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 - Krafft, B. A1 - Förster, Arnold A1 - Hart, A. van der A1 - Schäpers, T. T1 - Control of Aharonov-Bohm oscillations in a AlGaAs/GaAs ring by asymmetric and symmetric gate biasing JF - Physica E: Low-dimensional Systems and Nanostructures. 9 (2001), H. 4 Y1 - 2001 SN - 1386-9477 SP - 635 EP - 641 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 - Mennicken, Max A1 - Peter, Sophia Katharina A1 - Kaulen, Corinna A1 - Simon, Ulrich A1 - Karthäuser, Silvia T1 - Controlling the Electronic Contact at the Terpyridine/Metal Interface JF - The Journal of Physical Chemistry C Y1 - 2019 U6 - http://dx.doi.org/10.1021/acs.jpcc.9b05865 SN - 1932-7455 VL - 123 IS - 35 SP - 21367 EP - 21375 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 - Mourzina, Y. G. A1 - Schöning, Michael Josef A1 - Schubert, J. A1 - Zander, W. A1 - Legin, A. V. A1 - Vlasov, Y. G. A1 - Lüth, H. T1 - Copper, cadmium and thallium thin film sensors based on chalcogenide glasses JF - Analytica Chimica Acta. 433 (2001) Y1 - 2001 SN - 0378-4304 SP - 103 EP - 110 ER - TY - JOUR A1 - Krämer, Melina A1 - Pita, Marcos A1 - Zhou, Jian A1 - Ornatska, Maryna A1 - Poghossian, Arshak A1 - Schöning, Michael Josef A1 - Katz, Evgeny T1 - Coupling of Biocomputing Systems with Electronic Chips: Electronic Interface for Transduction of Biochemical Information JF - Journal of Physical Chemistry C: Nanomaterials and Interfaces. 113 (2009), H. 6 Y1 - 2009 SN - 1932-7455 SP - 2573 EP - 2579 PB - American Cemical Society CY - Washington, DC ER - TY - JOUR A1 - Molinnus, Denise A1 - Poghossian, Arshak A1 - Keusgen, Michael A1 - Katz, Evgeny A1 - Schöning, Michael Josef T1 - Coupling of Biomolecular Logic Gates with Electronic Transducers: From Single Enzyme Logic Gates to Sense/Act/Treat Chips JF - Electroanalysis N2 - The integration of biomolecular logic principles with electronic transducers allows designing novel digital biosensors with direct electrical output, logically triggered drug-release, and closed-loop sense/act/treat systems. This opens new opportunities for advanced personalized medicine in the context of theranostics. In the present work, we will discuss selected examples of recent developments in the field of interfacing enzyme logic gates with electrodes and semiconductor field-effect devices. Special attention is given to an enzyme OR/Reset logic gate based on a capacitive field-effect electrolyte-insulator-semiconductor sensor modified with a multi-enzyme membrane. Further examples are a digital adrenaline biosensor based on an AND logic gate with binary YES/NO output and an integrated closed-loop sense/act/treat system comprising an amperometric glucose sensor, a hydrogel actuator, and an insulin (drug) sensor. Y1 - 2017 U6 - http://dx.doi.org/10.1002/elan.201700208 SN - 1521-4109 VL - 29 IS - 8 SP - 1840 EP - 1849 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Tillmann, K. A1 - Förster, Arnold T1 - Critical dimensions for the formation of interfacial misfit dislocations of In0.6Ga0.4As islands on GaAs(001) JF - Thin Solid Films. 368 (2000), H. 1 Y1 - 2000 SN - 0040-6090 SP - 93 EP - 104 ER - TY - JOUR A1 - Poghossian, Arshak A1 - Thust, M. A1 - Schöning, Michael Josef A1 - Müller-Veggian, Mattea A1 - Kordos, P. A1 - Lüth, H. T1 - Cross-sensitivity of a capacitive penicillin sensor combined with a diffusion barrier JF - Sensors and Actuators B. 68 (2000), H. 1-3 Y1 - 2000 SN - 0925-4005 SP - 260 EP - 265 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 - Pinkenburg, Olaf A1 - Schiffels, Johannes A1 - Selmer, Thorsten T1 - Das CoLibry-Konzept – ein Werkzeugkasten für die Synthetische Biologie: Bioproduktion JF - BIOspektrum N2 - Regardless of size or destination, synthetic biology starts with com-parably small information units, which need to be combined and properly arranged in order to achieve a certain goal. This may be the de novo synthesis of individual genes from oligonucleotides, a shuffling of protein domains in order to create novel biocatalysts, the assembly of multiple enzyme encoding genes in metabolic pathway design, or strain development at the production stage. The CoLibry concept has been designed in order to close the gap between recombinant production of individual genes and genome editing. Y1 - 2016 U6 - http://dx.doi.org/10.1007/s12268-016-0734-8 VL - 22 IS - 6 SP - 593 EP - 595 PB - Springer CY - Berlin 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 - Schusser, Sebastian A1 - Menzel, S. A1 - Bäcker, Matthias A1 - Leinhos, Marcel A1 - Poghossian, Arshak A1 - Wagner, P. A1 - Schöning, Michael Josef T1 - Degradation of thin poly(lactic acid) films: characterization by capacitance-voltage, atomic force microscopy, scanning electron microscopy and contact-angle measurements JF - Electrochimica Acta Y1 - 2013 SN - 1873-3859 (E-Journal); 0013-4686 (Print) VL - Vol. 113 SP - 779 EP - 784 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Bäcker, Matthias A1 - Schusser, Sebastian A1 - Poghossian, Arshak A1 - Schöning, Michael Josef T1 - Degradationsverhalten bioabbaubarer Polymere : siliziumbasierte Sensorik zur Überwachung JF - GIT Labor-Fachzeitschrift Y1 - 2013 SN - 0016-3538 SP - 32 EP - 33 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Schöning, Michael Josef A1 - Schütz, S. T1 - Der Käfer/Chip-Sensor als bioelektronischer “Schnüffler” JF - Bioanalytik für Einsteiger : [Diabetes, Drogen und DNA] / Reinhard Renneberg. Darja Süßbier (Ill.). [Red.: Andreas Held ...] Y1 - 2009 SN - 978-3-8274-1831-9 SP - 204 EP - 205 PB - Spektrum, Akad. Verl. CY - Heidelberg ER - TY - JOUR A1 - Almajhdi, Fahad N. A1 - Senger, Tilo A1 - Amer, Haitham M. A1 - Gissmann, Lutz A1 - Öhlschläger, Peter T1 - Design of a highly effective therapeutic HPV16 E6/E7-specific DNA vaccine: optimization by different ways of sequence rearrangements (Shuffling) JF - PLOS one N2 - Persistent infection with the high-risk Human Papillomavirus type 16 (HPV 16) is the causative event for the development of cervical cancer and other malignant tumors of the anogenital tract and of the head and neck. Despite many attempts to develop therapeutic vaccines no candidate has entered late clinical trials. An interesting approach is a DNA based vaccine encompassing the nucleotide sequence of the E6 and E7 viral oncoproteins. Because both proteins are consistently expressed in HPV infected cells they represent excellent targets for immune therapy. Here we report the development of 8 DNA vaccine candidates consisting of differently rearranged HPV-16 E6 and E7 sequences within one molecule providing all naturally occurring epitopes but supposedly lacking transforming activity. The HPV sequences were fused to the J-domain and the SV40 enhancer in order to increase immune responses. We demonstrate that one out of the 8 vaccine candidates induces very strong cellular E6- and E7- specific cellular immune responses in mice and, as shown in regression experiments, efficiently controls growth of HPV 16 positive syngeneic tumors. This data demonstrates the potential of this vaccine candidate to control persistent HPV 16 infection that may lead to malignant disease. It also suggests that different sequence rearrangements influence the immunogenecity by an as yet unknown mechanism. Y1 - 2014 U6 - http://dx.doi.org/10.1371/journal.pone.0113461 SN - 1932-6203 VL - 11 IS - 9 PB - PLOS CY - San Francisco ER - TY - JOUR A1 - Poghossian, Arshak A1 - Schöning, Michael Josef T1 - Detecting Both Physical and (Bio-)Chemical Parameters by Means of ISFET Devices JF - Electroanalysis. 16 (2004), H. 22 Y1 - 2004 SN - 1040-0397 SP - 1863 EP - 1872 ER - TY - JOUR A1 - Welden, Melanie A1 - Severins, Robin A1 - Poghossian, Arshak A1 - Wege, Christina A1 - Bongaerts, Johannes A1 - Siegert, Petra A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Detection of acetoin and diacetyl by a tobacco mosaic virus-assisted field-effect biosensor JF - Chemosensors N2 - Acetoin and diacetyl have a major impact on the flavor of alcoholic beverages such as wine or beer. Therefore, their measurement is important during the fermentation process. Until now, gas chromatographic techniques have typically been applied; however, these require expensive laboratory equipment and trained staff, and do not allow for online monitoring. In this work, a capacitive electrolyte–insulator–semiconductor sensor modified with tobacco mosaic virus (TMV) particles as enzyme nanocarriers for the detection of acetoin and diacetyl is presented. The enzyme acetoin reductase from Alkalihalobacillus clausii DSM 8716ᵀ is immobilized via biotin–streptavidin affinity, binding to the surface of the TMV particles. The TMV-assisted biosensor is electrochemically characterized by means of leakage–current, capacitance–voltage, and constant capacitance measurements. In this paper, the novel biosensor is studied regarding its sensitivity and long-term stability in buffer solution. Moreover, the TMV-assisted capacitive field-effect sensor is applied for the detection of diacetyl for the first time. The measurement of acetoin and diacetyl with the same sensor setup is demonstrated. Finally, the successive detection of acetoin and diacetyl in buffer and in diluted beer is studied by tuning the sensitivity of the biosensor using the pH value of the measurement solution. Y1 - 2022 U6 - http://dx.doi.org/10.3390/chemosensors10060218 SN - 2227-9040 N1 - This article belongs to the Special Issue "Nanostructured Devices for Biochemical Sensing" VL - 10 IS - 6 PB - MDPI CY - Basel ER - TY - CHAP A1 - Molinnus, Denise A1 - Hardt, Gabriel A1 - Käver, Larissa A1 - Willenberg, Holger S. A1 - Poghossian, Arshak A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Detection of Adrenaline Based on Bioelectrocatalytical System to Support Tumor Diagnostic Technology T2 - MDPI Proceedings Y1 - 2017 U6 - http://dx.doi.org/10.3390/proceedings1040506 ER -