@article{HaegerGrankinWagner2023, author = {Haeger, Gerrit and Grankin, Alina and Wagner, Michaela}, title = {Construction of an Aspergillus oryzae triple amylase deletion mutant as a chassis to evaluate industrially relevant amylases using multiplex CRISPR/Cas9 editing technology}, series = {Applied Research}, journal = {Applied Research}, number = {Early View}, publisher = {Wiley-VCH}, issn = {2702-4288}, doi = {10.1002/appl.202200106}, pages = {1 -- 15}, year = {2023}, abstract = {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.}, language = {en} } @article{SchulteTiggesFoersterNikolovskietal.2022, author = {Schulte-Tigges, Joschua and F{\"o}rster, Marco and Nikolovski, Gjorgji and Reke, Michael and Ferrein, Alexander and Kaszner, Daniel and Matheis, Dominik and Walter, Thomas}, title = {Benchmarking of various LiDAR sensors for use in self-driving vehicles in real-world environments}, series = {Sensors}, volume = {22}, journal = {Sensors}, number = {19}, publisher = {MDPI}, address = {Basel}, issn = {1424-8220}, doi = {10.3390/s22197146}, pages = {20 Seiten}, year = {2022}, abstract = {Abstract In this paper, we report on our benchmark results of the LiDAR sensors Livox Horizon, Robosense M1, Blickfeld Cube, Blickfeld Cube Range, Velodyne Velarray H800, and Innoviz Pro. The idea was to test the sensors in different typical scenarios that were defined with real-world use cases in mind, in order to find a sensor that meet the requirements of self-driving vehicles. For this, we defined static and dynamic benchmark scenarios. In the static scenarios, both LiDAR and the detection target do not move during the measurement. In dynamic scenarios, the LiDAR sensor was mounted on the vehicle which was driving toward the detection target. We tested all mentioned LiDAR sensors in both scenarios, show the results regarding the detection accuracy of the targets, and discuss their usefulness for deployment in self-driving cars.}, language = {en} } @article{AkimbekovDigelTastambeketal.2022, author = {Akimbekov, Nuraly S. and Digel, Ilya and Tastambek, Kuanysh T. and Marat, Adel K. and Turaliyeva, Moldir A. and Kaiyrmanova, Gulzhan K.}, title = {Biotechnology of Microorganisms from Coal Environments: From Environmental Remediation to Energy Production}, series = {Biology}, volume = {11}, journal = {Biology}, number = {9}, publisher = {MDPI}, address = {Basel}, issn = {2079-7737}, doi = {10.3390/biology11091306}, pages = {47 Seiten}, year = {2022}, abstract = {It was generally believed that coal sources are not favorable as live-in habitats for microorganisms due to their recalcitrant chemical nature and negligible decomposition. However, accumulating evidence has revealed the presence of diverse microbial groups in coal environments and their significant metabolic role in coal biogeochemical dynamics and ecosystem functioning. The high oxygen content, organic fractions, and lignin-like structures of lower-rank coals may provide effective means for microbial attack, still representing a greatly unexplored frontier in microbiology. Coal degradation/conversion technology by native bacterial and fungal species has great potential in agricultural development, chemical industry production, and environmental rehabilitation. Furthermore, native microalgal species can offer a sustainable energy source and an excellent bioremediation strategy applicable to coal spill/seam waters. Additionally, the measures of the fate of the microbial community would serve as an indicator of restoration progress on post-coal-mining sites. This review puts forward a comprehensive vision of coal biodegradation and bioprocessing by microorganisms native to coal environments for determining their biotechnological potential and possible applications.}, language = {en} } @article{HoffmannUhlCeblinetal.2022, author = {Hoffmann, Andreas and Uhl, Matthias and Ceblin, Maximilian and Rohrbach, Felix and Bansmann, Joachim and Mallah, Marcel and Heuermann, Holger and Jacob, Timo and Kuehne, Alexander J.C.}, title = {Atmospheric pressure plasma-jet treatment of PAN-nonwovens—carbonization of nanofiber electrodes}, series = {C - Journal of Carbon Research}, volume = {8}, journal = {C - Journal of Carbon Research}, number = {3}, publisher = {MDPI}, address = {Basel}, issn = {2311-5629}, doi = {10.3390/c8030033}, pages = {8 Seiten}, year = {2022}, abstract = {Carbon nanofibers are produced from dielectric polymer precursors such as polyacrylonitrile (PAN). Carbonized nanofiber nonwovens show high surface area and good electrical conductivity, rendering these fiber materials interesting for application as electrodes in batteries, fuel cells, and supercapacitors. However, thermal processing is slow and costly, which is why new processing techniques have been explored for carbon fiber tows. Alternatives for the conversion of PAN-precursors into carbon fiber nonwovens are scarce. Here, we utilize an atmospheric pressure plasma jet to conduct carbonization of stabilized PAN nanofiber nonwovens. We explore the influence of various processing parameters on the conductivity and degree of carbonization of the converted nanofiber material. The precursor fibers are converted by plasma-jet treatment to carbon fiber nonwovens within seconds, by which they develop a rough surface making subsequent surface activation processes obsolete. The resulting carbon nanofiber nonwovens are applied as supercapacitor electrodes and examined by cyclic voltammetry and impedance spectroscopy. Nonwovens that are carbonized within 60 s show capacitances of up to 5 F g⁻¹.}, language = {en} } @article{VahidpourAlghazaliAkcaetal.2022, author = {Vahidpour, Farnoosh and Alghazali, Yousef and Akca, Sevilay and Hommes, Gregor and Sch{\"o}ning, Michael Josef}, title = {An Enzyme-Based Interdigitated Electrode-Type Biosensor for Detecting Low Concentrations of H₂O₂ Vapor/Aerosol}, series = {Chemosensors}, volume = {10}, journal = {Chemosensors}, number = {6}, publisher = {MDPI}, address = {Basel}, issn = {2227-9040}, doi = {10.3390/chemosensors10060202}, pages = {Arikel 202}, year = {2022}, abstract = {This work introduces a novel method for the detection of H₂O₂ vapor/aerosol of low concentrations, which is mainly applied in the sterilization of equipment in medical industry. Interdigitated electrode (IDE) structures have been fabricated by means of microfabrication techniques. A differential setup of IDEs was prepared, containing an active sensor element (active IDE) and a passive sensor element (passive IDE), where the former was immobilized with an enzymatic membrane of horseradish peroxidase that is selective towards H₂O₂. Changes in the IDEs' capacitance values (active sensor element versus passive sensor element) under H₂O₂ vapor/aerosol atmosphere proved the detection in the concentration range up to 630 ppm with a fast response time (<60 s). The influence of relative humidity was also tested with regard to the sensor signal, showing no cross-sensitivity. The repeatability assessment of the IDE biosensors confirmed their stable capacitive signal in eight subsequent cycles of exposure to H₂O₂ vapor/aerosol. Room-temperature detection of H₂O₂ vapor/aerosol with such miniaturized biosensors will allow a future three-dimensional, flexible mapping of aseptic chambers and help to evaluate sterilization assurance in medical industry.}, language = {en} } @article{PoghossianKarschuckWagneretal.2022, author = {Poghossian, Arshak and Karschuck, Tobias and Wagner, Patrick and Sch{\"o}ning, Michael Josef}, title = {Field-Effect Capacitors Decorated with Ligand-Stabilized Gold Nanoparticles: Modeling and Experiments}, series = {Biosensors}, volume = {12}, journal = {Biosensors}, number = {5}, publisher = {MDPI}, address = {Basel}, issn = {2079-6374}, doi = {10.3390/bios12050334}, pages = {Artikel 334}, year = {2022}, abstract = {Nanoparticles are recognized as highly attractive tunable materials for designing field-effect biosensors with enhanced performance. In this work, we present a theoretical model for electrolyte-insulator-semiconductor capacitors (EISCAP) decorated with ligand-stabilized charged gold nanoparticles. The charged AuNPs are taken into account as additional, nanometer-sized local gates. The capacitance-voltage (C-V) curves and constant-capacitance (ConCap) signals of the AuNP-decorated EISCAPs have been simulated. The impact of the AuNP coverage on the shift of the C-V curves and the ConCap signals was also studied experimentally on Al-p-Si-SiO₂ EISCAPs decorated with positively charged aminooctanethiol-capped AuNPs. In addition, the surface of the EISCAPs, modified with AuNPs, was characterized by scanning electron microscopy for different immobilization times of the nanoparticles.}, language = {en} } @article{PourshahidiAchtsnichtOffenhaeusseretal.2022, author = {Pourshahidi, Ali Mohammad and Achtsnicht, Stefan and Offenh{\"a}usser, Andreas and Krause, Hans-Joachim}, title = {Frequency Mixing Magnetic Detection Setup Employing Permanent Ring Magnets as a Static Offset Field Source}, series = {Sensors}, volume = {22}, journal = {Sensors}, number = {22}, editor = {Offenh{\"a}usser, Andreas}, publisher = {MDPI}, address = {Basel}, issn = {1424-8220}, doi = {10.3390/s22228776}, pages = {12 Seiten}, year = {2022}, abstract = {Frequency mixing magnetic detection (FMMD) has been explored for its applications in fields of magnetic biosensing, multiplex detection of magnetic nanoparticles (MNP) and the determination of core size distribution of MNP samples. Such applications rely on the application of a static offset magnetic field, which is generated traditionally with an electromagnet. Such a setup requires a current source, as well as passive or active cooling strategies, which directly sets a limitation based on the portability aspect that is desired for point of care (POC) monitoring applications. In this work, a measurement head is introduced that involves the utilization of two ring-shaped permanent magnets to generate a static offset magnetic field. A steel cylinder in the ring bores homogenizes the field. By variation of the distance between the ring magnets and of the thickness of the steel cylinder, the magnitude of the magnetic field at the sample position can be adjusted. Furthermore, the measurement setup is compared to the electromagnet offset module based on measured signals and temperature behavior.}, language = {en} } @article{KarschuckKaulenPoghossianetal.2021, author = {Karschuck, Tobias and Kaulen, Corinna and Poghossian, Arshak and Wagner, Patrick H. and Sch{\"o}ning, Michael Josef}, title = {Gold nanoparticle-modified capacitive field-effect sensors: Studying the surface density of nanoparticles and coupling of charged polyelectrolyte macromolecules}, series = {Electrochemical Science Advances}, volume = {2}, journal = {Electrochemical Science Advances}, number = {5}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {0938-5193}, doi = {10.1002/elsa.202100179}, pages = {10 Seiten}, year = {2021}, abstract = {The coupling of ligand-stabilized gold nanoparticles with field-effect devices offers new possibilities for label-free biosensing. In this work, we study the immobilization of aminooctanethiol-stabilized gold nanoparticles (AuAOTs) on the silicon dioxide surface of a capacitive field-effect sensor. The terminal amino group of the AuAOT is well suited for the functionalization with biomolecules. The attachment of the positively-charged AuAOTs on a capacitive field-effect sensor was detected by direct electrical readout using capacitance-voltage and constant capacitance measurements. With a higher particle density on the sensor surface, the measured signal change was correspondingly more pronounced. The results demonstrate the ability of capacitive field-effect sensors for the non-destructive quantitative validation of nanoparticle immobilization. In addition, the electrostatic binding of the polyanion polystyrene sulfonate to the AuAOT-modified sensor surface was studied as a model system for the label-free detection of charged macromolecules. Most likely, this approach can be transferred to the label-free detection of other charged molecules such as enzymes or antibodies.}, language = {en} } @article{HoffmannRohrbachUhletal.2022, author = {Hoffmann, Andreas and Rohrbach, Felix and Uhl, Matthias and Ceblin, Maximilian and Bauer, Thomas and Mallah, Marcel and Jacob, Timo and Heuermann, Holger and Kuehne, Alexander J. C.}, title = {Atmospheric pressure plasma-jet treatment of polyacrylonitrile-nonwovens—Stabilization and roll-to-roll processing}, series = {Journal of Applied Polymer Science}, volume = {139}, journal = {Journal of Applied Polymer Science}, number = {37}, publisher = {Wiley}, issn = {0021-8995 (Print)}, doi = {10.1002/app.52887}, pages = {1 -- 9}, year = {2022}, abstract = {Carbon nanofiber nonwovens represent a powerful class of materials with prospective application in filtration technology or as electrodes with high surface area in batteries, fuel cells, and supercapacitors. While new precursor-to-carbon conversion processes have been explored to overcome productivity restrictions for carbon fiber tows, alternatives for the two-step thermal conversion of polyacrylonitrile precursors into carbon fiber nonwovens are absent. In this work, we develop a continuous roll-to-roll stabilization process using an atmospheric pressure microwave plasma jet. We explore the influence of various plasma-jet parameters on the morphology of the nonwoven and compare the stabilized nonwoven to thermally stabilized samples using scanning electron microscopy, differential scanning calorimetry, and infrared spectroscopy. We show that stabilization with a non-equilibrium plasma-jet can be twice as productive as the conventional thermal stabilization in a convection furnace, while producing electrodes of comparable electrochemical performance.}, language = {en} } @article{FayyaziSardarThomasetal.2023, author = {Fayyazi, Mojgan and Sardar, Paramjotsingh and Thomas, Sumit Infent and Daghigh, Roonak and Jamali, Ali and Esch, Thomas and Kemper, Hans and Langari, Reza and Khayyam, Hamid}, title = {Artificial intelligence/machine learning in energy management systems, control, and optimization of hydrogen fuel cell vehicles}, volume = {15}, number = {6}, publisher = {MDPI}, address = {Basel}, doi = {10.3390/su15065249}, pages = {38}, year = {2023}, abstract = {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.}, language = {en} }