@article{HaegerGrankinWagner2023, author = {Haeger, Gerrit and Grankin, Alina and Wagner, Michaela}, title = {Construction of an Aspergillus oryzae triple amylase deletion mutant as a chassis to evaluate industrially relevant amylases using multiplex CRISPR/Cas9 editing technology}, series = {Applied Research}, journal = {Applied Research}, number = {Early View}, publisher = {Wiley-VCH}, issn = {2702-4288}, doi = {10.1002/appl.202200106}, pages = {1 -- 15}, year = {2023}, abstract = {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.}, language = {en} } @article{KrafftFoersterHartetal.2001, author = {Krafft, B. and F{\"o}rster, Arnold and Hart, A. van der and Sch{\"a}pers, T.}, title = {Control of Aharonov-Bohm oscillations in a AlGaAs/GaAs ring by asymmetric and symmetric gate biasing}, series = {Physica E: Low-dimensional Systems and Nanostructures. 9 (2001), H. 4}, journal = {Physica E: Low-dimensional Systems and Nanostructures. 9 (2001), H. 4}, isbn = {1386-9477}, pages = {635 -- 641}, year = {2001}, language = {en} } @article{ReisertGeisslerFlorkeetal.2011, author = {Reisert, Steffen and Geissler, H. and Florke, R. and Wagner, P. and Wagner, Torsten and Sch{\"o}ning, Michael Josef}, title = {Controlling aseptic sterilization processes by means of a multi-sensor system}, publisher = {IEEE}, address = {New York}, pages = {18 -- 22}, year = {2011}, language = {en} } @article{MennickenPeterKaulenetal.2019, author = {Mennicken, Max and Peter, Sophia Katharina and Kaulen, Corinna and Simon, Ulrich and Karth{\"a}user, Silvia}, title = {Controlling the Electronic Contact at the Terpyridine/Metal Interface}, series = {The Journal of Physical Chemistry C}, volume = {123}, journal = {The Journal of Physical Chemistry C}, number = {35}, issn = {1932-7455}, doi = {10.1021/acs.jpcc.9b05865}, pages = {21367 -- 21375}, year = {2019}, language = {en} } @article{HennemannKohlReisertetal.2013, author = {Hennemann, J{\"o}rg and Kohl, Claus-Dieter and Reisert, Steffen and Kirchner, Patrick and Sch{\"o}ning, Michael Josef}, title = {Copper oxide nanofibres for detection of hydrogen peroxide vapour at high concentrations}, series = {physica status solidi (a)}, volume = {210}, journal = {physica status solidi (a)}, number = {5}, publisher = {Wiley}, address = {Weinheim}, issn = {1862-6319}, doi = {10.1002/pssa.201200775}, pages = {859 -- 863}, year = {2013}, abstract = {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.}, language = {en} } @article{MourzinaSchoeningSchubertetal.2001, author = {Mourzina, Y. G. and Sch{\"o}ning, Michael Josef and Schubert, J. and Zander, W. and Legin, A. V. and Vlasov, Y. G. and L{\"u}th, H.}, title = {Copper, cadmium and thallium thin film sensors based on chalcogenide glasses}, series = {Analytica Chimica Acta. 433 (2001)}, journal = {Analytica Chimica Acta. 433 (2001)}, isbn = {0378-4304}, pages = {103 -- 110}, year = {2001}, language = {en} } @article{KraemerPitaZhouetal.2009, author = {Kr{\"a}mer, Melina and Pita, Marcos and Zhou, Jian and Ornatska, Maryna and Poghossian, Arshak and Sch{\"o}ning, Michael Josef and Katz, Evgeny}, title = {Coupling of Biocomputing Systems with Electronic Chips: Electronic Interface for Transduction of Biochemical Information}, series = {Journal of Physical Chemistry C: Nanomaterials and Interfaces. 113 (2009), H. 6}, journal = {Journal of Physical Chemistry C: Nanomaterials and Interfaces. 113 (2009), H. 6}, publisher = {American Cemical Society}, address = {Washington, DC}, isbn = {1932-7455}, pages = {2573 -- 2579}, year = {2009}, language = {en} } @article{MolinnusPoghossianKeusgenetal.2017, author = {Molinnus, Denise and Poghossian, Arshak and Keusgen, Michael and Katz, Evgeny and Sch{\"o}ning, Michael Josef}, title = {Coupling of Biomolecular Logic Gates with Electronic Transducers: From Single Enzyme Logic Gates to Sense/Act/Treat Chips}, series = {Electroanalysis}, volume = {29}, journal = {Electroanalysis}, number = {8}, publisher = {Wiley}, address = {Weinheim}, issn = {1521-4109}, doi = {10.1002/elan.201700208}, pages = {1840 -- 1849}, year = {2017}, abstract = {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.}, language = {en} } @article{TillmannFoerster2000, author = {Tillmann, K. and F{\"o}rster, Arnold}, title = {Critical dimensions for the formation of interfacial misfit dislocations of In0.6Ga0.4As islands on GaAs(001)}, series = {Thin Solid Films. 368 (2000), H. 1}, journal = {Thin Solid Films. 368 (2000), H. 1}, isbn = {0040-6090}, pages = {93 -- 104}, year = {2000}, language = {en} } @article{PoghossianThustSchoeningetal.2000, author = {Poghossian, Arshak and Thust, M. and Sch{\"o}ning, Michael Josef and M{\"u}ller-Veggian, Mattea and Kordos, P. and L{\"u}th, H.}, title = {Cross-sensitivity of a capacitive penicillin sensor combined with a diffusion barrier}, series = {Sensors and Actuators B. 68 (2000), H. 1-3}, journal = {Sensors and Actuators B. 68 (2000), H. 1-3}, isbn = {0925-4005}, pages = {260 -- 265}, year = {2000}, language = {en} }