systems is crucial. Therefore, the present study investigates the stabilization of water droplets in a highly non-polar oil phase using temperature responsive, soft and hydrophilic PNIPAM microgel particles (MGs) and solid and hydrophobic silica nanospheres (SNs) simultaneously. The SNs are about 20 times smaller than the MGs. In a multiscale approach the resulting emulsions are studied from the nanoscale particle properties over microscale droplet sizes to macroscopic observations. The synergy of the particles allows the stabilization of water-in-oil (w/o) emulsions, which was not possible with MGs alone, and offers a larger internal interface than the stabilization with SNs alone. Furthermore, the incorporation of hydrophilic MGs into a hydrophobic particle layer accelerates the emulsions sedimentation speed. Nevertheless, the droplets are still sufficiently protected against coalescence even in the sediment and can be redispersed by gentle shaking. Based on droplet size measurements and cryo-SEM studies we elaborate a model, which explains the found phenomena.

13.
Langmuir ; 36(40): 12010-12022, 2020 10 13.
Article in English | MEDLINE | ID: mdl-32938187

ABSTRACT

Lipase is one of the most important enzymes playing a key role in many biological and chemical processes, in particular for fat hydrolysis in living systems and technological applications such as food production, medicine, and biodiesel production. As lipase is soluble in water, the major hydrolysis process occurs at the water-oil interface, where lipase can get in contact with the oil. To provide optimum conditions, the emulsification of the oil is essential to provide a large interfacial area which is generally done by adding surfactants. However, the presence of surfactants can influence the lipase activity and also cause competitive adsorption, resulting in a removal of lipase from the interface or its conformational changes in the solution bulk. Here we have studied the dynamics of competitive adsorption and interfacial elasticity of mixed solutions containing lipase and the anionic surfactant sodium dodecyl sulfate (SDS) or the cationic surfactant cetyltrimethylammonium bromide (CTAB), respectively, at the water-air interface. The experiments were performed with a special coaxial double capillary setup for drop bulk-interface exchange developed for the drop profile analysis tensiometer PAT with two protocols: sequential and simultaneous adsorption of single components and mixed systems. The results in terms of dynamic surface tension and dilational viscoelasticity illustrate fast and complete desorption of a preadsorbed CTAB and SDS layers via subphase exchange with a buffer solution. In contrast, the preadsorbed lipase layer cannot be removed either by SDS or CTAB from the interface during drop bulk exchange with a buffer solution due to the unfolding process and conformation evolution of the protein molecules at the interface. In the opposite case, lipase can remove preadsorbed SDS and CTAB. The dynamic surface tension and viscoelasticity data measured before and after subphase exchange show joint adsorption of lipase and CTAB in the form of complexes, while SDS is adsorbed in competition with lipase. The results are in good correlation with the determined surface charges of the lipase gained by computational simulations which show a dominant negatively charged surface for lipase that can interact with the cationic CTAB while partial positively charged regions are observed for the interaction with the anionic SDS.

14.
J Colloid Interface Sci ; 573: 135-149, 2020 Aug 01.
Article in English | MEDLINE | ID: mdl-32278172

ABSTRACT

HYPOTHESIS: Pickering emulsions (PE) are becoming of increasing interest for catalytic multiphase processes. Ultrafiltration of PE is a promising procedure for catalyst recovery to enable continuous processes. Dispersing conditions during production of PE are expected to significantly influence PE characteristics, and control of these properties is essential for robust process design. However, while the impact of PE composition has been studied before, knowledge on dispersing conditions is surprisingly scarce. EXPERIMENTS: The influence of dispersing time, speed and emulsion volume during the preparation of PE with an UltraTurrax (2 dispersing tools) on the drop size distribution, rheology, stability and filtration was investigated. FINDINGS: In this first systematic study of PE preparation conditions, obtained Sauter mean diameters were correlated with energy density (R2 = 0.80), energy dissipation rate (R2 = 0.85) and tip speed (R2 = 0.86). All emulsions were stable for at least 10 weeks. With increasing tip speed (4-13 m/s), the dynamic viscosity first decreased, passed through a plateau value and then increased again. Filtration of concentrated PE was successful but strong membrane-particle-solvent interactions were revealed. This work contributes to a better understanding of PE properties that are essential for a sound application of PE in continuous multiphase catalysis.

15.
Membranes (Basel) ; 10(4)2020 Apr 04.
Article in English | MEDLINE | ID: mdl-32260462

ABSTRACT

: Membrane filtration of biological suspensions is frequently limited by fouling. This mechanism is well understood for ultrafiltration of activated sludge in membrane bioreactors. A rather young application of ultrafiltration is the recovery of nutrients from anaerobic digestates, e.g., from agricultural biogas plants. A process chain of solid/liquid separation, ultrafiltration, and reverse osmoses separates the digestate into different products: an organic N-P-fertilizer (solid digestate), a recirculate (UF retentate), a liquid N-K-fertilizer (RO retentate) and water. Despite the preceding particle removal, high crossflow velocities are required in the ultrafiltration step to overcome fouling. This leads to high operation costs of the ultrafiltration step and often limits the economical application of the complete process chain. In this study, under-stoichiometric ozone treatment of the ultrafiltration feed stream is investigated. Ozone treatment reduced the biopolymer concentration and apparent viscosity of different digestate centrates. Permeabilities of centrate treated with ozone were higher than without ozone treatment. In a laboratory test rig and in a pilot plant operated at the site of two full scale biogas plants, ultrafiltration flux could be improved by 50%-80% by ozonation. Nutrient concentrations in the fertilizer products were not affected by ozone treatment.

16.
Colloids Surf B Biointerfaces ; 185: 110580, 2020 Jan 01.
Article in English | MEDLINE | ID: mdl-31732392

ABSTRACT

The use of Pickering emulsions for biocatalytical applications has recently received increased attention in cases where hydrophobic reactants are involved. For process applications, knowledge of the emulsion's rheology is crucial for the fluid dynamical design of equipment and selection of operating conditions. Colloidal silica nanoparticle stabilized Pickering emulsions usually exhibit shear-thinning behavior caused by a complex particle-particle network. While this has been observed by many authors, no publication has yet dealt with the rheology of silica nanoparticle stabilized Pickering emulsions containing enzymes. Thus, the aim of this study was to investigate the impact of the commonly used biocatalyst lipase (type and concentration), the dispersed phase volume fraction and the silica particle concentration on the rheological behavior of water-in-oil Pickering emulsions. For this purpose, the impact of the named parameters on the viscosity curves were measured. Lipases reduced the viscosities and transferred the rheological behavior from shear-thinning to Newtonian, which might be due to interactions of the lipase molecules via the formation of intermolecular disulfide bonds, which disturb the hydrogen-bond based silica particle-particle network. However, by increasing the dispersed phase volume fraction or the silica particle concentration the rheological behavior of emulsions became again shear-thinning. This work will help to produce bioactive Pickering emulsions with tailor-made characteristics.


Subject(s)
Bacterial Proteins/chemistry , Colloids/chemistry , Emulsions , Lipase/chemistry , Nanoparticles/chemistry , Rheology , Silicon Dioxide/chemistry , Hydrophobic and Hydrophilic Interactions , Water/chemistry
17.
Bioengineering (Basel) ; 5(4)2018 Dec 04.
Article in English | MEDLINE | ID: mdl-30518117

ABSTRACT

Human adipose tissue-derived stromal/stem cells (hASCs) are a valuable source of cells for clinical applications, especially in the field of regenerative medicine. Therefore, it comes as no surprise that the interest in hASCs has greatly increased over the last decade. However, in order to use hASCs in clinically relevant numbers, in vitro expansion is required. Single-use stirred bioreactors in combination with microcarriers (MCs) have shown themselves to be suitable systems for this task. However, hASCs tend to be less robust, and thus, more shear sensitive than conventional production cell lines for therapeutic antibodies and vaccines (e.g., Chinese Hamster Ovary cells CHO, Baby Hamster Kidney cells BHK), for which these bioreactors were originally designed. Hence, the goal of this study was to investigate the influence of different shear stress levels on the growth of humane telomerase reversed transcriptase immortalized hASCs (hTERT-ASC) and aggregate formation in stirred single-use systems at the mL scale: the 125 mL (= SP100) and the 500 mL (= SP300) disposable Corning® spinner flask. Computational fluid dynamics (CFD) simulations based on an Euler⁻Euler and Euler⁻Lagrange approach were performed to predict the hydrodynamic stresses (0.06⁻0.87 Pa), the residence times (0.4⁻7.3 s), and the circulation times (1.6⁻16.6 s) of the MCs in different shear zones for different impeller speeds and the suspension criteria (Ns1u, Ns1). The numerical findings were linked to experimental data from cultivations studies to develop, for the first time, an unstructured, segregated mathematical growth model for hTERT-ASCs. While the 125 mL spinner flask with 100 mL working volume (SP100) provided up to 1.68.105 hTERT-ASC/cm² (= 0.63 × 106 living hTERT-ASCs/mL, EF 56) within eight days, the peak living cell density of the 500 mL spinner flask with 300 mL working volume (SP300) was 2.46 × 105 hTERT-ASC/cm² (= 0.88 × 106 hTERT-ASCs/mL, EF 81) and was achieved on day eight. Optimal cultivation conditions were found for Ns1u < N < Ns1, which corresponded to specific power inputs of 0.3⁻1.1 W/m³. The established growth model delivered reliable predictions for cell growth on the MCs with an accuracy of 76⁻96% for both investigated spinner flask types.

18.
J Biotechnol ; 266: 133-143, 2018 Jan 20.
Article in English | MEDLINE | ID: mdl-29208410

ABSTRACT

The application of biocalorimetry to fermentation processes offers advantageous insights, while being less complex compared to other, sophisticated PAT solutions. Although the general concept is established, calorimetric methods vary in detail. In this work, a special approach, called heat compensation calorimetry, was applied to an E. coli fed-batch process. Much work has been done for batch processes, proving the validity and accuracy of this calorimetric mode. However, the adaption of this strategy to fed-batch processes has some implications. In the first section of this work, batch fermentations were performed, comparing heat capacity calorimetry to the compensation mode. Both processes showed very good agreement by means of growth behavior. The heat related differences, e.g. temperature profiles, were obvious. In addition, the impact of the chosen mode on the calculation of in-process heat transfer coefficients was shown. Finally, a fed-batch fermentation was performed. The compensation mode was kept sufficiently, up to the point where the metabolic heat production accelerated strongly. Controller tuning was a neuralgic point, which would have needed further optimization under these conditions. Nevertheless, in the present work it was possible to realize a working compensation process while demonstrating critical aspects that must be considered when establishing such approach.


Subject(s)
Bioreactors , Escherichia coli/growth & development , Hot Temperature , Calorimetry
19.
Eng Life Sci ; 17(5): 500-511, 2017 05.
Article in English | MEDLINE | ID: mdl-28579937

ABSTRACT

Power input is an important engineering and scale-up/down criterion in stirred bioreactors. However, reliably measuring power input in laboratory-scale systems is still challenging. Even though torque measurements have proven to be suitable in pilot scale systems, sensor accuracy, resolution, and errors from relatively high levels of friction inside bearings can become limiting factors at smaller scales. An experimental setup for power input measurements was developed in this study by focusing on stainless steel and single-use bioreactors in the single-digit volume range. The friction losses inside the air bearings were effectively reduced to less than 0.5% of the measurement range of the torque meter. A comparison of dimensionless power numbers determined for a reference Rushton turbine stirrer (NP = 4.17 ± 0.14 for fully turbulent conditions) revealed good agreement with literature data. Hence, the power numbers of several reusable and single-use bioreactors could be determined over a wide range of Reynolds numbers between 100 and >104. Power numbers of between 0.3 and 4.5 (for Re = 104) were determined for the different systems. The rigid plastic vessels showed similar power characteristics to their reusable counterparts. Thus, it was demonstrated that the torque-based technique can be used to reliably measure power input in stirred reusable and single-use bioreactors at the laboratory scale.

20.
Eng Life Sci ; 17(12): 1234-1243, 2017 Dec.
Article in English | MEDLINE | ID: mdl-32624751

ABSTRACT

Single-use bioreactors are barely described by means of their heat transfer characteristics, although some of their properties might affect this process. Steady-state methods that use external heat sources enable precise investigations. One option, commonly present in stirred, stainless steel tanks, is to use adjustable electrical heaters. An alternative are exothermic chemical reactions that offer a higher flexibility and scalability. Here, the catalytic decay of hydrogen peroxide was considered a possible reaction, because of the high reaction enthalpy of -98.2 kJ/mole and its uncritical reaction products. To establish the reaction, a proper catalyst needed to be determined upfront. Three candidates were screened: catalase, iron(III)-nitrate and manganese(IV)-oxide. Whilst catalase showed strong inactivation kinetic and general instability and iron(III)-nitrate solution has a pH of 2, it was decided to use manganese(IV)-oxide for the bioreactor studies. First, a comparison between electrical and chemical power input in a benchtop glass bioreactor of 3.5 L showed good agreement. Afterwards the method was transferred to a 50 L stirred single-use bioreactor. The deviation in the final results was acceptable. The heat transfer coefficient for the electrical method was 242 W/m2/K, while the value achieved with the chemical differed by less than 5%. Finally, experiments were carried out in a 200 L single-use bioreactor proving the applicability of the chemical power input at technical relevant scales.

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