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New European Union (EU) regulations for UAS operations require an operational risk analysis, which includes an estimation of the potential danger of the UAS crashing. A key parameter for the potential ground risk is the kinetic impact energy of the UAS. The kinetic energy depends on the impact velocity of the UAS and, therefore, on the aerodynamic drag and the weight during free fall. Hence, estimating the impact energy of a UAS requires an accurate drag estimation of the UAS in that state. The paper at hand presents the aerodynamic drag estimation of small-scale multirotor UAS. Multirotor UAS of various sizes and configurations were analysed with a fully unsteady Reynolds-averaged Navier–Stokes approach. These simulations included different velocities and various fuselage pitch angles of the UAS. The results were compared against force measurements performed in a subsonic wind tunnel and provided good consistency. Furthermore, the influence of the UAS`s fuselage pitch angle as well as the influence of fixed and free spinning propellers on the aerodynamic drag was analysed. Free spinning propellers may increase the drag by up to 110%, depending on the fuselage pitch angle. Increasing the fuselage pitch angle of the UAS lowers the drag by 40% up to 85%, depending on the UAS. The data presented in this paper allow for increased accuracy of ground risk assessments.
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.
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.
Aspergillus oryzae is an industrially relevant organism for the secretory production of heterologous enzymes, especially amylases. The activities of potential heterologous amylases, however, cannot be quantified directly from the supernatant due to the high background activity of native α-amylase. This activity is caused by the gene products of amyA, amyB, and amyC. In this study, an in vitro CRISPR/Cas9 system was established in A. oryzae to delete these genes simultaneously. First, pyrG of A. oryzae NSAR1 was mutated by exploiting NHEJ to generate a counter-selection marker. Next, all amylase genes were deleted simultaneously by co-transforming a repair template carrying pyrG of Aspergillus nidulans and flanking sequences of amylase gene loci. The rate of obtained triple knock-outs was 47%. We showed that triple knockouts do not retain any amylase activity in the supernatant. The established in vitro CRISPR/Cas9 system was used to achieve sequence-specific knock-in of target genes. The system was intended to incorporate a single copy of the gene of interest into the desired host for the development of screening methods. Therefore, an integration cassette for the heterologous Fpi amylase was designed to specifically target the amyB locus. The site-specific integration rate of the plasmid was 78%, with exceptional additional integrations. Integration frequency was assessed via qPCR and directly correlated with heterologous amylase activity. Hence, we could compare the efficiency between two different signal peptides. In summary, we present a strategy to exploit CRISPR/Cas9 for gene mutation, multiplex knock-out, and the targeted knock-in of an expression cassette in A. oryzae. Our system provides straightforward strain engineering and paves the way for development of fungal screening systems.
Diese Bachelorarbeit befasst sich mit dem Corporate Design des aufstrebenden Startups "CoArea". Seine innovative Plattform vermittelt ungenutzte Flächen im urbanen Raum und bringt Flächenbedarf und -bereitstellung intelligent zusammen. Die Arbeit untersucht, wie das Erscheinungsbild von CoArea das Unternehmen als modern, nachhaltig und ressourcenschonend positioniert. Durch das neu gestaltete Corporate Design wird diese besondere Herangehensweise zur effizienten Nutzung begrenzter Ressourcen unserer Umwelt durch CoArea deutlich und nach außen getragen. CoArea strebt danach, die positive Entwicklung der Städte voranzutreiben und die soziale Nachhaltigkeit durch den Austausch und die Interaktion zwischen verschiedenen Menschen und Gemeinschaften zu fördern.
In order to reduce energy consumption of homes, it is important to make transparent which devices consume how much energy. However, power consumption is often only monitored aggregated at the house energy meter. Disaggregating this power consumption into the contributions of individual devices can be achieved using Machine Learning. Our work aims at making state of the art disaggregation algorithms accessibe for users of the open source home automation platform Home Assistant.
Die Bereitstellung von nachhaltig erzeugtem Wasserstoff als Energieträger und Rohstoff ist eine wichtige Schlüsseltechnologie sowohl als Ersatz für fossile Energieträger, aber auch als Produkt im Zusammenhang mit Kreislaufprozessen. In der Abwasserbehandlung bestehen verschiedene Möglichkeiten Wasserstoff herzustellen. Mehrere Wege, mögliche Synergien, aber auch deren Nachteile werden vorgestellt.
Preprint: Studies on the enzymatic reduction of levulinic acid using Chiralidon-R and Chiralidon-S
(2023)
The enzymatic reduction of levulinic acid by the chiral catalysts Chiralidon-R and Chiralidon-S which are commercially available superabsorbed alcohol dehydrogenases is described. The Chiralidon®-R/S reduces the levulinic acid to the (R,S)-4-hydroxy valeric acid and the (R)- or (S)- gamma-valerolactone.
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.