@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{RibitschHeumannKarletal.2012, author = {Ribitsch, D. and Heumann, S. and Karl, W. and Gerlach, J. and Leber, R. and Birner-Gruenberger, R. and Gruber, K. and Eiteljoerg, I. and Remler, P. and Siegert, Petra and Lange, J. and Maurer, Karl-Heinz and Berg, G. and Guebitz, G. M. and Schwab, H.}, title = {Extracellular serine proteases from Stenotrophomonas maltophilia: Screening, isolation and heterologous expression in E. coli}, series = {Journal of biotechnology}, volume = {157}, journal = {Journal of biotechnology}, number = {1}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1873-4863 (E-Journal); 0168-1656 (Print)}, doi = {10.1016/j.jbiotec.2011.09.025}, pages = {140 -- 147}, year = {2012}, abstract = {A large strain collection comprising antagonistic bacteria was screened for novel detergent proteases. Several strains displayed protease activity on agar plates containing skim milk but were inactive in liquid media. Encapsulation of cells in alginate beads induced protease production. Stenotrophomonas maltophilia emerged as best performer under washing conditions. For identification of wash-active proteases, four extracellular serine proteases called StmPr1, StmPr2, StmPr3 and StmPr4 were cloned. StmPr2 and StmPr4 were sufficiently overexpressed in E. coli. Expression of StmPr1 and StmPr3 resulted in unprocessed, insoluble protein. Truncation of most of the C-terminal domain which has been identified by enzyme modeling succeeded in expression of soluble, active StmPr1 but failed in case of StmPr3. From laundry application tests StmPr2 turned out to be a highly wash-active protease at 45 °C. Specific activity of StmPr2 determined with suc-l-Ala-l-Ala-l-Pro-l-Phe-p-nitroanilide as the substrate was 17 ± 2 U/mg. In addition we determined the kinetic parameters and cleavage preferences of protease StmPr2.}, language = {en} } @article{ReisertSchneiderGeissleretal.2013, author = {Reisert, Steffen and Schneider, Benno and Geissler, Hanno and Gompel, Matthias van and Wagner, Patrick and Sch{\"o}ning, Michael Josef}, title = {Multi-sensor chip for the investigation of different types of metal oxides for the detection of H2O2 in the ppm range}, series = {physica status solidi (a)}, volume = {210}, journal = {physica status solidi (a)}, number = {5}, publisher = {Wiley}, address = {Weinheim}, issn = {1862-6319}, pages = {898 -- 904}, year = {2013}, abstract = {In this work, a multi-sensor chip for the investigation of the sensing properties of different types of metal oxides towards hydrogen peroxide in the ppm range is presented. The fabrication process and physical characterization of the multi-sensor chip are described. Pure SnO2 and WO3 as well as Pd- and Pt-doped SnO2 films are characterized in terms of their sensitivity to H2O2. The sensing films have been prepared by drop-coating of water-dispensed nano-powders. A physical characterization, including scanning electron microscopy and X-ray diffraction analysis of the deposited metal-oxide films, was done. From the measurements in hydrogen peroxide atmosphere, it could be shown, that all of the tested metal oxide films are suitable for the detection of H2O2 in the ppm range. The highest sensitivity and reproducibility was achieved using Pt-doped SnO2. Calibration plot of a SnO2, WO3, Pt-, and Pd-doped SnO2 gas sensor for H2O2 concentrations in the ppm range.}, language = {en} } @article{ReisertHenkelSchneideretal.2010, author = {Reisert, Steffen and Henkel, Hartmut and Schneider, Andreas and Sch{\"a}fer, Daniel and Friedrich, Peter and Berger, J{\"o}rg and Sch{\"o}ning, Michael Josef}, title = {Development of a handheld sensor system for the online measurement of hydrogen peroxide in aseptic filling systems}, series = {Physica Status Solidi (A). 207 (2010), H. 4}, journal = {Physica Status Solidi (A). 207 (2010), H. 4}, isbn = {1862-6300}, pages = {913 -- 918}, year = {2010}, language = {en} } @article{ReisertHenkelSchneideretal.2009, author = {Reisert, Steffen and Henkel, H. and Schneider, A. and Sch{\"a}fer, D. and Friedrich, P. and Berger, J. and Sch{\"o}ning, Michael Josef}, title = {Entwicklung eines Handheld-Sensorsystems f{\"u}r die „On-line"-Messung der H2O2-Konzentration in aseptischen Entkeimungsprozessen}, series = {9. Dresdner Sensor-Symposium : Dresden, 07.-09. Dezember 2009 / Gerlach, Gerald ; Hauptmann, Peter [Hrsg.]}, journal = {9. Dresdner Sensor-Symposium : Dresden, 07.-09. Dezember 2009 / Gerlach, Gerald ; Hauptmann, Peter [Hrsg.]}, publisher = {TUDpress}, address = {Dresden}, isbn = {978-3-941298-44-6}, pages = {285 -- 288}, year = {2009}, language = {de} } @article{ReisertGeisslerFloerkeetal.2013, author = {Reisert, Steffen and Geissler, Hanno and Fl{\"o}rke, Rudolf and Weiler, Christian and Wagner, Patrick and Sch{\"o}ning, Michael Josef}, title = {Characterisation of aseptic sterilisation processes using an electronic nose}, series = {International journal of nanotechnology}, volume = {Vol. 10}, journal = {International journal of nanotechnology}, number = {No. 5-7}, publisher = {Inderscience Enterprises}, address = {Gen{\`e}ve}, issn = {1475-7435 (Print) 7141-8151 (Online)}, pages = {470 -- 484}, year = {2013}, language = {en} } @article{ReisertGeisslerFloerkeetal.2011, author = {Reisert, Steffen and Geissler, Hanno and Fl{\"o}rke, Rudolf and N{\"a}ther, Niko and Wagner, Patrick and Sch{\"o}ning, Michael Josef}, title = {Towards a multi-sensor system for the evaluation of aseptic processes employing hydrogen peroxide vapour (H2O2)}, series = {Physica status solidi (a) : applications and material science. 208 (2011), H. 6}, journal = {Physica status solidi (a) : applications and material science. 208 (2011), H. 6}, publisher = {Wiley}, address = {Weinheim}, isbn = {1862-6319}, pages = {1351 -- 1356}, year = {2011}, language = {en} } @article{ReisertGeisslerWeileretal.2015, author = {Reisert, Steffen and Geissler, H. and Weiler, C. and Wagner, P. and Sch{\"o}ning, Michael Josef}, title = {Multiple sensor-type system for monitoring the microbicidal effectiveness of aseptic sterilisation processes}, series = {Food control}, volume = {47}, journal = {Food control}, issn = {1873-7129 (E-Journal); 0956-7135 (Print)}, doi = {10.1016/j.foodcont.2014.07.063}, pages = {615 -- 622}, year = {2015}, abstract = {The present work describes a novel multiple sensor-type system for the real-time analysis of aseptic sterilisation processes employing gaseous hydrogen peroxide (H2O2) as a sterilant. The inactivation kinetics of Bacillus atrophaeus by gaseous H2O2 have been investigated by means of a methodical calibration experiment, taking into account the process variables H2O2 concentration, humidity and gas temperature. It has been found that the microbicidal effectiveness at H2O2 concentrations above 2\% v/v is largely determined by the concentration itself, while at lower H2O2 concentrations, the gas temperature and humidity play a leading role. Furthermore, the responses of different types of gas sensors towards the influencing factors of the sterilisation process have been analysed within the same experiment. Based on a correlation established between the inactivation kinetics and the sensor responses, a calorimetric H2O2 sensor and a metal-oxide semiconductor (MOX) sensor have been identified as possible candidates for monitoring the microbicidal effectiveness of aseptic sterilisation processes employing gaseous H2O2. Therefore, two linear models that describe the relationship between sensor response and microbicidal effectiveness have been proposed.}, 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{RachingerBauchStrittmatteretal.2013, author = {Rachinger, Michael and Bauch, Melanie and Strittmatter, Axel and Bongaerts, Johannes and Evers, Stefan and Maurer, Karl-Heinz and Daniel, Rolf and Liebl, Wolfgang and Liesegang, Heiko and Ehrenreich, Armin}, title = {Size unlimited markerless deletions by a transconjugative plasmid-system in Bacillus licheniformis}, series = {Journal of biotechnology}, volume = {Vol. 164}, journal = {Journal of biotechnology}, number = {Iss. 4}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1873-4863 (E-Journal); 0168-1656 (Print)}, pages = {365 -- 369}, year = {2013}, language = {en} }