04 Fakultät Energie-, Verfahrens- und Biotechnik
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Item Open Access Behavior of sulfur oxides in air and oxy-fuel combustion(2019) Spörl, Reinhold; Scheffknecht, Günter (Univ.-Prof. Dr. techn.)This thesis evaluates the behavior of sulfur oxides in pulverized fuel (PF) fired air and oxy-fuel systems. Sulfur oxides are responsible for certain operational problems and considerable gas cleaning requirements in air as well as oxy-fuel firing. A better understanding of the related issues will allow for a technical and economical optimization of the oxy-fuel combustion technology. A range of experimental investigations studying the stability and retention of sulfur oxides in ashes and deposits, acid gas (SO2, SO3, and HCl) control in air and oxy-fuel combustion by dry sorbent injection, and SO3 formation were conducted. The experimental work is in parts supported by theoretical considerations and thermodynamic equilibrium simulation. Studies for different coals and lignites showed that in practically relevant oxy-fuel configurations the exclusion of airborne N2 from combustion leads to an increase of the SO2 concentrations in oxy-fuel, compared to air firing, by a factor of about 3.4 to 4.2, referring to dry, and of about 2.9 to 3.5, when referring to wet flue gas conditions. The increased SO2 levels in oxy-fuel combustion are responsible for an increased stability of sulfates in oxy-fuel power boiler systems so that for example the decomposition temperature CaSO4 rises by about 50 to 80 °C, depending on flue gas atmospheres. The enhanced stability of sulfates in deposits at high temperatures when operating with increased SO2 levels was experimentally demonstrated. Compared to air firing, a considerable increase of the sulfur retention in the ash by 10 to 12 percentage points has been observed for oxy-fuel recycle combustion of Lusatian lignites. This leads to lower SO2 emissions and higher SO3 levels in process ashes and deposits. The results indicate that for fuels, such as the used lignites, the temperature level at which fouling by sulfatic deposits is problematic may be shifted to higher temperatures in oxy-fuel combustion and that the sintering of deposits by sulfation may be more pronounced. In contrast, in air and oxy-fuel combustion experiments with a hard coal with a low sulfur retention potential differences in the SO3 contents and degrees of sulfation of ashes and deposits were small. Besides higher SO3 contents and sulfation degrees, no other significant changes between the deposit samples from air and oxy-fuel combustion were identified. Experiments on dry sorbent injection in air and oxy-fuel mode showed that an increase of the average flue gas residence time in the furnace by flue gas recirculation and, to a lesser extent, the higher sulfate stability enhance the desulfurization efficiency in oxy-fuel recycle combustion considerably. SO2 capture efficiencies in oxy-fuel recycle combustion of 50 % to more than 80 % at moderate molar sulfur to calcium ratios between 1.7 and 2.9 were reached, when injecting CaCO3 and Ca(OH)2 together with the fuel or directly to the furnace. Under comparable injection conditions, the oxy-fuel performance was by as much as 29 percentage points higher than in air firing. Also an efficient SO3 and HCl control by DSI could be demonstrated. Experiments on formation of SO3 show that higher SO2 levels in oxy-fuel firing are the most important parameter responsible for the observed increase of the SO3 concentrations.Item Open Access Precision 3D‐printed cell scaffolds mimicking native tissue composition and mechanics(2020) Erben, Amelie; Hörning, Marcel; Hartmann, Bastian; Becke, Tanja; Eisler, Stephan A.; Southan, Alexander; Cranz, Séverine; Hayden, Oliver; Kneidinger, Nikolaus; Königshoff, Melanie; Lindner, Michael; Tovar, Günter E. M.; Burgstaller, Gerald; Clausen‐Schaumann, Hauke; Sudhop, Stefanie; Heymann, MichaelCellular dynamics are modeled by the 3D architecture and mechanics of the extracellular matrix (ECM) and vice versa. These bidirectional cell‐ECM interactions are the basis for all vital tissues, many of which have been investigated in 2D environments over the last decades. Experimental approaches to mimic in vivo cell niches in 3D with the highest biological conformity and resolution can enable new insights into these cell‐ECM interactions including proliferation, differentiation, migration, and invasion assays. Here, two‐photon stereolithography is adopted to print up to mm‐sized high‐precision 3D cell scaffolds at micrometer resolution with defined mechanical properties from protein‐based resins, such as bovine serum albumin or gelatin methacryloyl. By modifying the manufacturing process including two‐pass printing or post‐print crosslinking, high precision scaffolds with varying Young's moduli ranging from 7‐300 kPa are printed and quantified through atomic force microscopy. The impact of varying scaffold topographies on the dynamics of colonizing cells is observed using mouse myoblast cells and a 3D‐lung microtissue replica colonized with primary human lung fibroblast. This approach will allow for a systematic investigation of single‐cell and tissue dynamics in response to defined mechanical and bio‐molecular cues and is ultimately scalable to full organs.Item Open Access Untersuchung der Antriebsstrangdynamik in Windenergieanlagen(2020) Horch, JoachimDiese Arbeit beschäftigt sich damit die Stabilität und Funktionstüchtigkeit des Antriebsstranges einer Windenergieanlage der Größenordnung 10 MW zu untersuchen. Hierfür erfolgt der Aufbau eines Computermodells einer 10-MW-Windenergieanlage mithilfe des Mehrkörpersimulationsprogrammes SIMPACK. Weiterhin wird eine Parameterstudie durchgeführt, welche über eine Matlab-induzierte SIMPACK-Simulation speziell ausgewählte Parameter des Antriebsstranges variiert, Simulationen durchführt und so den Einfluss bestimmter Parameter, sowie Parameterkombinationen, auf die Stabilität des Antriebsstranges prüft. Auf diese Weise sollen Stabilitätskriterien für einen Antriebsstrang dieser Größenordnung ermittelt werden. Es erfolgen sowohl statische, als auch dynamische Untersuchungen.Item Open Access Construction of robust Escherichia coli strains for large-scale production(2022) Ziegler, Martin; Takors, Ralf (Prof. Dr.-Ing.)The biotechnical production of many fine chemicals, proteins or pharmaceuticals depends on large-scale microbial cultivations. Due to limited mixing, heterogeneities in process relevant parameters such as nutrient concentrations arise in such fermentations. Escherichia coli (E. coli) is a model organism frequently used in the biotechnological industry. If E. coli is cultivated under heterogeneous conditions, biological reactions of the microorganism result in reduced process performance. Since large-scale fermentations are not economically feasible in academic settings, scale-down reactors that mimic aforementioned heterogeneities are used to investigate heterogenous fermentations. Previous studies in scale-down reactors unraveled that, depending on the process strategy, the unstable supply of a limiting primary carbon or nitrogen source such as glucose or ammonium is one of the underlying causes of process performance loss. Low concentrations of glucose or ammonium elicit the stringent response as a biological starvation reaction which comprises extensive transcriptional reactions. In the first project that contributes to this thesis, the regulatory and transcriptional reactions of the strains E. coli MG1655 and E. coli SR to repeated exposure to ammonium starvation zones were examined in a scale-down reactor. The scale-down reactor followed a two-compartment approach and consisted of a stirred tank reactor and a plug-flow reactor simulating passage through a starvation zone. E. coli SR is a strain with modulated stringent response. It was observed that short-term starvation stimuli do not trigger this regulatory program in E. coli SR and the transcriptional reaction was noticeably reduced. Long-term adaptation of the strain to repeated cycles of limitation and starvation also clearly differed from E. coli MG1655. Despite lack of the stringent response, E. coli SR showed no deficits in the assimilation of the limiting ammonium or in biomass yield on ammonium. In the second project of this thesis, a series of deletion strains with robust phenotype against glucose starvation zones were constructed. Candidate genes were identified and successively removed from the genome of E. coli MG1655 by Recombineering. The fundamental growth parameters of the strains were determined in shaking flask fermentations and no noticeable differences compared to E. coli MG1655 were found. Chemostat cultivations in a scale-down reactor with glucose as the limiting nutrient source revealed that the final strain of the deletion series, E. coli RM214, had a significantly lower maintenance coefficient under heterogeneous conditions than E. coli MG1655. Moreover, in an exemplary heterologous protein productionscenario E. coli RM214 rhaB- pJOE4056.2_tetA proved to be more robust to heterogeneities and showed a significantly higher product yield than E. coli MG1655 rhaB- pJOE4056.2_tetA. In the third project of this thesis, the production of pyruvate in E. coli MG1655 by inhibition of pyruvate dehydrogenase through CRISPR interference was investigated. A central goal was to achieve the stable production in nitrogen-limited conditions. For this, different target sequences in the operon pdhR-aceEF-lpd were tested and the strains cultivated in shaking flask fermentations. All tested target sequences were generally suitable to trigger the accumulation of pyruvate. Combined CRISPR interference against two target sequences did not lead to an increased pyruvate yield in most cases. In addition, the strains E. coli MG1655 pdCas9 psgRNA_aceE_234 and E. coli MG1655 pdCas9 psgRNA_aceE_234_pdhR_329 were characterized in two phase fermentations in lab-scale reactors. The initial phase was an unlimited exponential growth phase and was followed by an ammonium-limited production phase. E. coli MG1655 pdCas9 psgRNA_aceE_234 only produced pyruvate during the exponential phase, and reuptake of pyruvate occurred in the second phase. In contrast, E. coli MG1655 pdCas9 psgRNA_aceE_234_pdhR_329 stably produced pyruvate during the exponential and the ammonium-limited phase and is a potential chassis strain for the growth-decoupled production of pyruvate derived bioproducts. The overarching research issues of the projects were the characterization of strains in heterogeneous conditions and the development of new strategies to improve their performance. The collected data leads me to conclude that the construction of robust microbial strains for large-scale applications is both expedient and feasible. Tailored genetic modifications are the method of choice to achieve this goal. Furthermore, suitable genetic constructs offer promising possibilities for the stable growth-decoupled production of chemicals in nitrogen-limited conditions.Item Open Access Messung und Modellierung der effektiven Wärmeleitfähigkeit von Dämmstoffschüttungen für vakuumwärmegedämmte Warmwasserspeicher(2022) Lang, Stephan; Spindler, Klaus (apl. Prof. Dr.-Ing.)Das Ziel dieser Arbeit ist es, hinsichtlich Dämmwirkung und Kosten geeignete schüttfähige Wärmedämmstoffe für doppelwandige Warmwasserspeicher mit Vakuumwärmedämmung zu identifizieren und eine Vorhersage über die effektive Wärmeleitfähigkeit in Abhängigkeit von Luftdruck und Temperatur treffen zu können. Zu diesem Zweck werden expandierte Perlite unterschiedlicher mittlerer Korngrößen und Mischungen aus expandierten Perliten und pyrogener Kieselsäure untersucht. Schwerpunkt der Untersuchungen ist die Ermittlung der effektiven Wärmeleitfähigkeit bei unterschiedlichen Luftdrücken und Temperaturen. Die effektive Wärmeleitfähigkeit wird in einer eigens entwickelten Versuchsanlage, nach einem stationären Messprinzip, bei Luftdrücken zwischen 0,001 mbar und Atmosphärendruck von (960 ± 20) mbar sowie bei Probenmitteltemperaturen zwischen -5 °C und 90 °C bestimmt. Die maximale relative Messunsicherheit der Versuchsanlage beträgt < 8 % bei den geringsten und < 3 % bei den höchsten gemessenen effektiven Wärmeleitfähigkeiten. Reine feinkörnige expandierte Perlite mit Dichten der Schüttungen ≥ 182 kg/m³ erreichen bei Luftdrücken ≤ 0,1 mbar und allen gemessenen Probenmitteltemperaturen die geringsten effektiven Wärmeleitfähigkeiten. Bei einer Probenmitteltemperatur von 48 °C liegen diese bei ≤ 4,84 mW/(m·K). Mischungen aus einem vergleichsweise grobkörnigen expandierten Perlit mit einer sehr feinkörnigen und feinporigen pyrogenen Kieselsäure können hingegen, bei technisch einfacher zu handhabenden Luftdrücken von > 1 mbar, z. T. geringere effektive Wärmeleitfähigkeiten erreichen als reine expandierte Perlite. Mischungen dieser Komponenten werden in dieser Arbeit erstmals in Form einer losen Schüttung bzgl. ihrer effektiven Wärmeleitfähigkeit untersucht. Bei einer Probenmitteltemperatur von 48 °C werden in diesem Luftdruckbereich effektive Wärmeleitfähigkeiten dieser Mischungen von > 12,17 mW/(m·K) gemessen. Anhand der gemessenen effektiven Wärmeleitfähigkeiten sowie weiterer Stoffeigenschaften, werden vollständig prädiktive analytische Modelle der effektiven Wärmeleitfähigkeit entwickelt. Das Modell für Mischungen gilt für ein ausgewähltes Stoffpaar, während das Modell für expandierte Perlite für beliebige ungemahlene expandierte Perlite verwendbar ist. Es handelt sich nach Kenntnis des Autors um das erste vollständig prädiktive analytische Modell der effektiven Wärmeleitfähigkeit für expandierte Perlite, welches auch die Kopplung von Gas- und Festkörperwärmeleitung berücksichtigt und für welches lediglich drei einfach und kostengünstig zu messende Größen bestimmt werden müssen. Diese Größen sind der volumengewichtet gemittelte Korndurchmesser, die mittlere Korndichte sowie die Dichte der Schüttung des expandierten Perlits. Aus den Messwerten der effektiven Wärmeleitfähigkeit wird ein Zusammenhang von volumengewichtet gemitteltem Korndurchmesser zur Kopplung von Gas- und Festkörperwärmeleitung deutlich, der für das Modell für expandierte Perlite verwendet wird. Mit den Modellen ist es nun möglich, ohne entsprechende Messungen, die effektive Wärmeleitfähigkeit von Mischungen aus einem expandierten Perlit und einer pyrogenen Kieselsäure sowie für beliebige ungemahlene expandierte Perlite, mit zufriedenstellender bis hoher Genauigkeit vorherzusagen.Item Open Access Development of novel bispecific antibodies for cancer therapy targeting the receptor tyrosine kinases HER4 and EGFR(2024) Kühl, Lennart; Kontermann, Roland E. (Prof. Dr.)In this study, novel mono- and bispecific antibodies targeting the ErbB receptor family members EGFR and HER4 were investigated. Dual targeting of EGFR and HER4 by a bispecific, tetravalent antibody comprising a novel, antagonistic HER4-targeting antibody showed inhibition of proliferation and migration for a HB-EGF-stimulated ovarian cancer cell line. No inhibitory effects in a breast cancer cell line expressing EGFR and HER4 indicated that successful dual targeting does not solely rely on target expression. The complexity of HER4 with its isoforms and their different signaling properties makes HER4 a challenging cancer target that needs further in-depth research. To overcome resistances based on escape mutations located in the epitopes of clinically approved antibodies, novel antagonistic EGFR-targeting antibodies binding to a different epitope were developed. This epitope was mapped to domain III of EGFR and binding to clinically relevant EGFR ectodomain mutations resulted in inhibition of EGFR signaling in stable cell lines used as test systems. Favorable activities in comparison to clinically approved antibodies regarding inhibition of EGFR signaling and proliferation were observed for cancer cell lines expressing the EGFR wildtype. Bispecific T-cell engagers can lead to a T-cell mediated target cell killing independent of intracellular downstream signaling in the cancer cell. One challenge for the applicability of T-cell engagers in solid tumors is to keep the balance between T-cell mediated tumor cell killing and severe side-effects caused by a systemic activation of the immune system. Studies on eleven different eIg-based formats for EGFR-binding T-cell engagers showed that valency, geometry, and size influenced their activity profile. Furthermore, one bivalent and one trivalent, bispecific format were investigated for two novel EGFR-targeting moieties. As these molecules bind to clinically relevant escape mutations located in the ectodomain of EGFR, they are expected to show activity in patients with an acquired resistance to approved EGFR-targeting antibodies. These molecules led to a robust T-cell mediated cytotoxicity of cancer cells expressing EGFR. Additionally, benefits regarding an EGFR-level dependent cytotoxicity were observed for reduced binding to EGFR. An initial in vivo study using surrogate molecules in a syngeneic mouse model showed reduction of tumor growth and prolonged survival for treatment with a trivalent, bispecific T-cell engager comprising a novel EGFR-binding moiety. Taken together, beneficial effects of the novel molecules may contribute to improved therapies for patients with both pre-existing and acquired resistances to EGFR-targeting antibodies.Item Open Access Insights into the structural and functional properties of the eukaryotic porin Tom40(2012) Gessmann, Dennis; Nußberger, Stephan (Prof. Dr.)Tom40 forms the preprotein conducting channel in the outer membrane of mitochondria enabling transport of up to 1500 different preproteins through an optimized pore environment. Moreover, Tom40 exhibits a voltage-dependent gating mechanism in terms of an ‘electrical switch’ making this eukaryotic beta-barrel a promising target for nanopore based applications. In this work, new bioinformatics methods were developed and verified through practical approaches to shed light on the structural elements of Tom40 facilitating its particular function in mitochondria. Based on these results, Tom40 proteins were designed with modified and optimized structural properties. TmSIP, a physical interaction model developed for TM beta-barrel proteins, was used to identify weakly stable regions in the TM domain of Tom40 from mammals and fungi. Three unfavorable beta-strands were determined for human Tom40A. Via CD and Trp-fluorescence spectroscopy it was shown that substitution of key amino acid residues in theses strands resulted in an improved resistance of the protein to chemical and thermal perturbations. Further, the mutated form of hTom40A was strictly found in its monomeric state. Equal improvements were gained for the apparent stability of Tom40 from Aspergillus fumigatus. Tom40 was isolated and purified in its native state from Neurospora crassa mitochondria. Time-limited proteolysis of native NcTom40 coupled to mass spectrometry revealed comparable protease-accessibility to VDAC isoform 1 from mammals suggesting a similar fold. Thus, a homology model of NcTom40 was developed on the basis of the solved mouse VDAC-1 crystal structure. It was found that Tom40 forms a 19-stranded beta-barrel with an N-terminal alpha-helix inside the pore. Further, a conserved ‘polar slide’ in the pore interior is possibly involved in preprotein translocation and a second conserved domain, termed ‘helix anchor region’, in arresting the helix inside the Tom40 pore. Based on the homology model of NcTom40, the structure and function of the N-terminal domain of Tom40 was addressed. Examination of the model structure revealed two different domains for the N-terminus, the inner-barrel and outer-barrel N-terminus. In vivo investigations showed that both parts prevent a heat-induced dysfunction of Tom40 in N. crassa mitochondria independently. By applying CD spectroscopy the predicted N-terminal alpha-helix could be allocated to the inner-barrel N-terminus. Further, in combination with Trp-fluorescence spectroscopy it was found that the N-terminal alpha-helix unfolds independently from the Tom40 beta-barrel, but is not necessary for pore stability or integrity. However, a conserved amino acid residue, Ile47 of NcTom40, in the inner-barrel N-terminus is essential for the structural integrity of the N-terminal alpha-helix. In conclusion, these results may offer a basis for future works on TM beta-barrel proteins with the aim to alter the structural properties in the absence of a high atomic resolution structure or an established knowledge of the biochemical and biophysical properties.Item Open Access Wege zur Ermittlung von Energieeffizienzpotenzialen von Informations- und Kommunikationstechnologien(Stuttgart : Universität Stuttgart, Institut für Energiewirtschaft und Rationelle Energieanwendung, 2020) Miller, Michael; Hufendiek, Kai (Prof. Dr.-Ing.)Item Open Access Enhanced processing of regrind as recycling material in single-screw extruders(2021) Thieleke, Philipp; Bonten, ChristianRegrind processing poses challenges for single-screw extruders due to the irregularly shaped particles. For grooved feed zones, the output is lessened by the reduction of bulk density in comparison to virgin material. Simultaneously, the melt temperature increases, reducing the extruder’s process window. Through experimental investigations on a test stand, a novel feed zone geometry (nominal diameter 35 mm) is developed. It aligns the regrind’s specific throughput with that of virgin material. The regrind processing window is essentially increased. As the solids conveying in the novel feed zone cannot be simulated with existing methods, numerical simulations using the discrete element method are performed. Since plastic deformation occurs in the novel feed zone geometry, a new hysteresis contact model is developed. In addition to spheres, the regrind and virgin particles are modeled as superquadrics to better approximate the irregular shape. The new contact model’s simulation results show excellent agreement with experimental compression tests. The throughput of the extruder simulations is considerably underestimated when using spheres to represent the real particles than when using irregularly shaped superquadrics. Corresponding advantages can be seen especially for virgin material.Item Open Access Global potentials and costs of synfuels via Fischer-Tropsch process(2023) Buchenberg, Patrick; Addanki, Thushara; Franzmann, David; Winkler, Christoph; Lippkau, Felix; Hamacher, Thomas; Kuhn, Philipp; Heinrichs, Heidi; Blesl, MarkusThis paper presents the potentials and costs of synthetic fuels (synfuels) produced by renewable energy via PEM water electrolysis and the subsequent Fischer-Tropsch process for the years 2020, 2030, 2040, and 2050 in selected countries across the globe. The renewable energy potential was determined by the open-source tool pyGRETA and includes photovoltaic, onshore wind, and biomass. Carbon dioxide is obtained from biomass and the atmosphere by direct air capture. The potentials and costs were determined by aggregating minimal cost energy systems for each location on a state level. Each linear energy system was modelled and optimised by the optimisation framework urbs. The analysis focused on decentralised and off-grid synthetic fuels’ production. The transportation costs were roughly estimated based on the distance to the nearest maritime port for export. The distribution infrastructure was not considered since the already-existing infrastructure for fossil fuels can be easily adopted. The results showed that large amounts of synthetic fuels are available for EUR 110/MWh (USD 203/bbl) mainly in Africa, Central and South America, as well as Australia for 2050. This corresponds to a cost reduction of more than half compared to EUR 250/MWh (USD 461/bbl) in 2020. The synfuels’ potentials follow the photovoltaic potentials because of the corresponding low levelised cost of electricity. Batteries are in particular used for photovoltaic-dominant locations, and transportation costs are low compared to production costs.