TY - JOUR A1 - Faßbender, F. A1 - Schmitt, G. A1 - Schöning, Michael Josef A1 - Lüth, H. A1 - Buß, G. A1 - Schultze, J. W. T1 - Optimization of passivation layers for corrosion protection of silicon-based microelectrode arrays JF - Sensors and Actuators B. 68 (2000), H. 1-3 Y1 - 2000 SN - 0925-4005 SP - 128 EP - 133 ER - TY - JOUR A1 - Falkenberg, Fabian A1 - Voß, Leonie A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - New robust subtilisins from halotolerant and halophilic Bacillaceae JF - Applied Microbiology and Biotechnology N2 - 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. KW - Biotechnological application KW - Bacillaceae KW - Subtilisin KW - Subtilases KW - Halotolerant protease Y1 - 2023 U6 - http://dx.doi.org/10.1007/s00253-023-12553-w SN - 1432-0614 N1 - Corresponding author: Petra Siegert VL - 107 SP - 3939 EP - 3954 PB - Springer Nature CY - Berlin ER - TY - JOUR A1 - Falkenberg, Fabian A1 - Rahba, Jade A1 - Fischer, David A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Biochemical characterization of a novel oxidatively stable, halotolerant, and high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101T JF - FEBS Open Bio N2 - 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. KW - Alkalihalobacillus okhensis KW - detergent protease KW - halotolerant protease KW - high-alkaline subtilisin KW - oxidative stable protease Y1 - 2022 U6 - http://dx.doi.org/10.1002/2211-5463.13457 SN - 2211-5463 N1 - Corresponding author: Petra Siegert VL - 12 IS - 10 SP - 1729 EP - 1746 PB - Wiley CY - Hoboken, NJ ER - TY - JOUR A1 - Falkenberg, Fabian A1 - Kohn, Sophie A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Biochemical characterisation of a novel broad pH spectrum subtilisin from Fictibacillus arsenicus DSM 15822ᵀ JF - FEBS Open Bio N2 - 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. KW - Bacillaceae KW - Biotechnological application KW - Broad pH spectrum KW - Subtilases KW - Subtilisin Y1 - 2023 U6 - http://dx.doi.org/10.1002/2211-5463.13701 SN - 2211-5463 N1 - Corresponding author: Petra Siegert VL - 13 IS - 11 SP - 2035 EP - 2046 PB - Wiley CY - Hoboken, NJ ER - TY - JOUR A1 - Falkenberg, Fabian A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Phylogenetic survey of the subtilase family and a data-mining-based search for new subtilisins from Bacillaceae JF - Frontiers in Microbiology N2 - 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. Y1 - 2022 U6 - http://dx.doi.org/10.3389/fmicb.2022.1017978 SN - 1664-302X VL - 2022 IS - 13 PB - Frontiers CY - Lausanne ER - TY - JOUR A1 - Ermolenko, Y.E. A1 - Vlasov, Y.G. A1 - Kolodnikov, V.V. A1 - Shabaldkin, D. A. A1 - Kloock, Joachim P. A1 - Schöning, Michael Josef T1 - Diffusion of radioactive tracers (204Tl, 110mAg) and ionic conductivity in membrane materials for the chemical sensors JF - 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.). Y1 - 2004 SN - 3893363629 N1 - Schriften des Forschungszentrums Jülich Reihe Allgemeines und Interdisziplinäres ; 3. International Conference on Nuclear and Radiochemistry ; (6, 2004, Aachen) SP - 483 EP - 485 PB - Forschungszentrum Jülich, Zentralbibliothek CY - Jülich ER - TY - JOUR A1 - Ermolenko, Y. E. A1 - Yoshinobu, T. A1 - Mourzina, Y. G. A1 - Vlasov, Y. G. A1 - Schöning, Michael Josef A1 - Iwasaki, H. T1 - Laserscanned transducer (LSST) as a multisensor system JF - Sensors and Actuators B. 103 (2004), H. 1-2 Y1 - 2004 SN - 0925-4005 SP - 457 EP - 462 ER - TY - JOUR A1 - Ermelenko, Y. A1 - Yoshinobu, T. A1 - Mourzina, Y. A1 - Schöning, Michael Josef A1 - Vlasov, Y. A1 - Iwasaki, H. T1 - The hybrid K+/Ca2+ sensor based on laser scanned silicon transducer for multi-component analysis JF - Proceedings of ICAS 2001, IUPAC [3rd] International Congress on Analytical Sciences 2001 : August 6 - 10, 2001, Waseda University, Tokyo Y1 - 2002 N1 - International Congress on Analytical Sciences <2001, Tokyo> ; International Union of Pure and Applied Chemistry. Pure and applied chemistry ; 73,10. ; Analytical sciences ; 17.2001, special issue SP - i777 EP - i780 PB - Japan Society for Analytical Chemistry CY - Tokyo ER - TY - CHAP A1 - Ermelenko, Y. A1 - Yoshinobu, T. A1 - Mourzina, Y. A1 - Schöning, Michael Josef A1 - Vlasov, Y. A1 - Iwasaki, H. T1 - A multisensor based on laser scanned silicon transducer (LSST): development and properties T2 - Eurosensors XVII : the 17th European Conference on Solid-State Transducers ; University of Minho, Guimarães, Portugal, September 21 - 24, 2003 Y1 - 2003 SP - 72 EP - 73 ER - TY - JOUR A1 - Ermelenko, Y. A1 - Yoshinobu, T. A1 - Mourzina, Y. A1 - Schöning, Michael Josef A1 - Furuichi, K. A1 - Levichev, S. A1 - Vlasov, Y. A1 - Iwasaki, H. T1 - The double K+/Ca2+ sensor based on laser scanned silicon transducer (LSST) for multicomponent analysis JF - Talanta. 59 (2003), H. 4 Y1 - 2003 SN - 0039-9140 SP - 785 EP - 795 ER - TY - JOUR A1 - Ermelenko, Y. A1 - Yoshinobu, T. A1 - Mourzina, Y. A1 - Furuichi, K. A1 - Levichev, S. A1 - Vlasov, Y. A1 - Schöning, Michael Josef A1 - Iwasaki, H. T1 - Lithium sensor based on the laser scanning semiconductor transducer JF - Analytica Chimica Acta. 459 (2002), H. 1 Y1 - 2002 SN - 0378-4304 SP - 1 EP - 9 ER - TY - JOUR A1 - Ermelenko, Y. A1 - Yoshinobu, T. A1 - Mourzina, Y. A1 - Furuichi, K. A1 - Iwasaki, H. A1 - Vlasov, Y. A1 - Schöning, Michael Josef T1 - Technology of photocurable polymeric membranes for integrated LAPS JF - 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 Y1 - 2001 N1 - Chemical sensors; 17.2001 Suppl. B; East Asian Conference on Chemical Sensors ; (5, 2001; Nagasaki) SP - 66 EP - 68 PB - Japan Association of Chemical Sensors CY - Kasuga, Fukuoka-ken ER - TY - JOUR A1 - Ermelenko, Y. A1 - Yoshinobu, T A1 - Mourzina, Y. A1 - Levichev, S. A1 - Furuichi, K. A1 - Vlasov, Y. A1 - Schöning, Michael Josef A1 - Iwasaki, H. T1 - Photocurable membranes for ion-selective light-addressable potentiometric sensors JF - Sensors and Actuators B. 85 (2002), H. 1-2 Y1 - 2002 SN - 0925-4005 SP - 79 EP - 85 ER - TY - JOUR A1 - Engelmann, Ulrich M. A1 - Simsek, Beril A1 - Shalaby, Ahmed A1 - Krause, Hans-Joachim T1 - Key contributors to signal generation in frequency mixing magnetic detection (FMMD): an in silico study JF - Sensors N2 - 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 Néel–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. KW - key performance indicators KW - magnetic biosensing KW - coupled Néel–Brownian relaxation dynamics KW - frequency mixing magnetic detection KW - magnetic relaxation KW - micromagnetic simulation KW - magnetic nanoparticles Y1 - 2024 U6 - http://dx.doi.org/10.3390/s24061945 SN - 1424-8220 N1 - This article belongs to the Special Issue "Advances in Magnetic Sensors and Their Applications" VL - 24 IS - 6 PB - MDPI CY - Basel ER - TY - JOUR A1 - Engelmann, Ulrich M. A1 - Pourshahidi, Mohammad Ali A1 - Shalaby, Ahmed A1 - Krause, Hans-Joachim T1 - Probing particle size dependency of frequency mixing magnetic detection with dynamic relaxation simulation JF - Journal of Magnetism and Magnetic Materials N2 - 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. Y1 - 2022 U6 - http://dx.doi.org/10.1016/j.jmmm.2022.169965 SN - 0304-8853 VL - 563 IS - In progress, Art. No. 169965 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Emons, H. A1 - Hüllenkremer, B. A1 - Schöning, Michael Josef T1 - Detection of metal ions in aqueous solutions by voltohmmetry JF - Fresenius' Journal of Analytical Chemistry. 369 (2001), H. 1 Y1 - 2001 SN - 0937-0633 SP - 42 EP - 46 ER - TY - JOUR A1 - Emons, H. A1 - Glück, O. A1 - Schöning, Michael Josef T1 - Voltohmmetry – An alternative detection principle at ultrathin metal electrodes in solution JF - Chemical and biological sensors and analytical methods : proceedings of the international symposium / Sensor, Physical Electrochemistry, and Organic and Biological Electrochemistry Divisions. Ed.: M. Butler Y1 - 2001 SN - 1-56677-351-2 N1 - International Symposium: Chemical and Biological Sensors and Analytical Methods ; (2 : 2001.) SP - 1 EP - 3 PB - Electrochemical Society CY - Pennington, NJ ER - TY - JOUR A1 - Emons, H. A1 - Glück, O. A1 - Hüllenkremer, B. A1 - Schöning, Michael Josef T1 - Voltohmmetry as an alternative detection method at polycrystalline metal film electrodes JF - Electroanalysis. 13 (2001), H. 8-9 Y1 - 2001 SN - 1040-0397 SP - 677 EP - 680 ER - TY - JOUR A1 - Emons, H. A1 - Baade, A. A1 - Schöning, Michael Josef T1 - Voltammetric determination of heavy metals in microvolumes of rain water JF - Electroanalysis. 12 (2000), H. 15 Y1 - 2000 SN - 1040-0397 SP - 1171 EP - 1176 ER - TY - JOUR A1 - Doll, Theodor A1 - Wagner, Torsten A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Engineering of functional interfaces / Theodor Doll ; Torsten Wagner ; Patrick Wagner ; Michael J. Schöning (eds.) JF - Physica status solidi (a) Y1 - 2016 U6 - http://dx.doi.org/10.1002/pssa.201670641 SN - 1862-6319 VL - 213 IS - 6 SP - 1393 EP - 1394 PB - Wiley-VCH CY - Weinheim ER -