TY - JOUR A1 - Molinnus, Denise A1 - Drinic, Aleksander A1 - Iken, Heiko A1 - Kröger, Nadja A1 - Zinser, Max A1 - Smeets, Ralf A1 - Köpf, Marius A1 - Kopp, Alexander A1 - Schöning, Michael Josef T1 - Towards a flexible electrochemical biosensor fabricated from biocompatible Bombyx mori silk JF - Biosensors and Bioelectronics Y1 - 2021 U6 - http://dx.doi.org/10.1016/j.bios.2021.113204 SN - 0956-5663 VL - 183 IS - Art. 113204 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Wert, Stefan A1 - Iken, Heiko A1 - Schöning, Michael Josef A1 - Matysik, Frank-Michael T1 - Development of a temperature‐pulse enhanced electrochemical glucose biosensor and characterization of its stability via scanning electrochemical microscopy JF - Electroanalysis N2 - Glucose oxidase (GOx) is an enzyme frequently used in glucose biosensors. As increased temperatures can enhance the performance of electrochemical sensors, we investigated the impact of temperature pulses on GOx that was drop-coated on flattened Pt microwires. The wires were heated by an alternating current. The sensitivity towards glucose and the temperature stability of GOx was investigated by amperometry. An up to 22-fold increase of sensitivity was observed. Spatially resolved enzyme activity changes were investigated via scanning electrochemical microscopy. The application of short (<100 ms) heat pulses was associated with less thermal inactivation of the immobilized GOx than long-term heating. Y1 - 2021 U6 - http://dx.doi.org/10.1002/elan.202100089 SN - 1521-4109 IS - Early View PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Yoshinobu, Tatsuo A1 - Schöning, Michael Josef T1 - Light-addressable potentiometric sensors (LAPS) for cell monitoring and biosensing JF - Current Opinion in Electrochemistry Y1 - 2021 U6 - http://dx.doi.org/10.1016/j.coelec.2021.100727 SN - 2451-9103 IS - In Press, Journal Pre-proof PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Givanoudi, Stella A1 - Cornelis, Peter A1 - Rasschaert, Geertrui A1 - Wackers, Gideon A1 - Iken, Heiko A1 - Rolka, David A1 - Yongabi, Derick A1 - Robbens, Johan A1 - Schöning, Michael Josef A1 - Heyndrickx, Marc A1 - Wagner, Patrick T1 - Selective Campylobacter detection and quantification in poultry: A sensor tool for detecting the cause of a common zoonosis at its source JF - Sensors and Actuators B: Chemical Y1 - 2021 U6 - http://dx.doi.org/10.1016/j.snb.2021.129484 SN - 0925-4005 IS - In Press, Journal Pre-proof SP - Article 129484 PB - Elsevier CY - Amsterdam ER - TY - THES A1 - Bronder, Thomas T1 - Label-free detection of tuberculosis DNA with capacitive field-effect biosensors Y1 - 2020 U6 - http://dx.doi.org/10.17192/z2021.0056 N1 - Dissertation, Universität, Marburg 2020 PB - Philipps-Universität Marburg CY - Marburg ER - TY - JOUR A1 - Jablonski, Melanie A1 - Münstermann, Felix A1 - Nork, Jasmina A1 - Molinnus, Denise A1 - Muschallik, Lukas A1 - Bongaerts, Johannes A1 - Wagner, Torsten A1 - Keusgen, Michael A1 - Siegert, Petra A1 - Schöning, Michael Josef T1 - Capacitive field‐effect biosensor applied for the detection of acetoin in alcoholic beverages and fermentation broths JF - physica status solidi (a) applications and materials science N2 - An acetoin biosensor based on a capacitive electrolyte–insulator–semiconductor (EIS) structure modified with the enzyme acetoin reductase, also known as butane-2,3-diol dehydrogenase (Bacillus clausii DSM 8716ᵀ), is applied for acetoin detection in beer, red wine, and fermentation broth samples for the first time. The EIS sensor consists of an Al/p-Si/SiO₂/Ta₂O₅ layer structure with immobilized acetoin reductase on top of the Ta₂O₅ transducer layer by means of crosslinking via glutaraldehyde. The unmodified and enzyme-modified sensors are electrochemically characterized by means of leakage current, capacitance–voltage, and constant capacitance methods, respectively. KW - acetoin KW - acetoin reductase KW - alcoholic beverages KW - biosensors KW - capacitive field-effect sensors Y1 - 2021 U6 - http://dx.doi.org/10.1002/pssa.202000765 SN - 1862-6319 VL - 218 IS - 13 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Jablonski, Melanie A1 - Poghossian, Arshak A1 - Severin, Robin A1 - Keusgen, Michael A1 - Wege, Christian A1 - Schöning, Michael Josef T1 - Capacitive Field-Effect Biosensor Studying Adsorption of Tobacco Mosaic Virus Particles JF - Micromachines N2 - Plant virus-like particles, and in particular, tobacco mosaic virus (TMV) particles, are increasingly being used in nano- and biotechnology as well as for biochemical sensing purposes as nanoscaffolds for the high-density immobilization of receptor molecules. The sensitive parameters of TMV-assisted biosensors depend, among others, on the density of adsorbed TMV particles on the sensor surface, which is affected by both the adsorption conditions and surface properties of the sensor. In this work, Ta₂O₅-gate field-effect capacitive sensors have been applied for the label-free electrical detection of TMV adsorption. The impact of the TMV concentration on both the sensor signal and the density of TMV particles adsorbed onto the Ta₂O₅-gate surface has been studied systematically by means of field-effect and scanning electron microscopy methods. In addition, the surface density of TMV particles loaded under different incubation times has been investigated. Finally, the field-effect sensor also demonstrates the label-free detection of penicillinase immobilization as model bioreceptor on TMV particles. KW - capacitive field-effect sensor KW - plant virus detection KW - tobacco mosaic virus (TMV) KW - TMV adsorption KW - Ta₂O₅ gate Y1 - 2021 U6 - http://dx.doi.org/10.3390/mi12010057 VL - 12 IS - 1 PB - MDPI CY - Basel ER - TY - JOUR A1 - Molinnus, Denise A1 - Iken, Heiko A1 - Johnen, Anna Lynn A1 - Richstein, Benjamin A1 - Hellmich, Lena A1 - Poghossian, Arshak A1 - Knoch, Joachim A1 - Schöning, Michael Josef T1 - Miniaturized pH-Sensitive Field-Effect Capacitors with Ultrathin Ta₂O₅ Films Prepared by Atomic Layer Deposition JF - physica status solidi (a) applications and materials science N2 - Miniaturized electrolyte–insulator–semiconductor capacitors (EISCAPs) with ultrathin gate insulators have been studied in terms of their pH-sensitive sensor characteristics: three different EISCAP systems consisting of Al–p-Si–Ta2O5(5 nm), Al–p-Si–Si3N4(1 or 2 nm)–Ta2O5 (5 nm), and Al–p-Si–SiO2(3.6 nm)–Ta2O5(5 nm) layer structures are characterized in buffer solution with different pH values by means of capacitance–voltage and constant capacitance method. The SiO2 and Si3N4 gate insulators are deposited by rapid thermal oxidation and rapid thermal nitridation, respectively, whereas the Ta2O5 film is prepared by atomic layer deposition. All EISCAP systems have a clear pH response, favoring the stacked gate insulators SiO2–Ta2O5 when considering the overall sensor characteristics, while the Si3N4(1 nm)–Ta2O5 stack delivers the largest accumulation capacitance (due to the lower equivalent oxide thickness) and a higher steepness in the slope of the capacitance–voltage curve among the studied stacked gate insulator systems. KW - atomic layer deposition KW - capacitive field-effect sensors KW - pH sensors KW - ultrathin gate insulators Y1 - 2022 U6 - http://dx.doi.org/10.1002/pssa.202100660 SN - 1862-6319 VL - 219 IS - 8 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Oliveira, Danilo A. A1 - Molinnus, Denise A1 - Beging, Stefan A1 - Siqueira Jr, José R. A1 - Schöning, Michael Josef T1 - Biosensor Based on Self-Assembled Films of Graphene Oxide and Polyaniline Using a Field-Effect Device Platform JF - physica status solidi (a) applications and materials science N2 - A new functionalization method to modify capacitive electrolyte–insulator–semiconductor (EIS) structures with nanofilms is presented. Layers of polyallylamine hydrochloride (PAH) and graphene oxide (GO) with the compound polyaniline:poly(2-acrylamido-2-methyl-1-propanesulfonic acid) (PANI:PAAMPSA) are deposited onto a p-Si/SiO2 chip using the layer-by-layer technique (LbL). Two different enzymes (urease and penicillinase) are separately immobilized on top of a five-bilayer stack of the PAH:GO/PANI:PAAMPSA-modified EIS chip, forming a biosensor for detection of urea and penicillin, respectively. Electrochemical characterization is performed by constant capacitance (ConCap) measurements, and the film morphology is characterized by atomic force microscopy (AFM) and scanning electron microscopy (SEM). An increase in the average sensitivity of the modified biosensors (EIS–nanofilm–enzyme) of around 15% is found in relation to sensors, only carrying the enzyme but without the nanofilm (EIS–enzyme). In this sense, the nanofilm acts as a stable bioreceptor onto the EIS chip improving the output signal in terms of sensitivity and stability. KW - capacitive electrolyte–insulator–semiconductor sensors KW - graphene oxide KW - layer-by-layer technique KW - nanomaterials KW - polyaniline Y1 - 2021 U6 - http://dx.doi.org/10.1002/pssa.202000747 SN - 1862-6319 N1 - Corresponding author: José R. Siqueira Jr & Michael J. Schöning VL - 218 IS - 13 SP - 1 EP - 9 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Welden, Rene A1 - Nagamine Komesu, Cindy A. A1 - Wagner, Patrick H. A1 - Schöning, Michael Josef A1 - Wagner, Torsten T1 - Photoelectrochemical enzymatic penicillin biosensor: A proof-of-concept experiment JF - Electrochemical Science Advances N2 - Photoelectrochemical (PEC) biosensors are a rather novel type of biosensors thatutilizelighttoprovideinformationaboutthecompositionofananalyte,enablinglight-controlled multi-analyte measurements. For enzymatic PEC biosensors,amperometric detection principles are already known in the literature. In con-trast, there is only a little information on H+-ion sensitive PEC biosensors. Inthis work, we demonstrate the detection of H+ions emerged by H+-generatingenzymes, exemplarily demonstrated with penicillinase as a model enzyme on atitanium dioxide photoanode. First, we describe the pH sensitivity of the sensorand study possible photoelectrocatalytic reactions with penicillin. Second, weshow the enzymatic PEC detection of penicillin. KW - enzymatic biosensor KW - penicillin KW - penicillinase KW - photoelectrochemistry KW - titanium dioxide photoanode Y1 - 2021 U6 - http://dx.doi.org/10.1002/elsa.202100131 SN - 2698-5977 N1 - Corresponding auhtor: Michael J. Schöning VL - 2 IS - 4 SP - 1 EP - 5 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Jablonski, Melanie A1 - Poghossian, Arshak A1 - Keusgen, Michael A1 - Wege, Christina A1 - Schöning, Michael Josef T1 - Detection of plant virus particles with a capacitive field-effect sensor JF - Analytical and Bioanalytical Chemistry N2 - Plant viruses are major contributors to crop losses and induce high economic costs worldwide. For reliable, on-site and early detection of plant viral diseases, portable biosensors are of great interest. In this study, a field-effect SiO2-gate electrolyte-insulator-semiconductor (EIS) sensor was utilized for the label-free electrostatic detection of tobacco mosaic virus (TMV) particles as a model plant pathogen. The capacitive EIS sensor has been characterized regarding its TMV sensitivity by means of constant-capacitance method. The EIS sensor was able to detect biotinylated TMV particles from a solution with a TMV concentration as low as 0.025 nM. A good correlation between the registered EIS sensor signal and the density of adsorbed TMV particles assessed from scanning electron microscopy images of the SiO2-gate chip surface was observed. Additionally, the isoelectric point of the biotinylated TMV particles was determined via zeta potential measurements and the influence of ionic strength of the measurement solution on the TMV-modified EIS sensor signal has been studied. KW - Plant virus KW - Capacitive field-effect sensor KW - Tobacco mosaic virus (TMV) KW - Label-free detection KW - Zeta potential Y1 - 2021 U6 - http://dx.doi.org/10.1007/s00216-021-03448-8 SN - 1618-2650 N1 - Corresponding authors: Arshak Poghossian & Michael J. Schöning VL - 413 SP - 5669 EP - 5678 PB - Springer Nature CY - Cham ER - TY - JOUR A1 - Poghossian, Arshak A1 - Karschuck, Tobias A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Field-Effect Capacitors Decorated with Ligand-Stabilized Gold Nanoparticles: Modeling and Experiments JF - Biosensors N2 - Nanoparticles are recognized as highly attractive tunable materials for designing field-effect biosensors with enhanced performance. In this work, we present a theoretical model for electrolyte-insulator-semiconductor capacitors (EISCAP) decorated with ligand-stabilized charged gold nanoparticles. The charged AuNPs are taken into account as additional, nanometer-sized local gates. The capacitance-voltage (C–V) curves and constant-capacitance (ConCap) signals of the AuNP-decorated EISCAPs have been simulated. The impact of the AuNP coverage on the shift of the C–V curves and the ConCap signals was also studied experimentally on Al–p-Si–SiO₂ EISCAPs decorated with positively charged aminooctanethiol-capped AuNPs. In addition, the surface of the EISCAPs, modified with AuNPs, was characterized by scanning electron microscopy for different immobilization times of the nanoparticles. KW - aminooctanethiol KW - nanoparticle coverage KW - capacitive model KW - gold nanoparticles KW - field-effect sensor KW - electrolyte-insulator-semiconductor capacitors Y1 - 2022 U6 - http://dx.doi.org/10.3390/bios12050334 SN - 2079-6374 N1 - This article belongs to the Special Issue "Biosensors in Nanotechnology" VL - 12 IS - 5 PB - MDPI CY - Basel ER - TY - JOUR A1 - Pourshahidi, Ali Mohammad A1 - Achtsnicht, Stefan A1 - Offenhäusser, Andreas A1 - Krause, Hans-Joachim ED - Offenhäusser, Andreas T1 - Frequency Mixing Magnetic Detection Setup Employing Permanent Ring Magnets as a Static Offset Field Source JF - Sensors N2 - Frequency mixing magnetic detection (FMMD) has been explored for its applications in fields of magnetic biosensing, multiplex detection of magnetic nanoparticles (MNP) and the determination of core size distribution of MNP samples. Such applications rely on the application of a static offset magnetic field, which is generated traditionally with an electromagnet. Such a setup requires a current source, as well as passive or active cooling strategies, which directly sets a limitation based on the portability aspect that is desired for point of care (POC) monitoring applications. In this work, a measurement head is introduced that involves the utilization of two ring-shaped permanent magnets to generate a static offset magnetic field. A steel cylinder in the ring bores homogenizes the field. By variation of the distance between the ring magnets and of the thickness of the steel cylinder, the magnitude of the magnetic field at the sample position can be adjusted. Furthermore, the measurement setup is compared to the electromagnet offset module based on measured signals and temperature behavior. KW - magnetic sensors KW - biosensors KW - frequency mixing magnetic detection KW - magnetic nanoparticles Y1 - 2022 U6 - http://dx.doi.org/10.3390/s22228776 SN - 1424-8220 VL - 22 IS - 22 PB - MDPI CY - Basel ER - TY - JOUR A1 - Karschuck, Tobias A1 - Kaulen, Corinna A1 - Poghossian, Arshak A1 - Wagner, Patrick H. A1 - Schöning, Michael Josef T1 - Gold nanoparticle-modified capacitive field-effect sensors: Studying the surface density of nanoparticles and coupling of charged polyelectrolyte macromolecules JF - Electrochemical Science Advances N2 - The coupling of ligand-stabilized gold nanoparticles with field-effect devices offers new possibilities for label-free biosensing. In this work, we study the immobilization of aminooctanethiol-stabilized gold nanoparticles (AuAOTs) on the silicon dioxide surface of a capacitive field-effect sensor. The terminal amino group of the AuAOT is well suited for the functionalization with biomolecules. The attachment of the positively-charged AuAOTs on a capacitive field-effect sensor was detected by direct electrical readout using capacitance-voltage and constant capacitance measurements. With a higher particle density on the sensor surface, the measured signal change was correspondingly more pronounced. The results demonstrate the ability of capacitive field-effect sensors for the non-destructive quantitative validation of nanoparticle immobilization. In addition, the electrostatic binding of the polyanion polystyrene sulfonate to the AuAOT-modified sensor surface was studied as a model system for the label-free detection of charged macromolecules. Most likely, this approach can be transferred to the label-free detection of other charged molecules such as enzymes or antibodies. KW - polystyrene sulfonate KW - gold nanoparticles KW - field-effect sensor KW - detection of charged macromolecules KW - capacitive EIS sensor Y1 - 2021 U6 - http://dx.doi.org/10.1002/elsa.202100179 SN - 0938-5193 VL - 2 IS - 5 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Vahidpour, Farnoosh A1 - Guthman, Eric A1 - Arreola, Julia A1 - Alghazali, Yousef H. M. A1 - Wagner, Torsten A1 - Schöning, Michael Josef T1 - Assessment of Various Process Parameters for Optimized Sterilization Conditions Using a Multi-Sensing Platform JF - Foods N2 - In this study, an online multi-sensing platform was engineered to simultaneously evaluate various process parameters of food package sterilization using gaseous hydrogen peroxide (H₂O₂). The platform enabled the validation of critical aseptic parameters. In parallel, one series of microbiological count reduction tests was performed using highly resistant spores of B. atrophaeus DSM 675 to act as the reference method for sterility validation. By means of the multi-sensing platform together with microbiological tests, we examined sterilization process parameters to define the most effective conditions with regards to the highest spore kill rate necessary for aseptic packaging. As these parameters are mutually associated, a correlation between different factors was elaborated. The resulting correlation indicated the need for specific conditions regarding the applied H₂O₂ gas temperature, the gas flow and concentration, the relative humidity and the exposure time. Finally, the novel multi-sensing platform together with the mobile electronic readout setup allowed for the online and on-site monitoring of the sterilization process, selecting the best conditions for sterility and, at the same time, reducing the use of the time-consuming and costly microbiological tests that are currently used in the food package industry. KW - spore kill rate KW - sterility KW - aseptic parameters KW - multi-sensing platform KW - gaseous hydrogen peroxide Y1 - 2022 U6 - http://dx.doi.org/10.3390/foods11050660 SN - 2304-8158 N1 - This article belongs to the Special Issue "Sensors and Biosensors Application for Food Industries" VL - 11 IS - 5 PB - MDPI CY - Basel ER - TY - JOUR A1 - Poghossian, Arshak A1 - Welden, Rene A1 - Buniatyan, Vahe V. A1 - Schöning, Michael Josef T1 - An Array of On-Chip Integrated, Individually Addressable Capacitive Field-Effect Sensors with Control Gate: Design and Modelling JF - Sensors N2 - The on-chip integration of multiple biochemical sensors based on field-effect electrolyte-insulator-semiconductor capacitors (EISCAP) is challenging due to technological difficulties in realization of electrically isolated EISCAPs on the same Si chip. In this work, we present a new simple design for an array of on-chip integrated, individually electrically addressable EISCAPs with an additional control gate (CG-EISCAP). The existence of the CG enables an addressable activation or deactivation of on-chip integrated individual CG-EISCAPs by simple electrical switching the CG of each sensor in various setups, and makes the new design capable for multianalyte detection without cross-talk effects between the sensors in the array. The new designed CG-EISCAP chip was modelled in so-called floating/short-circuited and floating/capacitively-coupled setups, and the corresponding electrical equivalent circuits were developed. In addition, the capacitance-voltage curves of the CG-EISCAP chip in different setups were simulated and compared with that of a single EISCAP sensor. Moreover, the sensitivity of the CG-EISCAP chip to surface potential changes induced by biochemical reactions was simulated and an impact of different parameters, such as gate voltage, insulator thickness and doping concentration in Si, on the sensitivity has been discussed. KW - equivalent circuit KW - multianalyte detection KW - control gate KW - on-chip integrated addressable EISCAP sensors KW - capacitive field-effect sensor Y1 - 2021 U6 - http://dx.doi.org/10.3390/s21186161 SN - 1424-8220 N1 - This article belongs to the Special Issue "Field-Effect Sensors: From pH Sensing to Biosensing" VL - 21 IS - 18 SP - 17 PB - MDPI CY - Basel ER - TY - JOUR A1 - Pourshahidi, Ali Mohammad A1 - Achtsnicht, Stefan A1 - Nambipareechee, Mrinal Murali A1 - Offenhäusser, Andreas A1 - Krause, Hans-Joachim T1 - Multiplex detection of magnetic beads using offset field dependent frequency mixing magnetic detection JF - Sensors N2 - Magnetic immunoassays employing Frequency Mixing Magnetic Detection (FMMD) have recently become increasingly popular for quantitative detection of various analytes. Simultaneous analysis of a sample for two or more targets is desirable in order to reduce the sample amount, save consumables, and save time. We show that different types of magnetic beads can be distinguished according to their frequency mixing response to a two-frequency magnetic excitation at different static magnetic offset fields. We recorded the offset field dependent FMMD response of two different particle types at frequencies ƒ₁ + n⋅ƒ₂, n = 1, 2, 3, 4 with ƒ₁ = 30.8 kHz and ƒ₂ = 63 Hz. Their signals were clearly distinguishable by the locations of the extremes and zeros of their responses. Binary mixtures of the two particle types were prepared with different mixing ratios. The mixture samples were analyzed by determining the best linear combination of the two pure constituents that best resembled the measured signals of the mixtures. Using a quadratic programming algorithm, the mixing ratios could be determined with an accuracy of greater than 14%. If each particle type is functionalized with a different antibody, multiplex detection of two different analytes becomes feasible. KW - colorization KW - multiplex detection KW - frequency mixing magnetic detection KW - magnetic nanoparticles Y1 - 2021 U6 - http://dx.doi.org/10.3390/s21175859 SN - 1424-8220 N1 - This article belongs to the Special Issue "Advanced Nanomaterial-Based Sensors for Biomedical Applications" VL - 21 IS - 17 PB - MDPI CY - Basel ER - TY - JOUR A1 - Haeger, Gerrit A1 - Grankin, Alina A1 - Wagner, Michaela T1 - Construction of an Aspergillus oryzae triple amylase deletion mutant as a chassis to evaluate industrially relevant amylases using multiplex CRISPR/Cas9 editing technology JF - Applied Research N2 - Aspergillus oryzae is an industrially relevant organism for the secretory production of heterologous enzymes, especially amylases. The activities of potential heterologous amylases, however, cannot be quantified directly from the supernatant due to the high background activity of native α-amylase. This activity is caused by the gene products of amyA, amyB, and amyC. In this study, an in vitro CRISPR/Cas9 system was established in A. oryzae to delete these genes simultaneously. First, pyrG of A. oryzae NSAR1 was mutated by exploiting NHEJ to generate a counter-selection marker. Next, all amylase genes were deleted simultaneously by co-transforming a repair template carrying pyrG of Aspergillus nidulans and flanking sequences of amylase gene loci. The rate of obtained triple knock-outs was 47%. We showed that triple knockouts do not retain any amylase activity in the supernatant. The established in vitro CRISPR/Cas9 system was used to achieve sequence-specific knock-in of target genes. The system was intended to incorporate a single copy of the gene of interest into the desired host for the development of screening methods. Therefore, an integration cassette for the heterologous Fpi amylase was designed to specifically target the amyB locus. The site-specific integration rate of the plasmid was 78%, with exceptional additional integrations. Integration frequency was assessed via qPCR and directly correlated with heterologous amylase activity. Hence, we could compare the efficiency between two different signal peptides. In summary, we present a strategy to exploit CRISPR/Cas9 for gene mutation, multiplex knock-out, and the targeted knock-in of an expression cassette in A. oryzae. Our system provides straightforward strain engineering and paves the way for development of fungal screening systems. KW - aspergillus KW - CRISPR/Cas9 KW - filamentous fungi KW - genome engineering Y1 - 2023 U6 - http://dx.doi.org/10.1002/appl.202200106 SN - 2702-4288 IS - Early View SP - 1 EP - 15 PB - Wiley-VCH ER - TY - JOUR A1 - Vahidpour, Farnoosh A1 - Alghazali, Yousef A1 - Akca, Sevilay A1 - Hommes, Gregor A1 - Schöning, Michael Josef T1 - An Enzyme-Based Interdigitated Electrode-Type Biosensor for Detecting Low Concentrations of H₂O₂ Vapor/Aerosol JF - Chemosensors N2 - This work introduces a novel method for the detection of H₂O₂ vapor/aerosol of low concentrations, which is mainly applied in the sterilization of equipment in medical industry. Interdigitated electrode (IDE) structures have been fabricated by means of microfabrication techniques. A differential setup of IDEs was prepared, containing an active sensor element (active IDE) and a passive sensor element (passive IDE), where the former was immobilized with an enzymatic membrane of horseradish peroxidase that is selective towards H₂O₂. Changes in the IDEs’ capacitance values (active sensor element versus passive sensor element) under H₂O₂ vapor/aerosol atmosphere proved the detection in the concentration range up to 630 ppm with a fast response time (<60 s). The influence of relative humidity was also tested with regard to the sensor signal, showing no cross-sensitivity. The repeatability assessment of the IDE biosensors confirmed their stable capacitive signal in eight subsequent cycles of exposure to H₂O₂ vapor/aerosol. Room-temperature detection of H₂O₂ vapor/aerosol with such miniaturized biosensors will allow a future three-dimensional, flexible mapping of aseptic chambers and help to evaluate sterilization assurance in medical industry. Y1 - 2022 U6 - http://dx.doi.org/10.3390/chemosensors10060202 SN - 2227-9040 N1 - This article belongs to the Special Issue "Bioinspired Chemical Sensors and Micro-Nano Devices" VL - 10 IS - 6 PB - MDPI CY - Basel ER - TY - JOUR A1 - Haeger, Gerrit A1 - Jolmes, Tristan A1 - Oyen, Sven A1 - Jaeger, Karl-Erich A1 - Bongaerts, Johannes A1 - Schörken, Ulrich A1 - Siegert, Petra T1 - Novel recombinant aminoacylase from Paraburkholderia monticola capable of N-acyl-amino acid synthesis JF - Applied Microbiology and Biotechnology N2 - N-Acyl-amino acids can act as mild biobased surfactants, which are used, e.g., in baby shampoos. However, their chemical synthesis needs acyl chlorides and does not meet sustainability criteria. Thus, the identification of biocatalysts to develop greener synthesis routes is desirable. We describe a novel aminoacylase from Paraburkholderia monticola DSM 100849 (PmAcy) which was identified, cloned, and evaluated for its N-acyl-amino acid synthesis potential. Soluble protein was obtained by expression in lactose autoinduction medium and co-expression of molecular chaperones GroEL/S. Strep-tag affinity purification enriched the enzyme 16-fold and yielded 15 mg pure enzyme from 100 mL of culture. Biochemical characterization revealed that PmAcy possesses beneficial traits for industrial application like high temperature and pH-stability. A heat activation of PmAcy was observed upon incubation at temperatures up to 80 °C. Hydrolytic activity of PmAcy was detected with several N-acyl-amino acids as substrates and exhibited the highest conversion rate of 773 U/mg with N-lauroyl-L-alanine at 75 °C. The enzyme preferred long-chain acyl-amino-acids and displayed hardly any activity with acetyl-amino acids. PmAcy was also capable of N-acyl-amino acid synthesis with good conversion rates. The best synthesis results were obtained with the cationic L-amino acids L-arginine and L-lysine as well as with L-leucine and L-phenylalanine. Exemplarily, L-phenylalanine was acylated with fatty acids of chain lengths from C8 to C18 with conversion rates of up to 75%. N-lauroyl-L-phenylalanine was purified by precipitation, and the structure of the reaction product was verified by LC–MS and NMR. KW - Chaperone KW - Biocatalysis KW - Aminoacylase KW - Acylation KW - Acyl-amino acids KW - Biosurfactants Y1 - 2024 U6 - http://dx.doi.org/10.1007/s00253-023-12868-8 SN - 1432-0614 N1 - Corresponding author: Petra Siegert IS - 108 PB - Springer CY - Berlin ER -