@inproceedings{BegingPoghossianMlyneketal.2010, author = {Beging, Stefan and Poghossian, Arshak and Mlynek, D. and Hataihimakul, S. and Pedraza, A. and Dhawan, S. and Laube, N. and Kleinen, L. and Baldsiefen, G. and Busch, H. and Sch{\"o}ning, Michael Josef}, title = {Ion-selective sensors for the determination of the risk of urinary stone formation}, series = {Micro- and Nanosystems in biochemical diagnosis : Principles and applications}, booktitle = {Micro- and Nanosystems in biochemical diagnosis : Principles and applications}, address = {Warsaw}, pages = {74 -- 80}, year = {2010}, language = {en} } @article{RibitschKarlBirnerGruenbergeretal.2010, author = {Ribitsch, D. and Karl, W. and Birner-Gruenberger, R. and Gruber, K. and Eiteljoerg, I. and Remler, P. and Wieland, S. and Siegert, Petra and Maurer, Karl-Heinz and Schwab, H.}, title = {C-terminal truncation of a metagenome-derived detergent protease for effective expression in E. coli}, series = {Journal of biotechnology}, volume = {150}, journal = {Journal of biotechnology}, number = {3}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1873-4863 (E-Journal); 0168-1656 (Print)}, doi = {10.1016/j.jbiotec.2010.09.947}, pages = {408 -- 416}, year = {2010}, abstract = {Recently, a new alkaline protease named HP70 showing highest homology to extracellular serine proteases of Stenotrophomonas maltophilia and Xanthomonas campestris was found in the course of a metagenome screening for detergent proteases (Niehaus et al., submitted for publication). Attempts to efficiently express the enzyme in common expression hosts had failed. This study reports on the realization of overexpression in Escherichia coli after structural modification of HP70. Modelling of HP70 resulted in a two-domain structure, comprising the catalytic domain and a C-terminal domain which includes about 100 amino acids. On the basis of the modelled structure the enzyme was truncated by deletion of most of the C-terminal domain yielding HP70-C477. This structural modification allowed effective expression of active enzyme using E. coli BL21-Gold as the host. Specific activity of HP70-C477 determined with suc-l-Ala-l-Ala-l-Pro-l-Phe-p-nitroanilide as the substrate was 30 ± 5 U/mg compared to 8 ± 1 U/mg of the native enzyme. HP70-C477 was most active at 40 °C and pH 7-11; these conditions are prerequisite for a potential application as detergent enzyme. Determination of kinetic parameters at 40 °C and pH = 9.5 resulted in KM = 0.23 ± 0.01 mM and kcat = 167.5 ± 3.6 s⁻¹. MS-analysis of peptide fragments obtained from incubation of HP70 and HP70-C477 with insulin B indicated that the C-terminal domain influences the cleavage preferences of the enzyme. Washing experiments confirmed the high potential of HP70-C477 as detergent protease.}, language = {en} } @article{DegeringEggertPulsetal.2010, author = {Degering, Christian and Eggert, Thorsten and Puls, Michael and Bongaerts, Johannes and Evers, Stefan and Maurer, Karl-Heinz and Jaeger, Karl-Erich}, title = {Optimization of protease secretion in Bacillus subtilis and Bacillus licheniformis by screening of homologous and herologous signal peptides}, series = {Applied and environmental microbiology}, volume = {76}, journal = {Applied and environmental microbiology}, number = {19}, publisher = {American Society for Microbiology}, address = {Washington, DC}, issn = {1098-5336 (E-Journal); 0003-6919 (Print); 0099-2240 (Print)}, doi = {10.1128/AEM.01146-10}, pages = {6370 -- 6378}, year = {2010}, abstract = {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.}, language = {en} } @article{MiyamotoKanekoMatsuoetal.2010, author = {Miyamoto, Ko-ichiro and Kaneko, Kazumi and Matsuo, Akira and Wagner, Torsten and Kanoh, Shin`ichiro and Sch{\"o}ning, Michael Josef and Yoshinobu, Tatsuo}, title = {Miniaturized chemical imaging sensor system using an OLED display panel}, series = {Procedia Engineering. 5 (2010)}, journal = {Procedia Engineering. 5 (2010)}, isbn = {1877-7058}, pages = {516 -- 519}, year = {2010}, language = {en} } @article{BohrnStuetzFleischeretal.2010, author = {Bohrn, U. and St{\"u}tz, E. and Fleischer, M. and Sch{\"o}ning, Michael Josef}, title = {Real-time detection of CO by eukaryotic cells}, series = {Procedia Engineering. 5 (2010)}, journal = {Procedia Engineering. 5 (2010)}, isbn = {1877-7058}, pages = {17 -- 20}, year = {2010}, language = {en} } @article{PoghossianWagnerSchoening2010, author = {Poghossian, Arshak and Wagner, H. and Sch{\"o}ning, Michael Josef}, title = {Automatisiertes „wafer level"-Testsystem zur Charakterisierung von siliziumbasierten Chemo- und Biosensoren}, series = {Sensoren und Messsysteme 2010 [Elektronische Ressource] : Vortr{\"a}ge der 15. ITG/GMA-Fachtagung vom 18. bis 19. Mai 2010 in N{\"u}rnberg / Informationstechnische Gesellschaft im VDE (ITG); VDI/VDE-Gesellschaft Mess- und Automatisierungstechnik (GMA)}, journal = {Sensoren und Messsysteme 2010 [Elektronische Ressource] : Vortr{\"a}ge der 15. ITG/GMA-Fachtagung vom 18. bis 19. Mai 2010 in N{\"u}rnberg / Informationstechnische Gesellschaft im VDE (ITG); VDI/VDE-Gesellschaft Mess- und Automatisierungstechnik (GMA)}, publisher = {VDE Verlag}, address = {Berlin}, isbn = {978-3-8007-3260-9}, pages = {89 -- 92}, year = {2010}, language = {de} } @article{WagnerMiyamotoSchoeningetal.2010, author = {Wagner, Torsten and Miyamoto, Ko-ichiro and Sch{\"o}ning, Michael Josef and Yoshinobu, Tatsuo}, title = {Novel combination of digital light processing (DLP) and light-addressable potentiometric sensors (LAPS) for flexible chemical imaging}, series = {Procedia Engineering. 5 (2010)}, journal = {Procedia Engineering. 5 (2010)}, isbn = {1877-7058}, pages = {520 -- 523}, year = {2010}, language = {en} } @article{KirchnerOberlaenderFriedrichetal.2010, author = {Kirchner, Patrick and Oberl{\"a}nder, Jan and Friedrich, Peter and Rysstad, G. and Berger, J. and Keusgen, M. and Sch{\"o}ning, Michael Josef}, title = {Realization of a calorimetric gas sensor on polyimide foil for applications in aseptic food industry}, series = {Procedia Engineering. 5 (2010)}, journal = {Procedia Engineering. 5 (2010)}, isbn = {1877-7058}, pages = {264 -- 267}, year = {2010}, language = {en} } @article{BiselliBaeckerPoghossianetal.2010, author = {Biselli, Manfred and B{\"a}cker, Matthias and Poghossian, Arshak and Sch{\"o}ning, Michael Josef and Schnitzler, Thomas and Zang, Werner and Wagner, P.}, title = {Entwicklung eines modularen festk{\"o}rperbasierten Sensorsystems f{\"u}r die {\"U}berwachung von Zellkulturfermenationen}, series = {Sensoren und Messsysteme 2010 [Elektronische Ressource] : Vortr{\"a}ge der 15. ITG/GMA-Fachtagung vom 18. bis 19. Mai 2010 in N{\"u}rnberg / Informationstechnische Gesellschaft im VDE (ITG); VDI/VDE-Gesellschaft Mess- und Automatisierungstechnik (GMA)}, journal = {Sensoren und Messsysteme 2010 [Elektronische Ressource] : Vortr{\"a}ge der 15. ITG/GMA-Fachtagung vom 18. bis 19. Mai 2010 in N{\"u}rnberg / Informationstechnische Gesellschaft im VDE (ITG); VDI/VDE-Gesellschaft Mess- und Automatisierungstechnik (GMA)}, publisher = {VDE Verlag}, address = {Berlin}, isbn = {978-3-8007-3260-9}, pages = {688 -- 691}, year = {2010}, language = {de} } @article{KirchnerSpelthahnSchoeningetal.2010, author = {Kirchner, Patrick and Spelthahn, H. and Sch{\"o}ning, Michael Josef and Henkel, H. and Schneider, A. and Friedrich, P. and Kolstad, J. and Berger, J.}, title = {Realisierung eines Polyimid-basierten kalorimetrischen Gassensors zur Inline-{\"U}berwachung der H2O2-Konzentration in aseptischen Abf{\"u}llanlagen}, series = {Sensoren und Messsysteme 2010 [Elektronische Ressource] : Vortr{\"a}ge der 15. ITG/GMA-Fachtagung vom 18. bis 19. Mai 2010 in N{\"u}rnberg / Informationstechnische Gesellschaft im VDE (ITG); VDI/VDE-Gesellschaft Mess- und Automatisierungstechnik (GMA)}, journal = {Sensoren und Messsysteme 2010 [Elektronische Ressource] : Vortr{\"a}ge der 15. ITG/GMA-Fachtagung vom 18. bis 19. Mai 2010 in N{\"u}rnberg / Informationstechnische Gesellschaft im VDE (ITG); VDI/VDE-Gesellschaft Mess- und Automatisierungstechnik (GMA)}, publisher = {VDE Verlag}, address = {Berlin}, isbn = {978-3-8007-3260-9}, pages = {607 -- 612}, year = {2010}, language = {de} }