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 - CHAP A1 - Wendorff, Marion A1 - Eggert, Thorsten A1 - Pohl, Martina A1 - Dresen, Carola A1 - Müller, Michael A1 - Jaeger, Karl-Erich A1 - Sprenger, Georg A. A1 - Schürmann, Melanie A1 - Schürmann, Martin A1 - Johnen, Sandra A1 - Sprenger, Gerda A1 - Sahm, Hermann A1 - Inoue, Tomoyuki A1 - Schörken, Ulrich A1 - Breittaupt, Holger A1 - Frölich, Bettina A1 - Heim, Petra A1 - Iding, Hans A1 - Juchem, Bettina A1 - Siegert, Petra A1 - Kula, Maria-Regina A1 - Weckbecker, Andrea A1 - Hummel, Werner A1 - Fessner, Wolf-Dieter A1 - Elling, Lothar A1 - Wolberg, Michael A1 - Bode, Silke A1 - Feldmann, Ralf A1 - Geilenkirchen, Petra A1 - Schubert, Thomas A1 - Walter, Lydia A1 - Dünnwald, Thomas A1 - Demir, Ayhan S. A1 - Kolter-Jung, Doris A1 - Nitsche, Adam A1 - Dünkelmann, Pascal A1 - Cosp, Annabel A1 - Lingen, Bettina T1 - Catalytic asymmetric synthesis : section 2.2 T2 - Asymmetric synthesis with chemical and biological methods / ed. by Dieter Enders ... Y1 - 2007 SN - 978-3-527-31473-7 SP - 298 EP - 413 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Siegert, Petra A1 - McLeish, Michael J. A1 - Baumann, Martin A1 - Iding, Hans A1 - Kneen, Malea M. A1 - Kenyon, George L. A1 - Pohl, Martina T1 - Exchanging the substrate specificities of pyruvate decarboxylase from Zymomonas mobilis and benzoylformate decarboxylase from Pseudomonas putida JF - Protein engineering, design, and selection : peds Y1 - 2005 SN - 1460-213X (E-Journal); 1741-0134 (E-Journal); 0269-2139 (Print); 1741-0126 (Print) VL - Vol. 18 IS - Iss. 7 SP - 345 EP - 357 ER - TY - CHAP A1 - Siegert, Petra A1 - Pohl, Martina A1 - Kneen, Malea M. A1 - Pogozheva, Irina D. A1 - Kenyon, George L. A1 - McLeish, Michael J. T1 - Exploring the substrate specificity of benzoylformate decarboxylase, pyruvate decarboxylase, and benzaldehyde lyase T2 - Thiamine : catalytic mechanisms in normal and disease states / ed. by Frank Jordan ... Y1 - 2004 SN - 0-8247-4062-9 SP - 275 EP - 290 PB - Dekker CY - New York, NY ER - TY - JOUR A1 - Dünkelmann, Pascal A1 - Kolter-Jung, Doris A1 - Nitsche, Adam A1 - Demir, Ayhan S. A1 - Siegert, Petra A1 - Lingen, Bettina A1 - Baumann, Martin A1 - Pohl, Martina A1 - Müller, Michael T1 - Development of a donor-acceptor concept for enzymatic cross-coupling reactions of adehydes : the first asymmetric cross-benzoin condensation JF - Journal of the American Chemical Society Y1 - 2002 SN - 1520-5126 (E-Journal); 0002-7863 (Print) VL - Vol. 124 SP - 12084 EP - 12085 ER - TY - JOUR A1 - Dünnwald, Thomas A1 - Demir, Ayhan S. A1 - Siegert, Petra A1 - Pohl, Martina A1 - Müller, Michael T1 - ChemInform Abstract: Enantioselective synthesis of (S)-2-Hydroxypropanone derivatives by Benzoylformate Decarboxylase Catalyzed C—C Bond Formation JF - Cheminform Y1 - 2001 SN - 1522-2667 (E-Journal); 0931-7597 (Print) VL - Vol. 32 IS - Iss. 4 SP - Publ. online ER - TY - JOUR A1 - Dünnwald, Thomas A1 - Demir, Ayhan S. A1 - Siegert, Petra A1 - Pohl, Martina A1 - Müller, Michael T1 - Enantioselective Synthesis of (S)-2-Hydroxypropanone Derivatives by Benzoylformate Decarboxylase Catalyzed C−C Bond Formation JF - European journal of organic chemistry Y1 - 2000 SN - 0365-5490 (E-Journal); 1099-0690 (E-Journal); 0075-4617 (Print); 0170-2041 (Print); 0947-3440 (Print); 1434-193X (Print); 1434-243X (Print) VL - Vol. 2000 IS - Iss. 11 SP - 2161 EP - 2170 ER - TY - CHAP A1 - Siegert, Petra A1 - Iding, Hans A1 - Baumann, Martin A1 - McLeish, Michael J. A1 - Kenyon, George L. A1 - Pohl, Martina T1 - Broadening of the substrate spectra of two ThDP-dependent decarboxylases using site-directed-mutagenesis T2 - Proceedings of the 4th International Congress on Biochemical Engineering : 17 and 18 February 2000, Stuttgart Y1 - 2000 SN - 3-8167-5570-4 SP - 38 EP - 42 ER - TY - JOUR A1 - Iding, Hans A1 - Dünnwald, Thomas A1 - Greiner, Lasse A1 - Liese, Andreas A1 - Müller, Michael A1 - Siegert, Petra A1 - Grötzinger, Joachim A1 - Demir, Ayhan S. A1 - Pohl, Martina T1 - Benzoylformate Decarboxylase from Pseudomonas putida as Stable Catalyst for the Synthesis of Chiral 2-Hydroxy Ketones JF - Chemistry - a European journal Y1 - 2000 SN - 1521-3765 (E-Journal); 0947-6539 (Print) VL - Vol. 6 IS - Iss. 8 SP - 1483 EP - 1495 ER - TY - JOUR A1 - Iding, Hans A1 - Siegert, Petra A1 - Mesch, K. A1 - Pohl, Martina T1 - Application of α-keto acid decarboxylases in biotransformations JF - Biochimica et biophysica acta (BBA) - Protein structure and molecular enzymology Y1 - 1998 SN - 1879-2588 (E-Journal); 0167-4838 (Print) VL - Vol. 1385 IS - Iss. 2 SP - 307 EP - 322 ER - TY - JOUR A1 - Pohl, Martina A1 - Siegert, Petra A1 - Mesch, K. A1 - Bruhn, H. A1 - Grötzinger, Joachim T1 - Active site mutants of pyruvate decarboxylase from Zymomonas mobilis : a site-directed mutagenesis study of L112, I472, I476, E473 and N482 JF - European journal of biochemistry Y1 - 1998 SN - 1432-1033 (E-Journal); 1742-4658 (E-Journal); 0014-2956 (Print); 1742-464X (Print) VL - Vol. 257 IS - Iss. 3 SP - 538 EP - 546 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 - Handtke, Stefan A1 - Volland, Sonja A1 - Methling, Karen A1 - Albrecht, Dirk A1 - Becher, Dörte A1 - Nehls, Jenny A1 - Bongaerts, Johannes A1 - Maurer, Karl-Heinz A1 - Lalk, Michael A1 - Liesegang, Heiko A1 - Voigt, Birgit A1 - Daniel, Rolf A1 - Hecker, Michael T1 - Cell physiology of the biotechnological relevant bacterium Bacillus pumilus - An omics-based approach JF - Journal of Biotechnology N2 - Members of the species Bacillus pumilus get more and more in focus of the biotechnological industry as potential new production strains. Based on exoproteome analysis, B. pumilus strain Jo2, possessing a high secretion capability, was chosen for an omics-based investigation. The proteome and metabolome of B. pumilus cells growing either in minimal or complex medium was analyzed. In total, 1542 proteins were identified in growing B. pumilus cells, among them 1182 cytosolic proteins, 297 membrane and lipoproteins and 63 secreted proteins. This accounts for about 43% of the 3616 proteins encoded in the B. pumilus Jo2 genome sequence. By using GC–MS, IP-LC/MS and H NMR methods numerous metabolites were analyzed and assigned to reconstructed metabolic pathways. In the genome sequence a functional secretion system including the components of the Sec- and Tat-secretion machinery was found. Analysis of the exoproteome revealed secretion of about 70 proteins with predicted secretion signals. In addition, selected production-relevant genome features such as restriction modification systems and NRPS clusters of B. pumilus Jo2 are discussed. Y1 - 2014 U6 - http://dx.doi.org/10.1016/j.jbiotec.2014.08.028 SN - 1873-4863 (E-Journal); 0168-1656 (Print) IS - 192(A) SP - 204 EP - 214 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Nachtrodt, Frederik A1 - Tietsch, Wolfgang A1 - Mostacci, Domiziano A1 - Scherer, Ulrich W. T1 - Set-up and first operation of a plasma oven for treatment of low level radioactive wastes JF - Nuclear technology and radiation protection Y1 - 2014 U6 - http://dx.doi.org/10.2298/NTRP140SS47N SN - 1451-3994 VL - 29 IS - Suppl. SP - 47 EP - 51 PB - VINČA Institute of Nuclear Sciences CY - Belgrad ER - TY - CHAP A1 - Frotscher, Ralf A1 - Goßmann, Matthias A1 - Raatschen, Hans-Jürgen A1 - Temiz Artmann, Aysegül A1 - Staat, Manfred T1 - Simulation of cardiac cell-seeded membranes using the edge-based smoothed FEM T2 - Shell and membrane theories in mechanics and biology. (Advanced structured materials ; 45) N2 - We present an electromechanically coupled Finite Element model for cardiac tissue. It bases on the mechanical model for cardiac tissue of Hunter et al. that we couple to the McAllister-Noble-Tsien electrophysiological model of purkinje fibre cells. The corresponding system of ordinary differential equations is implemented on the level of the constitutive equations in a geometrically and physically nonlinear version of the so-called edge-based smoothed FEM for plates. Mechanical material parameters are determined from our own pressure-deflection experimental setup. The main purpose of the model is to further examine the experimental results not only on mechanical but also on electrophysiological level down to ion channel gates. Moreover, we present first drug treatment simulations and validate the model with respect to the experiments. Y1 - 2015 SN - 978-3-319-02534-6 ; 978-3-319-02535-3 SP - 187 EP - 212 PB - Springer CY - Heidelberg 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 - Seifarth, Volker A1 - Goßmann, Matthias A1 - Grosse, J. O. A1 - Becker, C. A1 - Heschel, I. A1 - Artmann, Gerhard A1 - Temiz Artmann, Aysegül T1 - Development of a Bioreactor to Culture Tissue Engineered Ureters Based on the Application of Tubular OPTIMAIX 3D Scaffolds JF - Urologia Internationalis Y1 - 2015 U6 - http://dx.doi.org/10.1159/000368419 SN - 0042-1138 VL - 2015 IS - 95 SP - 106 EP - 113 PB - Karger CY - Basel ER - TY - JOUR A1 - Pilas, Johanna A1 - Iken, Heiko A1 - Selmer, Thorsten A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Development of a multi‐parameter sensor chip for the simultaneous detection of organic compounds in biogas processes JF - Physica status solidi (a) N2 - An enzyme-based multi-parameter biosensor is developed for monitoring the concentration of formate, d-lactate, and l-lactate in biological samples. The sensor is based on the specific dehydrogenation by an oxidized β-nicotinamide adenine dinucleotide (NAD+)-dependent dehydrogenase (formate dehydrogenase, d-lactic dehydrogenase, and l-lactic dehydrogenase, respectively) in combination with a diaphorase from Clostridium kluyveri (EC 1.8.1.4). The enzymes are immobilized on a platinum working electrode by cross-linking with glutaraldehyde (GA). The principle of the determination scheme in case of l-lactate is as follows: l-lactic dehydrogenase (l-LDH) converts l-lactate into pyruvate by reaction with NAD+. In the presence of hexacyanoferrate(III), the resulting reduced β-nicotinamide adenine dinucleotide (NADH) is then regenerated enzymatically by diaphorase. The electrochemical detection is based on the current generated by oxidation of hexacyanoferrate(II) at an applied potential of +0.3 V vs. an Ag/AgCl reference electrode. The biosensor will be electrochemically characterized in terms of linear working range and sensitivity. Additionally, the successful practical application of the sensor is demonstrated in an extract from maize silage. Y1 - 2015 U6 - http://dx.doi.org/10.1002/pssa.201431894 SN - 1862-6319 VL - 212 IS - 6 SP - 1306 EP - 1312 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Breuer, Lars A1 - Raue, Markus A1 - Kirschbaum, M. A1 - Mang, Thomas A1 - Schöning, Michael Josef A1 - Thoelen, R. A1 - Wagner, Torsten T1 - Light-controllable polymeric material based on temperature-sensitive hydrogels with incorporated graphene oxide JF - Physica status solidi (a) N2 - Poly(N-isopropylacrylamide) (PNIPAAm) hydrogel films with incorporated graphene oxide (GO) were developed and tested as light-stimulated actuators. GO dispersions were synthesized via Hummers method and characterized toward their optical properties and photothermal energy conversion. The hydrogels were prepared by means of photopolymerization. In addition, the influence of GO within the hydrogel network on the lower critical solution temperature (LCST) was investigated by differential scanning calorimetry (DSC). The optical absorbance and the response to illumination were determined as a function of GO concentration for thin hydrogel films. A proof of principle for the stimulation with light was performed. Y1 - 2015 U6 - http://dx.doi.org/10.1002/pssa.201431944 SN - 1862-6319 VL - 212 IS - 6 SP - 1368 EP - 1374 PB - Wiley CY - Weinheim 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 -