7 research outputs found
A dynamic method for the investigation of induced state metabolic capacities as a function of temperature
Kontrollumfang und Kontrolldichte bei Verbandsklagen im Umweltrecht:Rechtsvergleichende Studie unter Berücksichtigung des britischen und französischen Rechts
Bedeutung und Reichweite von verwaltungsgerichtlichen Umweltverbandsklagen gegen die Zulassung von Vorhaben mit Umweltauswirkungen und andere umweltbezogene behördliche Entscheidungen haben in Deutschland seit der Jahrtausendwende zugenommen. Maßgeblich dafür waren die Schaffung des Umwelt-Rechtsbehelfsgesetzes sowie der Einfluss der Aarhus-Konvention, unionsrechtlicher Rechtsakte und der Rechtsprechung des Europäischen Gerichtshofs. Der Autor untersucht die Bedeutung dieser rechtlichen Rahmenbedingungen für die verwaltungsgerichtliche Begründetheitsprüfung, insbesondere für die Kontrolldichte und somit für die Letztentscheidungsspielräume der Exekutive. Zu diesem Zweck werden Dogmatik und Praxis der gerichtlichen Begründetheitskontrolle in Deutschland, England und Frankreich analysiert. Dabei nimmt der Autor neben der Kontrolldichte auch die gerichtliche Sanktionierung von Verfahrensfehlern der Exekutive in den Blick. Die Arbeit schließt mit einer kritischen Analyse von Tendenzen und Vorschlägen zur Rücknahme der Kontrolldichte und Überlegungen für eine ausgewogene Reform des Rechtsschutzes der Umweltvereinigungen
An Integrated Downstream Process Development Strategy along QbD Principles
The development, optimization, and analysis of downstream processes are challenged by a high number of potentially critical process parameters that need to be investigated using lab-scale experiments. These process parameters are spread across multiple unit operations and potentially show interactions across unit operations. In this contribution, we present a novel strategy for bioprocess development that considers the risk of parameter interactions across unit operations for efficient experimental design. A novel risk assessment tool (interaction matrix) is introduced to the Quality by Design (QbD) workflow. Using this tool, the risk of interaction across unit operations is rated. Subsequently, a design of experiments (DoE) across unit operations is conducted that has the power to reveal multivariate interdependencies. The power of the presented strategy is demonstrated for protein isolation steps of an inclusion body process, focusing on the quality attribute inclusion body purity. The concentration of Triton X-100 in the course of inclusion body (IB) purification was shown to interact with the g-number of the subsequent centrifugation step. The presented strategy targets a holistic view on the process and allows handling of a high number of experimental parameters across unit operations using minimal experimental effort. It is generically applicable for process development along QbD principles
Fed-Batch Production of Bacterial Ghosts Using Dielectric Spectroscopy for Dynamic Process Control
The Bacterial Ghost (BG) platform technology evolved from a microbiological expression system incorporating the ϕX174 lysis gene E. E-lysis generates empty but structurally intact cell envelopes (BGs) from Gram-negative bacteria which have been suggested as candidate vaccines, immunotherapeutic agents or drug delivery vehicles. E-lysis is a highly dynamic and complex biological process that puts exceptional demands towards process understanding and control. The development of a both economic and robust fed-batch production process for BGs required a toolset capable of dealing with rapidly changing concentrations of viable biomass during the E-lysis phase. This challenge was addressed using a transfer function combining dielectric spectroscopy and soft-sensor based biomass estimation for monitoring the rapid decline of viable biomass during the E-lysis phase. The transfer function was implemented to a feed-controller, which followed the permittivity signal closely and was capable of maintaining a constant specific substrate uptake rate during lysis phase. With the described toolset, we were able to increase the yield of BG production processes by a factor of 8–10 when compared to currently used batch procedures reaching lysis efficiencies >98%. This provides elevated potentials for commercial application of the Bacterial Ghost platform technology
A novel method to recover inclusion body protein from recombinant E. coli fed-batch processes based on phage ΦX174-derived lysis protein E
Production of recombinant proteins as inclusion bodies is an important strategy in the production of technical enzymes and biopharmaceutical products. So far, protein from inclusion bodies has been recovered from the cell factory through mechanical or chemical disruption methods, requiring additional cost-intensive unit operations. We describe a novel method that is using a bacteriophage-derived lysis protein to directly recover inclusion body protein from Escherichia coli from high cell density fermentation process: The recombinant inclusion body product is expressed by using a mixed feed fed-batch process which allows expression tuning via adjusting the specific uptake rate of the inducing substrate. Then, bacteriophage ΦX174-derived lysis protein E is expressed to induce cell lysis. Inclusion bodies in empty cell envelopes are harvested via centrifugation of the fermentation broth. A subsequent solubilization step reveals the recombinant protein. The process was investigated by analyzing the impact of fermentation conditions on protein E-mediated cell lysis as well as cell lysis kinetics. Optimal cell lysis efficiencies of 99% were obtained with inclusion body titers of >2.0 g/l at specific growth rates higher 0.12 h−1 and inducer uptake rates below 0.125 g/(g × h). Protein E-mediated cell disruption showed a first-order kinetics with a kinetic constant of −0.8 ± 0.3 h−1. This alternative inclusion body protein isolation technique was compared to the one via high-pressure homogenization. SDS gel analysis showed 10% less protein impurities when cells had been disrupted via high-pressure homogenization, than when empty cell envelopes including inclusion bodies were investigated. Within this contribution, an innovative technology, tuning recombinant protein production and substituting cost-intensive mechanical cell disruption, is presented. We anticipate that the presented method will simplify and reduce the production costs of inclusion body processes to produce technical enzymes and biopharmaceutical products.Morphoplant GmbH, Bochum (RCPE)560356141
Ausschreibungen für erneuerbare Energien. Wissenschaftliche Empfehlungen: Studie im Auftrag des Bundesministeriums für Wirtschaft und Energie. Projektnummer: DESDE15240. 07. Juli 2015
Eine Grundlage für die Eckpunkte zur Ausschreibung für die Förderung erneuerbarer Energien sind die Zwischenergebnisse einer wissenschaftlichen Studie zum Ausschreibungsdesign erneuerbarer Energien. Der Bericht analysiert die Marktsituation der verschiedenen erneuerbaren Energien und spricht Empfehlungen für ein Ausschreibungsdesign aus. Er wurde von einem Forschungskonsortium, das sich aus Ecofys, Fraunhofer ISI, Consentec, dem Zentrum für Sonnenenergie- und Wasserstoff-Forschung Baden-Württemberg (ZSW), Takon und den Rechtsanwaltskanzleien Görg sowie BBG und Partner zusammensetzt, erarbeitet