TY - JOUR A1 - Muschallik, Lukas A1 - Molinnus, Denise A1 - Bongaerts, Johannes A1 - Pohl, Martina A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Siegert, Petra A1 - Selmer, Thorsten T1 - (R,R)-Butane-2,3-diol Dehydrogenase from Bacillus clausii DSM 8716T: Cloning and Expression of the bdhA-Gene, and Initial Characterization of Enzyme JF - Journal of Biotechnology N2 - The gene encoding a putative (R,R)-butane-2,3-diol dehydrogenase (bdhA) from Bacillus clausii DSM 8716T was isolated, sequenced and expressed in Escherichia coli. The amino acid sequence of the encoded protein is only distantly related to previously studied enzymes (identity 33–43%) and exhibited some uncharted peculiarities. An N-terminally StrepII-tagged enzyme variant was purified and initially characterized. The isolated enzyme catalyzed the (R)-specific oxidation of (R,R)- and meso-butane-2,3-diol to (R)- and (S)-acetoin with specific activities of 12 U/mg and 23 U/mg, respectively. Likewise, racemic acetoin was reduced with a specific activity of up to 115 U/mg yielding a mixture of (R,R)- and meso-butane-2,3-diol, while the enzyme reduced butane-2,3-dione (Vmax 74 U/mg) solely to (R,R)-butane-2,3-diol via (R)-acetoin. For these reactions only activity with the co-substrates NADH/NAD+ was observed. The enzyme accepted a selection of vicinal diketones, α-hydroxy ketones and vicinal diols as alternative substrates. Although the physiological function of the enzyme in B. clausii remains elusive, the data presented herein clearly demonstrates that the encoded enzyme is a genuine (R,R)-butane-2,3-diol dehydrogenase with potential for applications in biocatalysis and sensor development. Y1 - 2017 U6 - http://dx.doi.org/10.1016/j.jbiotec.2017.07.020 SN - 0168-1656 VL - 258 SP - 41 EP - 50 PB - Elsevier CY - Amsterdam ER - TY - PAT A1 - Banowski, Bernhard A1 - Waldmann-Laue, Marianne A1 - Wadle, Armin A1 - Siegert, Petra A1 - Sättler, Andreas T1 - 5-Lipoxigenase-Inhibitoren in Deodorantien und Antitranspirantien [Offenlegungsschrift] T1 - 5-Lipooxygenase inhibitors in deodorants and antiperspirants [Europäische Patentanmeldung] Y1 - 2004 SP - 1 EP - 12 PB - Deutsches Patent- und Markenamt / Europäisches Patentamt CY - München / Den Hague ER - TY - JOUR A1 - Haeger, Gerrit A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - A convenient ninhydrin assay in 96-well format for amino acid-releasing enzymes using an air-stable reagent JF - Analytical Biochemistry N2 - An improved and convenient ninhydrin assay for aminoacylase activity measurements was developed using the commercial EZ Nin™ reagent. Alternative reagents from literature were also evaluated and compared. The addition of DMSO to the reagent enhanced the solubility of Ruhemann's purple (RP). Furthermore, we found that the use of a basic, aqueous buffer enhances stability of RP. An acidic protocol for the quantification of lysine was developed by addition of glacial acetic acid. The assay allows for parallel processing in a 96-well format with measurements microtiter plates. Y1 - 2022 U6 - http://dx.doi.org/10.1016/j.ab.2022.114819 SN - 1096-0309 IS - 624 PB - Elsevier CY - Amsterdam ER - TY - RPRT A1 - Haeger, Gerrit A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Abschlussbericht Teil II: Eingehende Darstellung Neue biobasierte Lipopeptide aus nachhaltiger Produktion (LipoPep) Y1 - 2023 N1 - Förderkennzeichen: 13FH256PA6 Titel: FHprofUnt 2016: Neue biobasierte Lipopeptide aus nachhaltiger Produktion Laufzeit: 01.02.2019 – 31.10.2022 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 - PAT A1 - O'Connell, Timothy A1 - Hoven, Nina A1 - Siegert, Petra A1 - Maurer, Karl-Heinz T1 - Amadoriasen in Wasch- und Reinigungsmitteln [Offenlegungsschrift] T1 - Amadoriases in washing and cleaning products [Europäische Patentanmeldung / Internationale Patentanmeldung] Y1 - 2007 SP - 1 EP - 45 PB - Deutsches Patent- und Markenamt / Europäisches Patentamt / WIPO CY - München / Den Hague / Genf ER - TY - JOUR A1 - Brahma, Aischarya A1 - Musio, Biagia A1 - Ismayilova, Uliviya A1 - Nikbin, Nikzad A1 - Kamptmann, Sonja B. A1 - Siegert, Petra A1 - Jeromin, Günter Erich A1 - Ley, Steven A1 - Pohl, Martina T1 - An orthogonal biocatalytic approach for the safe generation and use of HCN in a multi-step continuous preparation of chiral O-acetylcyanohydrins JF - Synlett Y1 - 2015 U6 - http://dx.doi.org/10.1055/s-0035-1560644 SN - 0936-5214 (Print) ; 1437-2096 (e-Journal) IS - Publ. online 29.09.2015 PB - Thieme CY - Stuttgart 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 - PAT A1 - Banowski, Bernhard A1 - Wadle, Armin A1 - Siegert, Petra T1 - Arylsulfatase-Inhibitoren in Deodorantien und Antitranspirantien [Offenlegungsschrift] T1 - Arylsulfatase inhibitors in deodorants and antiperspirants [Europäische Patentanmeldung] Y1 - 2004 SP - 1 EP - 15 PB - Deutsches Patent- und Markenamt / Europäisches Patentamt CY - München / Den Hague ER - TY - PAT A1 - Banowski, Bernhard A1 - Wadle, Armin A1 - Siegert, Petra T1 - Arylsulfatase-Inhibitoren in Deodorantien und Antitranspirantien [Offenlegungsschrift] T1 - Reducing body odor, e.g. in men, comprises applying deodorant or antiperspirant composition containing arylsulfatase-inhibiting phenoxyphenol derivative to skin Y1 - 2003 SP - 1 EP - 20 PB - Deutsches Patent- und Markenamt CY - München ER - TY - PAT A1 - Banowski, Bernhard A1 - Hoffmann, Daniele A1 - Wadle, Armin A1 - Siegert, Petra A1 - Sättler, Andrea A1 - Gerke, Thomas T1 - Arylsulfatase-Inhibitoren in Deodorantien und Antitranspirantien [Offenlegungsschrift] T1 - Arylsufatas-inhibitors in deodorants and antiperspirants[Europäische Patentschrift / Österreichische Patentanmeldung / Polnische Patentanmeldung / Internationale Patentanmeldung] Y1 - 2002 SP - 1 EP - 22 PB - Deutsches Patent- und Markenamt / Polish Patent Office / WIPO CY - München / Warsaw / Genf 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 - PAT A1 - Bankowski, Bernhard A1 - Hoffmann, Daniele A1 - Wadle, Armin A1 - Siegert, Petra A1 - Sättler, Andrea A1 - Gerke, Thomas T1 - Beta-Glucuronidase-Inhibitoren in Deodorantien und Antitranspirantien [Offenlegungsschrift] T1 - Beta-Glucuronidase inhibitors for use in deodorants and antiperspirants [Europäische Patentanmeldung / Internationale Patentanmeldung] Y1 - 2003 SP - 1 EP - 24 PB - Deutsches Patent- und Markenamt / Europäisches Patentamt / WIPO CY - München / Den Hague / Genf 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 - 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 - 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 - Welden, Melanie A1 - Severins, Robin A1 - Poghossian, Arshak A1 - Wege, Christina A1 - Bongaerts, Johannes A1 - Siegert, Petra A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Detection of acetoin and diacetyl by a tobacco mosaic virus-assisted field-effect biosensor JF - Chemosensors N2 - Acetoin and diacetyl have a major impact on the flavor of alcoholic beverages such as wine or beer. Therefore, their measurement is important during the fermentation process. Until now, gas chromatographic techniques have typically been applied; however, these require expensive laboratory equipment and trained staff, and do not allow for online monitoring. In this work, a capacitive electrolyte–insulator–semiconductor sensor modified with tobacco mosaic virus (TMV) particles as enzyme nanocarriers for the detection of acetoin and diacetyl is presented. The enzyme acetoin reductase from Alkalihalobacillus clausii DSM 8716ᵀ is immobilized via biotin–streptavidin affinity, binding to the surface of the TMV particles. The TMV-assisted biosensor is electrochemically characterized by means of leakage–current, capacitance–voltage, and constant capacitance measurements. In this paper, the novel biosensor is studied regarding its sensitivity and long-term stability in buffer solution. Moreover, the TMV-assisted capacitive field-effect sensor is applied for the detection of diacetyl for the first time. The measurement of acetoin and diacetyl with the same sensor setup is demonstrated. Finally, the successive detection of acetoin and diacetyl in buffer and in diluted beer is studied by tuning the sensitivity of the biosensor using the pH value of the measurement solution. Y1 - 2022 U6 - http://dx.doi.org/10.3390/chemosensors10060218 SN - 2227-9040 N1 - This article belongs to the Special Issue "Nanostructured Devices for Biochemical Sensing" VL - 10 IS - 6 PB - MDPI CY - Basel ER - TY - JOUR A1 - Molinnus, Denise A1 - Bäcker, Matthias A1 - Siegert, Petra A1 - Willenberg, H. A1 - Poghossian, Arshak A1 - Keusgen, M. A1 - Schöning, Michael Josef T1 - Detection of Adrenaline Based on Substrate Recycling Amplification JF - Procedia Engineering N2 - An amperometric enzyme biosensor has been applied for the detection of adrenaline. The adrenaline biosensor has been prepared by modification of an oxygen electrode with the enzyme laccase that operates at a broad pH range between pH 3.5 to pH 8. The enzyme molecules were immobilized via cross-linking with glutaraldehyde. The sensitivity of the developed adrenaline biosensor in different pH buffer solutions has been studied. Y1 - 2015 U6 - http://dx.doi.org/10.1016/j.proeng.2015.08.708 SN - 1877-7058 N1 - Eurosensors 2015 VL - 120 SP - 540 EP - 543 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Molinnus, Denise A1 - Hardt, Gabriel A1 - Siegert, Petra A1 - Willenberg, Holger S. A1 - Poghossian, Arshak A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Detection of Adrenaline in Blood Plasma as Biomarker for Adrenal Venous Sampling JF - Electroanalysis N2 - An amperometric bi-enzyme biosensor based on substrate recycling principle for the amplification of the sensor signal has been developed for the detection of adrenaline in blood. Adrenaline can be used as biomarker verifying successful adrenal venous sampling procedure. The adrenaline biosensor has been realized via modification of a galvanic oxygen sensor with a bi-enzyme membrane combining a genetically modified laccase and a pyrroloquinoline quinone-dependent glucose dehydrogenase. The measurement conditions such as pH value and temperature were optimized to enhance the sensor performance. A high sensitivity and a low detection limit of about 0.5–1 nM adrenaline have been achieved in phosphate buffer at pH 7.4, relevant for measurements in blood samples. The sensitivity of the biosensor to other catecholamines such as noradrenaline, dopamine and dobutamine has been studied. Finally, the sensor has been successfully applied for the detection of adrenaline in human blood plasma. Y1 - 2018 U6 - http://dx.doi.org/10.1002/elan.201800026 SN - 1521-4109 VL - 30 IS - 5 SP - 937 EP - 942 PB - Wiley-VCH CY - Weinheim ER -