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Keywords
Ribitsch, Doris ; Heumann, Sonja ; Karl, Wolfgang ; Gerlach, Jochen ; Leber, Regina ; Birner-Gruenberger, Ruth ; Gruber, Karl ; Eiteljoerg, Inge ; Remler, Peter ; Siegert, Petra ; Lange, Jennifer ; Maurer, Karl-Heinz ; Berg, Gabriele ; Guebitz, G. M. ; Schwab, H.
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.
Reisert, Steffen ; Schneider, Benno ; Geissler, Hanno ; Gompel, Matthias van ; Wagner, Patrick ; Schöning, Michael Josef
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.
Reisert, Steffen ; Henkel, Hartmut ; Schneider, Andreas ; Schäfer, Daniel ; Friedrich, Peter ; Berger, Jörg ; Schöning, Michael Josef
A handheld sensor system for the online measurement of hydrogen peroxide (H2O2) in aseptic sterilisation processes has been developed. It is based on a calorimetric-type gas sensor that consists of a differential set-up of two temperature sensors, of which one is catalytically activated and the second one is passivated and used as reference. The sensor principle relies in detecting a rise in temperature on the active sensor due to the exothermic reaction of H2O2 on the catalytic surface. To characterise the sensor system towards H2O2 sensitivity and other influencing factors, measurements have been carried out both at an experimental set-up and a manufacturer's sterilisation machine. Physical sensor characterisation was done by means of the optical microscopy.
Reisert, Steffen ; Geissler, Hanno ; Flörke, Rudolf ; Weiler, Christian ; Wagner, Patrick ; Schöning, Michael Josef
Reisert, Steffen ; Geissler, Hanno ; Flörke, Rudolf ; Näther, Niko ; Wagner, Patrick ; Schöning, Michael Josef
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.
Reisert, Steffen ; Geissler, H. ; Florke, R. ; Wagner, P. ; Wagner, Torsten ; Schöning, Michael Josef
Size unlimited markerless deletions by a transconjugative plasmid-system in Bacillus licheniformis
(2013)
Rachinger, Michael ; Bauch, Melanie ; Strittmatter, Axel ; Bongaerts, Johannes ; Evers, Stefan ; Maurer, Karl-Heinz ; Daniel, Rolf ; Liebl, Wolfgang ; Liesegang, Heiko ; Ehrenreich, Armin
Magnetic detection structure for Lab-on-Chip applications based on the frequency mixing technique
(2018)
Rabehi, Amine ; Garlan, Benjamin ; Achtsnicht, Stefan ; Krause, Hans-Joachim ; Offenhäusser, Andreas ; Ngo, Kieu ; Neveu, Sophie ; Graff-Dubois, Stephanie ; Kokabi, Hamid
A magnetic frequency mixing technique with a set of miniaturized planar coils was investigated for use with a completely integrated Lab-on-Chip (LoC) pathogen sensing system. The system allows the detection and quantification of superparamagnetic beads. Additionally, in terms of magnetic nanoparticle characterization ability, the system can be used for immunoassays using the beads as markers. Analytical calculations and simulations for both excitation and pick-up coils are presented; the goal was to investigate the miniaturization of simple and cost-effective planar spiral coils. Following these calculations, a Printed Circuit Board (PCB) prototype was designed, manufactured, and tested for limit of detection, linear response, and validation of theoretical concepts. Using the magnetic frequency mixing technique, a limit of detection of 15 µg/mL of 20 nm core-sized nanoparticles was achieved without any shielding.
Frequency mixing magnetic detection (FMMD) has been widely utilized as a measurement technique in magnetic immunoassays. It can also be used for the characterization and distinction (also known as “colourization”) of different types of magnetic nanoparticles (MNPs) based on their core sizes. In a previous work, it was shown that the large particles contribute most of the FMMD signal. This leads to ambiguities in core size determination from fitting since the contribution of the small-sized particles is almost undetectable among the strong responses from the large ones. In this work, we report on how this ambiguity can be overcome by modelling the signal intensity using the Langevin model in thermodynamic equilibrium including a lognormal core size distribution fL(dc,d0,σ) fitted to experimentally measured FMMD data of immobilized MNPs. For each given median diameter d0, an ambiguous amount of best-fitting pairs of parameters distribution width σ and number of particles Np with R2 > 0.99 are extracted. By determining the samples’ total iron mass, mFe, with inductively coupled plasma optical emission spectrometry (ICP-OES), we are then able to identify the one specific best-fitting pair (σ, Np) one uniquely. With this additional externally measured parameter, we resolved the ambiguity in core size distribution and determined the parameters (d0, σ, Np) directly from FMMD measurements, allowing precise MNPs sample characterization.