TY - JOUR A1 - Jildeh, Zaid B. A1 - Oberländer, Jan A1 - Kirchner, Patrick A1 - Wagner, Patrick H. A1 - Schöning, Michael Josef T1 - Thermocatalytic Behavior of Manganese (IV) Oxide as Nanoporous Material on the Dissociation of a Gas Mixture Containing Hydrogen Peroxide JF - Nanomaterials N2 - In this article, we present an overview on the thermocatalytic reaction of hydrogen peroxide (H₂O₂) gas on a manganese (IV) oxide (MnO₂) catalytic structure. The principle of operation and manufacturing techniques are introduced for a calorimetric H₂O₂ gas sensor based on porous MnO₂. Results from surface analyses by X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM) of the catalytic material provide indication of the H₂O₂ dissociation reaction schemes. The correlation between theory and the experiments is documented in numerical models of the catalytic reaction. The aim of the numerical models is to provide further information on the reaction kinetics and performance enhancement of the porous MnO₂ catalyst. Y1 - 2018 U6 - http://dx.doi.org/10.3390/nano8040262 SN - 2079-4991 VL - 8 IS - 4 PB - MDPI CY - Basel ER - TY - JOUR A1 - Jildeh, Zaid B. A1 - Oberländer, Jan A1 - Kirchner, Patrick A1 - Keusgen, Michael A1 - Wagner, Patrick H. A1 - Schöning, Michael Josef T1 - Experimental and Numerical Analyzes of a Sensor Based on Interdigitated Electrodes for Studying Microbiological Alterations JF - physica status solidi (a): applications and materials science N2 - In this work, a cell-based biosensor to evaluate the sterilization efficacy of hydrogen peroxide vapor sterilization processes is characterized. The transducer of the biosensor is based on interdigitated gold electrodes fabricated on an inert glass substrate. Impedance spectroscopy is applied to evaluate the sensor behavior and the alteration of test microorganisms due to the sterilization process. These alterations are related to changes in relative permittivity and electrical conductivity of the bacterial spores. Sensor measurements are conducted with and without bacterial spores (Bacillus atrophaeus), as well as after an industrial sterilization protocol. Equivalent two-dimensional numerical models based on finite element method of the periodic finger structures of the interdigitated gold electrodes are designed and validated using COMSOL® Multiphysics software by the application of known dielectric properties. The validated models are used to compute the electrical properties at different sensor states (blank, loaded with spores, and after sterilization). As a final result, we will derive and tabulate the frequency-dependent electrical parameters of the spore layer using a novel model that combines experimental data with numerical optimization techniques. Y1 - 2018 U6 - http://dx.doi.org/10.1002/pssa.201700920 SN - 1862-6319 VL - 215 IS - 15 PB - Wiley-VCH CY - Weinheim 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 - Jildeh, Zaid B. A1 - Kirchner, Patrick A1 - Oberländer, Jan A1 - Kremers, Alexander A1 - Wagner, Torsten A1 - Wagner, Patrick H. A1 - Schöning, Michael Josef T1 - FEM-based modeling of a calorimetric gas sensor for hydrogen peroxide monitoring JF - physica status solidi a : applications and materials sciences N2 - A physically coupled finite element method (FEM) model is developed to study the response behavior of a calorimetric gas sensor. The modeled sensor serves as a monitoring device of the concentration of gaseous hydrogen peroxide (H2 O2) in a high temperature mixture stream in aseptic sterilization processes. The principle of operation of a calorimetric H2 O2 sensor is analyzed and the results of the numerical model have been validated by using previously published sensor experiments. The deviation in the results between the FEM model and experimental data are presented and discussed. Y1 - 2017 U6 - http://dx.doi.org/10.1002/pssa.201600912 SN - 1862-6319 IS - Early View PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Jildeh, Zaid B. A1 - Kirchner, Patrick A1 - Baltes, Klaus A1 - Wagner, Patrick H. A1 - Schöning, Michael Josef T1 - Development of an in-line evaporation unit for the production of gas mixtures containing hydrogen peroxide – numerical modeling and experimental results JF - International Journal of Heat and Mass Transfer N2 - Hydrogen peroxide (H2O2) is a typical surface sterilization agent for packaging materials used in the pharmaceutical, food and beverage industries. We use the finite-elements method to analyze the conceptual design of an in-line thermal evaporation unit to produce a heated gas mixture of air and evaporated H2O2 solution. For the numerical model, the required phase-transition variables of pure H2O2 solution and of the aerosol mixture are acquired from vapor-liquid equilibrium (VLE) diagrams derived from vapor-pressure formulations. This work combines homogeneous single-phase turbulent flow with heat-transfer physics to describe the operation of the evaporation unit. We introduce the apparent heat-capacity concept to approximate the non-isothermal phase-transition process of the H2O2-containing aerosol. Empirical and analytical functions are defined to represent the temperature- and pressure-dependent material properties of the aqueous H2O2 solution, the aerosol and the gas mixture. To validate the numerical model, the simulation results are compared to experimental data on the heating power required to produce the gas mixture. This shows good agreement with the deviations below 10%. Experimental observations on the formation of deposits due to the evaporation of stabilized H2O2 solution fits the prediction made from simulation results. Y1 - 2019 U6 - http://dx.doi.org/10.1016/j.ijheatmasstransfer.2019.118519 SN - 0017-9310 VL - 143 PB - Elsevier CY - Amsterdam 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 - 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 - 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 - 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 - 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 - 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 - Ismail, A.B.M. A1 - Harada, T. A1 - Yoshinobu, T. A1 - Iwasaki, H. A1 - Schöning, Michael Josef A1 - Lüth, H. T1 - Investigation of pulsed laser-deposited Al2O3 as a high pH-sensitive layer for LAPS-based biosensing applications JF - Sensors and Actuators B. 71 (2000), H. 3 Y1 - 2000 SN - 0925-4005 SP - 169 EP - 172 ER - TY - JOUR A1 - Ingebrandt, S. A1 - Han, Y. A1 - Nakamura, F. A1 - Poghossian, Arshak A1 - Schöning, Michael Josef A1 - Offenhäusser, A. T1 - Label-free detection of single nucleotide polymorphisms utilizing the differential transfer function of field-effect transistors JF - Biosensors and Bioelectronics. 22 (2007), H. 12 Y1 - 2007 SN - 0956-5663 SP - 2834 EP - 2840 ER - TY - JOUR A1 - Immel, Timo A1 - Grützke, Martin A1 - Späte, Anne-Katrin A1 - Groth, Ulrich A1 - Öhlschläger, Peter A1 - Huhn, Thomas T1 - Synthesis and X-ray structure analysis of a heptacoordinate titanium(IV)-bis-chelate with enhanced in vivo antitumor efficacy JF - Chemical Communications N2 - Chelate stabilization of a titanium(IV)–salan alkoxide by ligand exchange with 2,6-pyridinedicarboxylic acid (dipic) resulted in heptacoordinate complex 3 which is not redox-active, stable on silica gel and has increased aqueous stability. 3 is highly toxic in HeLa S3 and Hep G2 and has enhanced antitumor efficacy in a mouse cervical-cancer model. Y1 - 2012 U6 - http://dx.doi.org/10.1039/C2CC31624B SN - 1364-548X VL - 48 IS - 46 SP - 5790 EP - 5792 PB - Royal Society of Chemistry CY - Cambridge ER - TY - JOUR A1 - Iken, Heiko A1 - Kirsanov, D. A1 - Legin, A. A1 - Schöning, Michael Josef T1 - Novel Thin-Film Polymeric Materials for the Detection of Heavy Metals JF - Procedia Engineering N2 - A variety of transition metals, e.g., copper, zinc, cadmium, lead, etc. are widely used in industry as components for wires, coatings, alloys, batteries, paints and so on. The inevitable presence of transition metals in industrial processes implies the ambition of developing a proper analytical technique for their adequate monitoring. Most of these elements, especially lead and cadmium, are acutely toxic for biological organisms. Quantitative determination of these metals at low activity levels in different environmental and industrial samples is therefore a vital task. A promising approach to achieve an at-side or on-line monitoring on a miniaturized and cost efficient way is the combination of a common potentiometric sensor array with heavy metal-sensitive thin-film materials, like chalcogenide glasses and polymeric materials, respectively. Y1 - 2012 U6 - http://dx.doi.org/10.1016/j.proeng.2012.09.148 SN - 1877-7058 N1 - Part of special issue "26th European Conference on Solid-State Transducers, EUROSENSOR 2012" IS - 47 SP - 322 EP - 325 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 - Iken, Heiko A1 - Ahlborn, K. A1 - Gerlach, F. A1 - Vonau, W. A1 - Zander, W. A1 - Schubert, J. A1 - Schöning, Michael Josef T1 - Development of redox glasses and subsequent processing by means of pulsed laser deposition for realizing silicon-based thin-film sensors JF - Electrochimica acta Y1 - 2013 SN - 1873-3859 (E-Journal); 0013-4686 (Print) SP - Available online 30.8.2013 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Huck, Christina A1 - Schiffels, Johannes A1 - Herrera, Cony N. A1 - Schelden, Maximilian A1 - Selmer, Thorsten A1 - Poghossian, Arshak A1 - Baumann, Marcus A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Metabolic responses of Escherichia coli upon glucose pulses captured by a capacitive field-effect sensor JF - Physica Status Solidi (A) N2 - Living cells are complex biological systems transforming metabolites taken up from the surrounding medium. Monitoring the responses of such cells to certain substrate concentrations is a challenging task and offers possibilities to gain insight into the vitality of a community influenced by the growth environment. Cell-based sensors represent a promising platform for monitoring the metabolic activity and thus, the “welfare” of relevant organisms. In the present study, metabolic responses of the model bacterium Escherichia coli in suspension, layered onto a capacitive field-effect structure, were examined to pulses of glucose in the concentration range between 0.05 and 2 mM. It was found that acidification of the surrounding medium takes place immediately after glucose addition and follows Michaelis–Menten kinetic behavior as a function of the glucose concentration. In future, the presented setup can, therefore, be used to study substrate specificities on the enzymatic level and may as well be used to perform investigations of more complex metabolic responses. Conclusions and perspectives highlighting this system are discussed. Y1 - 2013 U6 - http://dx.doi.org/10.1002/pssa.201200900 SN - 0031-8965 VL - 210 IS - 5 SP - 926 EP - 931 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Huck, Christina A1 - Poghossian, Arshak A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Combined amperometric/field-effect sensor for the detection of dissolved hydrogen JF - Sensors and actuators B: Chemical N2 - Real-time and reliable monitoring of the biogas process is crucial for a stable and efficient operation of biogas production in order to avoid digester breakdowns. The concentration of dissolved hydrogen (H₂) represents one of the key parameters for biogas process control. In this work, a one-chip integrated combined amperometric/field-effect sensor for monitoring the dissolved H₂ concentration has been developed for biogas applications. The combination of two different transducer principles might allow a more accurate and reliable measurement of dissolved H₂ as an early warning indicator of digester failures. The feasibility of the approach has been demonstrated by simultaneous amperometric/field-effect measurements of dissolved H₂ concentrations in electrolyte solutions. Both, the amperometric and the field-effect transducer show a linear response behaviour in the H₂ concentration range from 0.1 to 3% (v/v) with a slope of 198.4 ± 13.7 nA/% (v/v) and 14.9 ± 0.5 mV/% (v/v), respectively. Y1 - 2012 U6 - http://dx.doi.org/10.1016/j.snb.2012.10.050 SN - 0925-4005 N1 - Part of special issue "Selected Papers from the 14th International Meeting on Chemical Sensors" VL - 187 SP - 168 EP - 173 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Huck, Christina A1 - Poghossian, Arshak A1 - Kerroumi, Iman A1 - Schusser, Sebastian A1 - Bäcker, Matthias A1 - Zander, Willi A1 - Schubert, Jürgen A1 - Buniatyan, Vahe V. A1 - Martirosyan, Norayr W. A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Multiparameter sensor chip with Barium Strontium Titanate as multipurpose material JF - Electroanalysis N2 - It is well known that biochemical and biotechnological processes are strongly dependent and affected by a variety of physico-chemical parameters such as pH value, temperature, pressure and electrolyte conductivity. Therefore, these quantities have to be monitored or controlled in order to guarantee a stable process operation, optimization and high yield. In this work, a sensor chip for the multiparameter detection of three physico-chemical parameters such as electrolyte conductivity, pH and temperature is realized using barium strontium titanate (BST) as multipurpose material. The chip integrates a capacitively coupled four-electrode electrolyte-conductivity sensor, a capacitive field-effect pH sensor and a thin-film Pt-temperature sensor. Due to the multifunctional properties of BST, it is utilized as final outermost coating layer of the processed sensor chip and serves as passivation and protection layer as well as pH-sensitive transducer material at the same time. The results of testing of the individual sensors of the developed multiparameter sensor chip are presented. In addition, a quasi-simultaneous multiparameter characterization of the sensor chip in buffer solutions with different pH value and electrolyte conductivity is performed. To study the sensor behavior and the suitability of BST as multifunctional material under harsh environmental conditions, the sensor chip was exemplarily tested in a biogas digestate. Y1 - 2014 U6 - http://dx.doi.org/10.1002/elan.201400076 SN - 1521-4109 (E-Journal); 1040-0397 (Print) VL - 26 IS - 5 SP - 980 EP - 987 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Huck, Christina A1 - Poghossian, Arshak A1 - Bäcker, Matthias A1 - Reisert, Steffen A1 - Kramer, Friederike A1 - Begoyan, Vardges K. A1 - Buniatyan, Vahe V. A1 - Schöning, Michael Josef T1 - Multi-parameter sensing using high-k oxide of barium strontium titanate JF - Physica status solidi (a) N2 - High-k perovskite oxide of barium strontium titanate (BST) represents a very attractive multi-functional transducer material for the development of (bio-)chemical sensors. In this work, a Si-based sensor chip containing Pt interdigitated electrodes covered with a thin BST layer (485 nm) has been developed for multi-parameter chemical sensing. The chip has been applied for the contactless measurement of the electrolyte conductivity, the detection of adsorbed charged macromolecules (positively charged polyelectrolytes of polyethylenimine) and the concentration of hydrogen peroxide (H2O2) vapor. The experimental results of functional testing of individual sensors are presented. The mechanism of the BST sensitivity to charged polyelectrolytes and H2O2 vapor has been proposed and discussed. Y1 - 2015 U6 - http://dx.doi.org/10.1002/pssa.201431911 SN - 1862-6319 VL - 212 IS - 6 SP - 1259 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Huck, Christina A1 - Poghossian, Arshak A1 - Bäcker, Matthias A1 - Chaudhuri, S. A1 - Zander, W. A1 - Schubert, J. A1 - Begoyan, V. K. A1 - Buniatyan, V. V. A1 - Wagner, P. A1 - Schöning, Michael Josef T1 - Capacitively coupled electrolyte-conductivity sensor based on high-k material of barium strontium titanate JF - Sensors and actuators. B: Chemical Y1 - 2014 U6 - http://dx.doi.org/10.1016/j.snb.2014.02.103 SN - 1873-3077 (E-Journal); 0925-4005 (Print) IS - 198 SP - 102 EP - 109 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Hoskens, R.C.P. A1 - Roer, T.G. van de A1 - Tolstikhin, V.I. A1 - Förster, Arnold T1 - Hot electron injection laser: vertically integrated transistor-laser structure for high-speed, low-chirp direct modulation JF - LEOS 2000 : 2000 IEEE annual meeting conference proceedings; 13th annual meeting; IEEE Lasers and Electro-Optics Society 2000 annual meeting; 13-16 November 2000, The Westin Rio Mar Beach, Rio Grande, Puerto Rico. Vol. 2 Y1 - 2000 SN - 0-7803-5947-X N1 - 2000 IEEE LEOS annual meeting SP - 444 EP - 445 PB - IEEE Service Center CY - Piscataway, NJ ER - TY - JOUR A1 - Hoskens, R. C. P. A1 - Tolstikhin, V.I. A1 - Förster, Arnold A1 - Roer, T.G. van de T1 - Vertically integrated transistor-laser structure, take 2 JF - WOCSDICE 2000, 24th Workshop on Compound Semiconductor Devices and Integrated Circuits held in Europe : May 29 - June 02, 2000, Aegean Sea, Greece. Y1 - 2000 SN - 0970311109 N1 - WOCSDICE ; (24, 2000) Workshop on Compound Semiconductor Devices and Integrated Circuits held in Europe ; (24 : ; 2000.05.29-06.02) PB - Univ. of Michigan CY - Ann Arbor, Mich. ER - TY - JOUR A1 - Honarvarfard, Elham A1 - Gamella, Maria A1 - Poghossian, Arshak A1 - Schöning, Michael Josef A1 - Katz, Evgeny T1 - An enzyme-based reversible Controlled NOT (CNOT) logic gate operating on a semiconductor transducer JF - Applied Materials Today N2 - An enzyme-based biocatalytic system mimicking operation of a logically reversible Controlled NOT (CNOT) gate has been interfaced with semiconductor electronic transducers. Electrolyte–insulator–semiconductor (EIS) structures have been used to transduce chemical changes produced by the enzyme system to an electronically readable capacitive output signal using field-effect features of the EIS device. Two enzymes, urease and esterase, were immobilized on the insulating interface of EIS structure producing local pH changes performing XOR logic operation controlled by various combinations of the input signals represented by urea and ethyl butyrate. Another EIS transducer was functionalized with esterase only, thus performing Identity (ID) logic operation for the ethyl butyrate input. Both semiconductor devices assembled in parallel operated as a logically reversible CNOT gate. The present system, despite its simplicity, demonstrated for the first time logically reversible function of the enzyme system transduced electronically with the semiconductor devices. The biomolecular realization of a CNOT gate interfaced with semiconductors is promising for integration into complex biomolecular networks and future biosensor/biomedical applications. KW - Electrolyte–insulator–semiconductor KW - Capacitive field-effect KW - CNOT KW - XOR KW - Enzyme logic gate Y1 - 2017 U6 - http://dx.doi.org/10.1016/j.apmt.2017.08.003 SN - 2352-9407 VL - 9 SP - 266 EP - 270 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Honarvarfard, Elham A1 - Gamella, Maria A1 - Channaveerappa, Devika A1 - Darie, Costel C. A1 - Poghossian, Arshak A1 - Schöning, Michael Josef A1 - Katz, Evgeny T1 - Electrochemically Stimulated Insulin Release from a Modified Graphene–functionalized Carbon Fiber Electrode JF - Electroanalysis N2 - A graphene-functionalized carbon fiber electrode was modified with adsorbed polyethylenimine to introduce amino functionalities and then with trigonelline and 4-carboxyphenylboronic acid covalently bound to the amino groups. The trigonelline species containing quarterized pyridine groups produced positive charge on the electrode surface regardless of the pH value, while the phenylboronic acid species were neutral below pH 8 and negatively charged above pH 9 (note that their pKa=8.4). The total charge on the monolayer-modified electrode was positive at the neutral pH and negative at pH > 9. Note that 4-carboxyphenylboronic acid was attached to the electrode surface in molar excess to trigonelline, thus allowing the negative charge to dominate on the electrode surface at basic pH. Negatively charged fluorescent dye-labeled insulin (insulin-FITC) was loaded on the modified electrode surface at pH 7.0 due to its electrostatic attraction to the positively charged interface. The local pH in close vicinity to the electrode surface was increased to ca. 9–10 due to consumption of H+ ions upon electrochemical reduction of oxygen proceeding at the potential of −1.0 V (vs. Ag/AgCl) applied on the modified electrode. The process resulted in recharging of the electrode surface to the negative value due to the formation of the negative charge on the phenylboronic acid groups, thus resulting in the electrostatic repulsion of insulin-FITC and stimulating its release from the electrode surface. The insulin release was characterized by fluorescence spectroscopy (using the FITC-labeled insulin), by electrochemical measurements on an iridium oxide, IrOx, electrode and by mass spectrometry. The graphene-functionalized carbon fiber electrode demonstrated significant advantages in the signal-stimulated insulin release comparing with the carbon fiber electrode without the graphene species. Y1 - 2017 U6 - http://dx.doi.org/10.1002/elan.201700095 SN - 1521-4109 VL - 29 IS - 6 SP - 1543 EP - 1553 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Hodel, U. A1 - Orzati, A. A1 - Marso, M. A1 - Homann, O. A1 - Fox, A. A1 - Hart, A. v. d. A1 - Förster, Arnold A1 - Kordos, P. A1 - Lüth, H. T1 - A novel InAlAs/InGaAs layer structure for monolithically integrated photoreceiver JF - Conference Proceedings: 2000 International Conference on Indium Phosphide and related materials Y1 - 2000 SN - 0-7803-6320-5 N1 - International Conference on Indium Phosphide and Related Materials <12, 2000, Williamsburg, Va.> SP - 466 EP - 469 PB - IEEE Service Center CY - Piscataway, NJ ER - TY - JOUR A1 - Hennemann, Jörg A1 - Kohl, Claus-Dieter A1 - Reisert, Steffen A1 - Kirchner, Patrick A1 - Schöning, Michael Josef T1 - Copper oxide nanofibres for detection of hydrogen peroxide vapour at high concentrations JF - physica status solidi (a) N2 - We present a sensor concept based on copper(II)oxide (CuO) nanofibres for the detection of hydrogen peroxide (H2O2) vapour in the percent per volume (% v/v) range. The fibres were produced by using the electrospinning technique. To avoid water condensation in the pores, the fibres were initially modified by an exposure to H2S to get an enclosed surface. By a thermal treatment at 350 °C the fibres were oxidised back to CuO. Thereby, the visible pores disappear which was verified by SEM analysis. The fibres show a decrease of resistance with increasing H2O2 concentration which is due to the fact that hydrogen peroxide is an oxidising gas and CuO a p-type semiconductor. The sensor shows a change of resistance within the minute range to the exposure until the maximum concentration of 6.9% v/v H2O2. At operating temperatures below 450 °C the corresponding sensor response to a concentration of 4.1% v/v increases. The sensor shows a good reproducibility of the signal at different measurements. CuO seems to be a suitable candidate for the detection of H2O2 vapour at high concentrations. Resistance behaviour of the sensor under exposure to H2O2 vapours between 2.3 and 6.9% v/v at an operating temperature of 450 °C. Y1 - 2013 U6 - http://dx.doi.org/10.1002/pssa.201200775 SN - 1862-6319 VL - 210 IS - 5 SP - 859 EP - 863 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Henken, F. E. A1 - Oosterhuis, K. A1 - Öhlschläger, Peter A1 - Bosch, L. A1 - Hooijberg, E. A1 - Haanen, J. B. A. G. A1 - Steenbergen, R. D. M. T1 - Preclinical safety evaluation of DNA vaccines encoding modified HPV16 E6 and E7 JF - Vaccine N2 - Persistent infection with high-risk human papillomaviruses (hrHPV) can result in the formation of anogenital cancers. As hrHPV proteins E6 and E7 are required for cancer initiation and maintenance, they are ideal targets for immunotherapeutic interventions. Previously, we have described the development of DNA vaccines for the induction of HPV16 E6 and E7 specific T cell immunity. These vaccines consist of ‘gene-shuffled’ (SH) versions of HPV16 E6 and E7 that were fused to Tetanus Toxin Fragment C domain 1 (TTFC) and were named TTFC-E6SH and TTFC-E7SH. Gene-shuffling was performed to avoid the risk of inducing malignant transformation at the vaccination site. Here, we describe the preclinical safety evaluation of these candidate vaccines by analysis of their transforming capacity in vitro using established murine fibroblasts (NIH 3T3 cells) and primary human foreskin keratinocytes (HFKs). We demonstrate that neither ectopic expression of TTFC-E6SH and TTFC-E7SH alone or in combination enabled NIH 3T3 cells to form colonies in soft agar. In contrast, expression of HPV16 E6WT and E7WT alone or in combination resulted in effective transformation. Similarly, retroviral transduction of HFKs from three independent donors with both TTFC-E6SH and TTFC-E7SH alone or in combination did not show any signs of immortalization. In contrast, the combined expression of E6WT and E7WT induced immortalization in HFKs from all donors. Based on these results we consider it justified to proceed to clinical evaluation of DNA vaccines encoding TTFC-E6SH and TTFC-E7SH in patients with HPV16 associated (pre)malignancies. Y1 - 2012 U6 - http://dx.doi.org/10.1016/j.vaccine.2012.04.013 SN - 0264-410X VL - 30 IS - 28 SP - 4259 EP - 4266 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Heine, A. A1 - Herrmann, G. A1 - Selmer, Thorsten A1 - Terwesten, F. A1 - Buckel, W. A1 - Reuter, K. T1 - High resolution crystal structure of clostridium propionicum β-Alanyl-CoA:Ammonia Lyase, a new member of the "Hot Dog Fold" protein superfamily JF - Proteins N2 - Clostridium propionicum is the only organism known to ferment β-alanine, a constituent of coenzyme A (CoA) and the phosphopantetheinyl prosthetic group of holo-acyl carrier protein. The first step in the fermentation is a CoA-transfer to β-alanine. Subsequently, the resulting β-alanyl-CoA is deaminated by the enzyme β-alanyl-CoA:ammonia lyase (Acl) to reversibly form ammonia and acrylyl-CoA. We have determined the crystal structure of Acl in its apo-form at a resolution of 0.97 Å as well as in complex with CoA at a resolution of 1.59 Å. The structures reveal that the enyzme belongs to a superfamily of proteins exhibiting a so called “hot dog fold” which is characterized by a five-stranded antiparallel β-sheet with a long α-helix packed against it. The functional unit of all “hot dog fold” proteins is a homodimer containing two equivalent substrate binding sites which are established by the dimer interface. In the case of Acl, three functional dimers combine to a homohexamer strongly resembling the homohexamer formed by YciA-like acyl-CoA thioesterases. Here, we propose an enzymatic mechanism based on the crystal structure of the Acl·CoA complex and molecular docking. Proteins 2014; 82:2041–2053. © 2014 Wiley Periodicals, Inc. Y1 - 2014 U6 - http://dx.doi.org/10.1002/prot.24557 SN - 1097-0134 (E-Journal); 0887-3585 (Print) VL - 82 IS - 9 SP - 2041 EP - 2053 PB - Wiley-Liss CY - New York ER - TY - JOUR A1 - Heiduschka, P. A1 - Romann, I. A1 - Ecken, H. A1 - Schöning, Michael Josef A1 - Schuhmann, W. A1 - Thanos, S. T1 - Defined adhesion and growth of neurones on artificial structured substrates JF - Scaling down in electrochemistry : electrochemical micro- and nanosystem technology ; proceedings of the 3rd International Symposium on Electrochemical Microsystem Technologies, Garmisch-Patenkirchen, Germany, 11 - 15 September 2000 / ed. by J. W. Schultz Y1 - 2001 SN - 0-08-044014-2 SP - 299 EP - 307 PB - Elsevier [u.a.] CY - Amsterdam [u.a.] ER - TY - JOUR A1 - Heiden, W. A1 - Turek, M. A1 - Schöning, Michael Josef T1 - TasteIT : Analyzing chemical sensor data using fuzzy logic Y1 - 2011 SN - 978-1-4244-9910-6 N1 - 2011 IEEE Workshop on Merging Fields of Computational Intelligence and Sensor Technology ; 11.-15. April 2011 Paris, France SP - 1 EP - 6 PB - IEEE CY - New York ER - TY - JOUR A1 - Heiden, W. A1 - Turek, M. A1 - Schöning, Michael Josef T1 - Analysis of chemical sensor data JF - Proceedings of the 4th Russian-German Workshop "Innovation Information Technologies: Theory and practice" : Ufa, Russia, April 8-13, 2011 / eds. Yupsova, Nafisa ... Y1 - 2011 SN - 978-5-4221-0159-7 SP - 76 EP - 81 PB - State Aviation Technical Univ. CY - Ufa ER - TY - JOUR A1 - Handtke, Stefan A1 - Volland, Sonja A1 - Methling, Karen A1 - Albrecht, Dirk A1 - Becher, Dörte A1 - Nehls, Jenny A1 - Bongaerts, Johannes A1 - Maurer, Karl-Heinz A1 - Lalk, Michael A1 - Liesegang, Heiko A1 - Voigt, Birgit A1 - Daniel, Rolf A1 - Hecker, Michael T1 - Cell physiology of the biotechnological relevant bacterium Bacillus pumilus - An omics-based approach JF - Journal of Biotechnology N2 - Members of the species Bacillus pumilus get more and more in focus of the biotechnological industry as potential new production strains. Based on exoproteome analysis, B. pumilus strain Jo2, possessing a high secretion capability, was chosen for an omics-based investigation. The proteome and metabolome of B. pumilus cells growing either in minimal or complex medium was analyzed. In total, 1542 proteins were identified in growing B. pumilus cells, among them 1182 cytosolic proteins, 297 membrane and lipoproteins and 63 secreted proteins. This accounts for about 43% of the 3616 proteins encoded in the B. pumilus Jo2 genome sequence. By using GC–MS, IP-LC/MS and H NMR methods numerous metabolites were analyzed and assigned to reconstructed metabolic pathways. In the genome sequence a functional secretion system including the components of the Sec- and Tat-secretion machinery was found. Analysis of the exoproteome revealed secretion of about 70 proteins with predicted secretion signals. In addition, selected production-relevant genome features such as restriction modification systems and NRPS clusters of B. pumilus Jo2 are discussed. Y1 - 2014 U6 - http://dx.doi.org/10.1016/j.jbiotec.2014.08.028 SN - 1873-4863 (E-Journal); 0168-1656 (Print) IS - 192(A) SP - 204 EP - 214 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Hamad, E. M. A1 - Bilatto, S. E. R. A1 - Adly, N. Y. A1 - Correa, D. S. A1 - Wolfrum, B. A1 - Schöning, Michael Josef A1 - Offenhäusser, A. A1 - Yakushenko, A. T1 - Inkjet printing of UV-curable adhesive and dielectric inks for microfluidic devices JF - Lab on a Chip N2 - Bonding of polymer-based microfluidics to polymer substrates still poses a challenge for Lab-On-a-Chip applications. Especially, when sensing elements are incorporated, patterned deposition of adhesives with curing at ambient conditions is required. Here, we demonstrate a fabrication method for fully printed microfluidic systems with sensing elements using inkjet and stereolithographic 3D-printing. Y1 - 2016 U6 - http://dx.doi.org/10.1039/C5LC01195G SN - 1473-0189 VL - 16 IS - 1 SP - 70 EP - 74 PB - Royal Society of Chemistry CY - Cambridge 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 - 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 - 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 - 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 - Haeger, Gerrit A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - A convenient ninhydrin assay in 96-well format for amino acid-releasing enzymes using an air-stable reagent JF - Analytical Biochemistry N2 - An improved and convenient ninhydrin assay for aminoacylase activity measurements was developed using the commercial EZ Nin™ reagent. Alternative reagents from literature were also evaluated and compared. The addition of DMSO to the reagent enhanced the solubility of Ruhemann's purple (RP). Furthermore, we found that the use of a basic, aqueous buffer enhances stability of RP. An acidic protocol for the quantification of lysine was developed by addition of glacial acetic acid. The assay allows for parallel processing in a 96-well format with measurements microtiter plates. Y1 - 2022 U6 - http://dx.doi.org/10.1016/j.ab.2022.114819 SN - 1096-0309 IS - 624 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Guo, Yuanyuan A1 - Seki, Kosuke A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Yoshinobu, Tatsuo T1 - Novel photoexcitation method for light-addressable potentiometric sensor with higher spatial resolution JF - Applied physics express : APEX N2 - A novel photoexcitation method for the light-addressable potentiometric sensor (LAPS) is proposed to achieve a higher spatial resolution of chemical images. The proposed method employs a combined light source that consists of a modulated light probe, which generates the alternating photocurrent signal, and a ring of constant illumination surrounding it. The constant illumination generates a sheath of carriers with increased concentration which suppresses the spread of photocarriers by enhanced recombination. A device simulation was carried out to verify the effect of constant illumination on the spatial resolution, which demonstrated that a higher spatial resolution can be obtained. Y1 - 2014 U6 - http://dx.doi.org/10.7567/APEX.7.067301 SN - 1882-0786 (E-Journa); 1882-0778 (Print) VL - 7 IS - 6 SP - 067301-4 PB - IOP CY - Bristol ER - TY - JOUR A1 - Guo, Yuanyuan A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Yoshinobu, Tatsuo T1 - Theoretical study and simulation of light-addressable potentiometric sensors JF - Physica status solidi (A) : applications and materials N2 - The light-addressable potentiometric sensor (LAPS) is a semiconductor-based potentiometric sensor using a light probe with an ability of detecting the concentration of biochemical species in a spatially resolved manner. As an important biomedical sensor, research has been conducted to improve its performance, for instance, to realize high-speed measurement. In this work, the idea of facilitating the device-level simulation, instead of using an equivalent-circuit model, is presented for detailed analysis and optimization of the performance of the LAPS. Both carrier distribution and photocurrent response have been simulated to provide new insight into both amplitude-mode and phase-mode operations of the LAPS. Various device parameters can be examined to effectively design and optimize the LAPS structures and setups for enhanced performance. Y1 - 2014 U6 - http://dx.doi.org/10.1002/pssa.201330354 SN - 0031-8965 VL - 211 IS - 6 SP - 1467 EP - 1472 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Guo, Yuanyuan A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Yoshinobu, Tatsuo T1 - Device simulation of the light-addressable potentiometric sensor for the investigation of the spatial resolution JF - Sensors and actuators B: Chemical N2 - As a semiconductor-based electrochemical sensor, the light-addressable potentiometric sensor (LAPS) can realize two dimensional visualization of (bio-)chemical reactions at the sensor surface addressed by localized illumination. Thanks to this imaging capability, various applications in biochemical and biomedical fields are expected, for which the spatial resolution is critically significant. In this study, therefore, the spatial resolution of the LAPS was investigated in detail based on the device simulation. By calculating the spatiotemporal change of the distributions of electrons and holes inside the semiconductor layer in response to a modulated illumination, the photocurrent response as well as the spatial resolution was obtained as a function of various parameters such as the thickness of the Si substrate, the doping concentration, the wavelength and the intensity of illumination. The simulation results verified that both thinning the semiconductor substrate and increasing the doping concentration could improve the spatial resolution, which were in good agreement with known experimental results and theoretical analysis. More importantly, new findings of interests were also obtained. As for the dependence on the wavelength of illumination, it was found that the known dependence was not always the case. When the Si substrate was thick, a longer wavelength resulted in a higher spatial resolution which was known by experiments. When the Si substrate was thin, however, a longer wavelength of light resulted in a lower spatial resolution. This finding was explained as an effect of raised concentration of carriers, which reduced the thickness of the space charge region. The device simulation was found to be helpful to understand the relationship between the spatial resolution and device parameters, to understand the physics behind it, and to optimize the device structure and measurement conditions for realizing higher performance of chemical imaging systems. Y1 - 2014 U6 - http://dx.doi.org/10.1016/j.snb.2014.08.016 SN - 1873-3077 (E-Journal); 0925-4005 (Print) VL - 204 SP - 659 EP - 665 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Gun, Jenny A1 - Schöning, Michael Josef A1 - Abouzar, Maryam H. A1 - Poghossian, Arshak A1 - Katz, Evgeny T1 - Field-Effect Nanoparticle-Based Glucose Sensor on a Chip: Amplification Effect of Coimmobilized Redox Species JF - Electroanalysis. 20 (2008), H. 16 Y1 - 2008 SN - 1521-4109 SP - 1748 EP - 1753 ER - TY - JOUR A1 - Gun, Jenny A1 - Rizkov, Dan A1 - Lev, Ovadia A1 - Abouzar, Maryam H. A1 - Poghossian, Arshak A1 - Schöning, Michael Josef T1 - Oxygen plasma-treated gold nanoparticle-based field-effect devices as transducer structures for bio-chemical sensing JF - Microchimica Acta. 164 (2008), H. 3-4 Y1 - 2008 SN - 1436-5073 SP - 395 EP - 404 ER - TY - JOUR A1 - Gun, Jenny A1 - Gutkin, Vitaly A1 - Lev, Ovadia A1 - Boyen, Hans-Gerd A1 - Saitner, Marc A1 - Wagner, Patrick A1 - Olieslaeger, Marc D´ A1 - Abouzar, Maryam H. A1 - Poghossian, Arshak A1 - Schöning, Michael Josef T1 - Tracing gold nanoparticle charge by electrolyte-insulator-semiconductor devices JF - Journal of Physical Chemistry C. 115 (2011), H. 11 Y1 - 2011 SN - 1932-7455 SP - 4439 EP - 4445 PB - American Cemical Society CY - Washington, DC ER - TY - JOUR A1 - Grinsven, Bart van A1 - Bon, Natalie vanden A1 - Strauven, Hannelore A1 - Grieten, Lars A1 - Murib, Mohammed A1 - Jiménez Monroy, Kathia L. A1 - Janssens, Stoffel D. A1 - Haenen, Ken A1 - Schöning, Michael Josef A1 - Vermeeren, Veronique A1 - Ameloot, Marcel A1 - Michiels, Luc A1 - Thoelen, Ronald A1 - Ceuninck, Ward de A1 - Wagner, Patrick T1 - Heat-Transfer Resistance at Solid-Liquid Interfaces: A Tool for The Detection of Single Nucleotide Polymorphisms in DNA. JF - ACS Nano N2 - In this article, we report on the heat-transfer resistance at interfaces as a novel, denaturation-based method to detect single-nucleotide polymorphisms in DNA. We observed that a molecular brush of double-stranded DNA grafted onto synthetic diamond surfaces does not notably affect the heat-transfer resistance at the solid-to-liquid interface. In contrast to this, molecular brushes of single-stranded DNA cause, surprisingly, a substantially higher heat-transfer resistance and behave like a thermally insulating layer. This effect can be utilized to identify ds-DNA melting temperatures via the switching from low- to high heat-transfer resistance. The melting temperatures identified with this method for different DNA duplexes (29 base pairs without and with built-in mutations) correlate nicely with data calculated by modeling. The method is fast, label-free (without the need for fluorescent or radioactive markers), allows for repetitive measurements, and can also be extended toward array formats. Reference measurements by confocal fluorescence microscopy and impedance spectroscopy confirm that the switching of heat-transfer resistance upon denaturation is indeed related to the thermal on-chip denaturation of DNA. Y1 - 2012 U6 - http://dx.doi.org/10.1021/nn300147e SN - 1936-086X VL - 6 IS - 3 SP - 2712 EP - 2721 PB - ACS Publications CY - Washington, DC ER - TY - JOUR A1 - Grinsven, B. van A1 - Bon, N. vanden A1 - Grieten, L. A1 - Murib, M. A1 - Janssen, S. D. A1 - Haenen, K. A1 - Schneider, E. A1 - Ingebrandt, E. A1 - Schöning, Michael Josef A1 - Vermeeren, V. A1 - Ameloot, M. A1 - Michiels, L. A1 - Thoelen, R. A1 - Ceuninck, W. de A1 - Wagner, P. T1 - Rapid assessment of the stability of DNA duplexes by impedimetric real-time monitoring of chemically induced denaturation JF - Lab on a Chip Y1 - 2011 SN - 1473-0197 VL - 11 IS - 9 SP - 1656 EP - 1663 PB - Royal Society of Chemistry (RSC) CY - Cambridge 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 - JOUR A1 - Ge, B. A1 - Meyer, T. A1 - Schöning, Michael Josef A1 - Wollenberger, U. A1 - Lisdat, F., T1 - Cytochrome c‘ from Chromatium vinosum on gold electrodes JF - Electrochemistry Communications. 2 (2000), H. 8 Y1 - 2000 SN - 1388-2481 SP - 557 EP - 561 ER - TY - JOUR A1 - Gasparyan, Ferdinand V. A1 - Poghossian, Arshak A1 - Vitusevich, Svetlana A. A1 - Petrychuk, Mykhaylo V. A1 - Sydoruk, Viktor A. A1 - Siqueira, José R. Jr. A1 - Oliveira, Osvaldo N. Jr. A1 - Offenhäusser, Andreas A1 - Schöning, Michael Josef T1 - Low-Frequency Noise in Field-Effect Devices Functionalized With Dendrimer/Carbon-Nanotube Multilayers JF - IEEE Sensors Journal. 11 (2011), H. 1 Y1 - 2011 SN - 1530-437X SP - 142 EP - 149 PB - IEEE CY - New York ER - TY - JOUR A1 - Gasparyan, F.V. A1 - Vitusevich, S.A. A1 - Offenhäusser, A. A1 - Schöning, Michael Josef T1 - Modified charge fluctuation noise model for electrolyte-insulator-semiconductor devices JF - Modern Physics Letters B (MPLB). 25 (2011), H. 11 Y1 - 2011 SN - 0217-9849 SP - 831 EP - 840 PB - World Scientific Publ. CY - Singapur 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 - TY - JOUR A1 - Gamella, Maria A1 - Zakharchenko, Andrey A1 - Guz, Nataliia A1 - Masi, Madeline A1 - Minko, Sergiy A1 - Kolpashchikov, Dmitry M. A1 - Iken, Heiko A1 - Poghossian, Arshak A1 - Schöning, Michael Josef A1 - Katz, Evgeny T1 - DNA computing system activated by electrochemically triggered DNA realease from a polymer-brush-modified electrode array JF - Electroanalysis N2 - An array of four independently wired indium tin oxide (ITO) electrodes was used for electrochemically stimulated DNA release and activation of DNA-based Identity, AND and XOR logic gates. Single-stranded DNA molecules were loaded on the mixed poly(N,N-dimethylaminoethyl methacrylate) (PDMAEMA)/poly(methacrylic acid) (PMAA) brush covalently attached to the ITO electrodes. The DNA deposition was performed at pH 5.0 when the polymer brush is positively charged due to protonation of tertiary amino groups in PDMAEMA, thus resulting in electrostatic attraction of the negatively charged DNA. By applying electrolysis at −1.0 V(vs. Ag/AgCl reference) electrochemical oxygen reduction resulted in the consumption of hydrogen ions and local pH increase near the electrode surface. The process resulted in recharging the polymer brush to the negative state due to dissociation of carboxylic groups of PMAA, thus repulsing the negatively charged DNA and releasing it from the electrode surface. The DNA release was performed in various combinations from different electrodes in the array assembly. The released DNA operated as input signals for activation of the Boolean logic gates. The developed system represents a step forward in DNA computing, combining for the first time DNA chemical processes with electronic input signals. Y1 - 2017 U6 - http://dx.doi.org/10.1002/elan.201600389 SN - 1521-4109 VL - 29 IS - 2 SP - 398 EP - 408 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Förster, Arnold A1 - Stock, J. A1 - Indlekofer, K. M. T1 - Perspectives of resonant tunneling diodes JF - Recent research developments in materials science & engineering/ 1,2 Y1 - 2002 SN - 81-7895-057-X N1 - Nebent.: Materials science & engineering SP - 527 EP - 556 PB - Transworld Research Network CY - Trivandrum, India ER - TY - JOUR A1 - Förster, Arnold A1 - Lepsa, M. I. A1 - Freundt, D. T1 - Hot electron injector Gunn diode for advanced driver assistance systems JF - Applied physics A: materials science and processing. 87 (2007), H. 3 Y1 - 2007 SN - 0947-8396 N1 - ISSN der E-Ausg.: 1432-0630 ; Special issue SP - 545 EP - 558 ER - TY - JOUR A1 - Förster, Arnold T1 - Resonant tunneling diodes operating as modern quantum transport devices JF - Proceedings of the Tenth International Workshop on the Physics of Semiconductor Devices : (December 14 - 18, 1999) / ed.: Vikram Kumar ... - Vol. 2 Y1 - 2000 SN - 81-7023-998-2 N1 - International Workshop on the Physics of Semiconductor Devices <10, 1999, Delhi> SP - 966 EP - 966 PB - Allied Publ. CY - New Delhi [u.a.] ER - TY - JOUR A1 - Furuichi, K. A1 - Yoshinobu, T. A1 - Ermelenko, T. A1 - Mourzina, Y. A1 - Iwasaki, H. A1 - Schöning, Michael Josef T1 - Simultaneous visualization of multiple ions by the chemical imaging sensor JF - Proceedings of the 5th East Asian Conference on Chemical Sensors; the 33rd Chemical Sensor Symposium : December 4 - 7, 2001, Huis Ten Bosch, Sasebo-shi, Nagasaki, Japan / Japan Association of Chemical Sensors; the Electrochemical Society of Japan Y1 - 2001 N1 - Chemical sensors; 17.2001 Suppl. B; East Asian Conference on Chemical Sensors ; (5, 2001; Nagasaki) SP - 399 EP - 401 PB - Japan Association of Chemical Sensors CY - Kasuga, Fukuoka-ken ER - TY - JOUR A1 - Figueroa-Miranda, Gabriela A1 - Feng, Lingyan A1 - Shiu, Simon Chi-Chin A1 - Dirkzwager, Roderick Marshall A1 - Cheung, Yee-Wai A1 - Tanner, Julian Alexander A1 - Schöning, Michael Josef A1 - Offenhäusser, Andreas A1 - Mayer, Dirk T1 - Aptamer-based electrochemical biosensor for highly sensitive and selective malaria detection with adjustable dynamic response range and reusability JF - Sensor and Actuators B: Chemical N2 - Malaria infection remains a significant risk for much of the population of tropical and subtropical areas, particularly in developing countries. Therefore, it is of high importance to develop sensitive, accurate and inexpensive malaria diagnosis tests. Here, we present a novel aptamer-based electrochemical biosensor (aptasensor) for malaria detection by impedance spectroscopy, through the specific recognition between a highly discriminatory DNA aptamer and its target Plasmodium falciparum lactate dehydrogenase (PfLDH). Interestingly, due to the isoelectric point (pI) of PfLDH, the aptasensor response showed an adjustable detection range based on the different protein net-charge at variable pH environments. The specific aptamer recognition allows sensitive protein detection with an expanded detection range and a low detection limit, as well as a high specificity for PfLDH compared to analogous proteins. The specific feasibility of the aptasensor is further demonstrated by detection of the target PfLDH in human serum. Furthermore, the aptasensor can be easily regenerated and thus applied for multiple usages. The robustness, sensitivity, and reusability of the presented aptasensor make it a promising candidate for point-of-care diagnostic systems. Y1 - 2018 U6 - http://dx.doi.org/10.1016/j.snb.2017.07.117 SN - 0925-4005 VL - 255 IS - P1 SP - 235 EP - 243 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Faßbender, F. A1 - Schmitt, G. A1 - Schöning, Michael Josef A1 - Lüth, H. A1 - Buß, G. A1 - Schultze, J. W. T1 - Optimization of passivation layers for corrosion protection of silicon-based microelectrode arrays JF - Sensors and Actuators B. 68 (2000), H. 1-3 Y1 - 2000 SN - 0925-4005 SP - 128 EP - 133 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 - Falkenberg, Fabian A1 - Rahba, Jade A1 - Fischer, David A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Biochemical characterization of a novel oxidatively stable, halotolerant, and high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101T JF - FEBS Open Bio N2 - Halophilic and halotolerant microorganisms represent a promising source of salt-tolerant enzymes suitable for various biotechnological applications where high salt concentrations would otherwise limit enzymatic activity. Considering the current growing enzyme market and the need for more efficient and new biocatalysts, the present study aimed at the characterization of a high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101T. The protease gene was cloned and expressed in Bacillus subtilis DB104. The recombinant protease SPAO with 269 amino acids belongs to the subfamily of high-alkaline subtilisins. The biochemical characteristics of purified SPAO were analyzed in comparison with subtilisin Carlsberg, Savinase, and BPN'. SPAO, a monomer with a molecular mass of 27.1 kDa, was active over a wide range of pH 6.0–12.0 and temperature 20–80 °C, optimally at pH 9.0–9.5 and 55 °C. The protease is highly oxidatively stable to hydrogen peroxide and retained 58% of residual activity when incubated at 10 °C with 5% (v/v) H2O2 for 1 h while stimulated at 1% (v/v) H2O2. Furthermore, SPAO was very stable and active at NaCl concentrations up to 5.0 m. This study demonstrates the potential of SPAO for biotechnological applications in the future. KW - Alkalihalobacillus okhensis KW - detergent protease KW - halotolerant protease KW - high-alkaline subtilisin KW - oxidative stable protease Y1 - 2022 U6 - http://dx.doi.org/10.1002/2211-5463.13457 SN - 2211-5463 N1 - Corresponding author: Petra Siegert VL - 12 IS - 10 SP - 1729 EP - 1746 PB - Wiley CY - Hoboken, NJ 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 - Falkenberg, Fabian A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Phylogenetic survey of the subtilase family and a data-mining-based search for new subtilisins from Bacillaceae JF - Frontiers in Microbiology N2 - The subtilase family (S8), a member of the clan SB of serine proteases are ubiquitous in all kingdoms of life and fulfil different physiological functions. Subtilases are divided in several groups and especially subtilisins are of interest as they are used in various industrial sectors. Therefore, we searched for new subtilisin sequences of the family Bacillaceae using a data mining approach. The obtained 1,400 sequences were phylogenetically classified in the context of the subtilase family. This required an updated comprehensive overview of the different groups within this family. To fill this gap, we conducted a phylogenetic survey of the S8 family with characterised holotypes derived from the MEROPS database. The analysis revealed the presence of eight previously uncharacterised groups and 13 subgroups within the S8 family. The sequences that emerged from the data mining with the set filter parameters were mainly assigned to the subtilisin subgroups of true subtilisins, high-alkaline subtilisins, and phylogenetically intermediate subtilisins and represent an excellent source for new subtilisin candidates. Y1 - 2022 U6 - http://dx.doi.org/10.3389/fmicb.2022.1017978 SN - 1664-302X VL - 2022 IS - 13 PB - Frontiers CY - Lausanne ER - TY - JOUR A1 - Ermolenko, Y.E. A1 - Vlasov, Y.G. A1 - Kolodnikov, V.V. A1 - Shabaldkin, D. A. A1 - Kloock, Joachim P. A1 - Schöning, Michael Josef T1 - Diffusion of radioactive tracers (204Tl, 110mAg) and ionic conductivity in membrane materials for the chemical sensors JF - Advances in nuclear and radiochemistry : extended abstracts of papers presented at the Sixth International Conference on Nuclear and Radiochemistry (NRC-6), 29 August to 3 September 2004, Aachen, Germany ; in cooperation with University of Cologne, GDCh, FECS, OECD-NEA and IAEA / Syed M. Qaim ... (eds.). Y1 - 2004 SN - 3893363629 N1 - Schriften des Forschungszentrums Jülich Reihe Allgemeines und Interdisziplinäres ; 3. International Conference on Nuclear and Radiochemistry ; (6, 2004, Aachen) SP - 483 EP - 485 PB - Forschungszentrum Jülich, Zentralbibliothek CY - Jülich ER - TY - JOUR A1 - Ermolenko, Y. E. A1 - Yoshinobu, T. A1 - Mourzina, Y. G. A1 - Vlasov, Y. G. A1 - Schöning, Michael Josef A1 - Iwasaki, H. T1 - Laserscanned transducer (LSST) as a multisensor system JF - Sensors and Actuators B. 103 (2004), H. 1-2 Y1 - 2004 SN - 0925-4005 SP - 457 EP - 462 ER - TY - JOUR A1 - Ermelenko, Y. A1 - Yoshinobu, T. A1 - Mourzina, Y. A1 - Schöning, Michael Josef A1 - Vlasov, Y. A1 - Iwasaki, H. T1 - The hybrid K+/Ca2+ sensor based on laser scanned silicon transducer for multi-component analysis JF - Proceedings of ICAS 2001, IUPAC [3rd] International Congress on Analytical Sciences 2001 : August 6 - 10, 2001, Waseda University, Tokyo Y1 - 2002 N1 - International Congress on Analytical Sciences <2001, Tokyo> ; International Union of Pure and Applied Chemistry. Pure and applied chemistry ; 73,10. ; Analytical sciences ; 17.2001, special issue SP - i777 EP - i780 PB - Japan Society for Analytical Chemistry CY - Tokyo ER - TY - JOUR A1 - Ermelenko, Y. A1 - Yoshinobu, T. A1 - Mourzina, Y. A1 - Schöning, Michael Josef A1 - Furuichi, K. A1 - Levichev, S. A1 - Vlasov, Y. A1 - Iwasaki, H. T1 - The double K+/Ca2+ sensor based on laser scanned silicon transducer (LSST) for multicomponent analysis JF - Talanta. 59 (2003), H. 4 Y1 - 2003 SN - 0039-9140 SP - 785 EP - 795 ER - TY - JOUR A1 - Ermelenko, Y. A1 - Yoshinobu, T. A1 - Mourzina, Y. A1 - Furuichi, K. A1 - Levichev, S. A1 - Vlasov, Y. A1 - Schöning, Michael Josef A1 - Iwasaki, H. T1 - Lithium sensor based on the laser scanning semiconductor transducer JF - Analytica Chimica Acta. 459 (2002), H. 1 Y1 - 2002 SN - 0378-4304 SP - 1 EP - 9 ER - TY - JOUR A1 - Ermelenko, Y. A1 - Yoshinobu, T. A1 - Mourzina, Y. A1 - Furuichi, K. A1 - Iwasaki, H. A1 - Vlasov, Y. A1 - Schöning, Michael Josef T1 - Technology of photocurable polymeric membranes for integrated LAPS JF - Proceedings of the 5th East Asian Conference on Chemical Sensors; the 33rd Chemical Sensor Symposium : December 4 - 7, 2001, Huis Ten Bosch, Sasebo-shi, Nagasaki, Japan / Japan Association of Chemical Sensors; the Electrochemical Society of Japan Y1 - 2001 N1 - Chemical sensors; 17.2001 Suppl. B; East Asian Conference on Chemical Sensors ; (5, 2001; Nagasaki) SP - 66 EP - 68 PB - Japan Association of Chemical Sensors CY - Kasuga, Fukuoka-ken ER - TY - JOUR A1 - Ermelenko, Y. A1 - Yoshinobu, T A1 - Mourzina, Y. A1 - Levichev, S. A1 - Furuichi, K. A1 - Vlasov, Y. A1 - Schöning, Michael Josef A1 - Iwasaki, H. T1 - Photocurable membranes for ion-selective light-addressable potentiometric sensors JF - Sensors and Actuators B. 85 (2002), H. 1-2 Y1 - 2002 SN - 0925-4005 SP - 79 EP - 85 ER - TY - JOUR A1 - Engelmann, Ulrich M. A1 - Simsek, Beril A1 - Shalaby, Ahmed A1 - Krause, Hans-Joachim T1 - Key contributors to signal generation in frequency mixing magnetic detection (FMMD): an in silico study JF - Sensors N2 - Frequency mixing magnetic detection (FMMD) is a sensitive and selective technique to detect magnetic nanoparticles (MNPs) serving as probes for binding biological targets. Its principle relies on the nonlinear magnetic relaxation dynamics of a particle ensemble interacting with a dual frequency external magnetic field. In order to increase its sensitivity, lower its limit of detection and overall improve its applicability in biosensing, matching combinations of external field parameters and internal particle properties are being sought to advance FMMD. In this study, we systematically probe the aforementioned interaction with coupled Néel–Brownian dynamic relaxation simulations to examine how key MNP properties as well as applied field parameters affect the frequency mixing signal generation. It is found that the core size of MNPs dominates their nonlinear magnetic response, with the strongest contributions from the largest particles. The drive field amplitude dominates the shape of the field-dependent response, whereas effective anisotropy and hydrodynamic size of the particles only weakly influence the signal generation in FMMD. For tailoring the MNP properties and parameters of the setup towards optimal FMMD signal generation, our findings suggest choosing large particles of core sizes dc > 25 nm nm with narrow size distributions (σ < 0.1) to minimize the required drive field amplitude. This allows potential improvements of FMMD as a stand-alone application, as well as advances in magnetic particle imaging, hyperthermia and magnetic immunoassays. KW - key performance indicators KW - magnetic biosensing KW - coupled Néel–Brownian relaxation dynamics KW - frequency mixing magnetic detection KW - magnetic relaxation KW - micromagnetic simulation KW - magnetic nanoparticles Y1 - 2024 U6 - http://dx.doi.org/10.3390/s24061945 SN - 1424-8220 N1 - This article belongs to the Special Issue "Advances in Magnetic Sensors and Their Applications" VL - 24 IS - 6 PB - MDPI CY - Basel ER - TY - JOUR A1 - Engelmann, Ulrich M. A1 - Pourshahidi, Mohammad Ali A1 - Shalaby, Ahmed A1 - Krause, Hans-Joachim T1 - Probing particle size dependency of frequency mixing magnetic detection with dynamic relaxation simulation JF - Journal of Magnetism and Magnetic Materials N2 - Biomedical applications of magnetic nanoparticles (MNP) fundamentally rely on the particles’ magnetic relaxation as a response to an alternating magnetic field. The magnetic relaxation complexly depends on the interplay of MNP magnetic and physical properties with the applied field parameters. It is commonly accepted that particle core size is a major contributor to signal generation in all the above applications, however, most MNP samples comprise broad distribution spanning nm and more. Therefore, precise knowledge of the exact contribution of individual core sizes to signal generation is desired for optimal MNP design generally for each application. Specifically, we present a magnetic relaxation simulation-driven analysis of experimental frequency mixing magnetic detection (FMMD) for biosensing to quantify the contributions of individual core size fractions towards signal generation. Applying our method to two different experimental MNP systems, we found the most dominant contributions from approx. 20 nm sized particles in the two independent MNP systems. Additional comparison between freely suspended and immobilized MNP also reveals insight in the MNP microstructure, allowing to use FMMD for MNP characterization, as well as to further fine-tune its applicability in biosensing. Y1 - 2022 U6 - http://dx.doi.org/10.1016/j.jmmm.2022.169965 SN - 0304-8853 VL - 563 IS - In progress, Art. No. 169965 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Emons, H. A1 - Hüllenkremer, B. A1 - Schöning, Michael Josef T1 - Detection of metal ions in aqueous solutions by voltohmmetry JF - Fresenius' Journal of Analytical Chemistry. 369 (2001), H. 1 Y1 - 2001 SN - 0937-0633 SP - 42 EP - 46 ER - TY - JOUR A1 - Emons, H. A1 - Glück, O. A1 - Schöning, Michael Josef T1 - Voltohmmetry – An alternative detection principle at ultrathin metal electrodes in solution JF - Chemical and biological sensors and analytical methods : proceedings of the international symposium / Sensor, Physical Electrochemistry, and Organic and Biological Electrochemistry Divisions. Ed.: M. Butler Y1 - 2001 SN - 1-56677-351-2 N1 - International Symposium: Chemical and Biological Sensors and Analytical Methods ; (2 : 2001.) SP - 1 EP - 3 PB - Electrochemical Society CY - Pennington, NJ ER - TY - JOUR A1 - Emons, H. A1 - Glück, O. A1 - Hüllenkremer, B. A1 - Schöning, Michael Josef T1 - Voltohmmetry as an alternative detection method at polycrystalline metal film electrodes JF - Electroanalysis. 13 (2001), H. 8-9 Y1 - 2001 SN - 1040-0397 SP - 677 EP - 680 ER - TY - JOUR A1 - Emons, H. A1 - Baade, A. A1 - Schöning, Michael Josef T1 - Voltammetric determination of heavy metals in microvolumes of rain water JF - Electroanalysis. 12 (2000), H. 15 Y1 - 2000 SN - 1040-0397 SP - 1171 EP - 1176 ER - TY - JOUR A1 - Doll, Theodor A1 - Wagner, Torsten A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Engineering of functional interfaces / Theodor Doll ; Torsten Wagner ; Patrick Wagner ; Michael J. Schöning (eds.) JF - Physica status solidi (a) Y1 - 2016 U6 - http://dx.doi.org/10.1002/pssa.201670641 SN - 1862-6319 VL - 213 IS - 6 SP - 1393 EP - 1394 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Deppe, Veronika Maria A1 - Klatte, Stephanie A1 - Bongaerts, Johannes A1 - Maurer, Karl-Heinz A1 - O'Connell, Timothy A1 - Meinhardt, Friedhelm T1 - Genetic control of Amadori product degradation in Bacillus subtilis via regulation of frlBONMD expression by FrlR JF - Applied and environmental microbiology Y1 - 2011 SN - 1098-5336 (E-Journal); 0003-6919 (Print); 0099-2240 (Print) VL - Vol. 77 IS - No. 9 SP - 2839 EP - 2846 PB - American Society of Mechanical Engineers (ASME) CY - New York ER - TY - JOUR A1 - Deppe, Veronika Maria A1 - Bongaerts, Johannes A1 - O'Connell, Timothy A1 - Maurer, Karl-Heinz A1 - Meinhardt, Friedhelm T1 - Enzymatic deglycation of Amadori products in bacteria JF - Applied microbiology and biotechnology Y1 - 2011 SN - 1432-0614 (E-Journal); 0171-1741 (Print); 0175-7598 (Print); 0340-2118 (Print) VL - Vol. 90 IS - Iss. 2 SP - 399 EP - 406 PB - Springer CY - Berlin ER - TY - JOUR A1 - Delle, Lotta E. A1 - Huck, Christina A1 - Bäcker, Matthias A1 - Müller, Frank A1 - Grandthyll, Samuel A1 - Jacobs, Karin A1 - Lilischkis, Rainer A1 - Vu, Xuan T. A1 - Schöning, Michael Josef A1 - Wagner, Patrick A1 - Thoelen, Roland A1 - Weil, Maryam A1 - Ingebrandt, Sven T1 - Impedimetric immunosensor for the detection of histamine based on reduced graphene oxide JF - Physica status solidi (a) Y1 - 2015 U6 - http://dx.doi.org/10.1002/pssa.201431863 SN - 1862-6319 VL - 212 IS - 6 SP - 1327 EP - 1334 PB - Wiley CY - Weinheim 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 - Darmo, J. A1 - Schäffer, F. A1 - Förster, Arnold A1 - Kordos, P. T1 - Beryllium doped low-temperature-grown MBE GaAs: material for photomixing in the THz frequency range JF - ASDAM 2000 : conference proceedings / edited by Jozef Osvald ... [et al.] Y1 - 2000 SN - 0780359399 N1 - International Conference on Advanced Semiconductor Devices and Microsystems ; (3rd : ; 2000 : ; Smolenice, Slovakia) SP - 147 EP - 150 PB - IEEE CY - Piscataway, NJ ER - TY - JOUR A1 - Dantism, Shahriar A1 - Takenaga, Shoko A1 - Wagner, Torsten A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Differential imaging of the metabolism of bacteria and eukaryotic cells based on light-addressable potentiometric sensors JF - Electrochimica Acta N2 - A light-addressable potentiometric sensor (LAPS) is a field-effect-based potentiometric sensor with an electrolyte/insulator/semiconductor (EIS) structure, which is able to monitor analyte concentrations of (bio-)chemical species in aqueous solutions in a spatially resolved way. Therefore, it is also an appropriate tool to record 2D-chemical images of concentration variations on the sensor surface. In the present work, two differential, LAPS-based measurement principles are introduced to determine the metabolic activity of Escherichia coli (E. coli) K12 and Chinese hamster ovary (CHO) cells as test microorganisms. Hereby, we focus on i) the determination of the extracellular acidification rate (ΔpH/min) after adding glucose solutions to the cell suspensions; and ii) recording the amplitude increase of the photocurrent (Iph) related to the produced acids from E. coli K12 bacteria and CHO cells on the sensor surface by 2D-chemical imaging. For this purpose, 3D-printed multi-chamber structures were developed and mounted on the planar sensor-chip surface to define four independent compartments, enabling differential measurements with varying cell concentrations. The differential concept allows eliminating unwanted drift effects and, with the four-chamber structures, measurements on the different cell concentrations were performed simultaneously, thus reducing also the overall measuring time. Y1 - 2017 U6 - http://dx.doi.org/10.1016/j.electacta.2017.05.196 SN - 0013-4686 VL - 246 SP - 234 EP - 241 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Dantism, Shahriar A1 - Takenaga, Shoko A1 - Wagner, Patrick A1 - Wagner, Torsten A1 - Schöning, Michael Josef T1 - Determination of the extracellular acidification of Escherichia coli K12 with a multi-​chamber-​based LAPS system JF - Physica status solidi (a) N2 - On-line monitoring of the metabolic activity of microorganisms involved in intermediate stages of biogas production plays an important role to avoid undesirable “down times” during the biogas production. In order to control this process, an on-chip differential measuring system based on the light-addressable potentiometric sensor (LAPS) principle combined with a 3D-printed multi-chamber structure has been realized. As a test microorganism, Escherichia coli K12 (E. coli K12) were used for cell-based measurements. Multi-chamber structures were developed to determine the metabolic activity of E. coli K12 in suspension for a different number of cells, responding to the addition of a constant or variable amount of glucose concentrations, enabling differential and simultaneous measurements. Y1 - 2016 U6 - http://dx.doi.org/10.1002/pssa.201533043 SN - 1862-6300 VL - 213 IS - 6 SP - 1479 EP - 1485 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Dantism, Shahriar A1 - Röhlen, Desiree A1 - Wagner, Torsten A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Optimization of Cell-Based Multi-Chamber LAPS Measurements Utilizing FPGA-Controlled Laser-Diode Modules JF - physica status solidi a : applications and materials sciences N2 - A light-addressable potentiometric sensor (LAPS) is a field-effect-based potentiometric device, which detects concentration changes of an analyte solution on the sensor surface in a spatially resolved way. It uses a light source to generate electron–hole pairs inside the semiconductor, which are separated in the depletion region due to an applied bias voltage across the sensor structure and hence, a surface-potential-dependent photocurrent can be read out. However, depending on the beam angle of the light source, scattering effects can occur, which influence the recorded signal in LAPS-based differential measurements. To solve this problem, a novel illumination unit based on a field programmable gate array (FPGA) consisting of 16 small-sized tunable infrared laser-diode modules (LDMs) is developed. Due to the improved focus of the LDMs with a beam angle of only 2 mrad, undesirable scattering effects are minimized. Escherichia coli (E. coli) K12 bacteria are used as a test microorganism to study the extracellular acidification on the sensor surface. Furthermore, a salt bridge chamber is built up and integrated with the LAPS system enabling multi-chamber differential measurements with a single Ag/AgCl reference electrode. Y1 - 2018 U6 - http://dx.doi.org/10.1002/pssa.201800058 SN - 1862-6319 VL - 215 IS - 15 SP - Article number 1800058 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Dantism, Shahriar A1 - Röhlen, Desiree A1 - Wagner, Torsten A1 - Wagner, P. A1 - Schöning, Michael Josef T1 - A LAPS-based differential sensor for parallelized metabolism monitoring of various bacteria JF - Sensors N2 - Monitoring the cellular metabolism of bacteria in (bio)fermentation processes is crucial to control and steer them, and to prevent undesired disturbances linked to metabolically inactive microorganisms. In this context, cell-based biosensors can play an important role to improve the quality and increase the yield of such processes. This work describes the simultaneous analysis of the metabolic behavior of three different types of bacteria by means of a differential light-addressable potentiometric sensor (LAPS) set-up. The study includes Lactobacillus brevis, Corynebacterium glutamicum, and Escherichia coli, which are often applied in fermentation processes in bioreactors. Differential measurements were carried out to compensate undesirable influences such as sensor signal drift, and pH value variation during the measurements. Furthermore, calibration curves of the cellular metabolism were established as a function of the glucose concentration or cell number variation with all three model microorganisms. In this context, simultaneous (bio)sensing with the multi-organism LAPS-based set-up can open new possibilities for a cost-effective, rapid detection of the extracellular acidification of bacteria on a single sensor chip. It can be applied to evaluate the metabolic response of bacteria populations in a (bio)fermentation process, for instance, in the biogas fermentation process. Y1 - 2019 U6 - http://dx.doi.org/10.3390/s19214692 SN - 1424-8220 VL - 19 IS - 21 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 - 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 - Dantism, S. A1 - Takenaga, S. A1 - Wagner, P. A1 - Wagner, Torsten A1 - Schöning, Michael Josef T1 - Light-addressable Potentiometric Sensor (LAPS) Combined with Multi-chamber Structures to Investigate the Metabolic Activity of Cells JF - Procedia Engineering N2 - LAPS are field-effect-based potentiometric sensors which are able to monitor analyte concentrations in a spatially resolved manner. Hence, a LAPS sensor system is a powerful device to record chemical imaging of the concentration of chemical species in an aqueous solution, chemical reactions, or the growth of cell colonies on the sensor surface, to record chemical images. In this work, multi-chamber 3D-printed structures made out of polymer (PP-ABS) were combined with LAPS chips to analyse differentially and simultaneously the metabolic activity of Escherichia coli K12 and Chinese hamster ovary (CHO) cells, and the responds of those cells to the addition of glucose solution. Y1 - 2015 U6 - http://dx.doi.org/10.1016/j.proeng.2015.08.647 SN - 1877-7058 N1 - Part of special issue "Eurosensors 2015" VL - 120 SP - 384 EP - 387 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Cornelis, Peter A1 - Givanoudi, Stella A1 - Yongabi, Derick A1 - Iken, Heiko A1 - Duwé, Sam A1 - Deschaume, Olivier A1 - Robbens, Johan A1 - Dedecker, Peter A1 - Bartic, Carmen A1 - Wübbenhorst, Michael A1 - Schöning, Michael Josef A1 - Heyndrickx, Marc A1 - Wagner, Patrick T1 - Sensitive and specific detection of E. coli using biomimetic receptors in combination with a modified heat-transfer method JF - Biosensors and Bioelectronics Y1 - 2019 U6 - http://dx.doi.org/10.1016/j.bios.2019.04.026 SN - 0956-5663 VL - 136 SP - 97 EP - 105 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Christiaens, P. A1 - Abouzar, Maryam H. A1 - Poghossian, Arshak A1 - Wagner, Torsten A1 - Bijnens, N. A1 - Williams, O. A. A1 - Daenen, M. A1 - Haenen, K. A1 - Schöning, Michael Josef A1 - Wagner, P. T1 - Nanocrystalline diamond-based field-effect capacitive pH sensor JF - Transducers '07 Eurosensors XXI : digest of technical papers ; the14th International Conference on Solid-State Sensors, Actuators and Microsystems, June 10-14, 2007, Lyon, France / Gilles Delapierre (Ed.) Y1 - 2007 SN - 1-4244-0841-5 N1 - Eurosensors 21, 2007, Lyon ; International Conference on Solid-State Sensors, Actuators and Microsystems 14, 2007, Lyon SP - 1891 EP - 1894 PB - IEEE CY - Piscataway ER - TY - JOUR A1 - Christiaens, P. A1 - Abouzar, Maryam H. A1 - Poghossian, Arshak A1 - Wagner, Torsten A1 - Bijnens, N. A1 - Williams, O. A. A1 - Daenen, M. A1 - Haenen, K. A1 - Douthéret, O. A1 - Haen, J. d´ A1 - Mekhalif, Z. A1 - Schöning, Michael Josef A1 - Wagner, P. T1 - pH sensitivity of nanocrystalline diamond films JF - Physica status solidi (A). 204 (2007), H. 9 Y1 - 2007 SN - 0031-8965 SP - 2925 EP - 2930 ER - TY - JOUR A1 - Bäcker, Matthias A1 - Raue, Markus A1 - Schusser, Sebastian A1 - Jeitner, C. A1 - Breuer, L. A1 - Wagner, P. A1 - Poghossian, Arshak A1 - Förster, Arnold A1 - Mang, Thomas A1 - Schöning, Michael Josef T1 - Microfluidic chip with integrated microvalves based on temperature- and pH-responsive hydrogel thin films JF - Physica Status Solidi (a) N2 - Two types of microvalves based on temperature-responsive poly(N-isopropylacrylamide) (PNIPAAm) and pH-responsive poly(sodium acrylate) (PSA) hydrogel films have been developed and tested. The PNIPAAm and PSA hydrogel films were prepared by means of in situ photopolymerization directly inside the fluidic channel of a microfluidic chip fabricated by combining Si and SU-8 technologies. The swelling/shrinking properties and height changes of the PNIPAAm and PSA films inside the fluidic channel were studied at temperatures of deionized water from 14 to 36 °C and different pH values (pH 3–12) of Titrisol buffer, respectively. Additionally, in separate experiments, the lower critical solution temperature (LCST) of the PNIPAAm hydrogel was investigated by means of a differential scanning calorimetry (DSC) and a surface plasmon resonance (SPR) method. Mass-flow measurements have shown the feasibility of the prepared hydrogel films to work as an on-chip integrated temperature- or pH-responsive microvalve capable to switch the flow channel on/off. Y1 - 2012 U6 - http://dx.doi.org/10.1002/pssa.201100763 SN - 1862-6319 VL - 209 IS - 5 SP - 839 EP - 845 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Bäcker, Matthias A1 - Rakowski, D. A1 - Poghossian, Arshak A1 - Biselli, Manfred A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Chip-based amperometric enzyme sensor system for monitoring of bioprocesses by flow-injection analysis JF - Journal of Biotechnology N2 - A microfluidic chip integrating amperometric enzyme sensors for the detection of glucose, glutamate and glutamine in cell-culture fermentation processes has been developed. The enzymes glucose oxidase, glutamate oxidase and glutaminase were immobilized by means of cross-linking with glutaraldehyde on platinum thin-film electrodes integrated within a microfluidic channel. The biosensor chip was coupled to a flow-injection analysis system for electrochemical characterization of the sensors. The sensors have been characterized in terms of sensitivity, linear working range and detection limit. The sensitivity evaluated from the respective peak areas was 1.47, 3.68 and 0.28 μAs/mM for the glucose, glutamate and glutamine sensor, respectively. The calibration curves were linear up to a concentration of 20 mM glucose and glutamine and up to 10 mM for glutamate. The lower detection limit amounted to be 0.05 mM for the glucose and glutamate sensor, respectively, and 0.1 mM for the glutamine sensor. Experiments in cell-culture medium have demonstrated a good correlation between the glutamate, glutamine and glucose concentrations measured with the chip-based biosensors in a differential-mode and the commercially available instrumentation. The obtained results demonstrate the feasibility of the realized microfluidic biosensor chip for monitoring of bioprocesses. Y1 - 2013 U6 - http://dx.doi.org/10.1016/j.jbiotec.2012.03.014 SN - 0168-1656 VL - 163 IS - 4 SP - 371 EP - 376 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Bäcker, Matthias A1 - Pouyeshman, S. A1 - Schnitzler, Thomas A1 - Poghossian, Arshak A1 - Wagner, Patrick A1 - Biselli, Manfred A1 - Schöning, Michael Josef T1 - A silicon-based multi-sensor chip for monitoring of fermentation processes JF - Physica status solidi (a) : applications and material science. 208 (2011), H. 6 Y1 - 2011 SN - 1862-6319 SP - 1364 EP - 1369 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Bäcker, Matthias A1 - Poghossian, Arshak A1 - Abouzar, Maryam H. A1 - Wenmackers, Sylvia A1 - Janssens, Stoffel D. A1 - Haenen, Ken A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Capacitive field-effect (bio-)chemical sensors based on nanocrystalline diamond films JF - Diamond Electronics and Bioelectronics — Fundamentals to Applications III, edited by P. Bergonzo, [u.a.] Y1 - 2010 N1 - MRS Proceedings Volume 1203 paper 1203-J17-31 ; Mater. Res. Soc. Sympos. Proc. Vol 1203 (2010) ; Materials Research Society SP - 1 EP - 6 ER - TY - JOUR A1 - Bäcker, Matthias A1 - Kramer, F. A1 - Huck, Christina A1 - Poghossian, Arshak A1 - Bratov, A. A1 - Abramova, N. A1 - Schöning, Michael Josef T1 - Planar and 3D interdigitated electrodes for biosensing applications: The impact of a dielectric barrier on the sensor properties JF - Physica Status Solidi (A) - Applications and Materials Science N2 - Planar and three-dimensional (3D) interdigitated electrodes (IDE) with electrode digits separated by an insulating barrier of different heights were electrochemically characterized and compared in terms of their sensing properties. Due to the impact of the surface resistance, both types of IDE structures display a non-linear behavior in low-ionic strength solutions. The experimental data were fitted to an electrical equivalent circuit and interpreted taking into account the surface-charge-governed properties. The effect of a charged polyelectrolyte layer electrostatically assembled onto the sensor surface on the surface resistance in solutions with different KCl concentration is studied. In case of the same electrode footprint, 3D-IDEs show a larger cell constant and a higher sensitivity to molecular adsorption than that of planar IDEs. The obtained results demonstrate the potential of 3D-IDEs as a new transducer structure for a direct label-free sensing of charged molecules. Y1 - 2014 U6 - http://dx.doi.org/10.1002/pssa.201330416 SN - 1521-396X (E-Journal); 1862-6319 (E-Journal); 0031-8965 (Print); 1862-6300 (Print) VL - 211 IS - 6 SP - 1357 EP - 1363 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Bäcker, Matthias A1 - Koch, Claudia A1 - Eiben, Sabine A1 - Geiger, Fania A1 - Eber, Fabian A1 - Gliemann, Hartmut A1 - Poghossian, Arshak A1 - Wege, Christina A1 - Schöning, Michael Josef T1 - Tobacco mosaic virus as enzyme nanocarrier for electrochemical biosensors JF - Sensors and Actuators B: Chemical N2 - The conjunction of (bio-)chemical recognition elements with nanoscale biological building blocks such as virus particles is considered as a very promising strategy for the creation of biohybrids opening novel opportunities for label-free biosensing. This work presents a new approach for the development of biosensors using tobacco mosaic virus (TMV) nanotubes or coat proteins (CPs) as enzyme nanocarriers. Sensor chips combining an array of Pt electrodes loaded with glucose oxidase (GOD)-modified TMV nanotubes or CP aggregates were used for amperometric detection of glucose as a model system for the first time. The presence of TMV nanotubes or CPs on the sensor surface allows binding of a high amount of precisely positioned enzymes without substantial loss of their activity, and may also ensure accessibility of their active centers for analyte molecules. Specific and efficient immobilization of streptavidin-conjugated GOD ([SA]-GOD) complexes on biotinylated TMV nanotubes or CPs was achieved via bioaffinity binding. These layouts were tested in parallel with glucose sensors with adsorptively immobilized [SA]-GOD, as well as [SA]-GOD crosslinked with glutardialdehyde, and came out to exhibit superior sensor performance. The achieved results underline a great potential of an integration of virus/biomolecule hybrids with electronic transducers for future applications in biosensorics and biochips. Y1 - 2017 U6 - http://dx.doi.org/10.1016/j.snb.2016.07.096 SN - 0925-4005 VL - 238 SP - 716 EP - 722 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Bäcker, Matthias A1 - Delle, L. A1 - Poghossian, Arshak A1 - Biselli, Manfred A1 - Zang, Werner A1 - Wagner, P. A1 - Schöning, Michael Josef T1 - Electrochemical sensor array for bioprocess monitoring JF - Electrochimica Acta (2011) Y1 - 2011 VL - 56 IS - 26 SP - 9673 EP - 9678 PB - Elsevier CY - Amsterdam ER -