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 - JOUR A1 - Selmer, Thorsten A1 - Yu, Lihua A1 - Blaser, Martin A1 - Andrei, Paula I. T1 - 4-Hydroxyphenylacetate decarboxylases: properties of a novel subclass of glycyl radical enzyme systems / Yu, L. ; Blaser, M. ; Andrei, PI. ; Pierik, AJ. Selmer, T. JF - Biochemistry. 31 (2006), H. 45 Y1 - 2006 N1 - PMID: 16878993 SP - 9584 EP - 9592 ER - TY - JOUR A1 - Schiffels, Johannes A1 - Selmer, Thorsten T1 - A flexible toolbox to study protein-assisted metalloenzyme assembly in vitro JF - Biotechnology and Bioengineering Y1 - 2015 U6 - http://dx.doi.org/10.1002/bit.25658 SN - 1097-0290 VL - 112 IS - 11 SP - 2360 EP - 2372 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Selmer, Thorsten A1 - Schmidt, Bernhard A1 - Ingendoh, Arnd A1 - Figura, Kurt von T1 - A novel amino acid modification in sulfatases that is defective in multiple sulfatase deficiency / Schmidt, Bernhard ; Selmer, Thorsten ; Ingendoh, Arnd ; Figurat, Kurt von JF - Cell. 82 (1995), H. 2 Y1 - 1995 SN - 0092-8674 SP - 271 EP - 278 ER - TY - JOUR A1 - Selmer, Thorsten A1 - Figura, Kurt von A1 - Schmidt, Bernhard A1 - Dierks, T. T1 - A novel protein modification generating an aldehyde group in sulfatases: its role in catalysis and disease / Figura, Kurt von ; Schmidt, Bernhard ; Selmer, Thorsten ; Dierks, Thomas JF - Bioessays. 20 (1998), H. 6 Y1 - 1998 SN - 1521-1878 SP - 505 EP - 510 ER - TY - JOUR A1 - Selmer, Thorsten A1 - Thamer, Wiebke A1 - Cirpus, Irina A1 - Hans, Marcus T1 - A two [4Fe-4S]-cluster-containing ferredoxin as an alternative electron donor for 2-hydroxyglutaryl-CoA dehydratase from Acidaminococcus fermentans / Thamer, Wiebke ; Cirpus, Irina ; Hans, Marcus ; Pierik, Antonio, J. ; Selmer, Thorsten ; Bill, Eckhard ; JF - Archives of Microbiology. 179 (2003), H. 3 Y1 - 2003 SN - 1432-072X SP - 197 EP - 204 ER - TY - JOUR A1 - Selmer, Thorsten A1 - Hetzel, Marc A1 - Brock, Matthias A1 - Pierik, Antonio J. T1 - Acryloyl-CoA reductase from Clostridium propionicum. An enzyme complex of propionyl-CoA dehydrogenase and electron-transferring flavoprotein / Hetzel, Marc ; Brock, Matthias ; Selmer, Thorsten, Pierik, Antonio J. ; Golding, Bernard T. ; Buckel, Wolfgang JF - European Journal of Biochemistry. 270 (2003), H. 5 Y1 - 2003 SN - 0014-2956 SP - 902 EP - 910 ER - TY - JOUR A1 - Selmer, Thorsten A1 - Brüser, Thomas A1 - Dahl, Christiane T1 - ADP Sulfurylase" from Thiobacillus denitrificans Is an Adenylylsulfate:Phosphate Adenylyltransferase and Belongs to a New Family of Nucleotidyltransferases / Brüser, Thomas ; Selmer, Thorsten ; Dahl, Christiane JF - Journal of Biological Chemistry. 275 (2000), H. 3 Y1 - 2000 SN - 1083-351X SP - 1691 EP - 1690 ER - TY - JOUR A1 - Schiffels, Johannes A1 - Pinkenburg, Olaf A1 - Schelden, Maximilian A1 - Aboulnaga, El-Hussiny A. A. A1 - Baumann, Marcus A1 - Selmer, Thorsten T1 - An innovative cloning platform enables large-scale production and maturation of an oxygen-tolerant [NiFe]-hydrogenase from cupriavidus necator in Escherichia coli JF - PLOS one. 2013 Y1 - 2013 U6 - http://dx.doi.org/10.1371/journal.pone.0068812 SN - 1932-6203 PB - Public Library of Science CY - San Francisco, California ER - TY - JOUR A1 - Pilas, Johanna A1 - Yazici, Y. A1 - Selmer, Thorsten A1 - Keusgen, M. A1 - Schöning, Michael Josef T1 - Application of a portable multi-analyte biosensor for organic acid determination in silage JF - Sensors N2 - Multi-analyte biosensors may offer the opportunity to perform cost-effective and rapid analysis with reduced sample volume, as compared to electrochemical biosensing of each analyte individually. This work describes the development of an enzyme-based biosensor system for multi-parametric determination of four different organic acids. The biosensor array comprises five working electrodes for simultaneous sensing of ethanol, formate, d-lactate, and l-lactate, and an integrated counter electrode. Storage stability of the biosensor was evaluated under different conditions (stored at +4 °C in buffer solution and dry at −21 °C, +4 °C, and room temperature) over a period of 140 days. After repeated and regular application, the individual sensing electrodes exhibited the best stability when stored at −21 °C. Furthermore, measurements in silage samples (maize and sugarcane silage) were conducted with the portable biosensor system. Comparison with a conventional photometric technique demonstrated successful employment for rapid monitoring of complex media. Y1 - 2018 U6 - http://dx.doi.org/10.3390/s18051470 SN - 1424-8220 VL - 18 IS - 5 PB - MDPI CY - Basel ER -