TY - JOUR A1 - Degering, Christian A1 - Eggert, Thorsten A1 - Puls, Michael A1 - Bongaerts, Johannes A1 - Evers, Stefan A1 - Maurer, Karl-Heinz A1 - Jaeger, Karl-Erich T1 - Optimization of protease secretion in Bacillus subtilis and Bacillus licheniformis by screening of homologous and herologous signal peptides JF - Applied and environmental microbiology N2 - Bacillus subtilis and Bacillus licheniformis are widely used for the large-scale industrial production of proteins. These strains can efficiently secrete proteins into the culture medium using the general secretion (Sec) pathway. A characteristic feature of all secreted proteins is their N-terminal signal peptides, which are recognized by the secretion machinery. Here, we have studied the production of an industrially important secreted protease, namely, subtilisin BPN′ from Bacillus amyloliquefaciens. One hundred seventy-three signal peptides originating from B. subtilis and 220 signal peptides from the B. licheniformis type strain were fused to this secretion target and expressed in B. subtilis, and the resulting library was analyzed by high-throughput screening for extracellular proteolytic activity. We have identified a number of signal peptides originating from both organisms which produced significantly increased yield of the secreted protease. Interestingly, we observed that levels of extracellular protease were improved not only in B. subtilis, which was used as the screening host, but also in two different B. licheniformis strains. To date, it is impossible to predict which signal peptide will result in better secretion and thus an improved yield of a given extracellular target protein. Our data show that screening a library consisting of homologous and heterologous signal peptides fused to a target protein can identify more-effective signal peptides, resulting in improved protein export not only in the original screening host but also in different production strains. Y1 - 2010 U6 - http://dx.doi.org/10.1128/AEM.01146-10 SN - 1098-5336 (E-Journal); 0003-6919 (Print); 0099-2240 (Print) VL - 76 IS - 19 SP - 6370 EP - 6378 PB - American Society for Microbiology CY - Washington, DC ER - TY - JOUR A1 - Arreola, Julio A1 - Mätzkow, Malte A1 - Durán, Marlena Palomar A1 - Greeff, Anton A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Optimization of the immobilization of bacterial spores on glass substrates with organosilanes JF - Physica status solidi (A) : Applications and materials science N2 - Spores can be immobilized on biosensors to function as sensitive recognition elements. However, the immobilization can affect the sensitivity and reproducibility of the sensor signal. In this work, three different immobilization strategies with organosilanes were optimized and characterized to immobilize Bacillus atrophaeus spores on glass substrates. Five different silanization parameters were investigated: nature of the solvent, concentration of the silane, silanization time, curing process, and silanization temperature. The resulting silane layers were resistant to a buffer solution (e.g., Ringer solution) with a polysorbate (e.g., Tween®80) and sonication. KW - silanization KW - organosilanes KW - immobilization KW - endospores KW - biosensors KW - Bacillus atrophaeus Y1 - 2016 U6 - http://dx.doi.org/10.1002/pssa.201532914 SN - 1862-6319 VL - 213 IS - 6 SP - 1463 EP - 1470 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Pita, Marcos A1 - Krämer, Melina A1 - Zouh, Jian A1 - Poghossian, Arshak A1 - Schöning, Michael Josef A1 - Fernandez, Victor M. A1 - Katz, Evgeny T1 - Optoelectronic Properties of Nanostructured Ensembles Controlled by Biomolecular Logic Systems JF - ACS Nano. 10 (2008), H. 2 Y1 - 2008 SN - 1936-086X SP - 2160 EP - 2166 ER - TY - JOUR A1 - Gun, Jenny A1 - Rizkov, Dan A1 - Lev, Ovadia A1 - Abouzar, Maryam H. A1 - Poghossian, Arshak A1 - Schöning, Michael Josef T1 - Oxygen plasma-treated gold nanoparticle-based field-effect devices as transducer structures for bio-chemical sensing JF - Microchimica Acta. 164 (2008), H. 3-4 Y1 - 2008 SN - 1436-5073 SP - 395 EP - 404 ER - TY - JOUR A1 - Schmitt, G. A1 - Faßbender, F. A1 - Lüth, H. A1 - Schöning, Michael Josef A1 - Schultze, J. W. A1 - Buß, G. T1 - Passivation and corrosion of microelectrode arrays JF - Materials and Corrosion. 51 (2000), H. 1 Y1 - 2000 SN - 0947-5117 SP - 20 EP - 25 ER - TY - JOUR A1 - Siqueira, José R. Jr. A1 - Abouzar, Maryam H. A1 - Poghossian, Arshak A1 - Zucolotto, Valtencir A1 - Oliveira, Osvaldo N. Jr. A1 - Schöning, Michael Josef T1 - Penicillin biosensor based on a capacitive field-effect structure functionalized with a dendrimer/carbon nanotube multilayer JF - Biosensors and Bioelectronics. 25 (2009), H. 2 Y1 - 2009 SN - 0956-5663 SP - 497 EP - 501 ER - TY - JOUR A1 - Poghossian, Arshak A1 - Yoshinobu, Tatsuo A1 - Simonis, A. A1 - Ecken, H. A1 - Lüth, Hans A1 - Schöning, Michael Josef T1 - Penicillin detection by means of field-effect based sensors: EnFET, capacitive EIS sensor or LAPS? JF - Sensors and Actuators B. 78 (2001), H. 1-3 Y1 - 2001 SN - 0925-4005 SP - 237 EP - 242 ER - TY - JOUR A1 - Poghossian, Arshak A1 - Yoshinobu, T. A1 - Simonis, A. A1 - Ecken, H. A1 - Lüth, H. A1 - Schöning, Michael Josef T1 - Penicillin detection by means of field-effect based sensors: EnFET, capacitive EIS sensor or LAPS? JF - Proceedings : Copenhagen, Denmark, 27 - 30 August 2000 / [ed.: R. de Reus ...] Y1 - 2000 SN - 87-89935-50-0 N1 - Eurosensors ; (14, 2000, København) ; Eurosensors ; (14 : ; 2000.08.27-30 : ; Copenhagen) ; European Conference on Solid-State Transducers ; (14 : ; 2000.08.27-30 : ; Copenhagen) SP - 27 EP - 30 PB - MIC, Mikroelektronik Centret CY - Lyngby, Denmark ER - TY - JOUR A1 - Poghossian, Arshak A1 - Thust, M. A1 - Schroth, P. A1 - Steffen, A. A1 - Lüth, H. A1 - Schöning, Michael Josef T1 - Penicillin detection by means of silicon-based field-effect structures JF - Sensors and Materials. 13 (2001), H. 4 Y1 - 2001 SN - 0392-2510 SP - 207 EP - 223 ER - TY - JOUR A1 - Koch, Claudia A1 - Poghossian, Arshak A1 - Schöning, Michael Josef A1 - Wege, Christian T1 - Penicillin Detection by Tobacco Mosaic Virus-Assisted Colorimetric Biosensors JF - Nanotheranostics N2 - The presentation of enzymes on viral scaffolds has beneficial effects such as an increased enzyme loading and a prolonged reusability in comparison to conventional immobilization platforms. Here, we used modified tobacco mosaic virus (TMV) nanorods as enzyme carriers in penicillin G detection for the first time. Penicillinase enzymes were conjugated with streptavidin and coupled to TMV rods by use of a bifunctional biotin-linker. Penicillinase-decorated TMV particles were characterized extensively in halochromic dye-based biosensing. Acidometric analyte detection was performed with bromcresol purple as pH indicator and spectrophotometry. The TMV-assisted sensors exhibited increased enzyme loading and strongly improved reusability, and higher analysis rates compared to layouts without viral adapters. They extended the half-life of the sensors from 4 - 6 days to 5 weeks and thus allowed an at least 8-fold longer use of the sensors. Using a commercial budget-priced penicillinase preparation, a detection limit of 100 µM penicillin was obtained. Initial experiments also indicate that the system may be transferred to label-free detection layouts. Y1 - 2018 U6 - http://dx.doi.org/10.7150/ntno.22114 SN - 2206-7418 VL - 2 IS - 2 SP - 184 EP - 196 PB - Ivyspring CY - Sydney ER -