TY - JOUR A1 - Ribitsch, Doris A1 - Karl, Wolfgang A1 - Birner-Gruenberger, R. A1 - Gruber, K. A1 - Eiteljoerg, I. A1 - Remler, Peter A1 - Wieland, S. A1 - Siegert, Petra A1 - Maurer, Karl-Heinz A1 - Schwab, H. T1 - C-terminal truncation of a metagenome-derived detergent protease for effective expression in E. coli JF - Journal of biotechnology N2 - 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. Y1 - 2010 U6 - https://doi.org/10.1016/j.jbiotec.2010.09.947 SN - 1873-4863 (E-Journal); 0168-1656 (Print) VL - 150 IS - 3 SP - 408 EP - 416 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Probst, M. A1 - Behbahani, Mehdi A1 - Borrmann, E. A1 - Elgeti, S. A1 - Nicolai, M. A1 - Behr, Marek T1 - Hemodynamic Modeling for Numerical Analysis and Design of Medical Devices Y1 - 2010 N1 - Posterpresentation ; NIC Symposium 2010 ; 24 - 25 February 2010 Jülich, Germany ER - TY - JOUR A1 - Behbahani, Mehdi A1 - Mai, A. A1 - Bergmann, B. A1 - Waluga, C. A1 - Behr, Marek A1 - Tran, L. A1 - Vonderstein, K. A1 - Mottaghy, K. T1 - Modeling and Numerical Simulation of Blood Damage Y1 - 2010 N1 - Posterpresentation ; Umbrella Symposium "Modelling and Simulation in Medicine, Engineering and Sciences", Forschungszentrum Jülich, January 18-20, 2010 ER - TY - JOUR A1 - Behbahani, Mehdi A1 - Probst, M. A1 - Mai, A. A1 - Behr, Marek A1 - Tran, L. A1 - Vonderstein, K. A1 - Mottaghy, K. T1 - Numerical Prediction of Blood Damage in Biomedical Devices Y1 - 2010 N1 - Posterpresentation ; Biomedica Congress, Aachen, March 17-18, 2010 ER - TY - JOUR A1 - Siqueira, Jose R. A1 - Maki, Rafael M. A1 - Paulovich, Fernando V. A1 - Werner, Frederik A1 - Poghossian, Arshak A1 - Oliveira, Maria C. F. de A1 - Zucolotto, Valtencir A1 - Oliveira, Osvaldo N. Jr. A1 - Schöning, Michael Josef T1 - Use of information visualization methods eliminating cross talk in multiple sensing units investigated for a light-addressable potentiometric sensor JF - Analytical Chemistry N2 - The integration of nanostructured films containing biomolecules and silicon-based technologies is a promising direction for reaching miniaturized biosensors that exhibit high sensitivity and selectivity. A challenge, however, is to avoid cross talk among sensing units in an array with multiple sensors located on a small area. In this letter, we describe an array of 16 sensing units of a light-addressable potentiometric sensor (LAPS), which was made with layer-by-layer (LbL) films of a poly(amidomine) dendrimer (PAMAM) and single-walled carbon nanotubes (SWNTs), coated with a layer of the enzyme penicillinase. A visual inspection of the data from constant-current measurements with liquid samples containing distinct concentrations of penicillin, glucose, or a buffer indicated a possible cross talk between units that contained penicillinase and those that did not. With the use of multidimensional data projection techniques, normally employed in information visualization methods, we managed to distinguish the results from the modified LAPS, even in cases where the units were adjacent to each other. Furthermore, the plots generated with the interactive document map (IDMAP) projection technique enabled the distinction of the different concentrations of penicillin, from 5 mmol L−1 down to 0.5 mmol L−1. Data visualization also confirmed the enhanced performance of the sensing units containing carbon nanotubes, consistent with the analysis of results for LAPS sensors. The use of visual analytics, as with projection methods, may be essential to handle a large amount of data generated in multiple sensor arrays to achieve high performance in miniaturized systems Y1 - 2010 U6 - https://doi.org/10.1021/ac9024076 SN - 0003-2700 VL - 82 IS - 1 SP - 61 EP - 65 PB - ACS Publications CY - Washington ER - TY - JOUR A1 - Grottke, O. A1 - Braunschweig, T. A1 - Philippen, B. A1 - Gatzweiler, Karl-Heinz A1 - Gronloh, N. A1 - Staat, Manfred A1 - Rossaint, R. A1 - Tolba, R. T1 - A new model for blunt liver injuries in the swine JF - European Surgical Research N2 - Background: To elaborate the impact of new haemostatic agents we developed an instrument for the pressure-controlled induction of blunt liver injuries in a porcine animal model. Materials and Methods: A dilutional coagulopathy of 80% of animal blood volume was induced in 9 anaesthetized pigs. Animals were randomly assigned to be injured with a force of 112 Newton (N) (n = 1), 224 ± 19 N (n = 4) or 355 ± 35 N (n = 4). The impact of injury was measured by blood loss, survival time and coagulation parameters. Liver histology was obtained to evaluate the degree of liver injury. Results: The profound haemodilution resulted in a significant alteration of all coagulation parameters. After inflicting the injury with 355 ± 35 N, both the survival time (30 ± 9 min; p = 0.006) and blood loss (68 ± 16 ml min–1, p = 0.002) were significantly different as compared to injuries with 224 ± 19 N (survival time: 76 ± 20 min, blood loss: 23 ± 4 ml min–1). In contrast, an injury with 112 N led to an insignificant blood loss of only 239 ml. Conclusion: We developed a pressure-controlled clamp that allows for the induction of blunt liver traumas with highly reproducible injuries with a positive correlation with blood loss and survival. Y1 - 2010 U6 - https://doi.org/10.1159/000265053 SN - 1421-9921 VL - 44 IS - 2 SP - 65 EP - 73 PB - Karger CY - Basel ER - TY - CHAP A1 - Lettini, Antonio A1 - Havermann, Marc A1 - Guidetti, Marco A1 - Fornaciari, Andrea T1 - Improved functionalities and energy saving potential on mobile machines combining electronics with flow sharing valve and variable displacement pump T2 - 7th International Fluid Power Conference - Vol. 3 - Aachen Efficiency through Fluid Power Workshop Proceedings Y1 - 2010 SN - 978-3-940565-92-1 N1 - IFK, 7, Internationales Fluidtechnisches Kolloquium, 7., Aachen, DE, 2010-03-22 - 2010-03-24 SP - 103 EP - 114 PB - Apprimus Verlag CY - Aachen ER -