Multienzymnetzwerke für die Feinchemie

Zusammenfassung: Die Produktion von Feinchemikalien beruht auf Prozessen mit mehreren Reaktionsschritten. Wir arbeiten an Konzepten für die Rekrutierung, Isolierung und Optimierung von in vitro-Enzymkaskaden aus Zellextrakten für die Synthese von komplexen Zucker

Klang – Farbe – Geschlecht – Sexualität : Diskursive Metaphorik nationaler Identität / Alterität in der Rezeptionsgeschichte der musikalischen Moderne am Beispiel des Komponisten Franz Schreker

Der österreichische Komponist Franz Schreker (1878 – 1934) zählte in der Weimarer Republik für kurze Zeit zu den erfolgreichsten Opernkomponisten deutscher Sprache. Bei genauerer Betrachtung der zeitgenössischen Aufführungs- und Kompositionskritiken über Schrekers Opernwerk zeigt sich, dass viele Kommentatoren den Komponisten auf der symbolhaften Ebene über nationsspezifische Geschlechterzuschreibungen zu einem ›undeutschen‹ bzw. ›effeminierten‹ Komponisten konstruierten. Schreker galt somit als Verkörperung eines inneren Anderen der deutschen Nation, der nicht nur ›feminisiert‹, sondern mit anderen Attributen der Devianz (Sexsismen, Rassismen, Pathologien) marginalisiert werden sollte. Anknüpfend an postmoderne Identitäts- und Nationalismustheorien untersucht diese Arbeit anhand eines diskursanalytischen Verfahrens am Beispiel der Schreker-Rezeption, welche Rolle die Kategorie Geschlecht auf dem Gebiet der Musik für die Stiftung der deutschen Nation im 19. und 20. Jahrhunderts gespielt hat. Sie leistet einen wichtigen Beitrag zum Verständnis historischer Konstruktionsprozesse nationaler Identität bzw. Alterität sowie den damit verbundenen diskursiven Imaginationen von ›Männlichkeit‹ versus ›Weiblichkeit‹ im deutschen Musikdiskurs. Dabei geht die Untersuchung zugleich auf (Dis-)Kontinuitäten dieser Rezeptionsgeschichte nach 1945 sowohl in der BRD als auch in DDR ein. Schließlich belegt die Arbeit, auf welche Weise Schreker selbst in seinem Spätwerk, namentlich im Christophorus, künstlerisch auf die ihm zugeschriebenen Bilder eines ›effeminierten‹ Komponisten reagiert, diese in die Vorstellung devianter ›Männlichkeit‹ positiv umdeutet und in seine eigenes konstitutives Selbstbild integriert. Somit lässt sich Schrekers später ästhetischer Stil als eine Form der Selbstermächtigung verstehen, mit welcher der Komponist auf der Ebene der Kunst eine subversive Gegenposition zu dem normativen Identitätsbegriff des Deutschen in der Musik entwickelt.In the early 20th century, the Austrian composer Franz Schreker (1878-1934) ranked among the most renowned opera composers in German-speaking countries. Upon closer consideration however, contemporary reviews of his works and of their performances illustrate how numerous critics constructed Schreker, on a symbolic level, as a “Non-German” and “effeminate” composer through attributions of gender and national bias. Thus Schreker – who, in the eyes of his critics, epitomized an inner “other” of the German nation – has seen himself not only effiminated but also marginalized through attributions of deviance (sexisms, racisms, pathologies) imposed on him. Building on postmodern theories of nation and identity while using the example of the reception of Schreker, the present study examines by means of discourse analysis the role of gender in 19th- and 20th-century music in light of the founding of a single German nation. Designed as a scientific contribution to understanding historical construction processes of national identity and alterity as well as discursive imaginations of “masculinity” and “femininity” related to them, it is not limited to critical reviews during the composer’s lifetime but does encompass the analysis of subsequent (dis-)continuities in post-war reception history both in the Federal Republic of Germany and in the German Democratic Republic. Essentially taking recourse to Christophorus, the study furthermore demonstrates how Schreker himself did not simply react to these attributions but positively redefined such imaginations of deviant masculinity and integrated them in his later work in an astoundingly creative way. The present thesis concludes with the finding that Schrekers later aesthetic style can be meticulously construed as a pattern of self-empowerment enabling the composer to develop a subversive counter-position to a normative notion of identity and Germanness in 20th-century music

Cross-Regulation of an Artificial Metalloenzyme

Cross-regulation of complex biochemical reaction networks is an essential feature of living systems. In a biomimetic spirit, we report on our efforts to program the temporal activation of an artificial metalloenzyme via cross-regulation by a natural enzyme. In the presence of urea, urease slowly releases ammonia that reversibly inhibits an artificial transfer hydrogenase. Addition of an acid, which acts as fuel, allows to maintain the system out of equilibrium

Intrinsic Thermal Sensing Controls Proteolysis of Yersinia Virulence Regulator RovA

Pathogens, which alternate between environmental reservoirs and a mammalian host, frequently use thermal sensing devices to adjust virulence gene expression. Here, we identify the Yersinia virulence regulator RovA as a protein thermometer. Thermal shifts encountered upon host entry lead to a reversible conformational change of the autoactivator, which reduces its DNA-binding functions and renders it more susceptible for proteolysis. Cooperative binding of RovA to its target promoters is significantly reduced at 37°C, indicating that temperature control of rovA transcription is primarily based on the autoregulatory loop. Thermally induced reduction of DNA-binding is accompanied by an enhanced degradation of RovA, primarily by the Lon protease. This process is also subject to growth phase control. Studies with modified/chimeric RovA proteins indicate that amino acid residues in the vicinity of the central DNA-binding domain are important for proteolytic susceptibility. Our results establish RovA as an intrinsic temperature-sensing protein in which thermally induced conformational changes interfere with DNA-binding capacity, and secondarily render RovA susceptible to proteolytic degradation

Visualization and Curve-Parameter Estimation Strategies for Efficient Exploration of Phenotype Microarray Kinetics

The Phenotype MicroArray (OmniLog® PM) system is able to simultaneously capture a large number of phenotypes by recording an organism's respiration over time on distinct substrates. This technique targets the object of natural selection itself, the phenotype, whereas previously addressed '-omics' techniques merely study components that finally contribute to it. The recording of respiration over time, however, adds a longitudinal dimension to the data. To optimally exploit this information, it must be extracted from the shapes of the recorded curves and displayed in analogy to conventional growth curves.The free software environment R was explored for both visualizing and fitting of PM respiration curves. Approaches using either a model fit (and commonly applied growth models) or a smoothing spline were evaluated. Their reliability in inferring curve parameters and confidence intervals was compared to the native OmniLog® PM analysis software. We consider the post-processing of the estimated parameters, the optimal classification of curve shapes and the detection of significant differences between them, as well as practically relevant questions such as detecting the impact of cultivation times and the minimum required number of experimental repeats.We provide a comprehensive framework for data visualization and parameter estimation according to user choices. A flexible graphical representation strategy for displaying the results is proposed, including 95% confidence intervals for the estimated parameters. The spline approach is less prone to irregular curve shapes than fitting any of the considered models or using the native PM software for calculating both point estimates and confidence intervals. These can serve as a starting point for the automated post-processing of PM data, providing much more information than the strict dichotomization into positive and negative reactions. Our results form the basis for a freely available R package for the analysis of PM data

Bottom-up construction of complex biomolecular systems with cell-free synthetic biology

Cell-free systems offer a promising approach to engineer biology since their open nature allows for well-controlled and characterized reaction conditions. In this review, we discuss the history and recent developments in engineering recombinant and crude extract systems, as well as breakthroughs in enabling technologies, that have facilitated increased throughput, compartmentalization, and spatial control of cell-free protein synthesis reactions. Combined with a deeper understanding of the cell-free systems themselves, these advances improve our ability to address a range of scientific questions. By mastering control of the cell-free platform, we will be in a position to construct increasingly complex biomolecular systems, and approach natural biological complexity in a bottom-up manner

Cell-free synthetic biology for in vitro prototype engineering

Cell-free transcription-translation is an expanding field in synthetic biology as a rapid prototyping platform for blueprinting the design of synthetic biological devices. Exemplar efforts include translation of prototype designs into medical test-kits for onsite identification of viruses (Zika, Ebola), whilst gene circuit cascades can be tested, debugged and re-designed within rapid turnover times. Coupled with mathematical modelling, this discipline lends itself towards the precision engineering of new synthetic life. The next stages of cell-free look set to unlock new microbial hosts that remain slow to engineer and unsuited to rapid iterative design cycles. It is hoped that the development of such systems will provide new tools to aid the transition from cell-free prototype designs to functioning synthetic genetic circuits and engineered natural product pathways in living cells