TY - GEN A1 - Sieker, T. A1 - Tippkötter, Nils A1 - Ulber, Roland T1 - Simultane Vorbehandlung, Hydrolyse und Fermentation bei der Nutzung von grüner Biomasse zur Produktion von Bioethanol T2 - Chemie Ingenieur Technik N2 - Gräser sind in der Lage, einen großen Teil der für eine biobasierte Wirtschaft benötigten Biomasse zur Verfügung zu stellen. Wie bei anderen lignocellulosehaltigen nachwachsenden Rohstoffen erfordert die Verwertung der im Gras enthaltenen Polysaccharide einen mehrstufigen Prozess aus Vorbehandlung, Hydrolyse und Fermentation. In Gräsern ist die Hemicellulose mitP henolcarbonsäuren wie Ferula- und p-Coumarsäure verestert, die die enzymatische Hydrolyse der Cellulose und Hemicellulose ebenso effektiv behindern wie Lignin. Anders als bei holzigen Rohstoffen ermöglicht dieser Aufbau aber eine enzymatische Vorbehandlung, mit der die Phenolcarbonsäuren abgespalten werden können. Da die bei der Vorbehandlung eingesetzten Enzyme in ihrer natürlichen Funktion synergistisch mit cellulytischen Enzymen zusammenarbeiten, besitzen sie ähnliche Optima wie die für die Hydrolyse der Polysaccharide eingesetzten Cellulasen und Hemicellulasen. Diese Eigenschaft ermöglicht die Integration von Vorbehandlung und Hydrolyse in einem einzigen Verfahrensschritt. Durch die Einführung der enzymatischen Vorbehandlung konnte das in der Literatur bekannte SSF-Verfahren für die Herstellung von Ethanol aus Gräsern um die Vorbehandlungsstufe erweitert werden. Das so realisierte simultaneous pretreatment, saccharification and fermentation (SPSF)-Verfahren stellt eine vollständige Integration der drei für die Nutzung von Lignocellulose nötigen Verfahrensschritte in der grünen Bioraffinerie dar. Y1 - 2010 U6 - https://doi.org/10.1002/cite.201050319 SN - 0009-286X N1 - ProcessNet-Jahrestagung 2010 und 28. DECHEMA-Jahrestagung der Biotechnologen, 21. - 23. September 2010, Eurogress Aachen VL - 82 IS - 9 SP - 1601 PB - Wiley-VCH CY - Weinheim ER - TY - PAT A1 - Siegert, Petra A1 - Spitz, Astrid A1 - Maurer, Karl-Heinz T1 - Neue Proteasen und Mittel enthaltend diese Proteasen [Offenlegungsschrift] T1 - Novel proteases and compositions comprising these proteases [Internationale Patentanmeldung] Y1 - 2010 SP - 1 EP - 31 PB - Deutsches Patentamt / WIPO CY - München / Genf ER - TY - PAT A1 - Siegert, Petra A1 - Spitz, Astrid A1 - Maurer, Karl-Heinz T1 - Wasch- und Reinigungsmittel enthaltend Proteasen aus Bacillus pumilus [Offenlegungsschrift] T1 - Detergents and cleaning agents containing proteases from bacillus pumilus [Europäische Patentschrift / Internationale Patentanmeldung] Y1 - 2010 SP - 1 EP - 20 PB - Deutsches Patentamt / Europäisches Patentamt / WIPO CY - München / Den Hague / Genf ER - TY - PAT A1 - Siegert, Petra A1 - Mussmann, Nina A1 - O'Connell, Timothy A1 - Maurer, Karl-Heinz T1 - Neue Proteasen und Mittel enthaltend diese Proteasen [Offenlegungsschrift] T1 - Novel proteases and means containing said proteases [Internationale Patentanmeldung] Y1 - 2010 SP - 1 EP - 30 PB - Deutsches Patent- und Markenamt / WIPO CY - München / Genf ER - TY - PAT A1 - Siegert, Petra A1 - Baumstark, Rebecca A1 - Kluin, Cornelia A1 - O'Connell, Timothy A1 - Maurer, Karl-Heinz A1 - Hellmuth, Hendrik T1 - Neue Proteasen und Mittel enthaltend diese Proteasen [Offenlegungsschrift] Y1 - 2010 SP - 1 EP - 30 PB - Deutsches Patent- und Markenamt CY - München ER - TY - CHAP A1 - Seibler, Jost A1 - Schwenk, Frieder T1 - Transgenic RNAi Applications in the Mouse T2 - Methods in Enzymology : Guide to Techniques in Mouse Development, Part B: Mouse Molecular Genetics. 2nd Edition Y1 - 2010 SN - 978-0-12-384880-2 N1 - Methods in Enzymology : Vol. 477 SP - 367 EP - 386 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Scheer, Nico A1 - Ross, Jillian A1 - Kapelyukh, Yury A1 - Rode, Anja A1 - Wolf, C. Roland T1 - In vivo responses of the human and murine pregnane X receptor to dexamethasone in mice JF - Drug Metabolism and Disposition N2 - Dexamethasone (DEX) is a potent and widely used anti-inflammatory and immunosuppressant glucocorticoid. It can bind and activate the pregnane X receptor (PXR), which plays a critical role as xenobiotic sensor in mammals to induce the expression of many enzymes, including cytochromes P450 in the CYP3A family. This induction results in its own metabolism. We have used a series of transgenic mouse lines, including a novel, improved humanized PXR line, to compare the induction profile of PXR-regulated drug-metabolizing enzymes after DEX administration, as well as looking at hepatic responses to rifampicin (RIF). The new humanized PXR model has uncovered further intriguing differences between the human and mouse receptors in that RIF only induced Cyp2b10 in the new humanized model. DEX was found to be a much more potent inducer of Cyp3a proteins in wild-type mice than in mice humanized for PXR. To assess whether PXR is involved in the detoxification of DEX in the liver, we analyzed the consequences of high doses of the glucocorticoid on hepatotoxicity on different PXR genetic backgrounds. We also studied these effects in an additional mouse model in which functional mouse Cyp3a genes have been deleted. These strains exhibited different sensitivities to DEX, indicating a protective role of the PXR and CYP3A proteins against the hepatotoxicity of this compound. Y1 - 2010 U6 - https://doi.org/10.1124/dmd.109.031872 SN - 1521-009X VL - 38 IS - 7 SP - 1046 EP - 1053 PB - ASPET CY - Bethesda ER - TY - JOUR A1 - Ross, Jillian A1 - Plummer, Simon M. A1 - Rode, Anja A1 - Scheer, Nico A1 - Bower, Conrad C. A1 - Vogel, Ortwin A1 - Henderson, Colin J. A1 - Wolf, C. Roland A1 - Elcombe, Clifford R. T1 - Human constitutive androstane receptor (CAR) and pregnane X receptor (PXR) support the hypertrophic but not the hyperplastic response to the murine nongenotoxic hepatocarcinogens phenobarbital and chlordane in vivo JF - Toxicological Sciences N2 - Mouse nongenotoxic hepatocarcinogens phenobarbital (PB) and chlordane induce hepatomegaly characterized by hypertrophy and hyperplasia. Increased cell proliferation is implicated in the mechanism of tumor induction. The relevance of these tumors to human health is unclear. The xenoreceptors, constitutive androstane receptors (CARs), and pregnane X receptor (PXR) play key roles in these processes. Novel “humanized” and knockout models for both receptors were developed to investigate potential species differences in hepatomegaly. The effects of PB (80 mg/kg/4 days) and chlordane (10 mg/kg/4 days) were investigated in double humanized PXR and CAR (huPXR/huCAR), double knockout PXR and CAR (PXRKO/CARKO), and wild-type (WT) C57BL/6J mice. In WT mice, both compounds caused increased liver weight, hepatocellular hypertrophy, and cell proliferation. Both compounds caused alterations to a number of cell cycle genes consistent with induction of cell proliferation in WT mice. However, these gene expression changes did not occur in PXRKO/CARKO or huPXR/huCAR mice. Liver hypertrophy without hyperplasia was demonstrated in the huPXR/huCAR animals in response to both compounds. Induction of the CAR and PXR target genes, Cyp2b10 and Cyp3a11, was observed in both WT and huPXR/huCAR mouse lines following treatment with PB or chlordane. In the PXRKO/CARKO mice, neither liver growth nor induction of Cyp2b10 and Cyp3a11 was seen following PB or chlordane treatment, indicating that these effects are CAR/PXR dependent. These data suggest that the human receptors are able to support the chemically induced hypertrophic responses but not the hyperplastic (cell proliferation) responses. At this time, we cannot be certain that hCAR and hPXR when expressed in the mouse can function exactly as the genes do when they are expressed in human cells. However, all parameters investigated to date suggest that much of their functionality is maintained. Y1 - 2010 U6 - https://doi.org/10.1093/toxsci/kfq118 SN - 1096-0929 VL - 116 IS - 2 SP - 452 EP - 466 PB - Oxford University Press CY - Oxford ER - 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 - 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 - CHAP A1 - Poth, Sebastian A1 - Monzon, Magaly A1 - Tippkötter, Nils A1 - Ulber, Roland T1 - Lignocellulosic biorefinery : process integration of hydrolysis and fermentation T2 - Proceedings / 11th European Workshop on Lignocellulosics and Pulp : August 16 - 19, 2010, Hamburg, Germany Y1 - 2010 SP - 65 EP - 68 PB - vTi CY - Hamburg ER -