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 - JOUR A1 - Falkenberg, Fabian A1 - Kohn, Sophie A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Biochemical characterisation of a novel broad pH spectrum subtilisin from Fictibacillus arsenicus DSM 15822ᵀ JF - FEBS Open Bio N2 - Subtilisins from microbial sources, especially from the Bacillaceae family, are of particular interest for biotechnological applications and serve the currently growing enzyme market as efficient and novel biocatalysts. Biotechnological applications include use in detergents, cosmetics, leather processing, wastewater treatment and pharmaceuticals. To identify a possible candidate for the enzyme market, here we cloned the gene of the subtilisin SPFA from Fictibacillus arsenicus DSM 15822ᵀ (obtained through a data mining-based search) and expressed it in Bacillus subtilis DB104. After production and purification, the protease showed a molecular mass of 27.57 kDa and a pI of 5.8. SPFA displayed hydrolytic activity at a temperature optimum of 80 °C and a very broad pH optimum between 8.5 and 11.5, with high activity up to pH 12.5. SPFA displayed no NaCl dependence but a high NaCl tolerance, with decreasing activity up to concentrations of 5 m NaCl. The stability enhanced with increasing NaCl concentration. Based on its substrate preference for 10 synthetic peptide 4-nitroanilide substrates with three or four amino acids and its phylogenetic classification, SPFA can be assigned to the subgroup of true subtilisins. Moreover, SPFA exhibited high tolerance to 5% (w/v) SDS and 5% H₂O₂ (v/v). The biochemical properties of SPFA, especially its tolerance of remarkably high pH, SDS and H₂O₂, suggest it has potential for biotechnological applications. KW - Bacillaceae KW - Biotechnological application KW - Broad pH spectrum KW - Subtilases KW - Subtilisin Y1 - 2023 U6 - http://dx.doi.org/10.1002/2211-5463.13701 SN - 2211-5463 N1 - Corresponding author: Petra Siegert VL - 13 IS - 11 SP - 2035 EP - 2046 PB - Wiley CY - Hoboken, NJ ER - TY - JOUR A1 - Falkenberg, Fabian A1 - Rahba, Jade A1 - Fischer, David A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Biochemical characterization of a novel oxidatively stable, halotolerant, and high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101T JF - FEBS Open Bio N2 - Halophilic and halotolerant microorganisms represent a promising source of salt-tolerant enzymes suitable for various biotechnological applications where high salt concentrations would otherwise limit enzymatic activity. Considering the current growing enzyme market and the need for more efficient and new biocatalysts, the present study aimed at the characterization of a high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101T. The protease gene was cloned and expressed in Bacillus subtilis DB104. The recombinant protease SPAO with 269 amino acids belongs to the subfamily of high-alkaline subtilisins. The biochemical characteristics of purified SPAO were analyzed in comparison with subtilisin Carlsberg, Savinase, and BPN'. SPAO, a monomer with a molecular mass of 27.1 kDa, was active over a wide range of pH 6.0–12.0 and temperature 20–80 °C, optimally at pH 9.0–9.5 and 55 °C. The protease is highly oxidatively stable to hydrogen peroxide and retained 58% of residual activity when incubated at 10 °C with 5% (v/v) H2O2 for 1 h while stimulated at 1% (v/v) H2O2. Furthermore, SPAO was very stable and active at NaCl concentrations up to 5.0 m. This study demonstrates the potential of SPAO for biotechnological applications in the future. KW - Alkalihalobacillus okhensis KW - detergent protease KW - halotolerant protease KW - high-alkaline subtilisin KW - oxidative stable protease Y1 - 2022 U6 - http://dx.doi.org/10.1002/2211-5463.13457 SN - 2211-5463 N1 - Corresponding author: Petra Siegert VL - 12 IS - 10 SP - 1729 EP - 1746 PB - Wiley CY - Hoboken, NJ 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 - JOUR A1 - Jablonski, Melanie A1 - Münstermann, Felix A1 - Nork, Jasmina A1 - Molinnus, Denise A1 - Muschallik, Lukas A1 - Bongaerts, Johannes A1 - Wagner, Torsten A1 - Keusgen, Michael A1 - Siegert, Petra A1 - Schöning, Michael Josef T1 - Capacitive field‐effect biosensor applied for the detection of acetoin in alcoholic beverages and fermentation broths JF - physica status solidi (a) applications and materials science N2 - An acetoin biosensor based on a capacitive electrolyte–insulator–semiconductor (EIS) structure modified with the enzyme acetoin reductase, also known as butane-2,3-diol dehydrogenase (Bacillus clausii DSM 8716ᵀ), is applied for acetoin detection in beer, red wine, and fermentation broth samples for the first time. The EIS sensor consists of an Al/p-Si/SiO₂/Ta₂O₅ layer structure with immobilized acetoin reductase on top of the Ta₂O₅ transducer layer by means of crosslinking via glutaraldehyde. The unmodified and enzyme-modified sensors are electrochemically characterized by means of leakage current, capacitance–voltage, and constant capacitance methods, respectively. KW - acetoin KW - acetoin reductase KW - alcoholic beverages KW - biosensors KW - capacitive field-effect sensors Y1 - 2021 U6 - http://dx.doi.org/10.1002/pssa.202000765 SN - 1862-6319 VL - 218 IS - 13 PB - Wiley-VCH CY - Weinheim 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 -