TY - JOUR A1 - Haeger, Gerrit A1 - Wirges, Jessika A1 - Tanzmann, Nicole A1 - Oyen, Sven A1 - Jolmes, Tristan A1 - Jaeger, Karl-Erich A1 - Schörken, Ulrich A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Chaperone assisted recombinant expression of a mycobacterial aminoacylase in Vibrio natriegens and Escherichia coli capable of N-lauroyl-L-amino acid synthesis JF - Microbial Cell Factories N2 - Background Aminoacylases are highly promising enzymes for the green synthesis of acyl-amino acids, potentially replacing the environmentally harmful Schotten-Baumann reaction. Long-chain acyl-amino acids can serve as strong surfactants and emulsifiers, with application in cosmetic industries. Heterologous expression of these enzymes, however, is often hampered, limiting their use in industrial processes. Results We identified a novel mycobacterial aminoacylase gene from Mycolicibacterium smegmatis MKD 8, cloned and expressed it in Escherichia coli and Vibrio natriegens using the T7 overexpression system. The recombinant enzyme was prone to aggregate as inclusion bodies, and while V. natriegens Vmax™ could produce soluble aminoacylase upon induction with isopropyl β-d-1-thiogalactopyranoside (IPTG), E. coli BL21 (DE3) needed autoinduction with lactose to produce soluble recombinant protein. We successfully conducted a chaperone co-expression study in both organisms to further enhance aminoacylase production and found that overexpression of chaperones GroEL/S enhanced aminoacylase activity in the cell-free extract 1.8-fold in V. natriegens and E. coli. Eventually, E. coli ArcticExpress™ (DE3), which co-expresses cold-adapted chaperonins Cpn60/10 from Oleispira antarctica, cultivated at 12 °C, rendered the most suitable expression system for this aminoacylase and exhibited twice the aminoacylase activity in the cell-free extract compared to E. coli BL21 (DE3) with GroEL/S co-expression at 20 °C. The purified aminoacylase was characterized based on hydrolytic activities, being most stable and active at pH 7.0, with a maximum activity at 70 °C, and stability at 40 °C and pH 7.0 for 5 days. The aminoacylase strongly prefers short-chain acyl-amino acids with smaller, hydrophobic amino acid residues. Several long-chain amino acids were fairly accepted in hydrolysis as well, especially N-lauroyl-L-methionine. To initially evaluate the relevance of this aminoacylase for the synthesis of N-acyl-amino acids, we demonstrated that lauroyl-methionine can be synthesized from lauric acid and methionine in an aqueous system. Conclusion Our results suggest that the recombinant enzyme is well suited for synthesis reactions and will thus be further investigated. KW - Acyl-amino acids KW - Inclusion bodies KW - Chaperone co-expression KW - Vibrio natriegens KW - Aminoacylase Y1 - 2023 U6 - https://doi.org/10.1186/s12934-023-02079-1 SN - 1475-2859 N1 - Corresponding author: Petra Siegert IS - 22 SP - Article number: 77 (2023) PB - Springer Nature ER - TY - JOUR A1 - Haeger, Gerrit A1 - Probst, Johanna A1 - Jaeger, Karl-Erich A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Novel aminoacylases from Streptomyces griseus DSM 40236 and their recombinant production in Streptomyces lividans JF - FEBS Open Bio N2 - Amino acid-based surfactants are valuable compounds for cosmetic formulations. The chemical synthesis of acyl-amino acids is conventionally performed by the Schotten-Baumann reaction using fatty acyl chlorides, but aminoacylases have also been investigated for use in biocatalytic synthesis with free fatty acids. Aminoacylases and their properties are diverse; they belong to different peptidase families and show differences in substrate specificity and biocatalytic potential. Bacterial aminoacylases capable of synthesis have been isolated from Burkholderia, Mycolicibacterium, and Streptomyces. Although several proteases and peptidases from S. griseus have been described, no aminoacylases from this species have been identified yet. In this study, we investigated two novel enzymes produced by S. griseus DSM 40236ᵀ . We identified and cloned the respective genes and recombinantly expressed an α-aminoacylase (EC 3.5.1.14), designated SgAA, and an ε-lysine acylase (EC 3.5.1.17), designated SgELA, in S. lividans TK23. The purified aminoacylase SgAA was biochemically characterized, focusing on its hydrolytic activity to determine temperature- and pH optima and stabilities. The aminoacylase could hydrolyze various acetyl-amino acids at the Nα -position with a broad specificity regarding the sidechain. Substrates with longer acyl chains, like lauroyl-amino acids, were hydrolyzed to a lesser extent. Purified aminoacylase SgELA specific for the hydrolysis of Nε -acetyl-L-lysine was unstable and lost its enzymatic activity upon storage for a longer period but could initially be characterized. The pH optimum of SgELA was pH 8.0. While synthesis of acyl-amino acids was not observed with SgELA, SgAA catalyzed the synthesis of lauroyl-methionine. KW - Streptomyces lividans KW - recombinant expression KW - Streptomyces griseus KW - ε-lysine acylase KW - α-aminoacylase Y1 - 2023 U6 - https://doi.org/10.1002/2211-5463.13723 SN - 2211-5463 N1 - Corresponding author: Petra Siegert VL - 13 IS - 12 SP - 2224 EP - 2238 PB - Wiley CY - Hoboken, NJ ER - TY - JOUR A1 - Haeger, Gerrit A1 - Jolmes, Tristan A1 - Oyen, Sven A1 - Jaeger, Karl-Erich A1 - Bongaerts, Johannes A1 - Schörken, Ulrich A1 - Siegert, Petra T1 - Novel recombinant aminoacylase from Paraburkholderia monticola capable of N-acyl-amino acid synthesis JF - Applied Microbiology and Biotechnology N2 - N-Acyl-amino acids can act as mild biobased surfactants, which are used, e.g., in baby shampoos. However, their chemical synthesis needs acyl chlorides and does not meet sustainability criteria. Thus, the identification of biocatalysts to develop greener synthesis routes is desirable. We describe a novel aminoacylase from Paraburkholderia monticola DSM 100849 (PmAcy) which was identified, cloned, and evaluated for its N-acyl-amino acid synthesis potential. Soluble protein was obtained by expression in lactose autoinduction medium and co-expression of molecular chaperones GroEL/S. Strep-tag affinity purification enriched the enzyme 16-fold and yielded 15 mg pure enzyme from 100 mL of culture. Biochemical characterization revealed that PmAcy possesses beneficial traits for industrial application like high temperature and pH-stability. A heat activation of PmAcy was observed upon incubation at temperatures up to 80 °C. Hydrolytic activity of PmAcy was detected with several N-acyl-amino acids as substrates and exhibited the highest conversion rate of 773 U/mg with N-lauroyl-L-alanine at 75 °C. The enzyme preferred long-chain acyl-amino-acids and displayed hardly any activity with acetyl-amino acids. PmAcy was also capable of N-acyl-amino acid synthesis with good conversion rates. The best synthesis results were obtained with the cationic L-amino acids L-arginine and L-lysine as well as with L-leucine and L-phenylalanine. Exemplarily, L-phenylalanine was acylated with fatty acids of chain lengths from C8 to C18 with conversion rates of up to 75%. N-lauroyl-L-phenylalanine was purified by precipitation, and the structure of the reaction product was verified by LC–MS and NMR. KW - Chaperone KW - Biocatalysis KW - Aminoacylase KW - Acylation KW - Acyl-amino acids KW - Biosurfactants Y1 - 2024 U6 - https://doi.org/10.1007/s00253-023-12868-8 SN - 1432-0614 N1 - Corresponding author: Petra Siegert IS - 108 PB - Springer CY - Berlin ER - TY - CHAP A1 - Göttsche, Joachim A1 - Korn, Michael A1 - Amato, Alexandre T1 - The Passivhaus concept for the Arabian Peninsula – An energetic-economical evaluation considering the thermal comfort T2 - QScience Proceedings: Vol 2015 N2 - The Passivhaus building standard is a concept developed for the realization of energy-efficient and economical buildings with a simultaneous high utilization comfort under European climate conditions. Major elements of the Passivhaus concept are a high thermal insulation of the external walls, the use of heat and/or solar shading glazing as well as an airtight building envelope in combination with energy-efficient technical building installations and heating or cooling generators, such as an efficient energy-recovery in the building air-conditioning. The objective of this research project is the inquiry to determine the parameters or constraints under which the Passivhaus concept can be implemented under the arid climate conditions in the Arabian Peninsula to achieve an energy-efficient and economical building with high utilization comfort. In cooperation between the Qatar Green Building Council (QGBC), Barwa Real Estate (BRE) and Kahramaa the first Passivhaus was constructed in Qatar and on the Arabian Peninsula in 2013. The Solar-Institut Jülich of Aachen University of Applied Science supports the Qatar Green Building Council with a dynamic building and equipment simulation of the Passivhaus and the neighbouring reference building. This includes simulation studies with different component configurations for the building envelope and different control strategies for heating or cooling systems as well as the air conditioning of buildings to find an energetic-economical optimum. Part of these analyses is the evaluation of the energy efficiency of the used energy recovery system in the Passivhaus air-conditioning and identification of possible energy-saving effects by the use of a bypass function integrated in the heat exchanger. In this way it is expected that on an annual basis the complete electricity demand of the building can be covered by the roof-integrated PV generator. Y1 - 2015 U6 - https://doi.org/10.5339/qproc.2015.qgbc.38 N1 - 10.5339/qproc.2015.qgbc.38 = abstract - Qatar Green Building Conference 2015 - The Vision, 38. ER - TY - JOUR A1 - Götten, Falk A1 - Finger, Felix A1 - Havermann, Marc A1 - Braun, Carsten A1 - Marino, M. A1 - Bil, C. T1 - Full configuration drag estimation of short-to-medium range fixed-wing UAVs and its impact on initial sizing optimization JF - CEAS Aeronautical Journal N2 - The paper presents the derivation of a new equivalent skin friction coefficient for estimating the parasitic drag of short-to-medium range fixed-wing unmanned aircraft. The new coefficient is derived from an aerodynamic analysis of ten different unmanned aircraft used for surveillance, reconnaissance, and search and rescue missions. The aircraft is simulated using a validated unsteady Reynolds-averaged Navier Stokes approach. The UAV’s parasitic drag is significantly influenced by the presence of miscellaneous components like fixed landing gears or electro-optical sensor turrets. These components are responsible for almost half of an unmanned aircraft’s total parasitic drag. The new equivalent skin friction coefficient accounts for these effects and is significantly higher compared to other aircraft categories. It is used to initially size an unmanned aircraft for a typical reconnaissance mission. The improved parasitic drag estimation yields a much heavier unmanned aircraft when compared to the sizing results using available drag data of manned aircraft. KW - Parasitic drag KW - UAV KW - CFD KW - Aircraft sizing Y1 - 2021 U6 - https://doi.org/10.1007/s13272-021-00522-w SN - 1869-5590 (Online) SN - 1869-5582 (Print) N1 - Corresponding author: Falk Götten VL - 12 SP - 589 EP - 603 PB - Springer CY - Berlin ER - TY - JOUR A1 - Grieger, Niklas A1 - Schwabedal, Justus T. C. A1 - Wendel, Stefanie A1 - Ritze, Yvonne A1 - Bialonski, Stephan T1 - Automated scoring of pre-REM sleep in mice with deep learning JF - Scientific Reports N2 - Reliable automation of the labor-intensive manual task of scoring animal sleep can facilitate the analysis of long-term sleep studies. In recent years, deep-learning-based systems, which learn optimal features from the data, increased scoring accuracies for the classical sleep stages of Wake, REM, and Non-REM. Meanwhile, it has been recognized that the statistics of transitional stages such as pre-REM, found between Non-REM and REM, may hold additional insight into the physiology of sleep and are now under vivid investigation. We propose a classification system based on a simple neural network architecture that scores the classical stages as well as pre-REM sleep in mice. When restricted to the classical stages, the optimized network showed state-of-the-art classification performance with an out-of-sample F1 score of 0.95 in male C57BL/6J mice. When unrestricted, the network showed lower F1 scores on pre-REM (0.5) compared to the classical stages. The result is comparable to previous attempts to score transitional stages in other species such as transition sleep in rats or N1 sleep in humans. Nevertheless, we observed that the sequence of predictions including pre-REM typically transitioned from Non-REM to REM reflecting sleep dynamics observed by human scorers. Our findings provide further evidence for the difficulty of scoring transitional sleep stages, likely because such stages of sleep are under-represented in typical data sets or show large inter-scorer variability. We further provide our source code and an online platform to run predictions with our trained network. Y1 - 2021 U6 - https://doi.org/10.1038/s41598-021-91286-0 SN - 2045-2322 N1 - Corresponding author: Stephan Bialonski VL - 11 IS - Art. 12245 PB - Springer Nature CY - London ER - TY - CHAP A1 - Gregorio, Fabrizio de A1 - Fatigati, Giovanni A1 - Kallweit, Stephan T1 - Tiltrotor airframe flow field characterization by SPIV T2 - 11th International Symposium on Partivle Image Velocimetry - PIV15 , Santa Barbara, California, Sept 14-16, 2015 Y1 - 2015 ER - TY - JOUR A1 - Grajewski, Matthias A1 - Kleefeld, Andreas T1 - Detecting and approximating decision boundaries in low-dimensional spaces JF - Numerical Algorithms N2 - A method for detecting and approximating fault lines or surfaces, respectively, or decision curves in two and three dimensions with guaranteed accuracy is presented. Reformulated as a classification problem, our method starts from a set of scattered points along with the corresponding classification algorithm to construct a representation of a decision curve by points with prescribed maximal distance to the true decision curve. Hereby, our algorithm ensures that the representing point set covers the decision curve in its entire extent and features local refinement based on the geometric properties of the decision curve. We demonstrate applications of our method to problems related to the detection of faults, to multi-criteria decision aid and, in combination with Kirsch’s factorization method, to solving an inverse acoustic scattering problem. In all applications we considered in this work, our method requires significantly less pointwise classifications than previously employed algorithms. KW - MCDA KW - Inverse scattering problem KW - Fault approximation KW - Fault detection Y1 - 2023 SN - 1572-9265 N1 - Corresponding author: Matthias Grajewski VL - 93 IS - 4 PB - Springer Science+Business Media CY - Dordrecht ER - TY - CHAP A1 - Gellert, Christoph A1 - Norda, Hannah A1 - Butenweg, Christoph T1 - Nonlinear behaviour of masonry under cyclic loading T2 - 7th European Conference on Structural Dynamics, EURODYN 2008 : 7-9 July 2008, Southampton Y1 - 2008 SP - 1 EP - 12 ER - TY - GEN A1 - Gebhardt, Andreas T1 - Short course on rapid prototyping N2 - Rapid Prototyping Technology: Types of models, rapid prototyping processes, prototyper Fundamentals of rapid prototyping Industrial rapid prototyping technology: Stereolithography, (Selective) laser sintering ((S)LS), Layer laminate manufacturing (LLM), Fused layer modeling (FLM), Three dimensional printing (3DP) KW - Rapid Prototyping KW - Rapid Prototyping Y1 - 2005 ER - TY - CHAP A1 - Gebhardt, Andreas T1 - Technology Diffusion through a Multi-Level Technology Transfer Infrastructure. Contribution to the 1st. All Africa Technology Diffusion Conference Boksburg, South Africa June 12th - 14th 2006 N2 - Table of contents 1. Introduction 2. Multi-level Technology Transfer Infrastructure 2.1 Level 1: University Education – Encourage the Idea of becoming an Entrepreneur 2.2 Level 2: Post Graduate Education – Improve your skills and focus it on a product family. 2.3 Level 3: Birth of a Company – Focus your skills on a product and a market segment. 2.4 Level 4: Ready to stand alone – Set up your own business 2.5 Level 5: Grow to be Strong – Develop your business 2.6 Level 6: Competitive and independent – Stay innovative. 3. Samples 3.1 Sample 1: Laser Processing and Consulting Centre, LBBZ 3.2 Sample 2: Prototyping Centre, CP 4. Funding - Waste money or even lost Money? 5. Conclusion KW - Technologietransfer KW - technology transfer KW - technology diffusion Y1 - 2006 ER - TY - JOUR A1 - Gebhardt, Andreas T1 - Generative Manufacturing of Ceramic Parts "Vision Rapid Prototyping" N2 - Table of Contents Introduction 1. Generative Manufacturing Processes 2. Classification of Generative Manufacturing Processes 3. Application of Generative Processes on the Fabrication of Ceramic Parts 3.1 Extrusion 3.2 3D-Printing 3.3 Sintering – Laser Sintering 3.4 Layer-Laminate Processes 3.5 Stereolithography (sometimes written: Stereo Lithography) 4. Layer Milling 5. Conclusion - Vision KW - Rapid prototyping KW - Rapid Technologie KW - Rapid Prototyping Y1 - 2006 ER - TY - CHAP A1 - Förster, Arnold A1 - Stock, Jürgen A1 - Montanari, Simone A1 - Lepsa, Mihail Ion A1 - Lüth, Hans T1 - Fabrication and characterisation of GaAs Gunn Diode Chips for applications at 77 GHz in automotive industry N2 - GaAs-based Gunn diodes with graded AlGaAs hot electron injector heterostructures have been developed under the special needs in automotive applications. The fabrication of the Gunn diode chips was based on total substrate removal and processing of integrated Au heat sinks. Especially, the thermal and RF behavior of the diodes have been analyzed by DC, impedance and S-parameter measurements. The electrical investigations have revealed the functionality of the hot electron injector. An optimized layer structure could fulfill the requirements in adaptive cruise control (ACC) systems at 77 GHz with typical output power between 50 and 90 mW. KW - Biosensor KW - Gunn diode KW - microwave generation KW - GaAs hot electron injector Y1 - 2006 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:hbz:a96-opus-1462 ER - TY - CHAP A1 - Frotscher, Ralf A1 - Staat, Manfred ED - Nithiarasu, Perumal T1 - Homogenization of a cardiac tissue construct T2 - CMBE15 : 4th International Conference on Computational & Mathematical Biomedical Engineering ; 29th June - 1st July 2015 ; École Normale Supérieure de Cachan ; Cachan (Paris), France Y1 - 2015 SN - 2227-9385 N1 - Konferenzband unter: http://www.compbiomed.net/getfile.php?type=12/site_documents&id=Proceedings_2227-9385_compressed.pdf SP - 645 EP - 648 PB - CMBE CY - [s.l.] ER - TY - CHAP A1 - Frotscher, Ralf A1 - Koch, Jan-Peter A1 - Raatschen, Hans-Jürgen A1 - Staat, Manfred ED - Onate, E. T1 - Evaluation of a computational model for drug action on cardiac tissue T2 - 11th World Congress on Computational Mechanics (WCCM XI) ; 5th European Conference on Computational Mechanics (ECCM V) ; 6th European Conference on Computational Fluid Dynamics (ECFD VI) ; July 20 - 25, 2014, Barcelona Y1 - 2014 SP - 1 EP - 12 ER - TY - CHAP A1 - Frotscher, Ralf A1 - Duong, Minh Tuan A1 - Staat, Manfred T1 - Simulating beating cardiomyocytes with electromechanical coupling T2 - II. International Conference on Biomedical Technology : 28-30 October 2015 Hannover, Germany / T. Lenarz, P. Wriggers (Eds.) Y1 - 2015 SP - 1 EP - 2 ER - TY - CHAP A1 - Fredebeul-Krein, Markus A1 - Haunert, Martina T1 - Assessing the effectiveness of price cap regimes for the regulation of fixed telephony markets: has it been successful in achieving competition? : preliminary draft / by Markus Fredebeul-Krein and Martina Haunert. 19th European Regional ITS Conference of the International Telecommunications Society, September 17–19, 2008 Rome, Italy N2 - 1. Introduction 2. Tariff regulation, rebalancing and price caps 3. Price cap regimes in selected European countries 4. Has price cap regulation been successful? 5. Regulatory implications for other countries KW - Telekommunikationsmarkt KW - Telekommunikationsmarkt Y1 - 2008 ER - TY - CHAP A1 - Fredebeul-Krein, Markus T1 - Time for revision: The regulation of Germany's next generation networks in the light of new economic and technological challenges on telecom markets : Presentation for the 18th Biennial conference of the International Telecommunications Society, June 24–27, 2008 Montreal, Canada N2 - On 1st January 1998, the German telecom market was fully liberalised. Since then genuine competition between market participants has developed, based on a comprehensive legal and regulatory framework that provides for safeguards against unfair competition and market power by Deutsche Telekom. Today, about 10 years after the liberalisation of the telecommunications sector a revision of this regulatory approach has become necessary because at least on three dimensions the situation is quite different from the one 10 years ago: First, with numerous established alternative operators in the market monopolies have been successfully challenged and competition introduced. Second, not only is Cable TV becoming in large parts of Germany a viable alternative for the provision of broadband services but also mobile services are becoming increasingly a substitute for fixed services. Last but not least there are important technological changes under way, requiring huge investments in infrastructure upgrades for next generation networks. In the light of these new developments the question is to which extent the current regulatory approach of severe ex-ante regulatory intervention is still appropriate. Is any part of the network of the former incumbent still a bottleneck? A more light handed regulatory approach might be the right response to this new situation. The paper is organised as follows: The first section will briefly examine the economic rationale for regulating network access. Based on the assumption that regulation is always necessary when bottlenecks exist regulatory principles for an efficient network access regime will be derived. The second section compares the situation of the German market in early 1998 with the one of today. Thereby three dimensions will be considered: the degree of competition, the potential for substitution and technological developments. The third section will define some requirements for the future regulation of telecom markets. Proposals will be elaborated how to ensure competitive telecom markets in the light of new economic and technological challenges. KW - Telekommunikationsmarkt KW - Telekommunikationsmarkt Y1 - 2008 ER - TY - CHAP A1 - Fredebeul-Krein, Markus T1 - Developing competitive broadband markets: Lessons learnt from German and European regulatory approaches on broad-band access N2 - A key feature of future broadband markets will be diversity of access technologies, meaning that numerous technologies will be exploited for broadband communication. Various factors will affect the success of these future broadband markets, the regulatory policy being one amongst others. So far, a coherent regulatory approach does not exist as to broadband markets. First results of policies so far suggest that less sector-specific regulation is likely to occur. Instead, regulators must ensure that access to networks and services of potentially dominant providers in a relevant broadband market will satisfy requirements for openness and non-discrimination. In this environment the future challenge of regulationg broadband markets will be to set the right incentives for investment into new infrastructures. This paper examines whether there is a need for the regulation of future broadband access markets an if yes, what is the appropriate regulatory tool to do so. Thereby the focus is on the analysis of European broadband markets and the regulatory approaches applied. The first section provides a description of the characteristics of future broadband markets. The second section discusses possible bottlenecks on broadband markets an their regulatory implications. The third section will examine regulatory issues concerning access to broadband networks in more detail. This will be done by comparing the regulatory approaches of European countries and the results in terms of bradband penetration. The final section will give key recommendations for a regulatory strategy on brandband access markets. KW - Telekommunikationsmarkt KW - Telekommunikationsmarkt Y1 - 2006 N1 - paper presented at 16th Biennial conference of the International Telecommunications Society. Information Communication Technology: Opportunities and Challenges for Telecommunications. June 12-16, 2006 Beijing, China. ER - TY - CHAP A1 - Fredebeul-Krein, Markus T1 - 10 years of liberalised telecom markets in Germany: Assessing the degree of competition on the markets for voice telephony N2 - Market data for the German telecom market shows that Deutsche Telekom as the former incumbent is constantly loosing shares on all arkets for voice telephony: the market for local calls, the market for long-distance calls and the market for international calls. At the same time prices decline steadily with the latest trend being that operators offer voice services free of charge, the costs of which are covered by a monthly subscription charge. Against this background the paper examines the state of policy and regulatory reform in the telecommunications sector in Germany almost 10 years after the liberalisation of the fixed telecommunications market. Thereby the focus is on the analysis of the competitive conditions that have been established on the German market for voice telephony services. If these retail markets are competitive, there might be a need to remove remaining regulatory provisions. In the new environment of converging markets the future challenge of regulating fixed telecom markets might be to ensure that access to the network and/or services of a potentially dominant provider in a relevant market will satisfy requirements for openness and non-discrimination. KW - Telekommunikationsmarkt KW - Telekommunikationsmarkt Y1 - 2007 N1 - paper presented at 18th European Regional ITS Conference of the International Telecommunications Society, September 2-4, 2007 Istanbul, Turkey ER -