TY - JOUR A1 - Ribitsch, D. A1 - Karl, W. A1 - Birner-Gruenberger, R. A1 - Gruber, K. A1 - Eiteljoerg, I. A1 - Remler, P. 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 - http://dx.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 - Ribitsch, D. A1 - Heumann, S. A1 - Karl, W. A1 - Gerlach, J. A1 - Leber, R. A1 - Birner-Gruenberger, R. A1 - Gruber, K. A1 - Eiteljoerg, I. A1 - Remler, P. A1 - Siegert, Petra A1 - Lange, J. A1 - Maurer, Karl-Heinz A1 - Berg, G. A1 - Guebitz, G. M. A1 - Schwab, H. T1 - Extracellular serine proteases from Stenotrophomonas maltophilia: Screening, isolation and heterologous expression in E. coli JF - Journal of biotechnology N2 - 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. KW - Alginate beads KW - Stenotrophomonas maltophilia KW - Detergent protease Y1 - 2012 U6 - http://dx.doi.org/10.1016/j.jbiotec.2011.09.025 SN - 1873-4863 (E-Journal); 0168-1656 (Print) VL - 157 IS - 1 SP - 140 EP - 147 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Reugels, Alexander M. A1 - Boggetti, Barbara A1 - Scheer, Nico A1 - Campos-Ortega, José A. T1 - Asymmetric localization of Numb:EGFP in dividing neuroepithelial cells during neurulation in Danio rerio JF - Developmental Dynamics Y1 - 2006 U6 - http://dx.doi.org/10.1002/dvdy.20699 SN - 1097-0177 VL - 235 IS - 4 SP - 934 EP - 948 ER - TY - JOUR A1 - Raupp, Sebastian M. A1 - Schmitt, Marcel A1 - Walz, Anna-Lena A1 - Diehm, Ralf A1 - Hummel, Helga A1 - Scharfer, Philip A1 - Schabel, Wilhelm T1 - Slot die stripe coating of low viscous fluids JF - Journal of Coatings Technology and Research N2 - Slot die coating is applied to deposit thin and homogenous films in roll-to-roll and sheet-to-sheet applications. The critical step in operation is to choose suitable process parameters within the process window. In this work, we investigate an upper limit for stripe coatings. This maximum film thickness is characterized by stripe merging which needs to be avoided in a stable process. It is shown that the upper limit reduces the process window for stripe coatings to a major extent. As a result, stripe coatings at large coating gaps and low viscosities are only possible for relatively thick films. Explaining the upper limit, a theory of balancing the side pressure in the gap region in the cross-web direction has been developed. Y1 - 2018 U6 - http://dx.doi.org/10.1007/s11998-017-0039-y SN - 1935-3804 VL - 15 IS - 5 SP - 899 EP - 911 PB - Springer ER - TY - JOUR A1 - Raue, Markus A1 - Wambach, M. A1 - Glöggler, S. A1 - Grefen, Dana A1 - Kaufmann, R. A1 - Abetz, C. A1 - Georgopanos, P. A1 - Handge, U. A. A1 - Mang, Thomas A1 - Blümich, B. A1 - Abetz, V. T1 - Investigation of historical hard rubber ornaments of Charles Goodyear JF - Macromolecular chemistry and physics Y1 - 2014 SN - 1022-1352 VL - Vol. 215 IS - No. 3 SP - 245 EP - 254 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Ratke, Lorenz A1 - Milow, Barbara A1 - Lisinski, Susanne A1 - Hoepfner, Sandra T1 - On an effect of fine ceramic particles on the structure of aerogels JF - Microgravity science and technology Y1 - 2014 U6 - http://dx.doi.org/10.1007/s12217-014-9380-2 SN - 0938-0108 ; 1875-0494 VL - 26 SP - 103 EP - 110 PB - Springer Nature CY - Heidelberg ER - TY - JOUR A1 - Rachinger, Michael A1 - Bauch, Melanie A1 - Strittmatter, Axel A1 - Bongaerts, Johannes A1 - Evers, Stefan A1 - Maurer, Karl-Heinz A1 - Daniel, Rolf A1 - Liebl, Wolfgang A1 - Liesegang, Heiko A1 - Ehrenreich, Armin T1 - Size unlimited markerless deletions by a transconjugative plasmid-system in Bacillus licheniformis JF - Journal of biotechnology Y1 - 2013 SN - 1873-4863 (E-Journal); 0168-1656 (Print) VL - Vol. 164 IS - Iss. 4 SP - 365 EP - 369 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Raab, Monika A1 - Kappel, Sven A1 - Krämer, Andrea A1 - Sanhaji, Mourad A1 - Matthess, Yves A1 - Kurunci-Csacsko, Elisabeth A1 - Calzada-Wack, Julia A1 - Rathkolb, Birgit A1 - Rosman, Jan A1 - Adler, Thure A1 - Busch, Dirk H. A1 - Esposito, Irene A1 - Fuchs, Helmut A1 - Gailus-Durner, Valérie A1 - Klingenspor, Martin A1 - Wolf, Eckhard A1 - Sänger, Nicole A1 - Prinz, Florian A1 - Hrabe de Angelis, Martin A1 - Seibler, Jost A1 - Yuan, Juping A1 - Bergmann, Martin A1 - Knecht, Rainald A1 - Kreft, Bertolt A1 - Strebhardt, Klaus T1 - Toxicity modelling of Plk1-targeted therapies in genetically engineered mice and cultured primary mammalian cells JF - Nature Communications Y1 - 2011 U6 - http://dx.doi.org/10.1038/ncomms1395 SN - 2041-1723 VL - 2 IS - 395 SP - 1 EP - 11 PB - Nature CY - London ER - TY - JOUR A1 - Prielmeier, Franz A1 - Woznyj, M. A1 - Lüdemann, H.-D. T1 - Pressure Dependence of the Melting and Self Diffusion in 2,2-Dimethylpropane, 2,2-Dimethylpropionitrile, and 2-Methylpropanol-2 / M. Woznyj, F. X. Prielmeier, H.-D. Lüdemann JF - Zeitschrift für Naturforschung A, Journal of Physical Sciences. 39 (1984) Y1 - 1984 SN - 0932-0784 SP - 800 ER - TY - JOUR A1 - Prielmeier, Franz A1 - Wick, Markus A1 - Nagatomo, Yasushi A1 - Frahm, Jens T1 - Alteration of Intracellular Metabolite Diffusion in Rat Brain In Vivo During Ischemia and Reperfusion / Markus Wick, Yasushi Nagatomo, Franz Prielmeier, Jens Frahm JF - Stroke. 26 (1995), H. 10 Y1 - 1995 SN - 0039-2499 SP - 1930 EP - 1934 ER - TY - JOUR A1 - Prielmeier, Franz A1 - Speedy, R. J. A1 - Vardag, T. A1 - Lang, E. W. T1 - Diffusion in simple fluids / R. J. Speedy; F. X. Prielmeier; T. Vardag; E. W. Lang; H.-D. Lüdemann JF - Molecular Physics. 66 (1989), H. 3 Y1 - 1989 SN - 0026-8976 SP - 577 EP - 590 ER - TY - JOUR A1 - Prielmeier, Franz A1 - Speedy, R. J. A1 - Lüdemann, H.-D. T1 - High Pressure NMR Self-Diffusion Studies on Supercooled Water JF - High Pressure Science and Technology Proceeding XI AIRAPT, Kiew. 1 Y1 - 1987 N1 - AIRAPT Conference / International Association for the Advancement of High Pressure Science and Technology SP - 75 ER - TY - JOUR A1 - Prielmeier, Franz A1 - Radkowitsch, H. A1 - Lang, E. W. A1 - Lüdemann, H.-D. T1 - Density dependence of the molecular dynamics of fluid CH3F and CF3H studied by NMR / H. Radkowitsch, F. X. Prielmeier, E. W. Lang and H.-D. Lüdemann JF - Physica B+C. 139-140 (1986) Y1 - 1986 SN - 0921-4526 SP - 96 EP - 99 ER - TY - JOUR A1 - Prielmeier, Franz A1 - Nagatomo, Yasushi A1 - Wick, Markus A1 - Frahm, Jens T1 - Dynamic monitoring of cerebral metabolites during and after transient global ischemia in rats by quantitative proton NMR spectroscopy in vivo / Yasushi Nagatomo, Markus Wick, Franz Prielmeier, Jens Frahm JF - NMR in Biomedicine. 8 (1995), H. 6 Y1 - 1995 SN - 1099-1492 SP - 265 EP - 270 ER - TY - JOUR A1 - Prielmeier, Franz A1 - Nagatomo, Yasushi A1 - Frahm, Jens T1 - Cerebral blood oxygenation in rat brain during hypoxic hypoxia. Quantitative MRI of effective transverse relaxation rates JF - Magnetic Resonance in Medicine. 31 (1994), H. 6 Y1 - 1994 SN - 0740-3194 SP - 678 EP - 681 ER - TY - JOUR A1 - Prielmeier, Franz A1 - Merboldt, K. D. A1 - Hanicke, W. A1 - Frahm, J. T1 - Dynamic high-resolution MR imaging of brain deoxygenation during transient anoxia in the anesthetized rat JF - Journal of cerebral blood flow and metabolism. 13 (1993), H. 5 Y1 - 1993 SN - 0271-678X SP - 889 EP - 894 ER - TY - JOUR A1 - Prielmeier, Franz A1 - Lüdemann, H.-D. T1 - Self diffusion in compressed liquid chloromethane, dichloromethane and trichloromethane / F. X. Prielmeier; H.-D. Lüdemann JF - Molecular Physics. 58 (1986), H. 3 Y1 - 1986 SN - 0026-8976 SP - 593 EP - 604 ER - TY - JOUR A1 - Prielmeier, Franz A1 - Lang, E. W. A1 - Speedy, R. J. A1 - Lüdemann, H.-D. T1 - Diffusion in supercooled water to 300 MPa JF - Physical Review Letters. 59 (1987), H. 10 Y1 - 1987 SN - 0031-9007 SP - 1128 EP - 1131 ER - TY - JOUR A1 - Prielmeier, Franz A1 - Lang, E. W. A1 - Speedy, R. J. A1 - Lüdemann, H.-D. T1 - The pressure Dependence of Self Diffusion in Supercooled Light and Heavy Water / F.X. Prielmeier, E .W. Lang, R. J. Speedy, H.-D. Lüdemann JF - Berichte der Bunsen-Gesellschaft für Physikalische Chemie. 92 (1988) Y1 - 1988 SN - 0005-9021 SP - 1111 ER - TY - JOUR A1 - Prielmeier, Franz A1 - Lang, E. W. A1 - Radkowitsch, H. A1 - Lüdemann, H.-D. T1 - High Pressure NMR Study of the Molecular Dynamics of Liquid methyl fluoride and deutero-methyl fluoride / Lang, E. W. ; Prielmeier, F. X. ; Radkowitsch, H. ; Lüdemann, H. D. JF - Berichte der Bunsen-Gesellschaft für Physikalische Chemie. 91 (1987), H. 10 Y1 - 1987 SN - 0005-9021 SP - 1017 EP - 1025 ER -