TY - JOUR A1 - Haeger, Gerrit A1 - Grankin, Alina A1 - Wagner, Michaela T1 - Construction of an Aspergillus oryzae triple amylase deletion mutant as a chassis to evaluate industrially relevant amylases using multiplex CRISPR/Cas9 editing technology JF - Applied Research N2 - 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. KW - aspergillus KW - CRISPR/Cas9 KW - filamentous fungi KW - genome engineering Y1 - 2023 U6 - https://doi.org/10.1002/appl.202200106 SN - 2702-4288 IS - Early View SP - 1 EP - 15 PB - Wiley-VCH ER - TY - JOUR A1 - Cheenakula, Dheeraja A1 - Griebel, Kai A1 - Montag, David A1 - Grömping, Markus ED - Huang, Xiaowu T1 - Concept development of a mainstream deammonification and comparison with conventional process in terms of energy, performance and economical construction perspectives JF - Frontiers in Microbiology N2 - Deammonification for nitrogen removal in municipal wastewater in temperate and cold climate zones is currently limited to the side stream of municipal wastewater treatment plants (MWWTP). This study developed a conceptual model of a mainstream deammonification plant, designed for 30,000 P.E., considering possible solutions corresponding to the challenging mainstream conditions in Germany. In addition, the energy-saving potential, nitrogen elimination performance and construction-related costs of mainstream deammonification were compared to a conventional plant model, having a single-stage activated sludge process with upstream denitrification. The results revealed that an additional treatment step by combining chemical precipitation and ultra-fine screening is advantageous prior the mainstream deammonification. Hereby chemical oxygen demand (COD) can be reduced by 80% so that the COD:N ratio can be reduced from 12 to 2.5. Laboratory experiments testing mainstream conditions of temperature (8–20°C), pH (6–9) and COD:N ratio (1–6) showed an achievable volumetric nitrogen removal rate (VNRR) of at least 50 gN/(m3∙d) for various deammonifying sludges from side stream deammonification systems in the state of North Rhine-Westphalia, Germany, where m3 denotes reactor volume. Assuming a retained Norganic content of 0.0035 kgNorg./(P.E.∙d) from the daily loads of N at carbon removal stage and a VNRR of 50 gN/(m3∙d) under mainstream conditions, a resident-specific reactor volume of 0.115 m3/(P.E.) is required for mainstream deammonification. This is in the same order of magnitude as the conventional activated sludge process, i.e., 0.173 m3/(P.E.) for an MWWTP of size class of 4. The conventional plant model yielded a total specific electricity demand of 35 kWh/(P.E.∙a) for the operation of the whole MWWTP and an energy recovery potential of 15.8 kWh/(P.E.∙a) through anaerobic digestion. In contrast, the developed mainstream deammonification model plant would require only a 21.5 kWh/(P.E.∙a) energy demand and result in 24 kWh/(P.E.∙a) energy recovery potential, enabling the mainstream deammonification model plant to be self-sufficient. The retrofitting costs for the implementation of mainstream deammonification in existing conventional MWWTPs are nearly negligible as the existing units like activated sludge reactors, aerators and monitoring technology are reusable. However, the mainstream deammonification must meet the performance requirement of VNRR of about 50 gN/(m3∙d) in this case. KW - anammox KW - energy efficiency KW - mainstream deammonification KW - nitrogen elimination KW - wastewater Y1 - 2023 U6 - https://doi.org/10.3389/fmicb.2023.1155235 SN - 1664-302X VL - 14 IS - 11155235 SP - 1 EP - 15 PB - Frontiers ER - TY - JOUR A1 - Waldvogel, Janice A1 - Freyler, Kathrin A1 - Helm, Michael A1 - Monti, Elena A1 - Stäudle, Benjamin A1 - Gollhofer, Albert A1 - Narici, Marco V. A1 - Ritzmann, Ramona A1 - Albracht, Kirsten T1 - Changes in gravity affect neuromuscular control, biomechanics, and muscle-tendon mechanics in energy storage and dissipation tasks JF - Journal of Applied Physiology N2 - This study evaluates neuromechanical control and muscle-tendon interaction during energy storage and dissipation tasks in hypergravity. During parabolic flights, while 17 subjects performed drop jumps (DJs) and drop landings (DLs), electromyography (EMG) of the lower limb muscles was combined with in vivo fascicle dynamics of the gastrocnemius medialis, two-dimensional (2D) kinematics, and kinetics to measure and analyze changes in energy management. Comparisons were made between movement modalities executed in hypergravity (1.8 G) and gravity on ground (1 G). In 1.8 G, ankle dorsiflexion, knee joint flexion, and vertical center of mass (COM) displacement are lower in DJs than in DLs; within each movement modality, joint flexion amplitudes and COM displacement demonstrate higher values in 1.8 G than in 1 G. Concomitantly, negative peak ankle joint power, vertical ground reaction forces, and leg stiffness are similar between both movement modalities (1.8 G). In DJs, EMG activity in 1.8 G is lower during the COM deceleration phase than in 1 G, thus impairing quasi-isometric fascicle behavior. In DLs, EMG activity before and during the COM deceleration phase is higher, and fascicles are stretched less in 1.8 G than in 1 G. Compared with the situation in 1 G, highly task-specific neuromuscular activity is diminished in 1.8 G, resulting in fascicle lengthening in both movement modalities. Specifically, in DJs, a high magnitude of neuromuscular activity is impaired, resulting in altered energy storage. In contrast, in DLs, linear stiffening of the system due to higher neuromuscular activity combined with lower fascicle stretch enhances the buffering function of the tendon, and thus the capacity to safely dissipate energy. KW - electromyography KW - locomotion KW - overload KW - stretch-shortening cycle KW - ultrasound Y1 - 2023 U6 - https://doi.org/10.1152/japplphysiol.00279.2022 SN - 1522-1601 (Onlineausgabe) SN - 8750-7587 (Druckausgabe) VL - 134 IS - 1 SP - 190 EP - 202 PB - American Physiological Society CY - Bethesda, Md. ER - TY - JOUR A1 - Pfaff, Raphael T1 - Braking distance prediction for vehicle consist in low-speed on-sight operation: a Monte Carlo approach JF - Railway Engineering Science N2 - The first and last mile of a railway journey, in both freight and transit applications, constitutes a high effort and is either non-productive (e.g. in the case of depot operations) or highly inefficient (e.g. in industrial railways). These parts are typically managed on-sight, i.e. with no signalling and train protection systems ensuring the freedom of movement. This is possible due to the rather short braking distances of individual vehicles and shunting consists. The present article analyses the braking behaviour of such shunting units. For this purpose, a dedicated model is developed. It is calibrated on published results of brake tests and validated against a high-definition model for low-speed applications. Based on this model, multiple simulations are executed to obtain a Monte Carlo simulation of the resulting braking distances. Based on the distribution properties and established safety levels, the risk of exceeding certain braking distances is evaluated and maximum braking distances are derived. Together with certain parameters of the system, these can serve in the design and safety assessment of driver assistance systems and automation of these processes. KW - Freight rail KW - Shunting KW - Braking curves KW - Brake set-up KW - Driver assistance system Y1 - 2023 U6 - https://doi.org/10.1007/s40534-023-00303-7 SN - 2662-4753 (eISSN) SN - 2662-4745 (Print) VL - 31 IS - 2 SP - 135 EP - 144 PB - SpringerOpen ER - TY - JOUR A1 - Timme, Michael T1 - Bauliche Veränderungen durch Wohnungseigentümer und die Zustimmung der Gemeinschaft : Zugleich eine Besprechung des BGH, Urt. v. 17.3.2023 – V ZR 140/22, MDR 2023, 619 JF - Monatsschrift für Deutsches Recht N2 - Durch das WEMoG (Gesetz v. 16.10.2020, BGBl. I 2187) hat der Gesetzgeber die Regelung des § 20 WEG eingeführt. Demnach ist auch für bauliche Veränderungen an Räumen und Flächen, die einem Sondernutzungsrecht unterliegen, die Zustimmung der Gemeinschaft notwendig. Der BGH hat nun mit Urt. v. 17.3.2023 –V ZR 140/22, MDR 2023, 619 deutlich gemacht, dass bei Fehlen eines entsprechenden Beschlusses, die bauliche Veränderung durch einen einzelnen Wohnungseigentümer nicht vorgenommen werden darf, sondern eine rechtswidrige Eigentumsbeeinträchtigung darstellt. Der folgende Beitrag nimmt die Entscheidung zum Anlass, um die neue Rechtslage durch § 20 WEG eingehend zu erläutern. Gleichzeitig werden die Folgen der aktuellen Entscheidung näher dargestellt. Y1 - 2023 SN - 2194-4202 (online) SN - 0340-1812 (print) VL - 77 IS - 13 SP - 815 EP - 818 PB - Verlag Dr. Otto Schmidt CY - Köln ER - TY - JOUR A1 - Fayyazi, Mojgan A1 - Sardar, Paramjotsingh A1 - Thomas, Sumit Infent A1 - Daghigh, Roonak A1 - Jamali, Ali A1 - Esch, Thomas A1 - Kemper, Hans A1 - Langari, Reza A1 - Khayyam, Hamid T1 - Artificial intelligence/machine learning in energy management systems, control, and optimization of hydrogen fuel cell vehicles N2 - 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. KW - optimization system KW - intelligent control KW - fuel cell vehicle KW - machine learning KW - artificial intelligence KW - intelligent energy management Y1 - 2023 U6 - https://doi.org/10.3390/su15065249 N1 - This article belongs to the Special Issue "Circular Economy and Artificial Intelligence" VL - 15 IS - 6 SP - 38 PB - MDPI CY - Basel ER - TY - JOUR A1 - Ayala, Rafael Ceja A1 - Harris, Isaac A1 - Kleefeld, Andreas A1 - Pallikarakis, Nikolaos T1 - Analysis of the transmission eigenvalue problem with two conductivity parameters JF - Applicable Analysis N2 - In this paper, we provide an analytical study of the transmission eigenvalue problem with two conductivity parameters. We will assume that the underlying physical model is given by the scattering of a plane wave for an isotropic scatterer. In previous studies, this eigenvalue problem was analyzed with one conductive boundary parameter whereas we will consider the case of two parameters. We prove the existence and discreteness of the transmission eigenvalues as well as study the dependence on the physical parameters. We are able to prove monotonicity of the first transmission eigenvalue with respect to the parameters and consider the limiting procedure as the second boundary parameter vanishes. Lastly, we provide extensive numerical experiments to validate the theoretical work. KW - Transmission Eigenvalues KW - Conductive Boundary Condition KW - Inverse Scattering Y1 - 2023 U6 - https://doi.org/10.1080/00036811.2023.2181167 SN - 0003-6811 PB - Taylor & Francis ER - TY - JOUR A1 - Emonts, Jessica A1 - Buyel, Johannes Felix T1 - An overview of descriptors to capture protein properties – Tools and perspectives in the context of QSAR modeling JF - Computational and Structural Biotechnology Journal N2 - Proteins are important ingredients in food and feed, they are the active components of many pharmaceutical products, and they are necessary, in the form of enzymes, for the success of many technical processes. However, production can be challenging, especially when using heterologous host cells such as bacteria to express and assemble recombinant mammalian proteins. The manufacturability of proteins can be hindered by low solubility, a tendency to aggregate, or inefficient purification. Tools such as in silico protein engineering and models that predict separation criteria can overcome these issues but usually require the complex shape and surface properties of proteins to be represented by a small number of quantitative numeric values known as descriptors, as similarly used to capture the features of small molecules. Here, we review the current status of protein descriptors, especially for application in quantitative structure activity relationship (QSAR) models. First, we describe the complexity of proteins and the properties that descriptors must accommodate. Then we introduce descriptors of shape and surface properties that quantify the global and local features of proteins. Finally, we highlight the current limitations of protein descriptors and propose strategies for the derivation of novel protein descriptors that are more informative. KW - Prediction of molecular features KW - Protein structure complexity KW - Quantitative structure activity relationship KW - Scalar parameters KW - Shape and surface properties Y1 - 2023 U6 - https://doi.org/10.1016/j.csbj.2023.05.022 SN - 2001-0370 (online-ressource) IS - 21 SP - 3234 EP - 3247 PB - Research Network of Computational and Structural Biotechnology CY - Gotenburg ER - TY - JOUR A1 - Kowalewski, Paul A1 - Bragard, Michael A1 - Hüning, Felix A1 - De Doncker, Rik W. T1 - An inexpensive Wiegand-sensor-based rotary encoder without rotating magnets for use in electrical drives JF - IEEE Transactions on Instrumentation and Measurement N2 - 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. KW - Rotary encoder KW - Wiegand sensor Y1 - 2023 U6 - https://doi.org/10.1109/TIM.2023.3326166 SN - 0018-9456 (Print) SN - 1557-9662 (Online) VL - 72 SP - 10 Seiten PB - IEEE CY - New York ER - TY - JOUR A1 - Gaigall, Daniel ED - AitSahlia, Farid T1 - Allocating and forecasting changes in risk JF - Journal of risk N2 - We consider time-dependent portfolios and discuss the allocation of changes in the risk of a portfolio to changes in the portfolio’s components. For this purpose we adopt established allocation principles. We also use our approach to obtain forecasts for changes in the risk of the portfolio’s components. To put the approach into practice we present an implementation based on the output of a simulation. Allocation is illustrated with an example portfolio in the context of Solvency II. The quality of the forecasts is investigated with an empirical study. KW - portfolio risk KW - allocation KW - forecast KW - covariance principle KW - conditional expectation principle Y1 - 2023 U6 - https://doi.org/10.21314/JOR.2022.048 SN - 1755-2842 SN - 1465-1211 VL - 25 IS - 3 SP - 1 EP - 24 PB - Infopro Digital Risk CY - London ER -