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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.
The eVTOL industry is a rapidly growing mass market expected to start in 2024. eVTOL compete, caused by their predicted missions, with ground-based transportation modes, including mainly passenger cars. Therefore, the automotive and classical aircraft design process is reviewed and compared to highlight advantages for eVTOL development. A special focus is on ergonomic comfort and safety. The need for further investigation of eVTOL’s crashworthiness is outlined by, first, specifying the relevance of passive safety via accident statistics and customer perception analysis; second, comparing the current state of regulation and certification; and third, discussing the advantages of integral safety and applying the automotive safety approach for eVTOL development. Integral safety links active and passive safety, while the automotive safety approach means implementing standardized mandatory full-vehicle crash tests for future eVTOL. Subsequently, possible crash impact conditions are analyzed, and three full-vehicle crash load cases are presented.
GHEtool is a Python package that contains all the functionalities needed to deal with borefield design. It is developed for both researchers and practitioners. The core of this package is the automated sizing of borefield under different conditions. The sizing of a borefield is typically slow due to the high complexity of the mathematical background. Because this tool has a lot of precalculated data, GHEtool can size a borefield in the order of tenths of milliseconds. This sizing typically takes the order of minutes. Therefore, this tool is suited for being implemented in typical workflows where iterations are required.
GHEtool also comes with a graphical user interface (GUI). This GUI is prebuilt as an exe-file because this provides access to all the functionalities without coding. A setup to install the GUI at the user-defined place is also implemented and available at: https://www.mech.kuleuven.be/en/tme/research/thermal_systems/tools/ghetool.
In general aviation, too, it is desirable to be able to operate existing internal combustion engines with fuels that produce less CO₂ than Avgas 100LL being widely used today It can be assumed that, in comparison, the fuels CNG, LPG or LNG, which are gaseous under normal conditions, produce significantly lower emissions. Necessary propulsion system adaptations were investigated as part of a research project at Aachen University of Applied Sciences.
This study analyses the expected utilization of an urban distribution grid under high penetration of photovoltaic and e-mobility with charging infrastructure on a residential level. The grid utilization and the corresponding power flow are evaluated, while varying the control strategies and photovoltaic installed capacity in different scenarios. Four scenarios are used to analyze the impact of e-mobility. The individual mobility demand is modelled based on the largest German studies on mobility “Mobilität in Deutschland”, which is carried out every 5 years. To estimate the ramp-up of photovoltaic generation, a potential analysis of the roof surfaces in the supply area is carried out via an evaluation of an open solar potential study. The photovoltaic feed-in time series is derived individually for each installed system in a resolution of 15 min. The residential consumption is estimated using historical smart meter data, which are collected in London between 2012 and 2014. For a realistic charging demand, each residential household decides daily on the state of charge if their vehicle requires to be charged. The resulting charging time series depends on the underlying behavior scenario. Market prices and mobility demand are therefore used as scenario input parameters for a utility function based on the current state of charge to model individual behavior. The aggregated electricity demand is the starting point of the power flow calculation. The evaluation is carried out for an urban region with approximately 3100 residents. The analysis shows that increased penetration of photovoltaics combined with a flexible and adaptive charging strategy can maximize PV usage and reduce the need for congestion-related intervention by the grid operator by reducing the amount of kWh charged from the grid by 30% which reduces the average price of a charged kWh by 35% to 14 ct/kWh from 21.8 ct/kWh without PV optimization. The resulting grid congestions are managed by implementing an intelligent price or control signal. The analysis took place using data from a real German grid with 10 subgrids. The entire software can be adapted for the analysis of different distribution grids and is publicly available as an open-source software library on GitHub.
This paper describes the realization of a novel neurocomputer which is based on the concepts of a coprocessor. In contrast to existing neurocomputers the main interest was the realization of a scalable, flexible system, which is capable of computing neural networks of arbitrary topology and scale, with full independence of special hardware from the software's point of view. On the other hand, computational power should be added, whenever needed and flexibly adapted to the requirements of the application. Hardware independence is achieved by a run time system which is capable of using all available computing power, including multiple host CPUs and an arbitrary number of neural coprocessors autonomously. The realization of arbitrary neural topologies is provided through the implementation of the elementary operations which can be found in most neural topologies.
Environmental emissions, global warming, and energy-related concerns have accelerated the advancements in conventional vehicles that primarily use internal combustion engines. Among the existing technologies, hydrogen fuel cell electric vehicles and fuel cell hybrid electric vehicles may have minimal contributions to greenhouse gas emissions and thus are the prime choices for environmental concerns. However, energy management in fuel cell electric vehicles and fuel cell hybrid electric vehicles is a major challenge. Appropriate control strategies should be used for effective energy management in these vehicles. On the other hand, there has been significant progress in artificial intelligence, machine learning, and designing data-driven intelligent controllers. These techniques have found much attention within the community, and state-of-the-art energy management technologies have been developed based on them. This manuscript reviews the application of machine learning and intelligent controllers for prediction, control, energy management, and vehicle to everything (V2X) in hydrogen fuel cell vehicles. The effectiveness of data-driven control and optimization systems are investigated to evolve, classify, and compare, and future trends and directions for sustainability are discussed.
On the applicability of several tests to models with not identically distributed random effects
(2023)
We consider Kolmogorov–Smirnov and Cramér–von-Mises type tests for testing central symmetry, exchangeability, and independence. In the standard case, the tests are intended for the application to independent and identically distributed data with unknown distribution. The tests are available for multivariate data and bootstrap procedures are suitable to obtain critical values. We discuss the applicability of the tests to random effects models, where the random effects are independent but not necessarily identically distributed and with possibly unknown distributions. Theoretical results show the adequacy of the tests in this situation. The quality of the tests in models with random effects is investigated by simulations. Empirical results obtained confirm the theoretical findings. A real data example illustrates the application.