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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.
Exposure to prolonged periods in microgravity is associated with deconditioning of the musculoskeletal system due to chronic changes in mechanical stimulation. Given astronauts will operate on the Lunar surface for extended periods of time, it is critical to quantify both external (e.g., ground reaction forces) and internal (e.g., joint reaction forces) loads of relevant movements performed during Lunar missions. Such knowledge is key to predict musculoskeletal deconditioning and determine appropriate exercise countermeasures associated with extended exposure to hypogravity.
This paper introduces an inexpensive Wiegand-sensor-based rotary encoder that avoids rotating magnets and is suitable for electrical-drive applications. So far, Wiegand-sensor-based encoders usually include a magnetic pole wheel with rotating permanent magnets. These encoders combine the disadvantages of an increased magnet demand and a limited maximal speed due to the centripetal force acting on the rotating magnets. The proposed approach reduces the total demand of permanent magnets drastically. Moreover, the rotating part is manufacturable from a single piece of steel, which makes it very robust and cheap. This work presents the theoretical operating principle of the proposed approach and validates its benefits on a hardware prototype. The presented proof-of-concept prototype achieves a mechanical resolution of 4.5 ° by using only 4 permanent magnets, 2Wiegand sensors and a rotating steel gear wheel with 20 teeth.
With the prevalence of glucosamine- and chondroitin-containing dietary supplements for people with osteoarthritis in the marketplace, it is important to have an accurate and reproducible analytical method for the quantitation of these compounds in finished products. NMR spectroscopic method based both on low- (80 MHz) and high- (500–600 MHz) field NMR instrumentation was established, compared and validated for the determination of chondroitin sulfate and glucosamine in dietary supplements. The proposed method was applied for analysis of 20 different dietary supplements. In the majority of cases, quantification results obtained on the low-field NMR spectrometer are similar to those obtained with high-field 500–600 MHz NMR devices. Validation results in terms of accuracy, precision, reproducibility, limit of detection and recovery demonstrated that the developed method is fit for purpose for the marketed products. The NMR method was extended to the analysis of methylsulfonylmethane, adulterant maltodextrin, acetate and inorganic ions. Low-field NMR can be a quicker and cheaper alternative to more expensive high-field NMR measurements for quality control of the investigated dietary supplements. High-field NMR instrumentation can be more favorable for samples with complex composition due to better resolution, simultaneously giving the possibility of analysis of inorganic species such as potassium and chloride.
The number of electronic vehicles increase steadily while the space for extending the charging infrastructure is limited. In particular in urban areas, where parking spaces in attractive areas are famous, opportunities to setup new charging stations is very limited. This leads to an overload of some very attractive charging stations and an underutilization of less attractive ones. Against this background, the paper at hand presents the design of an e-vehicle reservation system that aims at distributing the utilization of the charging infrastructure, particularly in urban areas. By applying a design science approach, the requirements for a reservation-based utilization approach are elicited and a model for a suitable distribution approach and its instantiation are developed. The artefact is evaluated by simulating the distribution effects based on data of real charging station utilizations.
Zwangsbelüftete Nasskühltürme haben im Gegensatz zur Trockenkühlung bei naßkaltem Wetter Nebelschwaden zur Folge. Dagegen ist bei Naßkühlung die spezifische Kühlleistung durch abgeführte Kondensationswärme höher als bei der Trockenkühlung. Hybridzellenkühltürme kombinieren beide Methoden, so daß ein Mischstrom beider Abluftströme die Wasserdampf-Sättigungsgrenze nicht überschreitet. Durch das Mischungsverhältnis kann man den gewünschten Sättigungsgrad einstellen. Je dichter dieser an der Sättigungsgrenze liegt, desto höher ist die Kühlleistung. Der von unten zugeführte Luftstrom der Naßkühlung und der seitlich zugeführte trockene Abluftstrom müssen sehr gut durchmischt werden, um über den gesamten Austrittsquerschnitt des Kühlturms die Sättigungsgrenze nicht zu überschreiten. In einem maßstabsgerechten Modell wurde der Mischungsgrad mit und ohne Einbauten untersucht. Über ein Raster von 10 mal 10 Punkten wurde die örtliche Temperaturverteilung ermittelt. Wärmebilanzen ergeben dann die Mischungsgüte in einer Ebene oberhalb der Zellenkrone. Während ohne Mischeinbauten der Trockenluftanteil in der Mitte des Querschnitts bei unter 15 % liegt erhöhen Einbauten den Trockenluftanteil auf 30 % bis über 40 %. Dabei wurde die Trockenluft auf jeder Kühlturmseite durch 4 konisch zulaufende, unten offene und oben geschlitzte Einbauten kanalisiert. Die Nassluft wurde durch eine im Querschnitt dreieckige Rinne in Richtung der Trockenluftauslässe umgelenkt. Im Raster leicht zu lokalisierende Abweichungen vom gewünschten Mittelwert zeigen Potential für die weitere Verbesserung der Einbauten.
Immunosorbent turnip vein clearing virus (TVCV) particles displaying the IgG-binding domains D and E of Staphylococcus aureus protein A (PA) on every coat protein (CP) subunit (TVCVPA) were purified from plants via optimized and new protocols. The latter used polyethylene glycol (PEG) raw precipitates, from which virions were selectively re-solubilized in reverse PEG concentration gradients. This procedure improved the integrity of both TVCVPA and the wild-type subgroup 3 tobamovirus. TVCVPA could be loaded with more than 500 IgGs per virion, which mediated the immunocapture of fluorescent dyes, GFP, and active enzymes. Bi-enzyme ensembles of cooperating glucose oxidase and horseradish peroxidase were tethered together on the TVCVPA carriers via a single antibody type, with one enzyme conjugated chemically to its Fc region, and the other one bound as a target, yielding synthetic multi-enzyme complexes. In microtiter plates, the TVCVPA-displayed sugar-sensing system possessed a considerably increased reusability upon repeated testing, compared to the IgG-bound enzyme pair in the absence of the virus. A high coverage of the viral adapters was also achieved on Ta2O5 sensor chip surfaces coated with a polyelectrolyte interlayer, as a prerequisite for durable TVCVPA-assisted electrochemical biosensing via modularly IgG-assembled sensor enzymes.
Hydrogen peroxide (H₂O₂), a strong oxidizer, is a commonly used sterilization agent employed during aseptic food processing and medical applications. To assess the sterilization efficiency with H₂O₂, bacterial spores are common microbial systems due to their remarkable robustness against a wide variety of decontamination strategies. Despite their widespread use, there is, however, only little information about the detailed time-resolved mechanism underlying the oxidative spore death by H₂O₂. In this work, we investigate chemical and morphological changes of individual Bacillus atrophaeus spores undergoing oxidative damage using optical sensing with trapping Raman microscopy in real-time. The time-resolved experiments reveal that spore death involves two distinct phases: (i) an initial phase dominated by the fast release of dipicolinic acid (DPA), a major spore biomarker, which indicates the rupture of the spore’s core; and (ii) the oxidation of the remaining spore material resulting in the subsequent fragmentation of the spores’ coat. Simultaneous observation of the spore morphology by optical microscopy corroborates these mechanisms. The dependence of the onset of DPA release and the time constant of spore fragmentation on H₂O₂ shows that the formation of reactive oxygen species from H₂O₂ is the rate-limiting factor of oxidative spore death.