@article{BaumannLaraHubbertenetal.1997, author = {Baumann, Marcus and Lara, R.J. and Hubberten, D.N. and Thomas, D. N.}, title = {Dissolved organic matter studies in enclosed systems: Application of hydophobic fractionation for the assessment of organic nitrogen dynamics / Lara, R.J. ; Hubberten, U. ; Thomas, D.N. ; Baumann, M.E.M. ; Kattner, G.}, series = {Journal of Marine Systems. 13 (1997), H. 1-4}, journal = {Journal of Marine Systems. 13 (1997), H. 1-4}, isbn = {0924-7963}, pages = {155 -- 161}, year = {1997}, language = {en} } @article{BaumannThomasGleitz1992, author = {Baumann, Marcus and Thomas, D. N. and Gleitz, M.}, title = {Efficiency of carbon assimilation and photoacclimation in a small unicellular Chaetoceros species from the Weddel Sea (Antarctica): Influence of temperature and irridiance / Thomas, D. ; Baumann, M.E.M. ; Gleitz, M.}, series = {Journal of Experimental Marine Biology and Ecology. 157 (1992), H. 2}, journal = {Journal of Experimental Marine Biology and Ecology. 157 (1992), H. 2}, isbn = {0022-0981}, pages = {195 -- 209}, year = {1992}, language = {en} } @book{Pettrak2014, author = {Pettrak, J{\"u}rgen}, title = {Nutzung nachwachsender Rohstoffe bei der Herstellung thermoplastischer Elastomere aus Folgeprodukten der Olefinmetathese}, publisher = {Attenkofer}, address = {Straubing}, isbn = {978-3-942742-35-1}, pages = {147 Seiten}, year = {2014}, language = {de} } @misc{SiekerTippkoetterUlberetal.2009, author = {Sieker, T. and Tippk{\"o}tter, Nils and Ulber, Roland and Bart, Hans-J{\"o}rg and Heinzle, E.}, title = {Nutzung von Silage zur fermentativen Produktion von Grund-und Feinchemikalien}, series = {Chemie Ingenieur Technik}, volume = {81}, journal = {Chemie Ingenieur Technik}, number = {8}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {0009-286X}, doi = {10.1002/cite.200950271}, pages = {1207}, year = {2009}, abstract = {Grasschnitt hat in Deutschland ein betr{\"a}chtliches Potenzial als nachwachsender Rohstoff. Da frischer Grasschnitt nur in den Sommermonaten zur Verf{\"u}gung steht und Gras bei der Lagerung verrottet, wird er unter anderem durch Silierung konserviert. W{\"a}hrend der Silierung wird ein Teil der wasserl{\"o}slichen Kohlenhydrate unter anaeroben Bedingungen zu Milchs{\"a}ure fermentiert. Die Kombination aus Luftabschluss und Ans{\"a}uerung bewirkt die Konservierung der Silage. Silage als weit verbreitetes landwirtschaftliches Erzeugnis ist somit ein potentieller, in großen Mengen verf{\"u}gbarer Lieferant f{\"u}r eine Vielzahl von Substraten f{\"u}r mikrobielle Fermentationen. Diese k{\"o}nnen entweder durch die Hydrolyse der in den Pflanzen enthaltenen Cellulosen und Hemicellulosen oder durch die Verwendung eines Silagepresssaftes nutzbar gemacht werden. Die zu entwickelnden Prozesse sollen die verbleibenden Kohlenhydrate, inklusive der Cellulose und Hemicellulose, sowie die Milchs{\"a}ure nutzen. Die in der Silage enthaltenen Zucker sollen zu Ethanol, Itakons{\"a}ure und Bernsteins{\"a}ure und die Milchs{\"a}ure zu 1,2-Propandiol umgesetzt werden. Anfallende Reststoffe wie Hydrolyser{\"u}ckst{\"a}nde, Presskuchen und Fermentationsr{\"u}ckst{\"a}nde sollen bei allen zu etablierenden Prozessen entweder als Viehfutter verwendet oder der Biogasproduktion zugef{\"u}hrt werden k{\"o}nnen, wodurch eine vollst{\"a}ndige stoffliche und energetische Nutzung der Silage erreicht wird.}, language = {de} } @article{SrivastavaSinghAggarwaletal.2010, author = {Srivastava, Alok and Singh, Virendra and Aggarwal, Pranav and Schneeweiss, F. and Scherer, Ulrich W. and Friedrich, W.}, title = {Optical studies of insulating polymers for radiation dose monitoring}, series = {Indian Journal of Pure and Applied Physics}, volume = {48}, journal = {Indian Journal of Pure and Applied Physics}, number = {11}, publisher = {Council Of Scientific And Industrial Research (CSIR), National Institute Of Science Communication and Policy Research (NIScPR)}, address = {New Delhi}, isbn = {0019-5596}, pages = {782 -- 786}, year = {2010}, abstract = {The optical study carried out on insulating polymers namely polyethyleneterephthalate (PET) and polyvinylchloride (PVC) has been described. The polymers are exposed to different radiation doses by exposing them to swift heavy ions of carbon (90 MeV), silicon (120 MeV) and nickel (100 MeV) which influence on their optical properties. The studies show that amongst the investigated polymers, PVC and PET have potential for application as dosimeter beyond a threshold dose which is strongly dependent on the nature of the material and the radiation type. The optical micrographs show a distinct change in colour of the sample with increase in radiation dose.}, language = {en} } @article{BodeBartschBoulikasetal.1998, author = {Bode, J{\"u}rgen and Bartsch, J{\"o}rg W. and Boulikas, Toulikas and Iber, Michaela and Mielke, Christian and Sch{\"u}beler, Dirk and Seibler, Jost and Benham, Craig}, title = {Transcription-promoting genomic sites in mammalia: their elucidation and architectural principles}, series = {Gene therapy \& molecular biology}, volume = {1}, journal = {Gene therapy \& molecular biology}, number = {1}, issn = {1529-9120}, pages = {1 -- 29}, year = {1998}, language = {en} } @article{ClaessenGrefenMangetal.2010, author = {Claessen, O. and Grefen, Dana and Mang, Thomas and Dikland, H. G. and Dikland, H. G. and Duin, M. van}, title = {Helle Fensterprofilmaterialien : Alterungsverhalten auf Basis von peroxidisch vernetztem EPDM}, series = {Kautschuk, Gummi, Kunststoffe : KGK}, volume = {63}, journal = {Kautschuk, Gummi, Kunststoffe : KGK}, number = {9}, publisher = {H{\"u}thig}, address = {Heidelberg}, isbn = {0948-3276}, pages = {350 -- 360}, year = {2010}, language = {de} } @misc{SiekerTippkoetterUlberetal.2010, author = {Sieker, T. and Tippk{\"o}tter, Nils and Ulber, Roland and Bart, Hans-J{\"o}rg and Heinzle, E.}, title = {Gr{\"u}ne Bioraffinerie: Ganzheitliche Nutzung von Grassilage f{\"u}r die Herstellung von Grund-und Feinchemikalien}, series = {Chemie Ingenieur Technik}, volume = {82}, journal = {Chemie Ingenieur Technik}, number = {9}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {0009-286X}, doi = {10.1002/cite.201050321}, pages = {1564}, year = {2010}, abstract = {Gras hat ein hohes Potenzial als nachwachsender Rohstoff. Bei ungeeigneter Lagerung verderben Gr{\"a}ser allerdings innerhalb weniger Tage. Dieser Nachteil kann durch die Silierung des Grasschnittes behoben werden. Eines der wichtigsten in der Silage enthaltenen Produkte ist die Milchs{\"a}ure. Um diese f{\"u}r weitere Aufarbeitungsschritte zug{\"a}nglich zu machen, wird aus der Silage ein Presssaft hergestellt. Die Milchs{\"a}ure wird aus einem Silagepresssaft mittels Extraktion durch ionische Fl{\"u}ssigkeiten isoliert. Dabei wird zum einen eine reine Milchs{\"a}ure hergestellt, die z. B. f{\"u}r die Herstellung von Polymilchs{\"a}ure genutzt werden kann. Zum anderen wird ein weniger aufgereinigter Extrakt gewonnen, der f{\"u}r die fermentative Produktion von L-Lysin und 1,2-Propandiol genutzt werden soll. Im Rahmen des Projekts erfolgt die gentechnische Optimierung von Corynebacterium glutamicum f{\"u}r die Umsetzung von Milchs{\"a}urezu L-Lysin. Die im nach der Pressung verbleibenden Presskuchen enthaltenen Grasfasern bestehen zu einem großen Teil aus Polysacchariden. Diese werden hydrolysiert und die dabei freigesetzten Zucker zu Grundchemikalien wie Ethanol oder Itakons{\"a}ure fermentiert. Im Rahmen einer vollst{\"a}ndigen Nutzung der Silage wird das Raffinat aus der Milchs{\"a}ureextraktion als Mediumsupplement in der Fermentation eingesetzt, was die Zugabe weiterer Medienbestandteile {\"u}berfl{\"u}ssig macht. Die R{\"u}ckst{\"a}nde der Hydrolysen und Fermentationen sollen dar{\"u}berhinaus f{\"u}r die Herstellung von Biogas genutzt werden.}, language = {de} } @article{KowollikSchnitzlerBisellietal.2010, author = {Kowollik, Silvia and Schnitzler, Thomas and Biselli, Manfred and Krueger, R. and Zang, Werner and Peuscher, A. and Schillberg, S. and Fischer, R.}, title = {Die Rolle des Respirationsquotienten in der Zellkulturfermentation}, series = {Chemie Ingenieur Technik}, volume = {82}, journal = {Chemie Ingenieur Technik}, number = {9}, publisher = {Wiley-VCH}, address = {Weinheim}, isbn = {Chemie Ingenieur Tec}, doi = {10.1002/cite.201050393}, pages = {1505 -- 1506}, year = {2010}, abstract = {In der biopharmazeutischen Industrie werden rekombinante Proteine und monoklonale Antik{\"o}rper in Zellkulturfermentationen produziert, da nur humane oder tierische Zelllinien {\"u}ber die F{\"a}higkeit der Glykosylierung verf{\"u}gen. Um hohe Produktausbeuten in ausgezeichneter Qualit{\"a}t zu erzielen, ist eine funktionst{\"u}chtige Prozesskontrolle unerl{\"a}sslich. Hierzu wurde in Kooperation mit der Firma Hitec Zang GmbH die HiSense� Pr{\"a}zisionsabgasanalytik entwickelt, die auf Basis der vollautomatischen Ermittlung des Respirationsquotienten (RQ; Verh{\"a}ltnis vonKohlendioxidbildungsrate (CER) zu Sauerstoffaufnahmerate (OTR)) einen Fermentationsprozess nicht-invasiv {\"u}berwacht. Der RQ kann in Hybridoma- und CHO-Zellen (s. Abb.) in sowohl serumhaltigen als auch serumfreien Medien erfolgreich ermittelt werden. Hier spiegeln die CER und die OTR das Wachstumsverhalten der kultivierten CHO-Zellen wider. Der RQ nimmt dabei Werte zwischen 0,9 und 1,2 an. Dies l{\"a}sst auf verschiedene Stoffwechselaktivit{\"a}ten schließen. Da die momentane industrielle Prozesskontrolle auf gemessenen Sauerstoffaufnahmeraten oder entsprechende Offline-Analytiken der Metaboliten basieren, soll durch die vollautomatische RQ-Ermittlung ein neues Verfahren zur Fermentations{\"u}berwachung etabliert werden. Bisher war diese, in bakteriellen Kultivierungen standardisierte Methode, aufgrund der schwierigen CER-Berechnung bei Zellkulturen keine ad{\"a}quate Alternative.}, language = {de} } @article{MangHodeniusSchmitzRodeetal.2009, author = {Mang, Thomas and Hodenius, Michael A. J. and Schmitz-Rode, Thomas and Baumann, Martin and Ivanova, Gergana and Wong, John Erik and Haulena, Friedhelm and Soenen, Stefaan J. H. and de Cuyper, Marcel}, title = {Absorption of 10-hydroxycamptothecin into the coat of magnetoliposomes / Hodenius, M. A. J. ; Schmitz-Rode, T. ; Baumann, M. ; Ivoanova, G. ; Wong, J. E. ; Mang, T. ; Haulena, F. ; Soenen, S. J. H. ; De Cuyper, M.}, series = {Colloids and Surfaces A: Physicochemical and Engineering Aspects. 343 (2009), H. 1-3}, journal = {Colloids and Surfaces A: Physicochemical and Engineering Aspects. 343 (2009), H. 1-3}, publisher = {Elsevier}, address = {Amsterdam}, isbn = {0927-7757}, pages = {20 -- 23}, year = {2009}, language = {en} } @article{BalakrishnanAndreiSelmerSelmeretal.2010, author = {Balakrishnan, Karthikeyan and Andrei-Selmer, Luminita-Cornelia and Selmer, Thorsten and Bacher, Michael and Dodel, Richard}, title = {Comparison of intravenous immunoglobulins for naturally occurring autoantibodies against amyloid-β}, series = {Journal of Alzheimer's Disease}, volume = {20}, journal = {Journal of Alzheimer's Disease}, number = {1}, publisher = {IOS Press}, address = {Amsterdam}, issn = {1387-2877}, doi = {10.3233/JAD-2010-1353}, pages = {135 -- 143}, year = {2010}, abstract = {Intravenous immunoglobulins (IVIG) are currently used for therapeutic purposes in autoimmune disorders. Recently, we demonstrated the presence of naturally occurring antibodies against amyloid- β (nAbs-Aβ) within the pool of IVIG. In this study, we compared different brands of IVIG for nAbs-Aβ and have found differences in the specificity of the nAbs-Aβ towards Aβ1-40 and Aβ1-42 . We analyzed the influence of a pH-shift over the course of antibody storage using ELISA and investigated antibody dimerization at acidic and neutral pH as well as differences in the IgG subclass distributions among the IVIG using both HPLC and a nephelometric assay. Furthermore, we investigated the epitope region of purified nAbs-Aβ. The differences found in Aβ specificity are not directly proportionate to the binding nature of these antibodies when administered in vivo. This information, however, may serve as a guide when choosing the commercial source of IVIG for therapeutic applications in Alzheimer's disease}, language = {en} } @misc{EngelBayerUlberetal.2018, author = {Engel, Mareike and Bayer, Hendrik and Ulber, Roland and Tippk{\"o}tter, Nils}, title = {Kommt es in Elektrofermentationen mit Clostridium acetobutylicum zu einer Eisenlimitierung?}, series = {Chemie Ingenieur Technik}, volume = {90}, journal = {Chemie Ingenieur Technik}, number = {9}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {0009-286X}, doi = {10.1002/cite.201855047}, pages = {1154}, year = {2018}, abstract = {Das strikt anaerobe Bakterium Clostridium acetobutylicum bildet die L{\"o}semittel Aceton, Butanol und Ethanol (ABE-Fermentation). Im Fall einer Eisenlimitierung kommt es zus{\"a}tzlich zu einer Riboflavinsekretion (RF), was durch die gelbe F{\"a}rbung des Kultur{\"u}berstands erkennbar ist. In dieser Arbeit wurde beobachtet, dass w{\"a}hrend Elektrofermentationen mit C. acetobutylicum bei -600 mV eine gelbe F{\"a}rbung auftritt. Es wurde deshalb untersucht, ob eine Eisenlimitierung im bio-elektrochemischen System (BES) vorliegt. Hierzu wurden die Flavinspezien bei Kultivierungen in Medien mit einer Eisenlimitierung bzw. mit ausreichend Eisen in Serumflaschen sowie im BES mit und ohne angelegtem Potenzial verglichen. In den Serumflaschenversuchen wurden RF und Flavinadenindinukleotid (FAD)-Konzentrationen von ‡ 20 mg L⁻¹ sowie Flavinmononukleotid (FMN)-Konzentrationen von ca. 5 mg L⁻¹ detektiert. Bei ausreichender Eisenverf{\"u}gbarkeit hingegen wurden in den Serumflaschen fast keine Flavine sekretiert. Im BES bei -600 mV hingegen wurde auch in diesen Kultur{\"u}berst{\"a}nden FMN und FAD (1-5 mg L⁻¹ ),jedoch kein RF gemessen. Diese Ergebnisse zeigen, dass die Flavinbildung im BES mit angelegtem Potenzial nicht mit der Flavinbildung unter Eisenlimitierung in Serumflaschen korreliert. Andere Faktoren f{\"u}r eine m{\"o}gliche Flavinbildung werden aktuell n{\"a}her betrachtet.}, language = {de} } @article{TixMollKrafftetal.2024, author = {Tix, Julian and Moll, Fabian and Krafft, Simone and Betsch, Matthias and Tippk{\"o}tter, Nils}, title = {Hydrogen production from enzymatic pretreated organic waste with thermotoga neapolitana}, series = {Energies}, volume = {17}, journal = {Energies}, number = {12}, publisher = {MDPI}, address = {Basel}, issn = {1996-1073}, doi = {10.3390/en17122938}, pages = {20 Seiten}, year = {2024}, abstract = {Biomass from various types of organic waste was tested for possible use in hydrogen production. The composition consisted of lignified samples, green waste, and kitchen scraps such as fruit and vegetable peels and leftover food. For this purpose, the enzymatic pretreatment of organic waste with a combination of five different hydrolytic enzymes (cellulase, amylase, glucoamylase, pectinase and xylase) was investigated to determine its ability to produce hydrogen (H2) with the hydrolyzate produced here. In course, the anaerobic rod-shaped bacterium T. neapolitana was used for H2 production. First, the enzymes were investigated using different substrates in preliminary experiments. Subsequently, hydrolyses were carried out using different types of organic waste. In the hydrolysis carried out here for 48 h, an increase in glucose concentration of 481\% was measured for waste loads containing starch, corresponding to a glucose concentration at the end of hydrolysis of 7.5 g·L-1. In the subsequent set fermentation in serum bottles, a H2 yield of 1.26 mmol H2 was obtained in the overhead space when Terrific Broth Medium with glucose and yeast extract (TBGY medium) was used. When hydrolyzed organic waste was used, even a H2 yield of 1.37 mmol could be achieved in the overhead space. In addition, a dedicated reactor system for the anaerobic fermentation of T. neapolitana to produce H2 was developed. The bioreactor developed here can ferment anaerobically with a very low loss of produced gas. Here, after 24 h, a hydrogen concentration of 83\% could be measured in the overhead space.}, language = {en} } @article{VarrialeHengsbachGuoetal.2024, author = {Varriale, Ludovica and Hengsbach, Jan-Niklas and Guo, Tianyi and Kuka, Katrin and Tippk{\"o}tter, Nils and Ulber, Roland}, title = {Sustainable production of lactic acid using a perennial ryegrass as feedstock—a comparative study of fermentation at the bench- and reactor-scale, and ensiling}, series = {Sustainability}, volume = {16}, journal = {Sustainability}, number = {18}, publisher = {MDPI}, address = {Basel}, issn = {2071-1050}, doi = {10.3390/su16188054}, year = {2024}, abstract = {Perennial ryegrass (Lolium perenne) is an underutilized lignocellulosic biomass that has several benefits such as high availability, renewability, and biomass yield. The grass press-juice obtained from the mechanical pretreatment can be used for the bio-based production of chemicals. Lactic acid is a platform chemical that has attracted consideration due to its broad area of applications. For this reason, the more sustainable production of lactic acid is expected to increase. In this work, lactic acid was produced using complex medium at the bench- and reactor scale, and the results were compared to those obtained using an optimized press-juice medium. Bench-scale fermentations were carried out in a pH-control system and lactic acid production reached approximately 21.84 ± 0.95 g/L in complex medium, and 26.61 ± 1.2 g/L in press-juice medium. In the bioreactor, the production yield was 0.91 ± 0.07 g/g, corresponding to a 1.4-fold increase with respect to the complex medium with fructose. As a comparison to the traditional ensiling process, the ensiling of whole grass fractions of different varieties harvested in summer and autumn was performed. Ensiling showed variations in lactic acid yields, with a yield up to 15.2\% dry mass for the late-harvested samples, surpassing typical silage yields of 6-10\% dry mass.}, language = {en} } @article{UndenBeckerBongaertsetal.1995, author = {Unden, Gottfried and Becker, S. and Bongaerts, Johannes and Holighaus, G. and Schirawski, Jan and Six, Simon}, title = {O2-sensing and O2-dependent gene regulation in facultatively anaerobic bacteria}, series = {Archives of microbiology}, volume = {Vol. 164}, journal = {Archives of microbiology}, number = {Iss. 2}, issn = {1432-072X (E-Journal); 0003-9276 (Print); 0302-8933 (Print)}, pages = {81 -- 90}, year = {1995}, language = {en} } @article{UndenBeckerBongaertsetal.1994, author = {Unden, Gottfried and Becker, S. and Bongaerts, Johannes and Schirawski, Jan and Six, Simon}, title = {Oxygen regulated gene expression in facultatively anaerobic bacteria}, series = {Antonie van Leeuwenhoek}, volume = {Vol. 66}, journal = {Antonie van Leeuwenhoek}, number = {Iss. 1-3}, issn = {0003-6072 (Print) ; 1572-9699 (online)}, pages = {3 -- 22}, year = {1994}, language = {en} } @article{ElbersPfefferFrieseletal.1993, author = {Elbers, Gereon and Pfeffer, H.-U. and Friesel, J. and Beier, Reinhold}, title = {Pilotuntersuchungen gem{\"a}ß dem Verordnungsentwurf nach \S 40, Abs. 2 des Bundesimmissionsschutzgesetzes / H.-U. Pfeffer, J. Friesel, G. Elbers, R. Beier, K. Ellermann}, series = {Drei Jahrzehnte Luftqualit{\"a}ts{\"u}berwachung : Vortr{\"a}ge und Berichte zum Kolloquium "Drei Jahrzehnte Luftqualit{\"a}ts{\"u}berwachung - Bilanz und Perspektiven" am 11. November 1993 in Essen / [Hrsg.: Landesanstalt f{\"u}r Immissionsschutz Nordrhein-Westfalen. Red. Bearb.: Robert Niesenhaus]}, journal = {Drei Jahrzehnte Luftqualit{\"a}ts{\"u}berwachung : Vortr{\"a}ge und Berichte zum Kolloquium "Drei Jahrzehnte Luftqualit{\"a}ts{\"u}berwachung - Bilanz und Perspektiven" am 11. November 1993 in Essen / [Hrsg.: Landesanstalt f{\"u}r Immissionsschutz Nordrhein-Westfalen. Red. Bearb.: Robert Niesenhaus]}, address = {Essen}, pages = {117}, year = {1993}, language = {de} } @article{RibitschHeumannKarletal.2012, author = {Ribitsch, Doris and Heumann, Sonja and Karl, Wolfgang and Gerlach, Jochen and Leber, R. and Birner-Gruenberger, R. and Gruber, K. and Eiteljoerg, I. and Remler, Peter and Siegert, Petra and Lange, J. and Maurer, Karl-Heinz and Berg, G. and Guebitz, G. M. and Schwab, H.}, title = {Extracellular serine proteases from Stenotrophomonas maltophilia: Screening, isolation and heterologous expression in E. coli}, series = {Journal of biotechnology}, volume = {157}, journal = {Journal of biotechnology}, number = {1}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1873-4863 (E-Journal); 0168-1656 (Print)}, doi = {10.1016/j.jbiotec.2011.09.025}, pages = {140 -- 147}, year = {2012}, abstract = {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.}, language = {en} } @article{RibitschKarlBirnerGruenbergeretal.2010, author = {Ribitsch, Doris and Karl, Wolfgang and Birner-Gruenberger, R. and Gruber, K. and Eiteljoerg, I. and Remler, Peter and Wieland, S. and Siegert, Petra and Maurer, Karl-Heinz and Schwab, H.}, title = {C-terminal truncation of a metagenome-derived detergent protease for effective expression in E. coli}, series = {Journal of biotechnology}, volume = {150}, journal = {Journal of biotechnology}, number = {3}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1873-4863 (E-Journal); 0168-1656 (Print)}, doi = {10.1016/j.jbiotec.2010.09.947}, pages = {408 -- 416}, year = {2010}, abstract = {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.}, language = {en} } @misc{BergBesslerGerlachetal.2009, author = {Berg, Gabriele and Bessler, Cornelius and Gerlach, Jochen and G{\"u}bitz, Georg and Heumann, Sonja and Karl, Wolfgang and Maurer, Karl-Heinz and Remler, Peter and Ribitsch, Doris and Schwab, Helmut and Siegert, Petra and Wieland, Susanne}, title = {Mittel enthaltend Proteasen aus Stenotrophomonas maltophilia [Offenlegungsschrift]}, publisher = {Deutsches Patent- und Markenamt / Europ{\"a}isches Patentamt / USPTO / WIPO}, address = {M{\"u}nchen / Den Hague / Washington / Genf}, pages = {1 -- 47}, year = {2009}, language = {de} }