TY - JOUR A1 - Ribitsch, D. A1 - Karl, W. A1 - Birner-Gruenberger, R. A1 - Gruber, K. A1 - Eiteljoerg, I. A1 - Remler, P. A1 - Wieland, S. A1 - Siegert, Petra A1 - Maurer, Karl-Heinz A1 - Schwab, H. T1 - C-terminal truncation of a metagenome-derived detergent protease for effective expression in E. coli JF - Journal of biotechnology N2 - Recently, a new alkaline protease named HP70 showing highest homology to extracellular serine proteases of Stenotrophomonas maltophilia and Xanthomonas campestris was found in the course of a metagenome screening for detergent proteases (Niehaus et al., submitted for publication). Attempts to efficiently express the enzyme in common expression hosts had failed. This study reports on the realization of overexpression in Escherichia coli after structural modification of HP70. Modelling of HP70 resulted in a two-domain structure, comprising the catalytic domain and a C-terminal domain which includes about 100 amino acids. On the basis of the modelled structure the enzyme was truncated by deletion of most of the C-terminal domain yielding HP70-C477. This structural modification allowed effective expression of active enzyme using E. coli BL21-Gold as the host. Specific activity of HP70-C477 determined with suc-l-Ala-l-Ala-l-Pro-l-Phe-p-nitroanilide as the substrate was 30 ± 5 U/mg compared to 8 ± 1 U/mg of the native enzyme. HP70-C477 was most active at 40 °C and pH 7–11; these conditions are prerequisite for a potential application as detergent enzyme. Determination of kinetic parameters at 40 °C and pH = 9.5 resulted in KM = 0.23 ± 0.01 mM and kcat = 167.5 ± 3.6 s⁻¹. MS-analysis of peptide fragments obtained from incubation of HP70 and HP70-C477 with insulin B indicated that the C-terminal domain influences the cleavage preferences of the enzyme. Washing experiments confirmed the high potential of HP70-C477 as detergent protease. Y1 - 2010 U6 - http://dx.doi.org/10.1016/j.jbiotec.2010.09.947 SN - 1873-4863 (E-Journal); 0168-1656 (Print) VL - 150 IS - 3 SP - 408 EP - 416 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Ribitsch, D. A1 - Heumann, S. A1 - Karl, W. A1 - Gerlach, J. A1 - Leber, R. A1 - Birner-Gruenberger, R. A1 - Gruber, K. A1 - Eiteljoerg, I. A1 - Remler, P. A1 - Siegert, Petra A1 - Lange, J. A1 - Maurer, Karl-Heinz A1 - Berg, G. A1 - Guebitz, G. M. A1 - Schwab, H. T1 - Extracellular serine proteases from Stenotrophomonas maltophilia: Screening, isolation and heterologous expression in E. coli JF - Journal of biotechnology N2 - A large strain collection comprising antagonistic bacteria was screened for novel detergent proteases. Several strains displayed protease activity on agar plates containing skim milk but were inactive in liquid media. Encapsulation of cells in alginate beads induced protease production. Stenotrophomonas maltophilia emerged as best performer under washing conditions. For identification of wash-active proteases, four extracellular serine proteases called StmPr1, StmPr2, StmPr3 and StmPr4 were cloned. StmPr2 and StmPr4 were sufficiently overexpressed in E. coli. Expression of StmPr1 and StmPr3 resulted in unprocessed, insoluble protein. Truncation of most of the C-terminal domain which has been identified by enzyme modeling succeeded in expression of soluble, active StmPr1 but failed in case of StmPr3. From laundry application tests StmPr2 turned out to be a highly wash-active protease at 45 °C. Specific activity of StmPr2 determined with suc-l-Ala-l-Ala-l-Pro-l-Phe-p-nitroanilide as the substrate was 17 ± 2 U/mg. In addition we determined the kinetic parameters and cleavage preferences of protease StmPr2. KW - Alginate beads KW - Stenotrophomonas maltophilia KW - Detergent protease Y1 - 2012 U6 - http://dx.doi.org/10.1016/j.jbiotec.2011.09.025 SN - 1873-4863 (E-Journal); 0168-1656 (Print) VL - 157 IS - 1 SP - 140 EP - 147 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Reisert, Steffen A1 - Schneider, Benno A1 - Geissler, Hanno A1 - Gompel, Matthias van A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Multi-sensor chip for the investigation of different types of metal oxides for the detection of H2O2 in the ppm range JF - physica status solidi (a) N2 - In this work, a multi-sensor chip for the investigation of the sensing properties of different types of metal oxides towards hydrogen peroxide in the ppm range is presented. The fabrication process and physical characterization of the multi-sensor chip are described. Pure SnO2 and WO3 as well as Pd- and Pt-doped SnO2 films are characterized in terms of their sensitivity to H2O2. The sensing films have been prepared by drop-coating of water-dispensed nano-powders. A physical characterization, including scanning electron microscopy and X-ray diffraction analysis of the deposited metal-oxide films, was done. From the measurements in hydrogen peroxide atmosphere, it could be shown, that all of the tested metal oxide films are suitable for the detection of H2O2 in the ppm range. The highest sensitivity and reproducibility was achieved using Pt-doped SnO2. Calibration plot of a SnO2, WO3, Pt-, and Pd-doped SnO2 gas sensor for H2O2 concentrations in the ppm range. Y1 - 2013 SN - 1862-6319 VL - 210 IS - 5 SP - 898 EP - 904 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Reisert, Steffen A1 - Henkel, Hartmut A1 - Schneider, Andreas A1 - Schäfer, Daniel A1 - Friedrich, Peter A1 - Berger, Jörg A1 - Schöning, Michael Josef T1 - Development of a handheld sensor system for the online measurement of hydrogen peroxide in aseptic filling systems JF - Physica Status Solidi (A). 207 (2010), H. 4 Y1 - 2010 SN - 1862-6300 N1 - Special Issue: Engineering of Functional Interfaces EnFI 2009 SP - 913 EP - 918 ER - TY - JOUR A1 - Reisert, Steffen A1 - Henkel, H. A1 - Schneider, A. A1 - Schäfer, D. A1 - Friedrich, P. A1 - Berger, J. A1 - Schöning, Michael Josef T1 - Entwicklung eines Handheld-Sensorsystems für die „On-line“-Messung der H2O2-Konzentration in aseptischen Entkeimungsprozessen JF - 9. Dresdner Sensor-Symposium : Dresden, 07.-09. Dezember 2009 / Gerlach, Gerald ; Hauptmann, Peter [Hrsg.] Y1 - 2009 SN - 978-3-941298-44-6 N1 - Dresdner Sensor-Symposium ; (9, 2009, Dresden) SP - 285 EP - 288 PB - TUDpress CY - Dresden ER - TY - JOUR A1 - Reisert, Steffen A1 - Geissler, Hanno A1 - Flörke, Rudolf A1 - Weiler, Christian A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Characterisation of aseptic sterilisation processes using an electronic nose JF - International journal of nanotechnology Y1 - 2013 SN - 1475-7435 (Print) 7141-8151 (Online) VL - Vol. 10 IS - No. 5-7 SP - 470 EP - 484 PB - Inderscience Enterprises CY - Genève ER - TY - JOUR A1 - Reisert, Steffen A1 - Geissler, Hanno A1 - Flörke, Rudolf A1 - Näther, Niko A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Towards a multi-sensor system for the evaluation of aseptic processes employing hydrogen peroxide vapour (H2O2) JF - Physica status solidi (a) : applications and material science. 208 (2011), H. 6 Y1 - 2011 SN - 1862-6319 SP - 1351 EP - 1356 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Reisert, Steffen A1 - Geissler, H. A1 - Weiler, C. A1 - Wagner, P. A1 - Schöning, Michael Josef T1 - Multiple sensor-type system for monitoring the microbicidal effectiveness of aseptic sterilisation processes JF - Food control N2 - The present work describes a novel multiple sensor-type system for the real-time analysis of aseptic sterilisation processes employing gaseous hydrogen peroxide (H2O2) as a sterilant. The inactivation kinetics of Bacillus atrophaeus by gaseous H2O2 have been investigated by means of a methodical calibration experiment, taking into account the process variables H2O2 concentration, humidity and gas temperature. It has been found that the microbicidal effectiveness at H2O2 concentrations above 2% v/v is largely determined by the concentration itself, while at lower H2O2 concentrations, the gas temperature and humidity play a leading role. Furthermore, the responses of different types of gas sensors towards the influencing factors of the sterilisation process have been analysed within the same experiment. Based on a correlation established between the inactivation kinetics and the sensor responses, a calorimetric H2O2 sensor and a metal-oxide semiconductor (MOX) sensor have been identified as possible candidates for monitoring the microbicidal effectiveness of aseptic sterilisation processes employing gaseous H2O2. Therefore, two linear models that describe the relationship between sensor response and microbicidal effectiveness have been proposed. Y1 - 2015 U6 - http://dx.doi.org/10.1016/j.foodcont.2014.07.063 SN - 1873-7129 (E-Journal); 0956-7135 (Print) VL - 47 SP - 615 EP - 622 ER - TY - JOUR A1 - Reisert, Steffen A1 - Geissler, H. A1 - Florke, R. A1 - Wagner, P. A1 - Wagner, Torsten A1 - Schöning, Michael Josef T1 - Controlling aseptic sterilization processes by means of a multi-sensor system Y1 - 2011 N1 - 2011 IEEE Workshop on Merging Fields of Computational Intelligence and Sensor Technology ; 11.-15. April 2011 Paris, France SP - 18 EP - 22 PB - IEEE CY - New York ER - TY - JOUR A1 - Rachinger, Michael A1 - Bauch, Melanie A1 - Strittmatter, Axel A1 - Bongaerts, Johannes A1 - Evers, Stefan A1 - Maurer, Karl-Heinz A1 - Daniel, Rolf A1 - Liebl, Wolfgang A1 - Liesegang, Heiko A1 - Ehrenreich, Armin T1 - Size unlimited markerless deletions by a transconjugative plasmid-system in Bacillus licheniformis JF - Journal of biotechnology Y1 - 2013 SN - 1873-4863 (E-Journal); 0168-1656 (Print) VL - Vol. 164 IS - Iss. 4 SP - 365 EP - 369 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Rabehi, Amine A1 - Garlan, Benjamin A1 - Achtsnicht, Stefan A1 - Krause, Hans-Joachim A1 - Offenhäusser, Andreas A1 - Ngo, Kieu A1 - Neveu, Sophie A1 - Graff-Dubois, Stephanie A1 - Kokabi, Hamid T1 - Magnetic detection structure for Lab-on-Chip applications based on the frequency mixing technique JF - Sensors N2 - A magnetic frequency mixing technique with a set of miniaturized planar coils was investigated for use with a completely integrated Lab-on-Chip (LoC) pathogen sensing system. The system allows the detection and quantification of superparamagnetic beads. Additionally, in terms of magnetic nanoparticle characterization ability, the system can be used for immunoassays using the beads as markers. Analytical calculations and simulations for both excitation and pick-up coils are presented; the goal was to investigate the miniaturization of simple and cost-effective planar spiral coils. Following these calculations, a Printed Circuit Board (PCB) prototype was designed, manufactured, and tested for limit of detection, linear response, and validation of theoretical concepts. Using the magnetic frequency mixing technique, a limit of detection of 15 µg/mL of 20 nm core-sized nanoparticles was achieved without any shielding. KW - Lab-on-Chip KW - magnetic sensing KW - frequency mixing KW - superparamagnetic nanoparticles KW - magnetic beads Y1 - 2018 U6 - http://dx.doi.org/10.3390/s18061747 SN - 1424-8220 VL - 18 IS - 6 PB - MDPI CY - Basel ER - TY - JOUR A1 - Pourshahidi, Ali Mohammad A1 - Engelmann, Ulrich M. A1 - Offenhäusser, Andreas A1 - Krause, Hans-Joachim T1 - Resolving ambiguities in core size determination of magnetic nanoparticles from magnetic frequency mixing data JF - Journal of Magnetism and Magnetic Materials N2 - Frequency mixing magnetic detection (FMMD) has been widely utilized as a measurement technique in magnetic immunoassays. It can also be used for the characterization and distinction (also known as “colourization”) of different types of magnetic nanoparticles (MNPs) based on their core sizes. In a previous work, it was shown that the large particles contribute most of the FMMD signal. This leads to ambiguities in core size determination from fitting since the contribution of the small-sized particles is almost undetectable among the strong responses from the large ones. In this work, we report on how this ambiguity can be overcome by modelling the signal intensity using the Langevin model in thermodynamic equilibrium including a lognormal core size distribution fL(dc,d0,σ) fitted to experimentally measured FMMD data of immobilized MNPs. For each given median diameter d0, an ambiguous amount of best-fitting pairs of parameters distribution width σ and number of particles Np with R2 > 0.99 are extracted. By determining the samples’ total iron mass, mFe, with inductively coupled plasma optical emission spectrometry (ICP-OES), we are then able to identify the one specific best-fitting pair (σ, Np) one uniquely. With this additional externally measured parameter, we resolved the ambiguity in core size distribution and determined the parameters (d0, σ, Np) directly from FMMD measurements, allowing precise MNPs sample characterization. Y1 - 2022 U6 - http://dx.doi.org/10.1016/j.jmmm.2022.169969 SN - 0304-8853 VL - 563 IS - In progress, Art. No. 169969 PB - Elsevier CY - Amsterdam 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 - 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 - Polen, T. A1 - Krämer, Marco A1 - Bongaerts, Johannes A1 - Wubbolts, Marcel A1 - Wendisch, V. F. T1 - The global gene expression response of Escherichia coli to L-phenylalanine JF - Journal of biotechnology Y1 - 2005 SN - 1873-4863 (E-Journal); 0168-1656 (Print) VL - Vol. 115 IS - Iss. 3 SP - 221 EP - 237 ER - TY - JOUR A1 - Poghossian, Arshak A1 - Yoshinobu, Tatsuo A1 - Simonis, A. A1 - Ecken, H. A1 - Lüth, Hans A1 - Schöning, Michael Josef T1 - Penicillin detection by means of field-effect based sensors: EnFET, capacitive EIS sensor or LAPS? JF - Sensors and Actuators B. 78 (2001), H. 1-3 Y1 - 2001 SN - 0925-4005 SP - 237 EP - 242 ER - TY - JOUR A1 - Poghossian, Arshak A1 - Yoshinobu, Tatsuo A1 - Schöning, Michael Josef T1 - Flow-velocity microsensors based on semiconductor field-effect structures JF - Sensors. 3 (2003), H. 7 Y1 - 2003 SN - 1424-8220 SP - 202 EP - 212 ER - TY - JOUR A1 - Poghossian, Arshak A1 - Yoshinobu, T. A1 - Simonis, A. A1 - Ecken, H. A1 - Lüth, H. A1 - Schöning, Michael Josef T1 - Penicillin detection by means of field-effect based sensors: EnFET, capacitive EIS sensor or LAPS? JF - Proceedings : Copenhagen, Denmark, 27 - 30 August 2000 / [ed.: R. de Reus ...] Y1 - 2000 SN - 87-89935-50-0 N1 - Eurosensors ; (14, 2000, København) ; Eurosensors ; (14 : ; 2000.08.27-30 : ; Copenhagen) ; European Conference on Solid-State Transducers ; (14 : ; 2000.08.27-30 : ; Copenhagen) SP - 27 EP - 30 PB - MIC, Mikroelektronik Centret CY - Lyngby, Denmark ER - TY - JOUR A1 - Poghossian, Arshak A1 - Werner, Frederik A1 - Buniatyan, V. V. A1 - Wagner, Torsten A1 - Miamoto, K. A1 - Yoshinobu, T. A1 - Schöning, Michael Josef T1 - Towards addressability of light-addressable potentiometric sensors: Shunting effect of non-illuminated region and cross-talk JF - Sensor and Actuators B: Chemical N2 - The LAPS (light-addressable potentiometric sensor) platform is one of the most attractive approaches for chemical and biological sensing with many applications ranging from pH and ion/analyte concentration measurements up to cell metabolism detection and chemical imaging. However, although it is generally accepted that LAPS measurements are spatially resolved, the light-addressability feature of LAPS devices has not been discussed in detail so far. In this work, an extended electrical equivalent-circuit model of the LAPS has been presented, which takes into account possible cross-talk effects due to the capacitive coupling of the non-illuminated region. A shunting effect of the non-illuminated area on the measured photocurrent and addressability of LAPS devices has been studied. It has been shown, that the measured photocurrent will be determined not only by the local interfacial potential in the illuminated region but also by possible interfacial potential changes in the non-illuminated region, yielding cross-talk effects. These findings were supported by the experimental investigations of a penicillin-sensitive multi-spot LAPS and a metal-insulator-semiconductor LAPS as model systems. Y1 - 2017 U6 - http://dx.doi.org/10.1016/j.snb.2017.01.047 SN - 0925-4005 IS - 244 SP - 1071 EP - 1079 PB - Elsevier CY - Amsterdam 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 -