@article{DelleHuckBaeckeretal.2015, author = {Delle, Lotta E. and Huck, Christina and B{\"a}cker, Matthias and M{\"u}ller, Frank and Grandthyll, Samuel and Jacobs, Karin and Lilischkis, Rainer and Vu, Xuan T. and Sch{\"o}ning, Michael Josef and Wagner, Patrick and Thoelen, Roland and Weil, Maryam and Ingebrandt, Sven}, title = {Impedimetric immunosensor for the detection of histamine based on reduced graphene oxide}, series = {Physica status solidi (a)}, volume = {212}, journal = {Physica status solidi (a)}, number = {6}, publisher = {Wiley}, address = {Weinheim}, issn = {1862-6319}, doi = {10.1002/pssa.201431863}, pages = {1327 -- 1334}, year = {2015}, language = {en} } @article{DeppeBongaertsO'Connelletal.2011, author = {Deppe, Veronika Maria and Bongaerts, Johannes and O'Connell, Timothy and Maurer, Karl-Heinz and Meinhardt, Friedhelm}, title = {Enzymatic deglycation of Amadori products in bacteria}, series = {Applied microbiology and biotechnology}, volume = {Vol. 90}, journal = {Applied microbiology and biotechnology}, number = {Iss. 2}, publisher = {Springer}, address = {Berlin}, issn = {1432-0614 (E-Journal); 0171-1741 (Print); 0175-7598 (Print); 0340-2118 (Print)}, pages = {399 -- 406}, year = {2011}, language = {en} } @article{DeppeKlatteBongaertsetal.2011, author = {Deppe, Veronika Maria and Klatte, Stephanie and Bongaerts, Johannes and Maurer, Karl-Heinz and O'Connell, Timothy and Meinhardt, Friedhelm}, title = {Genetic control of Amadori product degradation in Bacillus subtilis via regulation of frlBONMD expression by FrlR}, series = {Applied and environmental microbiology}, volume = {Vol. 77}, journal = {Applied and environmental microbiology}, number = {No. 9}, publisher = {American Society of Mechanical Engineers (ASME)}, address = {New York}, issn = {1098-5336 (E-Journal); 0003-6919 (Print); 0099-2240 (Print)}, pages = {2839 -- 2846}, year = {2011}, language = {en} } @article{DollWagnerWagneretal.2016, author = {Doll, Theodor and Wagner, Torsten and Wagner, Patrick and Sch{\"o}ning, Michael Josef}, title = {Engineering of functional interfaces / Theodor Doll ; Torsten Wagner ; Patrick Wagner ; Michael J. Sch{\"o}ning (eds.)}, series = {Physica status solidi (a)}, volume = {213}, journal = {Physica status solidi (a)}, number = {6}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1862-6319}, doi = {10.1002/pssa.201670641}, pages = {1393 -- 1394}, year = {2016}, language = {en} } @article{EmonsBaadeSchoening2000, author = {Emons, H. and Baade, A. and Sch{\"o}ning, Michael Josef}, title = {Voltammetric determination of heavy metals in microvolumes of rain water}, series = {Electroanalysis. 12 (2000), H. 15}, journal = {Electroanalysis. 12 (2000), H. 15}, isbn = {1040-0397}, pages = {1171 -- 1176}, year = {2000}, language = {en} } @article{EmonsGlueckHuellenkremeretal.2001, author = {Emons, H. and Gl{\"u}ck, O. and H{\"u}llenkremer, B. and Sch{\"o}ning, Michael Josef}, title = {Voltohmmetry as an alternative detection method at polycrystalline metal film electrodes}, series = {Electroanalysis. 13 (2001), H. 8-9}, journal = {Electroanalysis. 13 (2001), H. 8-9}, isbn = {1040-0397}, pages = {677 -- 680}, year = {2001}, language = {en} } @article{EmonsGlueckSchoening2001, author = {Emons, H. and Gl{\"u}ck, O. and Sch{\"o}ning, Michael Josef}, title = {Voltohmmetry - An alternative detection principle at ultrathin metal electrodes in solution}, series = {Chemical and biological sensors and analytical methods : proceedings of the international symposium / Sensor, Physical Electrochemistry, and Organic and Biological Electrochemistry Divisions. Ed.: M. Butler}, journal = {Chemical and biological sensors and analytical methods : proceedings of the international symposium / Sensor, Physical Electrochemistry, and Organic and Biological Electrochemistry Divisions. Ed.: M. Butler}, publisher = {Electrochemical Society}, address = {Pennington, NJ}, isbn = {1-56677-351-2}, pages = {1 -- 3}, year = {2001}, language = {en} } @article{EmonsHuellenkremerSchoening2001, author = {Emons, H. and H{\"u}llenkremer, B. and Sch{\"o}ning, Michael Josef}, title = {Detection of metal ions in aqueous solutions by voltohmmetry}, series = {Fresenius' Journal of Analytical Chemistry. 369 (2001), H. 1}, journal = {Fresenius' Journal of Analytical Chemistry. 369 (2001), H. 1}, isbn = {0937-0633}, pages = {42 -- 46}, year = {2001}, language = {en} } @article{EngelmannPourshahidiShalabyetal.2022, author = {Engelmann, Ulrich M. and Pourshahidi, Mohammad Ali and Shalaby, Ahmed and Krause, Hans-Joachim}, title = {Probing particle size dependency of frequency mixing magnetic detection with dynamic relaxation simulation}, series = {Journal of Magnetism and Magnetic Materials}, volume = {563}, journal = {Journal of Magnetism and Magnetic Materials}, number = {In progress, Art. No. 169965}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0304-8853}, doi = {10.1016/j.jmmm.2022.169965}, year = {2022}, abstract = {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.}, language = {en} } @article{EngelmannSimsekShalabyetal.2024, author = {Engelmann, Ulrich M. and Simsek, Beril and Shalaby, Ahmed and Krause, Hans-Joachim}, title = {Key contributors to signal generation in frequency mixing magnetic detection (FMMD): an in silico study}, series = {Sensors}, volume = {24}, journal = {Sensors}, number = {6}, publisher = {MDPI}, address = {Basel}, issn = {1424-8220}, doi = {10.3390/s24061945}, pages = {Artikel 1945}, year = {2024}, abstract = {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 N{\´e}el-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.}, language = {en} } @article{ErmelenkoYoshinobuMourzinaetal.2002, author = {Ermelenko, Y. and Yoshinobu, T and Mourzina, Y. and Levichev, S. and Furuichi, K. and Vlasov, Y. and Sch{\"o}ning, Michael Josef and Iwasaki, H.}, title = {Photocurable membranes for ion-selective light-addressable potentiometric sensors}, series = {Sensors and Actuators B. 85 (2002), H. 1-2}, journal = {Sensors and Actuators B. 85 (2002), H. 1-2}, isbn = {0925-4005}, pages = {79 -- 85}, year = {2002}, language = {en} } @article{ErmelenkoYoshinobuMourzinaetal.2001, author = {Ermelenko, Y. and Yoshinobu, T. and Mourzina, Y. and Furuichi, K. and Iwasaki, H. and Vlasov, Y. and Sch{\"o}ning, Michael Josef}, title = {Technology of photocurable polymeric membranes for integrated LAPS}, series = {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}, journal = {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}, publisher = {Japan Association of Chemical Sensors}, address = {Kasuga, Fukuoka-ken}, pages = {66 -- 68}, year = {2001}, language = {en} } @article{ErmelenkoYoshinobuMourzinaetal.2002, author = {Ermelenko, Y. and Yoshinobu, T. and Mourzina, Y. and Furuichi, K. and Levichev, S. and Vlasov, Y. and Sch{\"o}ning, Michael Josef and Iwasaki, H.}, title = {Lithium sensor based on the laser scanning semiconductor transducer}, series = {Analytica Chimica Acta. 459 (2002), H. 1}, journal = {Analytica Chimica Acta. 459 (2002), H. 1}, issn = {0378-4304}, pages = {1 -- 9}, year = {2002}, language = {en} } @article{ErmelenkoYoshinobuMourzinaetal.2003, author = {Ermelenko, Y. and Yoshinobu, T. and Mourzina, Y. and Sch{\"o}ning, Michael Josef and Furuichi, K. and Levichev, S. and Vlasov, Y. and Iwasaki, H.}, title = {The double K+/Ca2+ sensor based on laser scanned silicon transducer (LSST) for multicomponent analysis}, series = {Talanta. 59 (2003), H. 4}, journal = {Talanta. 59 (2003), H. 4}, isbn = {0039-9140}, pages = {785 -- 795}, year = {2003}, language = {en} } @article{ErmelenkoYoshinobuMourzinaetal.2002, author = {Ermelenko, Y. and Yoshinobu, T. and Mourzina, Y. and Sch{\"o}ning, Michael Josef and Vlasov, Y. and Iwasaki, H.}, title = {The hybrid K+/Ca2+ sensor based on laser scanned silicon transducer for multi-component analysis}, series = {Proceedings of ICAS 2001, IUPAC [3rd] International Congress on Analytical Sciences 2001 : August 6 - 10, 2001, Waseda University, Tokyo}, journal = {Proceedings of ICAS 2001, IUPAC [3rd] International Congress on Analytical Sciences 2001 : August 6 - 10, 2001, Waseda University, Tokyo}, publisher = {Japan Society for Analytical Chemistry}, address = {Tokyo}, pages = {i777 -- i780}, year = {2002}, language = {en} } @inproceedings{ErmelenkoYoshinobuMourzinaetal.2003, author = {Ermelenko, Y. and Yoshinobu, T. and Mourzina, Y. and Sch{\"o}ning, Michael Josef and Vlasov, Y. and Iwasaki, H.}, title = {A multisensor based on laser scanned silicon transducer (LSST): development and properties}, series = {Eurosensors XVII : the 17th European Conference on Solid-State Transducers ; University of Minho, Guimar{\~a}es, Portugal, September 21 - 24, 2003}, booktitle = {Eurosensors XVII : the 17th European Conference on Solid-State Transducers ; University of Minho, Guimar{\~a}es, Portugal, September 21 - 24, 2003}, pages = {72 -- 73}, year = {2003}, language = {en} } @article{ErmolenkoYoshinobuMourzinaetal.2004, author = {Ermolenko, Y. E. and Yoshinobu, T. and Mourzina, Y. G. and Vlasov, Y. G. and Sch{\"o}ning, Michael Josef and Iwasaki, H.}, title = {Laserscanned transducer (LSST) as a multisensor system}, series = {Sensors and Actuators B. 103 (2004), H. 1-2}, journal = {Sensors and Actuators B. 103 (2004), H. 1-2}, isbn = {0925-4005}, pages = {457 -- 462}, year = {2004}, language = {en} } @article{ErmolenkoVlasovKolodnikovetal.2004, author = {Ermolenko, Y.E. and Vlasov, Y.G. and Kolodnikov, V.V. and Shabaldkin, D. A. and Kloock, Joachim P. and Sch{\"o}ning, Michael Josef}, title = {Diffusion of radioactive tracers (204Tl, 110mAg) and ionic conductivity in membrane materials for the chemical sensors}, series = {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.).}, journal = {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.).}, publisher = {Forschungszentrum J{\"u}lich, Zentralbibliothek}, address = {J{\"u}lich}, isbn = {3893363629}, pages = {483 -- 485}, year = {2004}, language = {en} } @article{FalkenbergBottBongaertsetal.2022, author = {Falkenberg, Fabian and Bott, Michael and Bongaerts, Johannes and Siegert, Petra}, title = {Phylogenetic survey of the subtilase family and a data-mining-based search for new subtilisins from Bacillaceae}, series = {Frontiers in Microbiology}, volume = {2022}, journal = {Frontiers in Microbiology}, number = {13}, publisher = {Frontiers}, address = {Lausanne}, issn = {1664-302X}, doi = {10.3389/fmicb.2022.1017978}, pages = {Artikel 13:1017978}, year = {2022}, abstract = {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.}, language = {en} } @article{FalkenbergKohnBottetal.2023, author = {Falkenberg, Fabian and Kohn, Sophie and Bott, Michael and Bongaerts, Johannes and Siegert, Petra}, title = {Biochemical characterisation of a novel broad pH spectrum subtilisin from Fictibacillus arsenicus DSM 15822ᵀ}, series = {FEBS Open Bio}, volume = {13}, journal = {FEBS Open Bio}, number = {11}, publisher = {Wiley}, address = {Hoboken, NJ}, issn = {2211-5463}, doi = {10.1002/2211-5463.13701}, pages = {2035 -- 2046}, year = {2023}, abstract = {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.}, language = {en} } @article{FalkenbergRahbaFischeretal.2022, author = {Falkenberg, Fabian and Rahba, Jade and Fischer, David and Bott, Michael and Bongaerts, Johannes and Siegert, Petra}, title = {Biochemical characterization of a novel oxidatively stable, halotolerant, and high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101T}, series = {FEBS Open Bio}, volume = {12}, journal = {FEBS Open Bio}, number = {10}, publisher = {Wiley}, address = {Hoboken, NJ}, issn = {2211-5463}, doi = {10.1002/2211-5463.13457}, pages = {1729 -- 1746}, year = {2022}, abstract = {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.}, language = {en} } @article{FalkenbergVossBottetal.2023, author = {Falkenberg, Fabian and Voß, Leonie and Bott, Michael and Bongaerts, Johannes and Siegert, Petra}, title = {New robust subtilisins from halotolerant and halophilic Bacillaceae}, series = {Applied Microbiology and Biotechnology}, volume = {107}, journal = {Applied Microbiology and Biotechnology}, publisher = {Springer Nature}, address = {Berlin}, issn = {1432-0614}, doi = {10.1007/s00253-023-12553-w}, pages = {3939 -- 3954}, year = {2023}, abstract = {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.}, language = {en} } @article{FassbenderSchmittSchoeningetal.2000, author = {Faßbender, F. and Schmitt, G. and Sch{\"o}ning, Michael Josef and L{\"u}th, H. and Buß, G. and Schultze, J. W.}, title = {Optimization of passivation layers for corrosion protection of silicon-based microelectrode arrays}, series = {Sensors and Actuators B. 68 (2000), H. 1-3}, journal = {Sensors and Actuators B. 68 (2000), H. 1-3}, isbn = {0925-4005}, pages = {128 -- 133}, year = {2000}, language = {en} } @article{FigueroaMirandaFengShiuetal.2018, author = {Figueroa-Miranda, Gabriela and Feng, Lingyan and Shiu, Simon Chi-Chin and Dirkzwager, Roderick Marshall and Cheung, Yee-Wai and Tanner, Julian Alexander and Sch{\"o}ning, Michael Josef and Offenh{\"a}usser, Andreas and Mayer, Dirk}, title = {Aptamer-based electrochemical biosensor for highly sensitive and selective malaria detection with adjustable dynamic response range and reusability}, series = {Sensor and Actuators B: Chemical}, volume = {255}, journal = {Sensor and Actuators B: Chemical}, number = {P1}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0925-4005}, doi = {10.1016/j.snb.2017.07.117}, pages = {235 -- 243}, year = {2018}, abstract = {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.}, language = {en} } @article{FuruichiYoshinobuErmelenkoetal.2001, author = {Furuichi, K. and Yoshinobu, T. and Ermelenko, T. and Mourzina, Y. and Iwasaki, H. and Sch{\"o}ning, Michael Josef}, title = {Simultaneous visualization of multiple ions by the chemical imaging sensor}, series = {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}, journal = {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}, publisher = {Japan Association of Chemical Sensors}, address = {Kasuga, Fukuoka-ken}, pages = {399 -- 401}, year = {2001}, language = {en} } @article{Foerster2000, author = {F{\"o}rster, Arnold}, title = {Resonant tunneling diodes operating as modern quantum transport devices}, series = {Proceedings of the Tenth International Workshop on the Physics of Semiconductor Devices : (December 14 - 18, 1999) / ed.: Vikram Kumar ... - Vol. 2}, journal = {Proceedings of the Tenth International Workshop on the Physics of Semiconductor Devices : (December 14 - 18, 1999) / ed.: Vikram Kumar ... - Vol. 2}, publisher = {Allied Publ.}, address = {New Delhi [u.a.]}, isbn = {81-7023-998-2}, pages = {966 -- 966}, year = {2000}, language = {en} } @article{FoersterLepsaFreundt2007, author = {F{\"o}rster, Arnold and Lepsa, M. I. and Freundt, D.}, title = {Hot electron injector Gunn diode for advanced driver assistance systems}, series = {Applied physics A: materials science and processing. 87 (2007), H. 3}, journal = {Applied physics A: materials science and processing. 87 (2007), H. 3}, isbn = {0947-8396}, pages = {545 -- 558}, year = {2007}, language = {en} } @article{FoersterStockIndlekofer2002, author = {F{\"o}rster, Arnold and Stock, J. and Indlekofer, K. M.}, title = {Perspectives of resonant tunneling diodes}, series = {Recent research developments in materials science \& engineering/ 1,2}, journal = {Recent research developments in materials science \& engineering/ 1,2}, publisher = {Transworld Research Network}, address = {Trivandrum, India}, isbn = {81-7895-057-X}, pages = {527 -- 556}, year = {2002}, language = {en} } @inproceedings{FoersterStockMontanarietal.2006, author = {F{\"o}rster, Arnold and Stock, J{\"u}rgen and Montanari, Simone and Lepsa, Mihail Ion and L{\"u}th, Hans}, title = {Fabrication and characterisation of GaAs Gunn Diode Chips for applications at 77 GHz in automotive industry}, url = {http://nbn-resolving.de/urn:nbn:de:hbz:a96-opus-1462}, year = {2006}, abstract = {GaAs-based Gunn diodes with graded AlGaAs hot electron injector heterostructures have been developed under the special needs in automotive applications. The fabrication of the Gunn diode chips was based on total substrate removal and processing of integrated Au heat sinks. Especially, the thermal and RF behavior of the diodes have been analyzed by DC, impedance and S-parameter measurements. The electrical investigations have revealed the functionality of the hot electron injector. An optimized layer structure could fulfill the requirements in adaptive cruise control (ACC) systems at 77 GHz with typical output power between 50 and 90 mW.}, subject = {Biosensor}, language = {en} } @article{GamellaZakharchenkoGuzetal.2017, author = {Gamella, Maria and Zakharchenko, Andrey and Guz, Nataliia and Masi, Madeline and Minko, Sergiy and Kolpashchikov, Dmitry M. and Iken, Heiko and Poghossian, Arshak and Sch{\"o}ning, Michael Josef and Katz, Evgeny}, title = {DNA computing system activated by electrochemically triggered DNA realease from a polymer-brush-modified electrode array}, series = {Electroanalysis}, volume = {29}, journal = {Electroanalysis}, number = {2}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1521-4109}, doi = {10.1002/elan.201600389}, pages = {398 -- 408}, year = {2017}, abstract = {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.}, language = {en} } @article{GasparyanPoghossianVitusevichetal.2009, author = {Gasparyan, F. V. and Poghossian, Arshak and Vitusevich, S. A. and Petrychuk, M. V. and Sydoruk, V. A. and Surmalyan, A. V. and Siqueira, J. R. and Oliveira, O. N. and Offenh{\"a}usser, A. and Sch{\"o}ning, Michael Josef}, title = {Low Frequency Noise In Electrolyte-Gate Field-Effect Devices Functionalized With Dendrimer/Carbon-Nanotube Multilayers}, series = {Noise and fluctuations : 20th International Conference on Noise and Fluctuations, ICNF 2009, Pisa, Italy, 14 - 19 June 2009 / ed. Massimo Macucci; Giovanni Basso}, journal = {Noise and fluctuations : 20th International Conference on Noise and Fluctuations, ICNF 2009, Pisa, Italy, 14 - 19 June 2009 / ed. Massimo Macucci; Giovanni Basso}, publisher = {American Inst. of Physics}, address = {Melville, NY}, isbn = {9780735406650}, pages = {133 -- 136}, year = {2009}, language = {en} } @article{GasparyanVitusevichOffenhaeusseretal.2011, author = {Gasparyan, F.V. and Vitusevich, S.A. and Offenh{\"a}usser, A. and Sch{\"o}ning, Michael Josef}, title = {Modified charge fluctuation noise model for electrolyte-insulator-semiconductor devices}, series = {Modern Physics Letters B (MPLB). 25 (2011), H. 11}, journal = {Modern Physics Letters B (MPLB). 25 (2011), H. 11}, publisher = {World Scientific Publ.}, address = {Singapur}, isbn = {0217-9849}, pages = {831 -- 840}, year = {2011}, language = {en} } @article{GasparyanPoghossianVitusevichetal.2011, author = {Gasparyan, Ferdinand V. and Poghossian, Arshak and Vitusevich, Svetlana A. and Petrychuk, Mykhaylo V. and Sydoruk, Viktor A. and Siqueira, Jos{\´e} R. Jr. and Oliveira, Osvaldo N. Jr. and Offenh{\"a}usser, Andreas and Sch{\"o}ning, Michael Josef}, title = {Low-Frequency Noise in Field-Effect Devices Functionalized With Dendrimer/Carbon-Nanotube Multilayers}, series = {IEEE Sensors Journal. 11 (2011), H. 1}, journal = {IEEE Sensors Journal. 11 (2011), H. 1}, publisher = {IEEE}, address = {New York}, isbn = {1530-437X}, pages = {142 -- 149}, year = {2011}, language = {en} } @article{GeMeyerSchoeningetal.2000, author = {Ge, B. and Meyer, T. and Sch{\"o}ning, Michael Josef and Wollenberger, U. and Lisdat, F.,}, title = {Cytochrome c' from Chromatium vinosum on gold electrodes}, series = {Electrochemistry Communications. 2 (2000), H. 8}, journal = {Electrochemistry Communications. 2 (2000), H. 8}, isbn = {1388-2481}, pages = {557 -- 561}, year = {2000}, language = {en} } @article{GivanoudiCornelisRasschaertetal.2021, author = {Givanoudi, Stella and Cornelis, Peter and Rasschaert, Geertrui and Wackers, Gideon and Iken, Heiko and Rolka, David and Yongabi, Derick and Robbens, Johan and Sch{\"o}ning, Michael Josef and Heyndrickx, Marc and Wagner, Patrick}, title = {Selective Campylobacter detection and quantification in poultry: A sensor tool for detecting the cause of a common zoonosis at its source}, series = {Sensors and Actuators B: Chemical}, journal = {Sensors and Actuators B: Chemical}, number = {In Press, Journal Pre-proof}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0925-4005}, doi = {10.1016/j.snb.2021.129484}, pages = {Article 129484}, year = {2021}, language = {en} } @article{GrinsvenBonGrietenetal.2011, author = {Grinsven, B. van and Bon, N. vanden and Grieten, L. and Murib, M. and Janssen, S. D. and Haenen, K. and Schneider, E. and Ingebrandt, E. and Sch{\"o}ning, Michael Josef and Vermeeren, V. and Ameloot, M. and Michiels, L. and Thoelen, R. and Ceuninck, W. de and Wagner, P.}, title = {Rapid assessment of the stability of DNA duplexes by impedimetric real-time monitoring of chemically induced denaturation}, series = {Lab on a Chip}, volume = {11}, journal = {Lab on a Chip}, number = {9}, publisher = {Royal Society of Chemistry (RSC)}, address = {Cambridge}, isbn = {1473-0197}, pages = {1656 -- 1663}, year = {2011}, language = {en} } @article{GrinsvenBonStrauvenetal.2012, author = {Grinsven, Bart van and Bon, Natalie vanden and Strauven, Hannelore and Grieten, Lars and Murib, Mohammed and Jim{\´e}nez Monroy, Kathia L. and Janssens, Stoffel D. and Haenen, Ken and Sch{\"o}ning, Michael Josef and Vermeeren, Veronique and Ameloot, Marcel and Michiels, Luc and Thoelen, Ronald and Ceuninck, Ward de and Wagner, Patrick}, title = {Heat-Transfer Resistance at Solid-Liquid Interfaces: A Tool for The Detection of Single Nucleotide Polymorphisms in DNA.}, series = {ACS Nano}, volume = {6}, journal = {ACS Nano}, number = {3}, publisher = {ACS Publications}, address = {Washington, DC}, issn = {1936-086X}, doi = {10.1021/nn300147e}, pages = {2712 -- 2721}, year = {2012}, abstract = {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.}, language = {en} } @article{GunGutkinLevetal.2011, author = {Gun, Jenny and Gutkin, Vitaly and Lev, Ovadia and Boyen, Hans-Gerd and Saitner, Marc and Wagner, Patrick and Olieslaeger, Marc D´ and Abouzar, Maryam H. and Poghossian, Arshak and Sch{\"o}ning, Michael Josef}, title = {Tracing gold nanoparticle charge by electrolyte-insulator-semiconductor devices}, series = {Journal of Physical Chemistry C. 115 (2011), H. 11}, journal = {Journal of Physical Chemistry C. 115 (2011), H. 11}, publisher = {American Cemical Society}, address = {Washington, DC}, isbn = {1932-7455}, pages = {4439 -- 4445}, year = {2011}, language = {en} } @article{GunRizkovLevetal.2008, author = {Gun, Jenny and Rizkov, Dan and Lev, Ovadia and Abouzar, Maryam H. and Poghossian, Arshak and Sch{\"o}ning, Michael Josef}, title = {Oxygen plasma-treated gold nanoparticle-based field-effect devices as transducer structures for bio-chemical sensing}, series = {Microchimica Acta. 164 (2008), H. 3-4}, journal = {Microchimica Acta. 164 (2008), H. 3-4}, isbn = {1436-5073}, pages = {395 -- 404}, year = {2008}, language = {en} } @article{GunSchoeningAbouzaretal.2008, author = {Gun, Jenny and Sch{\"o}ning, Michael Josef and Abouzar, Maryam H. and Poghossian, Arshak and Katz, Evgeny}, title = {Field-Effect Nanoparticle-Based Glucose Sensor on a Chip: Amplification Effect of Coimmobilized Redox Species}, series = {Electroanalysis. 20 (2008), H. 16}, journal = {Electroanalysis. 20 (2008), H. 16}, isbn = {1521-4109}, pages = {1748 -- 1753}, year = {2008}, language = {en} } @article{GuoMiyamotoWagneretal.2014, author = {Guo, Yuanyuan and Miyamoto, Ko-ichiro and Wagner, Torsten and Sch{\"o}ning, Michael Josef and Yoshinobu, Tatsuo}, title = {Theoretical study and simulation of light-addressable potentiometric sensors}, series = {Physica status solidi (A) : applications and materials}, volume = {211}, journal = {Physica status solidi (A) : applications and materials}, number = {6}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {0031-8965}, doi = {10.1002/pssa.201330354}, pages = {1467 -- 1472}, year = {2014}, abstract = {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.}, language = {en} } @article{GuoMiyamotoWagneretal.2014, author = {Guo, Yuanyuan and Miyamoto, Ko-ichiro and Wagner, Torsten and Sch{\"o}ning, Michael Josef and Yoshinobu, Tatsuo}, title = {Device simulation of the light-addressable potentiometric sensor for the investigation of the spatial resolution}, series = {Sensors and actuators B: Chemical}, volume = {204}, journal = {Sensors and actuators B: Chemical}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1873-3077 (E-Journal); 0925-4005 (Print)}, doi = {10.1016/j.snb.2014.08.016}, pages = {659 -- 665}, year = {2014}, abstract = {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.}, language = {en} } @article{GuoSekiMiyamotoetal.2014, author = {Guo, Yuanyuan and Seki, Kosuke and Miyamoto, Ko-ichiro and Wagner, Torsten and Sch{\"o}ning, Michael Josef and Yoshinobu, Tatsuo}, title = {Novel photoexcitation method for light-addressable potentiometric sensor with higher spatial resolution}, series = {Applied physics express : APEX}, volume = {7}, journal = {Applied physics express : APEX}, number = {6}, publisher = {IOP}, address = {Bristol}, issn = {1882-0786 (E-Journa); 1882-0778 (Print)}, doi = {10.7567/APEX.7.067301}, pages = {067301-4}, year = {2014}, abstract = {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.}, language = {en} } @article{HaegerBongaertsSiegert2022, author = {Haeger, Gerrit and Bongaerts, Johannes and Siegert, Petra}, title = {A convenient ninhydrin assay in 96-well format for amino acid-releasing enzymes using an air-stable reagent}, series = {Analytical Biochemistry}, journal = {Analytical Biochemistry}, number = {624}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1096-0309}, doi = {10.1016/j.ab.2022.114819}, pages = {Artikel 114819}, year = {2022}, abstract = {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.}, language = {en} } @techreport{HaegerBongaertsSiegert2023, author = {Haeger, Gerrit and Bongaerts, Johannes and Siegert, Petra}, title = {Abschlussbericht Teil II: Eingehende Darstellung Neue biobasierte Lipopeptide aus nachhaltiger Produktion (LipoPep)}, pages = {17Seiten}, year = {2023}, language = {de} } @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{HaegerJolmesOyenetal.2024, author = {Haeger, Gerrit and Jolmes, Tristan and Oyen, Sven and Jaeger, Karl-Erich and Bongaerts, Johannes and Sch{\"o}rken, Ulrich and Siegert, Petra}, title = {Novel recombinant aminoacylase from Paraburkholderia monticola capable of N-acyl-amino acid synthesis}, series = {Applied Microbiology and Biotechnology}, journal = {Applied Microbiology and Biotechnology}, number = {108}, publisher = {Springer}, address = {Berlin}, issn = {1432-0614}, doi = {10.1007/s00253-023-12868-8}, pages = {14 Seiten}, year = {2024}, abstract = {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.}, language = {en} } @article{HaegerProbstJaegeretal.2023, author = {Haeger, Gerrit and Probst, Johanna and Jaeger, Karl-Erich and Bongaerts, Johannes and Siegert, Petra}, title = {Novel aminoacylases from Streptomyces griseus DSM 40236 and their recombinant production in Streptomyces lividans}, series = {FEBS Open Bio}, volume = {13}, journal = {FEBS Open Bio}, number = {12}, publisher = {Wiley}, address = {Hoboken, NJ}, issn = {2211-5463}, doi = {10.1002/2211-5463.13723}, pages = {2224 -- 2238}, year = {2023}, abstract = {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.}, language = {en} } @article{HaegerWirgesTanzmannetal.2023, author = {Haeger, Gerrit and Wirges, Jessika and Tanzmann, Nicole and Oyen, Sven and Jolmes, Tristan and Jaeger, Karl-Erich and Sch{\"o}rken, Ulrich and Bongaerts, Johannes and Siegert, Petra}, title = {Chaperone assisted recombinant expression of a mycobacterial aminoacylase in Vibrio natriegens and Escherichia coli capable of N-lauroyl-L-amino acid synthesis}, series = {Microbial Cell Factories}, journal = {Microbial Cell Factories}, number = {22}, publisher = {Springer Nature}, issn = {1475-2859}, doi = {10.1186/s12934-023-02079-1}, pages = {Article number: 77 (2023)}, year = {2023}, abstract = {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.}, language = {en} } @article{HamadBilattoAdlyetal.2016, author = {Hamad, E. M. and Bilatto, S. E. R. and Adly, N. Y. and Correa, D. S. and Wolfrum, B. and Sch{\"o}ning, Michael Josef and Offenh{\"a}usser, A. and Yakushenko, A.}, title = {Inkjet printing of UV-curable adhesive and dielectric inks for microfluidic devices}, series = {Lab on a Chip}, volume = {16}, journal = {Lab on a Chip}, number = {1}, publisher = {Royal Society of Chemistry}, address = {Cambridge}, issn = {1473-0189}, doi = {10.1039/C5LC01195G}, pages = {70 -- 74}, year = {2016}, abstract = {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.}, language = {en} }