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 - Molinnus, Denise A1 - Muschallik, Lukas A1 - Gonzalez, Laura Osorio A1 - Bongaerts, Johannes A1 - Wagner, Torsten A1 - Selmer, Thorsten A1 - Siegert, Petra A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Development and characterization of a field-effect biosensor for the detection of acetoin JF - Biosensors and Bioelectronics N2 - A capacitive electrolyte-insulator-semiconductor (EIS) field-effect biosensor for acetoin detection has been presented for the first time. The EIS sensor consists of a layer structure of Al/p-Si/SiO₂/Ta₂O₅/enzyme acetoin reductase. The enzyme, also referred to as butane-2,3-diol dehydrogenase from B. clausii DSM 8716T, has been recently characterized. The enzyme catalyzes the (R)-specific reduction of racemic acetoin to (R,R)- and meso-butane-2,3-diol, respectively. Two different enzyme immobilization strategies (cross-linking by using glutaraldehyde and adsorption) have been studied. Typical biosensor parameters such as optimal pH working range, sensitivity, hysteresis, linear concentration range and long-term stability have been examined by means of constant-capacitance (ConCap) mode measurements. Furthermore, preliminary experiments have been successfully carried out for the detection of acetoin in diluted white wine samples. Y1 - 2018 U6 - http://dx.doi.org/10.1016/j.bios.2018.05.023 VL - 115 SP - 1 EP - 6 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Miyamoto, Koichiro A1 - Seki, Kosuke A1 - Suto, Takeyuki A1 - Werner, Frederik A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Yoshinobu, Tatsuo T1 - Improved spatial resolution of the chemical imaging sensor with a hybrid illumination that suppresses lateral diffusion of photocarriers JF - Sensor and Actuators B: Chemical N2 - The chemical imaging sensor is a semiconductor-based chemical sensor capable of visualizing pH and ion distributions. The spatial resolution depends on the lateral diffusion of photocarriers generated by illumination of the semiconductor substrate. In this study, two types of optical setups, one based on a bundle of optical fibers and the other based on a binocular tube head, were developed to project a hybrid illumination of a modulated light beam and a ring-shaped constant illumination onto the sensor plate. An improved spatial resolution was realized by the ring-shaped constant illumination, which suppressed lateral diffusion of photocarriers by enhanced recombination due to the increased carrier concentration. Y1 - 2018 U6 - http://dx.doi.org/10.1016/j.snb.2018.07.016 SN - 0925-4005 VL - 273 SP - 1328 EP - 1333 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Miyamoto, Ko-ichiro A1 - Yu, Bing A1 - Isoda, Hiroko A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Yoshinobu, Tatsuo T1 - Visualization of the recovery process of defects in a cultured cell layer by chemical imaging sensor JF - Sensors and Actuators B: Chemical N2 - The chemical imaging sensor is a field-effect sensor which is able to visualize both the distribution of ions (in LAPS mode) and the distribution of impedance (in SPIM mode) in the sample. In this study, a novel cell assay is proposed, in which the chemical imaging sensor operated in SPIM mode is applied to monitor the recovery of defects in a cell layer brought into proximity of the sensing surface. A reduced impedance at a defect formed artificially in a cell layer was successfully visualized in a photocurrent image. The cell layer was cultured over two weeks, during which the temporal change of the photocurrent distribution corresponding to the recovery of the defect was observed. Y1 - 2016 U6 - http://dx.doi.org/10.1016/j.snb.2016.04.018 SN - 0925-4005 VL - 236 SP - 965 EP - 969 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Miyamoto, Ko-ichiro A1 - Yoshida, Midori A1 - Sakai, Taito A1 - Matsuzaka, Atsushi A1 - Wagner, Torsten A1 - Kanoh, Sanoh A1 - Yoshinobu, Tatsuo A1 - Schöning, Michael Josef T1 - Differential setup of light-addressable potentiometric sensor with an enzyme reactor in a flow channel JF - Japanese Journal of Applied Physics. 50 (2011) Y1 - 2011 SN - 0021-4922 SP - 04DL08-1 EP - 04DL08-5 PB - Japan Society of Applied Physics CY - Bristol ER - TY - JOUR A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Yoshinobu, Tatsuo A1 - Kanoh, Shin`ichiro A1 - Schöning, Michael Josef T1 - Phase-mode LAPS and its application to chemical imaging JF - Sensors and Actuators B: Chemical. 154 (2011), H. 1 Y1 - 2011 SN - 1873-3077 SP - 28 EP - 32 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Yoshinobu, Tatsuo A1 - Kanoh, Shin`ichiro A1 - Schöning, Michael Josef T1 - Phase-mode operation of FDM-LAPS JF - Sensor letters Y1 - 2011 SN - 1546-1971 VL - 9 IS - 2 SP - 691 EP - 694 PB - American Scientific Publishers CY - Stevenson Ranch, Calif. ER - TY - JOUR A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Mimura, Shuhei A1 - Kanoh, Shiníchiro A1 - Yoshinobu, Tatsuo A1 - Schöning, Michael Josef T1 - Constant-phase-mode operation of the light-addressable potentiometric sensor JF - Sensors and Actuators B: Chemical. 154 (2011), H. 2 Y1 - 2011 SN - 1873-3077 N1 - EUROSENSORS XXIII SP - 119 EP - 123 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Mimura, Shuhei A1 - Kanoh, Shin`ichiro A1 - Yoshinobu, Tatsuo A1 - Schöning, Michael Josef T1 - Constant-phase-mode operation of the light-addressable potentiometric sensor JF - Procedia Chemistry. 1 (2009), H. 1 Y1 - 2009 SN - 1876-6196 N1 - Proceedings of the Eurosensors XXIII conference ; Eurosensors 23 SP - 1487 EP - 1490 PB - Elsevier CY - Amsterdam ER - TY - CHAP A1 - Miyamoto, Ko-ichiro A1 - Suto, Takeyuki A1 - Werner, Frederik A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Yoshinobu, Tatsuo T1 - Restraining the Diffusion of Photocarriers to Improve the Spatial Resolution of the Chemical Imaging Sensor T2 - MDPI Proceedings Y1 - 2017 U6 - http://dx.doi.org/10.3390/proceedings1040477 N1 - Eurosensors 2017 Conference, Paris, France, 3–6 September 2017 VL - 1 IS - 4 ER -