TY - JOUR A1 - Schusser, Sebastian A1 - Leinhos, Marcel A1 - Bäcker, Matthias A1 - Poghossian, Arshak A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Impedance spectroscopy: A tool for real-time in situ monitoring of the degradation of biopolymers JF - Physica Status Solidi (A) N2 - Investigation of the degradation kinetics of biodegradable polymers is essential for the development of implantable biomedical devices with predicted biodegradability. In this work, an impedimetric sensor has been applied for real-time and in situ monitoring of degradation processes of biopolymers. The sensor consists of two platinum thin-film electrodes covered by a polymer film to be studied. The benchmark biomedical polymer poly(D,L-lactic acid) (PDLLA) was used as a model system. PDLLA films were deposited on the sensor structure from a polymer solution by using the spin-coating method. The degradation kinetics of PDLLA films have been studied in alkaline solutions of pH 9 and 12 by means of an impedance spectroscopy (IS) method. Any changes in a polymer capacitance/resistance induced by water uptake and/or polymer degradation will modulate the global impedance of the polymer-covered sensor that can be used as an indicator of the polymer degradation. The degradation rate can be evaluated from the time-dependent impedance spectra. As expected, a faster degradation has been observed for PDLLA films exposed to pH 12 solution. Y1 - 2013 U6 - http://dx.doi.org/10.1002/pssa.201200941 SN - 1521-396X ; 0031-8965 VL - 210 IS - 5 SP - 905 EP - 910 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Itabashi, Akinori A1 - Kosaka, Naoki A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Schöning, Michael Josef T1 - High-speed chemical imaging system based on front-side-illuminated LAPS JF - Sensors and actuators B: Chemical N2 - The chemical imaging sensor is a semiconductor-based chemical sensor that can visualize the spatial distribution of specific ions on the sensing surface. The conventional chemical imaging system based on the light-addressable potentiometric sensor (LAPS), however, required a long time to obtain a chemical image, due to the slow mechanical scan of a single light beam. For high-speed imaging, a plurality of light beams modulated at different frequencies can be employed to measure the ion concentrations simultaneously at different locations on the sensor plate by frequency division multiplex (FDM). However, the conventional measurement geometry of back-side illumination limited the bandwidth of the modulation frequency required for FDM measurement, because of the low-pass filtering characteristics of carrier diffusion in the Si substrate. In this study, a high-speed chemical imaging system based on front-side-illuminated LAPS was developed, which achieved high-speed spatiotemporal recording of pH change at a rate of 70 frames per second. Y1 - 2013 U6 - http://dx.doi.org/10.1016/j.snb.2013.03.016 SN - 1873-3077 VL - 182 SP - 315 EP - 321 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Werner, Frederik A1 - Wagner, Torsten A1 - Yoshinobu, Tatsuo A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Frequency behaviour of light-addressable potentiometric sensors JF - Physica Status Solidi (A) N2 - Light-addressable potentiometric sensors (LAPS) are semiconductor-based potentiometric sensors, with the advantage to detect the concentration of a chemical species in a liquid solution above the sensor surface in a spatially resolved manner. The addressing is achieved by a modulated and focused light source illuminating the semiconductor and generating a concentration-depending photocurrent. This work introduces a LAPS set-up that is able to monitor the electrical impedance in addition to the photocurrent. The impedance spectra of a LAPS structure, with and without illumination, as well as the frequency behaviour of the LAPS measurement are investigated. The measurements are supported by electrical equivalent circuits to explain the impedance and the LAPS-frequency behaviour. The work investigates the influence of different parameters on the frequency behaviour of the LAPS. Furthermore, the phase shift of the photocurrent, the influence of the surface potential as well as the changes of the sensor impedance will be discussed. Y1 - 2013 U6 - http://dx.doi.org/10.1002/pssa.201200929 SN - 1521-396X ; 0031-8965 VL - 210 IS - 5 SP - 884 EP - 891 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Miyamoto, Ko-ichiro A1 - Ichimura, Hiroki A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Yoshinobu, Tatsuo T1 - Chemical imaging of the concentration profile of ion diffusion in a microfluidic channel JF - Sensors and actuators. B: Chemical N2 - The chemical imaging sensor is a device to visualize the spatial distribution of chemical species based on the principle of LAPS (light-addressable potentiometric sensor), which is a field-effect chemical sensor based on semiconductor. In this study, the chemical imaging sensor has been applied to investigate the ion profile of laminar flows in a microfluidic channel. The chemical images (pH maps) were collected in a Y-shaped microfluidic channel while injecting HCl and NaCl solutions into two branches. From the chemical images, it was clearly observed that the injected solutions formed laminar flows in the channel. In addition, ion diffusion across the laminar flows was observed, and the diffusion coefficient could be derived by fitting the pH profiles to the Fick's equation. Y1 - 2013 U6 - http://dx.doi.org/10.1016/j.snb.2013.04.057 SN - 1873-3077 (E-Journal); 0925-4005 (Print) N1 - Part of special issue "Selected Papers from the 26th European Conference on Solid-State Transducers" VL - 189 SP - 240 EP - 245 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Yoshinobu, Tatsuo A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Schöning, Michael Josef T1 - Field-effect sensors combined with the scanned light pulse technique: from artificial olfactory images to chemical imaging technologies JF - Chemosensors N2 - The artificial olfactory image was proposed by Lundström et al. in 1991 as a new strategy for an electronic nose system which generated a two-dimensional mapping to be interpreted as a fingerprint of the detected gas species. The potential distribution generated by the catalytic metals integrated into a semiconductor field-effect structure was read as a photocurrent signal generated by scanning light pulses. The impact of the proposed technology spread beyond gas sensing, inspiring the development of various imaging modalities based on the light addressing of field-effect structures to obtain spatial maps of pH distribution, ions, molecules, and impedance, and these modalities have been applied in both biological and non-biological systems. These light-addressing technologies have been further developed to realize the position control of a faradaic current on the electrode surface for localized electrochemical reactions and amperometric measurements, as well as the actuation of liquids in microfluidic devices. KW - visualization KW - light-addressing technologies KW - scanned light pulse technique KW - field-effect structure KW - MOS KW - metal-oxide-semiconductor structure KW - catalytic metal KW - electronic nose KW - gas sensor KW - artificial olfactory image Y1 - 2024 U6 - http://dx.doi.org/10.3390/chemosensors12020020 SN - 2227-9040 N1 - This article belongs to the Special Issue "An Exciting Journey of Chemical Sensors and Biosensors: A Theme Issue in Honor of Professor Ingemar Lundström" Corresponding author: Tatsuo Yoshinobu, Michael J. Schöning VL - 12 IS - 2 PB - MDPI CY - Basel ER - TY - JOUR A1 - Dantism, Shahriar A1 - Röhlen, Desiree A1 - Selmer, Thorsten A1 - Wagner, Torsten A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Quantitative differential monitoring of the metabolic activity of Corynebacterium glutamicum cultures utilizing a light-addressable potentiometric sensor system JF - Biosensors and Bioelectronics Y1 - 2019 U6 - http://dx.doi.org/10.1016/j.bios.2019.111332 VL - 139 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Iken, Heiko A1 - Bronder, Thomas A1 - Goretzki, Alexander A1 - Kriesel, Jana A1 - Ahlborn, Kristina A1 - Gerlach, Frank A1 - Vonau, Winfried A1 - Zander, Willi A1 - Schubert, Jürgen A1 - Schöning, Michael Josef T1 - Development of a Combined pH- and Redox-Sensitive Bi-Electrode Glass Thin-Film Sensor JF - physica status solidi a : applications and materials sciences Y1 - 2019 U6 - http://dx.doi.org/10.1002/pssa.201900114 SN - 1862-6319 VL - 216 IS - 12 SP - 1 EP - 8 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Özsoylu, Dua A1 - Kizildag, Sefa A1 - Schöning, Michael Josef A1 - Wagner, Torsten T1 - Effect of plasma treatment on the sensor properties of a light‐addressable potentiometric sensor (LAPS) JF - physica status solidi a : applications and materials sciences N2 - A light-addressable potentiometric sensor (LAPS) is a field-effect-based (bio-) chemical sensor, in which a desired sensing area on the sensor surface can be defined by illumination. Light addressability can be used to visualize the concentration and spatial distribution of the target molecules, e.g., H+ ions. This unique feature has great potential for the label-free imaging of the metabolic activity of living organisms. The cultivation of those organisms needs specially tailored surface properties of the sensor. O2 plasma treatment is an attractive and promising tool for rapid surface engineering. However, the potential impacts of the technique are carefully investigated for the sensors that suffer from plasma-induced damage. Herein, a LAPS with a Ta2O5 pH-sensitive surface is successfully patterned by plasma treatment, and its effects are investigated by contact angle and scanning LAPS measurements. The plasma duration of 30 s (30 W) is found to be the threshold value, where excessive wettability begins. Furthermore, this treatment approach causes moderate plasma-induced damage, which can be reduced by thermal annealing (10 min at 300 °C). These findings provide a useful guideline to support future studies, where the LAPS surface is desired to be more hydrophilic by O2 plasma treatment. Y1 - 2019 U6 - http://dx.doi.org/10.1002/pssa.201900259 SN - 1862-6319 N1 - Corresponding author: Torsten Wagner VL - 216 IS - 20 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Dantism, Shahriar A1 - Röhlen, Desiree A1 - Dahmen, Markus A1 - Wagner, Torsten A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - LAPS-based monitoring of metabolic responses of bacterial cultures in a paper fermentation broth JF - Sensors and Actuators B: Chemical N2 - As an alternative renewable energy source, methane production in biogas plants is gaining more and more attention. Biomass in a bioreactor contains different types of microorganisms, which should be considered in terms of process-stability control. Metabolically inactive microorganisms within the fermentation process can lead to undesirable, time-consuming and cost-intensive interventions. Hence, monitoring of the cellular metabolism of bacterial populations in a fermentation broth is crucial to improve the biogas production, operation efficiency, and sustainability. In this work, the extracellular acidification of bacteria in a paper-fermentation broth is monitored after glucose uptake, utilizing a differential light-addressable potentiometric sensor (LAPS) system. The LAPS system is loaded with three different model microorganisms (Escherichia coli, Corynebacterium glutamicum, and Lactobacillus brevis) and the effect of the fermentation broth at different process stages on the metabolism of these bacteria is studied. In this way, different signal patterns related to the metabolic response of microorganisms can be identified. By means of calibration curves after glucose uptake, the overall extracellular acidification of bacterial populations within the fermentation process can be evaluated. Y1 - 2020 U6 - http://dx.doi.org/10.1016/j.snb.2020.128232 SN - 0925-4005 VL - 320 IS - Art. 128232 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Mennicken, Max A1 - Peter, Sophia K. A1 - Kaulen, Corinna A1 - Simon, Ulrich A1 - Karthäuser, Silvia T1 - Transport through Redox-Active Ru-Terpyridine Complexes Integrated in Single Nanoparticle Devices JF - The Journal of Physical Chemistry C N2 - Transition metal complexes are electrofunctional molecules due to their high conductivity and their intrinsic switching ability involving a metal-to-ligand charge transfer. Here, a method is presented to contact reliably a few to single redox-active Ru-terpyridine complexes in a CMOS compatible nanodevice and preserve their electrical functionality. Using hybrid materials from 14 nm gold nanoparticles (AuNP) and bis-{4′-[4-(mercaptophenyl)-2,2′:6′,2″-terpyridine]}-ruthenium(II) complexes a device size of 30² nm² inclusive nanoelectrodes is achieved. Moreover, this method bears the opportunity for further downscaling. The Ru-complex AuNP devices show symmetric and asymmetric current versus voltage curves with a hysteretic characteristic in two well separated conductance ranges. By theoretical approximations based on the single-channel Landauer model, the charge transport through the formed double-barrier tunnel junction is thoroughly analyzed and its sensibility to the molecule/metal contact is revealed. It can be verified that tunneling transport through the HOMO is the main transport mechanism while decoherent hopping transport is present to a minor extent. Y1 - 2020 U6 - http://dx.doi.org/10.1021/acs.jpcc.9b11716 SN - 1932-7455 VL - 124 IS - 8 SP - 4881 EP - 4889 PB - ACS Publications CY - Washington, DC ER - TY - JOUR A1 - Pilas, Johanna A1 - Selmer, Thorsten A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Screen-printed carbon electrodes modified with graphene oxide for the design of a reagent-free NAD+-dependent biosensor array JF - Analytical Chemistry Y1 - 2019 U6 - http://dx.doi.org/10.1021/acs.analchem.9b04481 VL - 91 IS - 23 SP - 15293 EP - 15299 PB - ACS Publications CY - Washington ER - TY - JOUR A1 - Jildeh, Zaid B. A1 - Kirchner, Patrick A1 - Oberländer, Jan A1 - Vahidpour, Farnoosh A1 - Wagner, Patrick H. A1 - Schöning, Michael Josef T1 - Development of a package-sterilization process for aseptic filling machines: A numerical approach and validation for surface treatment with hydrogen peroxide JF - Sensor and Actuators A: Physical N2 - Within the present work a sterilization process by a heated gas mixture that contains hydrogen peroxide (H₂O₂) is validated by experiments and numerical modeling techniques. The operational parameters that affect the sterilization efficacy are described alongside the two modes of sterilization: gaseous and condensed H₂O₂. Measurements with a previously developed H₂O₂ gas sensor are carried out to validate the applied H₂O₂ gas concentration during sterilization. We performed microbiological tests at different H₂O₂ gas concentrations by applying an end-point method to carrier strips, which contain different inoculation loads of Geobacillus stearothermophilus spores. The analysis of the sterilization process of a pharmaceutical glass vial is performed by numerical modeling. The numerical model combines heat- and advection-diffusion mass transfer with vapor–pressure equations to predict the location of condensate formation and the concentration of H₂O₂ at the packaging surfaces by changing the gas temperature. For a sterilization process of 0.7 s, a H₂O₂ gas concentration above 4% v/v is required to reach a log-count reduction above six. The numerical results showed the location of H₂O₂ condensate formation, which decreases with increasing sterilant-gas temperature. The model can be transferred to different gas nozzle- and packaging geometries to assure the absence of H₂O₂ residues. Y1 - 2020 U6 - http://dx.doi.org/10.1016/j.sna.2019.111691 SN - 0924-4247 VL - 303 IS - 111691 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Muschallik, Lukas A1 - Molinnus, Denise A1 - Jablonski, Melanie A1 - Kipp, Carina Ronja A1 - Bongaerts, Johannes A1 - Pohl, Martina A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Selmer, Thorsten A1 - Siegert, Petra T1 - Synthesis of α-hydroxy ketones and vicinal (R, R)-diols by Bacillus clausii DSM 8716ᵀ butanediol dehydrogenase JF - RSC Advances N2 - α-hydroxy ketones (HK) and 1,2-diols are important building blocks for fine chemical synthesis. Here, we describe the R-selective 2,3-butanediol dehydrogenase from B. clausii DSM 8716ᵀ (BcBDH) that belongs to the metal-dependent medium chain dehydrogenases/reductases family (MDR) and catalyzes the selective asymmetric reduction of prochiral 1,2-diketones to the corresponding HK and, in some cases, the reduction of the same to the corresponding 1,2-diols. Aliphatic diketones, like 2,3-pentanedione, 2,3-hexanedione, 5-methyl-2,3-hexanedione, 3,4-hexanedione and 2,3-heptanedione are well transformed. In addition, surprisingly alkyl phenyl dicarbonyls, like 2-hydroxy-1-phenylpropan-1-one and phenylglyoxal are accepted, whereas their derivatives with two phenyl groups are not substrates. Supplementation of Mn²⁺ (1 mM) increases BcBDH's activity in biotransformations. Furthermore, the biocatalytic reduction of 5-methyl-2,3-hexanedione to mainly 5-methyl-3-hydroxy-2-hexanone with only small amounts of 5-methyl-2-hydroxy-3-hexanone within an enzyme membrane reactor is demonstrated. Y1 - 2020 U6 - http://dx.doi.org/10.1039/D0RA02066D SN - 2046-2069 VL - 10 SP - 12206 EP - 12216 PB - Royal Society of Chemistry (RSC) CY - Cambridge ER - TY - JOUR A1 - Welden, Rene A1 - Schöning, Michael Josef A1 - Wagner, Patrick H. A1 - Wagner, Torsten T1 - Light-Addressable Electrodes for Dynamic and Flexible Addressing of Biological Systems and Electrochemical Reactions JF - Sensors N2 - In this review article, we are going to present an overview on possible applications of light-addressable electrodes (LAE) as actuator/manipulation devices besides classical electrode structures. For LAEs, the electrode material consists of a semiconductor. Illumination with a light source with the appropiate wavelength leads to the generation of electron-hole pairs which can be utilized for further photoelectrochemical reaction. Due to recent progress in light-projection technologies, highly dynamic and flexible illumination patterns can be generated, opening new possibilities for light-addressable electrodes. A short introduction on semiconductor–electrolyte interfaces with light stimulation is given together with electrode-design approaches. Towards applications, the stimulation of cells with different electrode materials and fabrication designs is explained, followed by analyte-manipulation strategies and spatially resolved photoelectrochemical deposition of different material types. Y1 - 2020 U6 - http://dx.doi.org/10.3390/s20061680 SN - 1424-8220 VL - 20 IS - 6 PB - MDPI CY - Basel ER - TY - JOUR A1 - Jildeh, Zaid B. A1 - Wagner, Patrick H. A1 - Schöning, Michael Josef T1 - Sterilization of Objects, Products, and Packaging Surfaces and Their Characterization in Different Fields of Industry: The Status in 2020 JF - physica status solidi (a) applications and materials science N2 - The treatment method to deactivate viable microorganisms from objects or products is termed sterilization. There are multiple forms of sterilization, each intended to be applied for a specific target, which depends on—but not limited to—the thermal, physical, and chemical stability of that target. Herein, an overview on the currently used sterilization processes in the global market is provided. Different sterilization techniques are grouped under a category that describes the method of treatment: radiation (gamma, electron beam, X-ray, and ultraviolet), thermal (dry and moist heat), and chemical (ethylene oxide, ozone, chlorine dioxide, and hydrogen peroxide). For each sterilization process, the typical process parameters as defined by regulations and the mode of antimicrobial activity are summarized. Finally, the recommended microorganisms that are used as biological indicators to validate sterilization processes in accordance with the rules that are established by various regulatory agencies are summarized. KW - bioburdens KW - sterility tests KW - sterilization efficacy KW - sterilization methods KW - validation methods Y1 - 2021 U6 - http://dx.doi.org/10.1002/pssa.202000732 SN - 1862-6319 N1 - Corresponding author: Michael J. Schöning VL - 218 IS - 13 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Molinnus, Denise A1 - Drinic, Aleksander A1 - Iken, Heiko A1 - Kröger, Nadja A1 - Zinser, Max A1 - Smeets, Ralf A1 - Köpf, Marius A1 - Kopp, Alexander A1 - Schöning, Michael Josef T1 - Towards a flexible electrochemical biosensor fabricated from biocompatible Bombyx mori silk JF - Biosensors and Bioelectronics Y1 - 2021 U6 - http://dx.doi.org/10.1016/j.bios.2021.113204 SN - 0956-5663 VL - 183 IS - Art. 113204 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Wert, Stefan A1 - Iken, Heiko A1 - Schöning, Michael Josef A1 - Matysik, Frank-Michael T1 - Development of a temperature‐pulse enhanced electrochemical glucose biosensor and characterization of its stability via scanning electrochemical microscopy JF - Electroanalysis N2 - Glucose oxidase (GOx) is an enzyme frequently used in glucose biosensors. As increased temperatures can enhance the performance of electrochemical sensors, we investigated the impact of temperature pulses on GOx that was drop-coated on flattened Pt microwires. The wires were heated by an alternating current. The sensitivity towards glucose and the temperature stability of GOx was investigated by amperometry. An up to 22-fold increase of sensitivity was observed. Spatially resolved enzyme activity changes were investigated via scanning electrochemical microscopy. The application of short (<100 ms) heat pulses was associated with less thermal inactivation of the immobilized GOx than long-term heating. Y1 - 2021 U6 - http://dx.doi.org/10.1002/elan.202100089 SN - 1521-4109 IS - Early View PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Yoshinobu, Tatsuo A1 - Schöning, Michael Josef T1 - Light-addressable potentiometric sensors (LAPS) for cell monitoring and biosensing JF - Current Opinion in Electrochemistry Y1 - 2021 U6 - http://dx.doi.org/10.1016/j.coelec.2021.100727 SN - 2451-9103 IS - In Press, Journal Pre-proof PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Givanoudi, Stella A1 - Cornelis, Peter A1 - Rasschaert, Geertrui A1 - Wackers, Gideon A1 - Iken, Heiko A1 - Rolka, David A1 - Yongabi, Derick A1 - Robbens, Johan A1 - Schöning, Michael Josef A1 - Heyndrickx, Marc A1 - Wagner, Patrick T1 - Selective Campylobacter detection and quantification in poultry: A sensor tool for detecting the cause of a common zoonosis at its source JF - Sensors and Actuators B: Chemical Y1 - 2021 U6 - http://dx.doi.org/10.1016/j.snb.2021.129484 SN - 0925-4005 IS - In Press, Journal Pre-proof SP - Article 129484 PB - Elsevier CY - Amsterdam ER - TY - THES A1 - Bronder, Thomas T1 - Label-free detection of tuberculosis DNA with capacitive field-effect biosensors Y1 - 2020 U6 - http://dx.doi.org/10.17192/z2021.0056 N1 - Dissertation, Universität, Marburg 2020 PB - Philipps-Universität Marburg CY - Marburg ER - TY - JOUR A1 - Jablonski, Melanie A1 - Münstermann, Felix A1 - Nork, Jasmina A1 - Molinnus, Denise A1 - Muschallik, Lukas A1 - Bongaerts, Johannes A1 - Wagner, Torsten A1 - Keusgen, Michael A1 - Siegert, Petra A1 - Schöning, Michael Josef T1 - Capacitive field‐effect biosensor applied for the detection of acetoin in alcoholic beverages and fermentation broths JF - physica status solidi (a) applications and materials science N2 - An acetoin biosensor based on a capacitive electrolyte–insulator–semiconductor (EIS) structure modified with the enzyme acetoin reductase, also known as butane-2,3-diol dehydrogenase (Bacillus clausii DSM 8716ᵀ), is applied for acetoin detection in beer, red wine, and fermentation broth samples for the first time. The EIS sensor consists of an Al/p-Si/SiO₂/Ta₂O₅ layer structure with immobilized acetoin reductase on top of the Ta₂O₅ transducer layer by means of crosslinking via glutaraldehyde. The unmodified and enzyme-modified sensors are electrochemically characterized by means of leakage current, capacitance–voltage, and constant capacitance methods, respectively. KW - acetoin KW - acetoin reductase KW - alcoholic beverages KW - biosensors KW - capacitive field-effect sensors Y1 - 2021 U6 - http://dx.doi.org/10.1002/pssa.202000765 SN - 1862-6319 VL - 218 IS - 13 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Jablonski, Melanie A1 - Poghossian, Arshak A1 - Severin, Robin A1 - Keusgen, Michael A1 - Wege, Christian A1 - Schöning, Michael Josef T1 - Capacitive Field-Effect Biosensor Studying Adsorption of Tobacco Mosaic Virus Particles JF - Micromachines N2 - Plant virus-like particles, and in particular, tobacco mosaic virus (TMV) particles, are increasingly being used in nano- and biotechnology as well as for biochemical sensing purposes as nanoscaffolds for the high-density immobilization of receptor molecules. The sensitive parameters of TMV-assisted biosensors depend, among others, on the density of adsorbed TMV particles on the sensor surface, which is affected by both the adsorption conditions and surface properties of the sensor. In this work, Ta₂O₅-gate field-effect capacitive sensors have been applied for the label-free electrical detection of TMV adsorption. The impact of the TMV concentration on both the sensor signal and the density of TMV particles adsorbed onto the Ta₂O₅-gate surface has been studied systematically by means of field-effect and scanning electron microscopy methods. In addition, the surface density of TMV particles loaded under different incubation times has been investigated. Finally, the field-effect sensor also demonstrates the label-free detection of penicillinase immobilization as model bioreceptor on TMV particles. KW - capacitive field-effect sensor KW - plant virus detection KW - tobacco mosaic virus (TMV) KW - TMV adsorption KW - Ta₂O₅ gate Y1 - 2021 U6 - http://dx.doi.org/10.3390/mi12010057 VL - 12 IS - 1 PB - MDPI CY - Basel ER - TY - JOUR A1 - Molinnus, Denise A1 - Iken, Heiko A1 - Johnen, Anna Lynn A1 - Richstein, Benjamin A1 - Hellmich, Lena A1 - Poghossian, Arshak A1 - Knoch, Joachim A1 - Schöning, Michael Josef T1 - Miniaturized pH-Sensitive Field-Effect Capacitors with Ultrathin Ta₂O₅ Films Prepared by Atomic Layer Deposition JF - physica status solidi (a) applications and materials science N2 - Miniaturized electrolyte–insulator–semiconductor capacitors (EISCAPs) with ultrathin gate insulators have been studied in terms of their pH-sensitive sensor characteristics: three different EISCAP systems consisting of Al–p-Si–Ta2O5(5 nm), Al–p-Si–Si3N4(1 or 2 nm)–Ta2O5 (5 nm), and Al–p-Si–SiO2(3.6 nm)–Ta2O5(5 nm) layer structures are characterized in buffer solution with different pH values by means of capacitance–voltage and constant capacitance method. The SiO2 and Si3N4 gate insulators are deposited by rapid thermal oxidation and rapid thermal nitridation, respectively, whereas the Ta2O5 film is prepared by atomic layer deposition. All EISCAP systems have a clear pH response, favoring the stacked gate insulators SiO2–Ta2O5 when considering the overall sensor characteristics, while the Si3N4(1 nm)–Ta2O5 stack delivers the largest accumulation capacitance (due to the lower equivalent oxide thickness) and a higher steepness in the slope of the capacitance–voltage curve among the studied stacked gate insulator systems. KW - atomic layer deposition KW - capacitive field-effect sensors KW - pH sensors KW - ultrathin gate insulators Y1 - 2022 U6 - http://dx.doi.org/10.1002/pssa.202100660 SN - 1862-6319 VL - 219 IS - 8 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Oliveira, Danilo A. A1 - Molinnus, Denise A1 - Beging, Stefan A1 - Siqueira Jr, José R. A1 - Schöning, Michael Josef T1 - Biosensor Based on Self-Assembled Films of Graphene Oxide and Polyaniline Using a Field-Effect Device Platform JF - physica status solidi (a) applications and materials science N2 - A new functionalization method to modify capacitive electrolyte–insulator–semiconductor (EIS) structures with nanofilms is presented. Layers of polyallylamine hydrochloride (PAH) and graphene oxide (GO) with the compound polyaniline:poly(2-acrylamido-2-methyl-1-propanesulfonic acid) (PANI:PAAMPSA) are deposited onto a p-Si/SiO2 chip using the layer-by-layer technique (LbL). Two different enzymes (urease and penicillinase) are separately immobilized on top of a five-bilayer stack of the PAH:GO/PANI:PAAMPSA-modified EIS chip, forming a biosensor for detection of urea and penicillin, respectively. Electrochemical characterization is performed by constant capacitance (ConCap) measurements, and the film morphology is characterized by atomic force microscopy (AFM) and scanning electron microscopy (SEM). An increase in the average sensitivity of the modified biosensors (EIS–nanofilm–enzyme) of around 15% is found in relation to sensors, only carrying the enzyme but without the nanofilm (EIS–enzyme). In this sense, the nanofilm acts as a stable bioreceptor onto the EIS chip improving the output signal in terms of sensitivity and stability. KW - capacitive electrolyte–insulator–semiconductor sensors KW - graphene oxide KW - layer-by-layer technique KW - nanomaterials KW - polyaniline Y1 - 2021 U6 - http://dx.doi.org/10.1002/pssa.202000747 SN - 1862-6319 N1 - Corresponding author: José R. Siqueira Jr & Michael J. Schöning VL - 218 IS - 13 SP - 1 EP - 9 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Welden, Rene A1 - Nagamine Komesu, Cindy A. A1 - Wagner, Patrick H. A1 - Schöning, Michael Josef A1 - Wagner, Torsten T1 - Photoelectrochemical enzymatic penicillin biosensor: A proof-of-concept experiment JF - Electrochemical Science Advances N2 - Photoelectrochemical (PEC) biosensors are a rather novel type of biosensors thatutilizelighttoprovideinformationaboutthecompositionofananalyte,enablinglight-controlled multi-analyte measurements. For enzymatic PEC biosensors,amperometric detection principles are already known in the literature. In con-trast, there is only a little information on H+-ion sensitive PEC biosensors. Inthis work, we demonstrate the detection of H+ions emerged by H+-generatingenzymes, exemplarily demonstrated with penicillinase as a model enzyme on atitanium dioxide photoanode. First, we describe the pH sensitivity of the sensorand study possible photoelectrocatalytic reactions with penicillin. Second, weshow the enzymatic PEC detection of penicillin. KW - enzymatic biosensor KW - penicillin KW - penicillinase KW - photoelectrochemistry KW - titanium dioxide photoanode Y1 - 2021 U6 - http://dx.doi.org/10.1002/elsa.202100131 SN - 2698-5977 N1 - Corresponding auhtor: Michael J. Schöning VL - 2 IS - 4 SP - 1 EP - 5 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Jablonski, Melanie A1 - Poghossian, Arshak A1 - Keusgen, Michael A1 - Wege, Christina A1 - Schöning, Michael Josef T1 - Detection of plant virus particles with a capacitive field-effect sensor JF - Analytical and Bioanalytical Chemistry N2 - Plant viruses are major contributors to crop losses and induce high economic costs worldwide. For reliable, on-site and early detection of plant viral diseases, portable biosensors are of great interest. In this study, a field-effect SiO2-gate electrolyte-insulator-semiconductor (EIS) sensor was utilized for the label-free electrostatic detection of tobacco mosaic virus (TMV) particles as a model plant pathogen. The capacitive EIS sensor has been characterized regarding its TMV sensitivity by means of constant-capacitance method. The EIS sensor was able to detect biotinylated TMV particles from a solution with a TMV concentration as low as 0.025 nM. A good correlation between the registered EIS sensor signal and the density of adsorbed TMV particles assessed from scanning electron microscopy images of the SiO2-gate chip surface was observed. Additionally, the isoelectric point of the biotinylated TMV particles was determined via zeta potential measurements and the influence of ionic strength of the measurement solution on the TMV-modified EIS sensor signal has been studied. KW - Plant virus KW - Capacitive field-effect sensor KW - Tobacco mosaic virus (TMV) KW - Label-free detection KW - Zeta potential Y1 - 2021 U6 - http://dx.doi.org/10.1007/s00216-021-03448-8 SN - 1618-2650 N1 - Corresponding authors: Arshak Poghossian & Michael J. Schöning VL - 413 SP - 5669 EP - 5678 PB - Springer Nature CY - Cham ER - TY - JOUR A1 - Poghossian, Arshak A1 - Karschuck, Tobias A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Field-Effect Capacitors Decorated with Ligand-Stabilized Gold Nanoparticles: Modeling and Experiments JF - Biosensors N2 - 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. KW - aminooctanethiol KW - nanoparticle coverage KW - capacitive model KW - gold nanoparticles KW - field-effect sensor KW - electrolyte-insulator-semiconductor capacitors Y1 - 2022 U6 - http://dx.doi.org/10.3390/bios12050334 SN - 2079-6374 N1 - This article belongs to the Special Issue "Biosensors in Nanotechnology" VL - 12 IS - 5 PB - MDPI CY - Basel ER - TY - JOUR A1 - Pourshahidi, Ali Mohammad A1 - Achtsnicht, Stefan A1 - Offenhäusser, Andreas A1 - Krause, Hans-Joachim ED - Offenhäusser, Andreas T1 - Frequency Mixing Magnetic Detection Setup Employing Permanent Ring Magnets as a Static Offset Field Source JF - Sensors N2 - 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. KW - magnetic sensors KW - biosensors KW - frequency mixing magnetic detection KW - magnetic nanoparticles Y1 - 2022 U6 - http://dx.doi.org/10.3390/s22228776 SN - 1424-8220 VL - 22 IS - 22 PB - MDPI CY - Basel ER - TY - JOUR A1 - Karschuck, Tobias A1 - Kaulen, Corinna A1 - Poghossian, Arshak A1 - Wagner, Patrick H. A1 - Schöning, Michael Josef T1 - Gold nanoparticle-modified capacitive field-effect sensors: Studying the surface density of nanoparticles and coupling of charged polyelectrolyte macromolecules JF - Electrochemical Science Advances N2 - 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. KW - polystyrene sulfonate KW - gold nanoparticles KW - field-effect sensor KW - detection of charged macromolecules KW - capacitive EIS sensor Y1 - 2021 U6 - http://dx.doi.org/10.1002/elsa.202100179 SN - 0938-5193 VL - 2 IS - 5 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Vahidpour, Farnoosh A1 - Guthman, Eric A1 - Arreola, Julia A1 - Alghazali, Yousef H. M. A1 - Wagner, Torsten A1 - Schöning, Michael Josef T1 - Assessment of Various Process Parameters for Optimized Sterilization Conditions Using a Multi-Sensing Platform JF - Foods N2 - In this study, an online multi-sensing platform was engineered to simultaneously evaluate various process parameters of food package sterilization using gaseous hydrogen peroxide (H₂O₂). The platform enabled the validation of critical aseptic parameters. In parallel, one series of microbiological count reduction tests was performed using highly resistant spores of B. atrophaeus DSM 675 to act as the reference method for sterility validation. By means of the multi-sensing platform together with microbiological tests, we examined sterilization process parameters to define the most effective conditions with regards to the highest spore kill rate necessary for aseptic packaging. As these parameters are mutually associated, a correlation between different factors was elaborated. The resulting correlation indicated the need for specific conditions regarding the applied H₂O₂ gas temperature, the gas flow and concentration, the relative humidity and the exposure time. Finally, the novel multi-sensing platform together with the mobile electronic readout setup allowed for the online and on-site monitoring of the sterilization process, selecting the best conditions for sterility and, at the same time, reducing the use of the time-consuming and costly microbiological tests that are currently used in the food package industry. KW - spore kill rate KW - sterility KW - aseptic parameters KW - multi-sensing platform KW - gaseous hydrogen peroxide Y1 - 2022 U6 - http://dx.doi.org/10.3390/foods11050660 SN - 2304-8158 N1 - This article belongs to the Special Issue "Sensors and Biosensors Application for Food Industries" VL - 11 IS - 5 PB - MDPI CY - Basel ER - TY - JOUR A1 - Poghossian, Arshak A1 - Welden, Rene A1 - Buniatyan, Vahe V. A1 - Schöning, Michael Josef T1 - An Array of On-Chip Integrated, Individually Addressable Capacitive Field-Effect Sensors with Control Gate: Design and Modelling JF - Sensors N2 - The on-chip integration of multiple biochemical sensors based on field-effect electrolyte-insulator-semiconductor capacitors (EISCAP) is challenging due to technological difficulties in realization of electrically isolated EISCAPs on the same Si chip. In this work, we present a new simple design for an array of on-chip integrated, individually electrically addressable EISCAPs with an additional control gate (CG-EISCAP). The existence of the CG enables an addressable activation or deactivation of on-chip integrated individual CG-EISCAPs by simple electrical switching the CG of each sensor in various setups, and makes the new design capable for multianalyte detection without cross-talk effects between the sensors in the array. The new designed CG-EISCAP chip was modelled in so-called floating/short-circuited and floating/capacitively-coupled setups, and the corresponding electrical equivalent circuits were developed. In addition, the capacitance-voltage curves of the CG-EISCAP chip in different setups were simulated and compared with that of a single EISCAP sensor. Moreover, the sensitivity of the CG-EISCAP chip to surface potential changes induced by biochemical reactions was simulated and an impact of different parameters, such as gate voltage, insulator thickness and doping concentration in Si, on the sensitivity has been discussed. KW - equivalent circuit KW - multianalyte detection KW - control gate KW - on-chip integrated addressable EISCAP sensors KW - capacitive field-effect sensor Y1 - 2021 U6 - http://dx.doi.org/10.3390/s21186161 SN - 1424-8220 N1 - This article belongs to the Special Issue "Field-Effect Sensors: From pH Sensing to Biosensing" VL - 21 IS - 18 SP - 17 PB - MDPI CY - Basel ER - TY - JOUR A1 - Pourshahidi, Ali Mohammad A1 - Achtsnicht, Stefan A1 - Nambipareechee, Mrinal Murali A1 - Offenhäusser, Andreas A1 - Krause, Hans-Joachim T1 - Multiplex detection of magnetic beads using offset field dependent frequency mixing magnetic detection JF - Sensors N2 - Magnetic immunoassays employing Frequency Mixing Magnetic Detection (FMMD) have recently become increasingly popular for quantitative detection of various analytes. Simultaneous analysis of a sample for two or more targets is desirable in order to reduce the sample amount, save consumables, and save time. We show that different types of magnetic beads can be distinguished according to their frequency mixing response to a two-frequency magnetic excitation at different static magnetic offset fields. We recorded the offset field dependent FMMD response of two different particle types at frequencies ƒ₁ + n⋅ƒ₂, n = 1, 2, 3, 4 with ƒ₁ = 30.8 kHz and ƒ₂ = 63 Hz. Their signals were clearly distinguishable by the locations of the extremes and zeros of their responses. Binary mixtures of the two particle types were prepared with different mixing ratios. The mixture samples were analyzed by determining the best linear combination of the two pure constituents that best resembled the measured signals of the mixtures. Using a quadratic programming algorithm, the mixing ratios could be determined with an accuracy of greater than 14%. If each particle type is functionalized with a different antibody, multiplex detection of two different analytes becomes feasible. KW - colorization KW - multiplex detection KW - frequency mixing magnetic detection KW - magnetic nanoparticles Y1 - 2021 U6 - http://dx.doi.org/10.3390/s21175859 SN - 1424-8220 N1 - This article belongs to the Special Issue "Advanced Nanomaterial-Based Sensors for Biomedical Applications" VL - 21 IS - 17 PB - MDPI CY - Basel ER - 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 - http://dx.doi.org/10.1002/appl.202200106 SN - 2702-4288 IS - Early View SP - 1 EP - 15 PB - Wiley-VCH ER - TY - JOUR A1 - Vahidpour, Farnoosh A1 - Alghazali, Yousef A1 - Akca, Sevilay A1 - Hommes, Gregor A1 - Schöning, Michael Josef T1 - An Enzyme-Based Interdigitated Electrode-Type Biosensor for Detecting Low Concentrations of H₂O₂ Vapor/Aerosol JF - Chemosensors N2 - 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. Y1 - 2022 U6 - http://dx.doi.org/10.3390/chemosensors10060202 SN - 2227-9040 N1 - This article belongs to the Special Issue "Bioinspired Chemical Sensors and Micro-Nano Devices" VL - 10 IS - 6 PB - MDPI CY - Basel ER - TY - JOUR A1 - Haeger, Gerrit A1 - Jolmes, Tristan A1 - Oyen, Sven A1 - Jaeger, Karl-Erich A1 - Bongaerts, Johannes A1 - Schörken, Ulrich A1 - Siegert, Petra T1 - Novel recombinant aminoacylase from Paraburkholderia monticola capable of N-acyl-amino acid synthesis JF - Applied Microbiology and Biotechnology N2 - N-Acyl-amino acids can act as mild biobased surfactants, which are used, e.g., in baby shampoos. However, their chemical synthesis needs acyl chlorides and does not meet sustainability criteria. Thus, the identification of biocatalysts to develop greener synthesis routes is desirable. We describe a novel aminoacylase from Paraburkholderia monticola DSM 100849 (PmAcy) which was identified, cloned, and evaluated for its N-acyl-amino acid synthesis potential. Soluble protein was obtained by expression in lactose autoinduction medium and co-expression of molecular chaperones GroEL/S. Strep-tag affinity purification enriched the enzyme 16-fold and yielded 15 mg pure enzyme from 100 mL of culture. Biochemical characterization revealed that PmAcy possesses beneficial traits for industrial application like high temperature and pH-stability. A heat activation of PmAcy was observed upon incubation at temperatures up to 80 °C. Hydrolytic activity of PmAcy was detected with several N-acyl-amino acids as substrates and exhibited the highest conversion rate of 773 U/mg with N-lauroyl-L-alanine at 75 °C. The enzyme preferred long-chain acyl-amino-acids and displayed hardly any activity with acetyl-amino acids. PmAcy was also capable of N-acyl-amino acid synthesis with good conversion rates. The best synthesis results were obtained with the cationic L-amino acids L-arginine and L-lysine as well as with L-leucine and L-phenylalanine. Exemplarily, L-phenylalanine was acylated with fatty acids of chain lengths from C8 to C18 with conversion rates of up to 75%. N-lauroyl-L-phenylalanine was purified by precipitation, and the structure of the reaction product was verified by LC–MS and NMR. KW - Chaperone KW - Biocatalysis KW - Aminoacylase KW - Acylation KW - Acyl-amino acids KW - Biosurfactants Y1 - 2024 U6 - http://dx.doi.org/10.1007/s00253-023-12868-8 SN - 1432-0614 N1 - Corresponding author: Petra Siegert IS - 108 PB - Springer CY - Berlin ER - TY - JOUR A1 - Haeger, Gerrit A1 - Wirges, Jessika A1 - Tanzmann, Nicole A1 - Oyen, Sven A1 - Jolmes, Tristan A1 - Jaeger, Karl-Erich A1 - Schörken, Ulrich A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Chaperone assisted recombinant expression of a mycobacterial aminoacylase in Vibrio natriegens and Escherichia coli capable of N-lauroyl-L-amino acid synthesis JF - Microbial Cell Factories N2 - 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. KW - Acyl-amino acids KW - Inclusion bodies KW - Chaperone co-expression KW - Vibrio natriegens KW - Aminoacylase Y1 - 2023 U6 - http://dx.doi.org/10.1186/s12934-023-02079-1 SN - 1475-2859 N1 - Corresponding author: Petra Siegert IS - 22 SP - Article number: 77 (2023) PB - Springer Nature ER - TY - JOUR A1 - Janus, Kevin Alexander A1 - Achtsnicht, Stefan A1 - Tempel, Laura A1 - Drinic, Aleksaner A1 - Kopp, Alexander A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Influence of fibroin membrane composition and curing parameters on the performance of a biodegradable enzymatic biosensor manufactured from Silicon-Free Carbon JF - Physica status solidi : pss. A, Applications and materials science N2 - Herein, fibroin, polylactide (PLA), and carbon are investigated for their suitability as biocompatible and biodegradable materials for amperometric biosensors. For this purpose, screen-printed carbon electrodes on the biodegradable substrates fibroin and PLA are modified with a glucose oxidase membrane and then encapsulated with the biocompatible material Ecoflex. The influence of different curing parameters of the carbon electrodes on the resulting biosensor characteristics is studied. The morphology of the electrodes is investigated by scanning electron microscopy, and the biosensor performance is examined by amperometric measurements of glucose (0.5–10 mM) in phosphate buffer solution, pH 7.4, at an applied potential of 1.2 V versus a Ag/AgCl reference electrode. Instead of Ecoflex, fibroin, PLA, and wound adhesive are tested as alternative encapsulation compounds: a series of swelling tests with different fibroin compositions, PLA, and Ecoflex has been performed before characterizing the most promising candidates by chronoamperometry. Therefore, the carbon electrodes are completely covered with the particular encapsulation material. Chronoamperometric measurements with H2O2 concentrations between 0.5 and 10 mM enable studying the leakage current behavior. KW - amperometric biosensors KW - biocompatible KW - biodegradabl KW - encapsulation materials KW - fibroin Y1 - 2023 U6 - http://dx.doi.org/10.1002/pssa.202300081 SN - 1862-6300 (Print) SN - 1862-6319 (Online) N1 - Corresponding author: Michael J. Schöning VL - 220 IS - 22 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Achtsnicht, Stefan A1 - Pourshahidi, Ali Mohammad A1 - Offenhäusser, Andreas A1 - Krause, Hans-Joachim T1 - Multiplex detection of different magnetic beads using frequency scanning in magnetic frequency mixing technique JF - Sensors N2 - In modern bioanalytical methods, it is often desired to detect several targets in one sample within one measurement. Immunological methods including those that use superparamagnetic beads are an important group of techniques for these applications. The goal of this work is to investigate the feasibility of simultaneously detecting different superparamagnetic beads acting as markers using the magnetic frequency mixing technique. The frequency of the magnetic excitation field is scanned while the lower driving frequency is kept constant. Due to the particles’ nonlinear magnetization, mixing frequencies are generated. To record their amplitude and phase information, a direct digitization of the pickup-coil’s signal with subsequent Fast Fourier Transformation is performed. By synchronizing both magnetic beads using frequency scanning in magnetic frequency mixing technique magnetic fields, a stable phase information is gained. In this research, it is shown that the amplitude of the dominant mixing component is proportional to the amount of superparamagnetic beads inside a sample. Additionally, it is shown that the phase does not show this behaviour. Excitation frequency scans of different bead types were performed, showing different phases, without correlation to their diverse amplitudes. Two commercially available beads were selected and a determination of their amount in a mixture is performed as a demonstration for multiplex measurements. KW - frequency mixing magnetic detection KW - magnetic sandwich immunoassay KW - multiparametric immunoassays Y1 - 2019 U6 - http://dx.doi.org/10.3390/s19112599 SN - 1424-8220 VL - 19 IS - 11 PB - MDPI CY - Basel ER - TY - JOUR A1 - Falkenberg, Fabian A1 - Voß, Leonie A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - New robust subtilisins from halotolerant and halophilic Bacillaceae JF - Applied Microbiology and Biotechnology N2 - The aim of the present study was the characterisation of three true subtilisins and one phylogenetically intermediate subtilisin from halotolerant and halophilic microorganisms. Considering the currently growing enzyme market for efficient and novel biocatalysts, data mining is a promising source for novel, as yet uncharacterised enzymes, especially from halophilic or halotolerant Bacillaceae, which offer great potential to meet industrial needs. Both halophilic bacteria Pontibacillus marinus DSM 16465ᵀ and Alkalibacillus haloalkaliphilus DSM 5271ᵀ and both halotolerant bacteria Metabacillus indicus DSM 16189 and Litchfieldia alkalitelluris DSM 16976ᵀ served as a source for the four new subtilisins SPPM, SPAH, SPMI and SPLA. The protease genes were cloned and expressed in Bacillus subtilis DB104. Purification to apparent homogeneity was achieved by ethanol precipitation, desalting and ion-exchange chromatography. Enzyme activity could be observed between pH 5.0–12.0 with an optimum for SPPM, SPMI and SPLA around pH 9.0 and for SPAH at pH 10.0. The optimal temperature for SPMI and SPLA was 70 °C and for SPPM and SPAH 55 °C and 50 °C, respectively. All proteases showed high stability towards 5% (w/v) SDS and were active even at NaCl concentrations of 5 M. The four proteases demonstrate potential for future biotechnological applications. KW - Biotechnological application KW - Bacillaceae KW - Subtilisin KW - Subtilases KW - Halotolerant protease Y1 - 2023 U6 - http://dx.doi.org/10.1007/s00253-023-12553-w SN - 1432-0614 N1 - Corresponding author: Petra Siegert VL - 107 SP - 3939 EP - 3954 PB - Springer Nature CY - Berlin ER - TY - JOUR A1 - Achtsnicht, Stefan A1 - Tödter, Julia A1 - Niehues, Julia A1 - Telöken, Matthias A1 - Offenhäusser, Andreas A1 - Krause, Hans-Joachim A1 - Schröper, Florian T1 - 3D printed modular immunofiltration columns for frequency mixing-based multiplex magnetic immunodetection JF - Sensors N2 - For performing point-of-care molecular diagnostics, magnetic immunoassays constitute a promising alternative to established enzyme-linked immunosorbent assays (ELISA) because they are fast, robust and sensitive. Simultaneous detection of multiple biomolecular targets from one body fluid sample is desired. The aim of this work is to show that multiplex magnetic immunodetection based on magnetic frequency mixing by means of modular immunofiltration columns prepared for different targets is feasible. By calculations of the magnetic response signal, the required spacing between the modules was determined. Immunofiltration columns were manufactured by 3D printing and antibody immobilization was performed in a batch approach. It was shown experimentally that two different target molecules in a sample solution could be individually detected in a single assaying step with magnetic measurements of the corresponding immobilization filters. The arrangement order of the filters and of a negative control did not influence the results. Thus, a simple and reliable approach to multi-target magnetic immunodetection was demonstrated. Y1 - 2019 U6 - http://dx.doi.org/10.3390/s19010148 SN - 1424-8220 VL - 19 IS - 1 PB - MDPI CY - Basel ER - TY - JOUR A1 - Achtsnicht, Stefan A1 - Schönenborn, Kristina A1 - Offenhäusser, Andreas A1 - Krause, Hans-Joachim T1 - Measurement of the magnetophoretic velocity of different superparamagnetic beads JF - Journal of Magnetism and Magnetic Materials N2 - The movement of magnetic beads due to a magnetic field gradient is of great interest in different application fields. In this report we present a technique based on a magnetic tweezers setup to measure the velocity factor of magnetically actuated individual superparamagnetic beads in a fluidic environment. Several beads can be tracked simultaneously in order to gain and improve statistics. Furthermore we show our results for different beads with hydrodynamic diameters between 200 and 1000 nm from diverse manufacturers. These measurement data can, for example, be used to determine design parameters for a magnetic separation system, like maximum flow rate and minimum separation time, or to select suitable beads for fixed experimental requirements. KW - magnetophoretic velocity KW - superparamagnetic bead KW - magnetic tweezers KW - magnetic separation KW - magnetic actuation Y1 - 2019 U6 - http://dx.doi.org/10.1016/j.jmmm.2018.10.066 SN - 0304-8853 VL - 477 IS - 1 SP - 244 EP - 248 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Rabehi, Amine A1 - Garlan, Benjamin A1 - Achtsnicht, Stefan A1 - Krause, Hans-Joachim A1 - Offenhäusser, Andreas A1 - Ngo, Kieu A1 - Neveu, Sophie A1 - Graff-Dubois, Stephanie A1 - Kokabi, Hamid T1 - Magnetic detection structure for Lab-on-Chip applications based on the frequency mixing technique JF - Sensors N2 - A magnetic frequency mixing technique with a set of miniaturized planar coils was investigated for use with a completely integrated Lab-on-Chip (LoC) pathogen sensing system. The system allows the detection and quantification of superparamagnetic beads. Additionally, in terms of magnetic nanoparticle characterization ability, the system can be used for immunoassays using the beads as markers. Analytical calculations and simulations for both excitation and pick-up coils are presented; the goal was to investigate the miniaturization of simple and cost-effective planar spiral coils. Following these calculations, a Printed Circuit Board (PCB) prototype was designed, manufactured, and tested for limit of detection, linear response, and validation of theoretical concepts. Using the magnetic frequency mixing technique, a limit of detection of 15 µg/mL of 20 nm core-sized nanoparticles was achieved without any shielding. KW - Lab-on-Chip KW - magnetic sensing KW - frequency mixing KW - superparamagnetic nanoparticles KW - magnetic beads Y1 - 2018 U6 - http://dx.doi.org/10.3390/s18061747 SN - 1424-8220 VL - 18 IS - 6 PB - MDPI CY - Basel ER - TY - JOUR A1 - Bertz, Morten A1 - Molinnus, Denise A1 - Schöning, Michael Josef A1 - Homma, Takayuki T1 - Real-time monitoring of H₂O₂ sterilization on individual bacillus atrophaeus spores by optical sensing with trapping Raman spectroscopy JF - Chemosensors N2 - 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. KW - DPA (dipicolinic acid) KW - sterilization KW - Bacillus atrophaeus spores KW - optical trapping KW - Raman spectroscopy KW - optical sensor setup Y1 - 2023 U6 - http://dx.doi.org/10.3390/chemosensors11080445 SN - 2227-9040 N1 - This article belongs to the Special Issue "Biosensors and Chemical Sensors for Food and Healthcare Monitoring—Celebrating the 10th Anniversary" VL - 8 IS - 11 PB - MDPI CY - Basel ER - TY - JOUR A1 - Wendlandt, Tim A1 - Koch, Claudia A1 - Britz, Beate A1 - Liedek, Anke A1 - Schmidt, Nora A1 - Werner, Stefan A1 - Gleba, Yuri A1 - Vahidpour, Farnoosh A1 - Welden, Melanie A1 - Poghossian, Arshak A1 - Schöning, Michael Josef T1 - Facile Purification and Use of Tobamoviral Nanocarriers for Antibody-Mediated Display of a Two-Enzyme System JF - Viruses N2 - 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. KW - biosensor KW - horseradish peroxidase (HRP) KW - glucose oxidase (GOx) KW - enzyme cascade KW - turnip vein clearing virus (TVCV) KW - tobacco mosaic virus (TMV) Y1 - 2023 U6 - http://dx.doi.org/doi.org/10.3390/v15091951 SN - 1999-4915 N1 - This article belongs to the Special Issue "Tobamoviruses 2023" VL - 9 IS - 15 PB - MDPI CY - Basel ER - TY - JOUR A1 - Falkenberg, Fabian A1 - Kohn, Sophie A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Biochemical characterisation of a novel broad pH spectrum subtilisin from Fictibacillus arsenicus DSM 15822ᵀ JF - FEBS Open Bio N2 - Subtilisins from microbial sources, especially from the Bacillaceae family, are of particular interest for biotechnological applications and serve the currently growing enzyme market as efficient and novel biocatalysts. Biotechnological applications include use in detergents, cosmetics, leather processing, wastewater treatment and pharmaceuticals. To identify a possible candidate for the enzyme market, here we cloned the gene of the subtilisin SPFA from Fictibacillus arsenicus DSM 15822ᵀ (obtained through a data mining-based search) and expressed it in Bacillus subtilis DB104. After production and purification, the protease showed a molecular mass of 27.57 kDa and a pI of 5.8. SPFA displayed hydrolytic activity at a temperature optimum of 80 °C and a very broad pH optimum between 8.5 and 11.5, with high activity up to pH 12.5. SPFA displayed no NaCl dependence but a high NaCl tolerance, with decreasing activity up to concentrations of 5 m NaCl. The stability enhanced with increasing NaCl concentration. Based on its substrate preference for 10 synthetic peptide 4-nitroanilide substrates with three or four amino acids and its phylogenetic classification, SPFA can be assigned to the subgroup of true subtilisins. Moreover, SPFA exhibited high tolerance to 5% (w/v) SDS and 5% H₂O₂ (v/v). The biochemical properties of SPFA, especially its tolerance of remarkably high pH, SDS and H₂O₂, suggest it has potential for biotechnological applications. KW - Bacillaceae KW - Biotechnological application KW - Broad pH spectrum KW - Subtilases KW - Subtilisin Y1 - 2023 U6 - http://dx.doi.org/10.1002/2211-5463.13701 SN - 2211-5463 N1 - Corresponding author: Petra Siegert VL - 13 IS - 11 SP - 2035 EP - 2046 PB - Wiley CY - Hoboken, NJ ER - TY - JOUR A1 - Morais, Paulo V. A1 - Suman, Pedro H. A1 - Schöning, Michael Josef A1 - Siqueira Junior, José R. A1 - Orlandi, Marcelo O. T1 - Layer-by-layer film based on Sn₃O₄ nanobelts as sensing units to detect heavy metals using a capacitive field-effect sensor platform JF - Chemosensors N2 - Lead and nickel, as heavy metals, are still used in industrial processes, and are classified as “environmental health hazards” due to their toxicity and polluting potential. The detection of heavy metals can prevent environmental pollution at toxic levels that are critical to human health. In this sense, the electrolyte–insulator–semiconductor (EIS) field-effect sensor is an attractive sensing platform concerning the fabrication of reusable and robust sensors to detect such substances. This study is aimed to fabricate a sensing unit on an EIS device based on Sn₃O₄ nanobelts embedded in a polyelectrolyte matrix of polyvinylpyrrolidone (PVP) and polyacrylic acid (PAA) using the layer-by-layer (LbL) technique. The EIS-Sn₃O₄ sensor exhibited enhanced electrochemical performance for detecting Pb²⁺ and Ni²⁺ ions, revealing a higher affinity for Pb²⁺ ions, with sensitivities of ca. 25.8 mV/decade and 2.4 mV/decade, respectively. Such results indicate that Sn₃O₄ nanobelts can contemplate a feasible proof-of-concept capacitive field-effect sensor for heavy metal detection, envisaging other future studies focusing on environmental monitoring. KW - Sn₃O₄ KW - nanobelts KW - field-effect sensor KW - LbL films KW - heavy metals Y1 - 2023 U6 - http://dx.doi.org/10.3390/chemosensors11080436 SN - 2227-9040 N1 - This article belongs to the Special Issue The Application of Electrochemical Sensors or Biosensors Based on Nanomaterials VL - 11 IS - 8 PB - MDPI CY - Basel ER - TY - JOUR A1 - Janus, Kevin Alexander A1 - Achtsnicht, Stefan A1 - Drinic, Aleksander A1 - Kopp, Alexander A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Transient magnesium-based thin-film temperature sensor on a flexible, bioabsorbable substrate for future medical applications JF - Applied Research N2 - In this work, the bioabsorbable materials, namely fibroin, polylactide acid (PLA), magnesium and magnesium oxide are investigated for their application as transient, resistive temperature detectors (RTD). For this purpose, a thin-film magnesium-based meander-like electrode is deposited onto a flexible, bioabsorbable substrate (fibroin or PLA) and encapsulated (passivated) by additional magnesium oxide layers on top and below the magnesium-based electrode. The morphology of different layered RTDs is analyzed by scanning electron microscopy. The sensor performance and lifetime of the RTD is characterized both under ambient atmospheric conditions between 30°C and 43°C, and wet tissue-like conditions with a constant temperature regime of 37°C. The latter triggers the degradation process of the magnesium-based layers. The 3-layers RTDs on a PLA substrate could achieve a lifetime of 8.5 h. These sensors also show the best sensor performance under ambient atmospheric conditions with a mean sensitivity of 0.48 Ω/°C ± 0.01 Ω/°C. KW - Silk fibroin KW - Polylactide acid KW - Bioabsorbable KW - Resistive temperature detector Y1 - 2023 U6 - http://dx.doi.org/10.1002/appl.202300102 SN - 2702-4288 (Print) N1 - Corresponding author: Michael Josef Schöning IS - Accepted manuscript PB - Wiley-VCH ER - TY - JOUR A1 - Karschuck, Tobias A1 - Schmidt, Stefan A1 - Achtsnicht, Stefan A1 - Poghossian, Arshak A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Multiplexing system for automated characterization of a capacitive field-effect sensor array JF - Physica Status Solidi A N2 - In comparison to single-analyte devices, multiplexed systems for a multianalyte detection offer a reduced assay time and sample volume, low cost, and high throughput. Herein, a multiplexing platform for an automated quasi-simultaneous characterization of multiple (up to 16) capacitive field-effect sensors by the capacitive–voltage (C–V) and the constant-capacitance (ConCap) mode is presented. The sensors are mounted in a newly designed multicell arrangement with one common reference electrode and are electrically connected to the impedance analyzer via the base station. A Python script for the automated characterization of the sensors executes the user-defined measurement protocol. The developed multiplexing system is tested for pH measurements and the label-free detection of ligand-stabilized, charged gold nanoparticles. KW - Capacitive field-effect sensor KW - Gold nanoparticles KW - Label-free detection KW - Multicell KW - Multiplexing Y1 - 2023 U6 - http://dx.doi.org/10.1002/pssa.202300265 SN - 1862-6300 (Print) SN - 1862-6319 (Online) N1 - Corresponding author: Michael Josef Schöning VL - 220 IS - 22 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Haeger, Gerrit A1 - Probst, Johanna A1 - Jaeger, Karl-Erich A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Novel aminoacylases from Streptomyces griseus DSM 40236 and their recombinant production in Streptomyces lividans JF - FEBS Open Bio N2 - 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. KW - Streptomyces lividans KW - recombinant expression KW - Streptomyces griseus KW - ε-lysine acylase KW - α-aminoacylase Y1 - 2023 U6 - http://dx.doi.org/10.1002/2211-5463.13723 SN - 2211-5463 N1 - Corresponding author: Petra Siegert VL - 13 IS - 12 SP - 2224 EP - 2238 PB - Wiley CY - Hoboken, NJ ER - TY - JOUR A1 - Karschuck, Tobias A1 - Poghossian, Arshak A1 - Ser, Joey A1 - Tsokolakyan, Astghik A1 - Achtsnicht, Stefan A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Capacitive model of enzyme-modified field-effect biosensors: Impact of enzyme coverage JF - Sensors and Actuators B: Chemical N2 - Electrolyte-insulator-semiconductor capacitors (EISCAP) belong to field-effect sensors having an attractive transducer architecture for constructing various biochemical sensors. In this study, a capacitive model of enzyme-modified EISCAPs has been developed and the impact of the surface coverage of immobilized enzymes on its capacitance-voltage and constant-capacitance characteristics was studied theoretically and experimentally. The used multicell arrangement enables a multiplexed electrochemical characterization of up to sixteen EISCAPs. Different enzyme coverages have been achieved by means of parallel electrical connection of bare and enzyme-covered single EISCAPs in diverse combinations. As predicted by the model, with increasing the enzyme coverage, both the shift of capacitance-voltage curves and the amplitude of the constant-capacitance signal increase, resulting in an enhancement of analyte sensitivity of the EISCAP biosensor. In addition, the capability of the multicell arrangement with multi-enzyme covered EISCAPs for sequentially detecting multianalytes (penicillin and urea) utilizing the enzymes penicillinase and urease has been experimentally demonstrated and discussed. KW - Field-effect biosensor KW - Capacitive model KW - Enzyme coverage KW - Multianalyte detection KW - Penicillin Y1 - 2024 U6 - http://dx.doi.org/10.1016/j.snb.2024.135530 SN - 0925-4005 (Print) SN - 1873-3077 (Online) N1 - Corresponding Author: Michael J. Schöning VL - 408 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Degering, Christian A1 - Eggert, Thorsten A1 - Puls, Michael A1 - Bongaerts, Johannes A1 - Evers, Stefan A1 - Maurer, Karl-Heinz A1 - Jaeger, Karl-Erich T1 - Optimization of protease secretion in Bacillus subtilis and Bacillus licheniformis by screening of homologous and herologous signal peptides JF - Applied and environmental microbiology N2 - Bacillus subtilis and Bacillus licheniformis are widely used for the large-scale industrial production of proteins. These strains can efficiently secrete proteins into the culture medium using the general secretion (Sec) pathway. A characteristic feature of all secreted proteins is their N-terminal signal peptides, which are recognized by the secretion machinery. Here, we have studied the production of an industrially important secreted protease, namely, subtilisin BPN′ from Bacillus amyloliquefaciens. One hundred seventy-three signal peptides originating from B. subtilis and 220 signal peptides from the B. licheniformis type strain were fused to this secretion target and expressed in B. subtilis, and the resulting library was analyzed by high-throughput screening for extracellular proteolytic activity. We have identified a number of signal peptides originating from both organisms which produced significantly increased yield of the secreted protease. Interestingly, we observed that levels of extracellular protease were improved not only in B. subtilis, which was used as the screening host, but also in two different B. licheniformis strains. To date, it is impossible to predict which signal peptide will result in better secretion and thus an improved yield of a given extracellular target protein. Our data show that screening a library consisting of homologous and heterologous signal peptides fused to a target protein can identify more-effective signal peptides, resulting in improved protein export not only in the original screening host but also in different production strains. Y1 - 2010 U6 - http://dx.doi.org/10.1128/AEM.01146-10 SN - 1098-5336 (E-Journal); 0003-6919 (Print); 0099-2240 (Print) VL - 76 IS - 19 SP - 6370 EP - 6378 PB - American Society for Microbiology CY - Washington, DC ER - TY - JOUR A1 - Achtsnicht, Stefan A1 - Neuendorf, Christian A1 - Faßbender, Tobias A1 - Nölke, Greta A1 - Offenhäusser, Andreas A1 - Krause, Hans-Joachim A1 - Schröper, Florian T1 - Sensitive and rapid detection of cholera toxin subunit B using magnetic frequency mixing detection JF - Plos One N2 - Cholera is a life-threatening disease caused by the cholera toxin (CT) as produced by some Vibrio cholerae serogroups. In this research we present a method which directly detects the toxin’s B subunit (CTB) in drinking water. For this purpose we performed a magnetic sandwich immunoassay inside a 3D immunofiltration column. We used two different commercially available antibodies to capture CTB and for binding to superparamagnetic beads. ELISA experiments were performed to select the antibody combination. The beads act as labels for the magnetic frequency mixing detection technique. We show that the limit of detection depends on the type of magnetic beads. A nonlinear Hill curve was fitted to the calibration measurements by means of a custom-written python software. We achieved a sensitive and rapid detection of CTB within a broad concentration range from 0.2 ng/ml to more than 700 ng/ml. Y1 - 2019 U6 - http://dx.doi.org/10.1371/journal.pone.0219356 SN - 1932-6203 VL - 14 IS - 7 PB - Plos CY - San Francisco ER - TY - JOUR A1 - Schroeter, Rebecca A1 - Hoffmann, Tamara A1 - Voigt, Birgit A1 - Meyer, Hanna A1 - Bleisteiner, Monika A1 - Muntel, Jan A1 - Jürgen, Britta A1 - Albrecht, Dirk A1 - Becher, Dörte A1 - Lalk, Michael A1 - Evers, Stefan A1 - Bongaerts, Johannes A1 - Maurer, Karl-Heinz A1 - Putzer, Harald A1 - Hecker, Michael A1 - Schweder, Thomas A1 - Bremer, Erhard T1 - Stress responses of the industrial workhorse Bacillus licheniformis to osmotic challenges JF - PLoS ONE N2 - The Gram-positive endospore-forming bacterium Bacillus licheniformis can be found widely in nature and it is exploited in industrial processes for the manufacturing of antibiotics, specialty chemicals, and enzymes. Both in its varied natural habitats and in industrial settings, B. licheniformis cells will be exposed to increases in the external osmolarity, conditions that trigger water efflux, impair turgor, cause the cessation of growth, and negatively affect the productivity of cell factories in biotechnological processes. We have taken here both systems-wide and targeted physiological approaches to unravel the core of the osmostress responses of B. licheniformis. Cells were suddenly subjected to an osmotic upshift of considerable magnitude (with 1 M NaCl), and their transcriptional profile was then recorded in a time-resolved fashion on a genome-wide scale. A bioinformatics cluster analysis was used to group the osmotically up-regulated genes into categories that are functionally associated with the synthesis and import of osmostress-relieving compounds (compatible solutes), the SigB-controlled general stress response, and genes whose functional annotation suggests that salt stress triggers secondary oxidative stress responses in B. licheniformis. The data set focusing on the transcriptional profile of B. licheniformis was enriched by proteomics aimed at identifying those proteins that were accumulated by the cells through increased biosynthesis in response to osmotic stress. Furthermore, these global approaches were augmented by a set of experiments that addressed the synthesis of the compatible solutes proline and glycine betaine and assessed the growth-enhancing effects of various osmoprotectants. Combined, our data provide a blueprint of the cellular adjustment processes of B. licheniformis to both sudden and sustained osmotic stress. Y1 - 2014 U6 - http://dx.doi.org/10.1371/journal.pone.0080956 SN - 1932-6203 VL - 8 IS - 11 PB - PLOS CY - San Francisco ER - TY - JOUR A1 - Almajhdi, Fahad N. A1 - Senger, Tilo A1 - Amer, Haitham M. A1 - Gissmann, Lutz A1 - Öhlschläger, Peter T1 - Design of a highly effective therapeutic HPV16 E6/E7-specific DNA vaccine: optimization by different ways of sequence rearrangements (Shuffling) JF - PLOS one N2 - Persistent infection with the high-risk Human Papillomavirus type 16 (HPV 16) is the causative event for the development of cervical cancer and other malignant tumors of the anogenital tract and of the head and neck. Despite many attempts to develop therapeutic vaccines no candidate has entered late clinical trials. An interesting approach is a DNA based vaccine encompassing the nucleotide sequence of the E6 and E7 viral oncoproteins. Because both proteins are consistently expressed in HPV infected cells they represent excellent targets for immune therapy. Here we report the development of 8 DNA vaccine candidates consisting of differently rearranged HPV-16 E6 and E7 sequences within one molecule providing all naturally occurring epitopes but supposedly lacking transforming activity. The HPV sequences were fused to the J-domain and the SV40 enhancer in order to increase immune responses. We demonstrate that one out of the 8 vaccine candidates induces very strong cellular E6- and E7- specific cellular immune responses in mice and, as shown in regression experiments, efficiently controls growth of HPV 16 positive syngeneic tumors. This data demonstrates the potential of this vaccine candidate to control persistent HPV 16 infection that may lead to malignant disease. It also suggests that different sequence rearrangements influence the immunogenecity by an as yet unknown mechanism. Y1 - 2014 U6 - http://dx.doi.org/10.1371/journal.pone.0113461 SN - 1932-6203 VL - 11 IS - 9 PB - PLOS CY - San Francisco ER - TY - JOUR A1 - Gasparyan, F. V. A1 - Poghossian, Arshak A1 - Vitusevich, S. A. A1 - Petrychuk, M. V. A1 - Sydoruk, V. A. A1 - Surmalyan, A. V. A1 - Siqueira, J. R. A1 - Oliveira, O. N. A1 - Offenhäusser, A. A1 - Schöning, Michael Josef T1 - Low Frequency Noise In Electrolyte-Gate Field-Effect Devices Functionalized With Dendrimer/Carbon-Nanotube Multilayers JF - Noise and fluctuations : 20th International Conference on Noise and Fluctuations, ICNF 2009, Pisa, Italy, 14 - 19 June 2009 / ed. Massimo Macucci; Giovanni Basso Y1 - 2009 SN - 9780735406650 N1 - AIP conference proceedings ; 1129 ; International Conference on Noise and Fluctuations ; (20, 2009, Pisa) SP - 133 EP - 136 PB - American Inst. of Physics CY - Melville, NY ER -