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 - https://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 - Jossek, Ralf A1 - Bongaerts, Johannes A1 - Sprenger, Georg A. T1 - Characterization of a new feedback-resistant 3-deoxy-D-arabinoheptulosonate 7-phosphate synthase AroF of Escherichia coli JF - FEMS microbiology letters Y1 - 2001 SN - 1574-6968 VL - Vol. 202 IS - Iss. 1 SP - 145 EP - 148 ER - TY - JOUR A1 - Wilming, Anja A1 - Begemann, Jens A1 - Kuhne, Stefan A1 - Regestein, Lars A1 - Bongaerts, Johannes A1 - Evers, Stefan A1 - Maurer, Karl-Heinz A1 - Büchs, Jochen T1 - Metabolic studies of γ-polyglutamic acid production in Bacillus licheniformis by small-scale continuous cultivations JF - Biochemical engineering journal Y1 - 2013 SN - 1873-295X (E-Journal); 1369-703X (Print) VL - Vol. 73 SP - 29 EP - 37 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Seifarth, Volker A1 - Grosse, Joachim O. A1 - Grossmann, Matthias A1 - Janke, Heinz Peter A1 - Arndt, Patrick A1 - Koch, Sabine A1 - Epple, Matthias A1 - Artmann, Gerhard A1 - Temiz Artmann, Aysegül T1 - Mechanical induction of bi-directional orientation of primary porcine bladder smooth muscle cells in tubular fibrin-poly(vinylidene fluoride) scaffolds for ureteral and urethral repair using cyclic and focal balloon catheter stimulation JF - Journal of Biomaterials Applications Y1 - 2017 U6 - https://doi.org/10.1177/0885328217723178 SN - 1530-8022 VL - 32 IS - 3 SP - 321 EP - 330 PB - Sage CY - London ER - TY - JOUR A1 - Seifarth, Volker A1 - Goßmann, Matthias A1 - Grosse, J. O. A1 - Becker, C. A1 - Heschel, I. A1 - Artmann, Gerhard A1 - Temiz Artmann, Aysegül T1 - Development of a Bioreactor to Culture Tissue Engineered Ureters Based on the Application of Tubular OPTIMAIX 3D Scaffolds JF - Urologia Internationalis Y1 - 2015 U6 - https://doi.org/10.1159/000368419 SN - 0042-1138 VL - 2015 IS - 95 SP - 106 EP - 113 PB - Karger CY - Basel ER - TY - JOUR A1 - Scheele, Sandra A1 - Oertel, Dan A1 - Bongaerts, Johannes A1 - Evers, Stefan A1 - Hellmuth, Hendrik A1 - Maurer, Karl-Heinz A1 - Bott, Michael A1 - Freudl, Roland T1 - Secretory production of an FAD cofactor-containing cytosolic enzyme (sorbitol–xylitol oxidase from Streptomyces coelicolor) using the twin-arginine translocation (Tat) pathway of Corynebacterium glutamicum JF - Microbial biotechnology Y1 - 2013 SN - 1751-7915 SP - 202 EP - 206 PB - Wiley-Blackwell CY - Oxford 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 - https://doi.org/10.1016/j.bios.2018.05.023 VL - 115 SP - 1 EP - 6 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Schroeter, Rebecca A1 - Hoffmann, Tamara A1 - Voigt, Birgit A1 - Meyer, Hanna A1 - Bleisteiner, Monika A1 - Muntel, Jan A1 - Jürgen, Britta A1 - Albrecht, Dirk A1 - Becher, Dörte A1 - Lalk, Michael A1 - Evers, Stefan A1 - Bongaerts, Johannes A1 - Maurer, Karl-Heinz A1 - Putzer, Harald A1 - Hecker, Michael A1 - Schweder, Thomas A1 - Bremer, Erhard T1 - Stress responses of the industrial workhorse Bacillus licheniformis to osmotic challenges JF - PLoS ONE N2 - The Gram-positive endospore-forming bacterium Bacillus licheniformis can be found widely in nature and it is exploited in industrial processes for the manufacturing of antibiotics, specialty chemicals, and enzymes. Both in its varied natural habitats and in industrial settings, B. licheniformis cells will be exposed to increases in the external osmolarity, conditions that trigger water efflux, impair turgor, cause the cessation of growth, and negatively affect the productivity of cell factories in biotechnological processes. We have taken here both systems-wide and targeted physiological approaches to unravel the core of the osmostress responses of B. licheniformis. Cells were suddenly subjected to an osmotic upshift of considerable magnitude (with 1 M NaCl), and their transcriptional profile was then recorded in a time-resolved fashion on a genome-wide scale. A bioinformatics cluster analysis was used to group the osmotically up-regulated genes into categories that are functionally associated with the synthesis and import of osmostress-relieving compounds (compatible solutes), the SigB-controlled general stress response, and genes whose functional annotation suggests that salt stress triggers secondary oxidative stress responses in B. licheniformis. The data set focusing on the transcriptional profile of B. licheniformis was enriched by proteomics aimed at identifying those proteins that were accumulated by the cells through increased biosynthesis in response to osmotic stress. Furthermore, these global approaches were augmented by a set of experiments that addressed the synthesis of the compatible solutes proline and glycine betaine and assessed the growth-enhancing effects of various osmoprotectants. Combined, our data provide a blueprint of the cellular adjustment processes of B. licheniformis to both sudden and sustained osmotic stress. Y1 - 2014 U6 - https://doi.org/10.1371/journal.pone.0080956 SN - 1932-6203 VL - 8 IS - 11 PB - PLOS CY - San Francisco ER - TY - JOUR A1 - Voigt, Birgit A1 - Schroeter, Rebecca A1 - Jürgen, Britta A1 - Albrecht, Dirk A1 - Evers, Stefan A1 - Bongaerts, Johannes A1 - Maurer, Karl-Heinz A1 - Schweder, Thomas A1 - Hecker, Michael T1 - The response of Bacillus licheniformis to heat and ethanol stress and the role of the SigB regulon JF - Proteomics Y1 - 2013 SN - 1615-9861 (E-Journal); 1615-9853 (Print) VL - Vol. 13 IS - Iss. 14 SP - 2140 EP - 2146 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Schmitz, M. A1 - Hirsch, E. A1 - Bongaerts, Johannes A1 - Takors, Ralf T1 - Pulse experiments as a prerequisite for the quantification of in vivo enzyme kinetics in aromatic amino acid pathway of Eschericia coli JF - Biotechnology progress Y1 - 2002 SN - 1520-6033 (E-Journal); 8756-7938 (Print) VL - Vol. 18 IS - Iss. 5 SP - 935 EP - 941 ER - TY - JOUR A1 - Müller, Ulrike A1 - Bongaerts, Johannes A1 - Bovenberg, Roel A1 - Jossek, Ralf A1 - Krämer, Marco A1 - Linnemann, J. A1 - Müschen, S. A1 - Ritterbecks, S. A1 - Sprenger, G. A1 - Wubbolts, Marcel T1 - Metabolic engineering to produce fine chemicals in Escherichia coli JF - Mededelingen van de Faculteit Landbouwwetenschappen, Rijksuniversiteit Gent Y1 - 2001 SN - 0035-533x VL - 66 (3a) SP - 215 EP - 217 ER - TY - JOUR A1 - Handtke, Stefan A1 - Volland, Sonja A1 - Methling, Karen A1 - Albrecht, Dirk A1 - Becher, Dörte A1 - Nehls, Jenny A1 - Bongaerts, Johannes A1 - Maurer, Karl-Heinz A1 - Lalk, Michael A1 - Liesegang, Heiko A1 - Voigt, Birgit A1 - Daniel, Rolf A1 - Hecker, Michael T1 - Cell physiology of the biotechnological relevant bacterium Bacillus pumilus - An omics-based approach JF - Journal of Biotechnology N2 - Members of the species Bacillus pumilus get more and more in focus of the biotechnological industry as potential new production strains. Based on exoproteome analysis, B. pumilus strain Jo2, possessing a high secretion capability, was chosen for an omics-based investigation. The proteome and metabolome of B. pumilus cells growing either in minimal or complex medium was analyzed. In total, 1542 proteins were identified in growing B. pumilus cells, among them 1182 cytosolic proteins, 297 membrane and lipoproteins and 63 secreted proteins. This accounts for about 43% of the 3616 proteins encoded in the B. pumilus Jo2 genome sequence. By using GC–MS, IP-LC/MS and H NMR methods numerous metabolites were analyzed and assigned to reconstructed metabolic pathways. In the genome sequence a functional secretion system including the components of the Sec- and Tat-secretion machinery was found. Analysis of the exoproteome revealed secretion of about 70 proteins with predicted secretion signals. In addition, selected production-relevant genome features such as restriction modification systems and NRPS clusters of B. pumilus Jo2 are discussed. Y1 - 2014 U6 - https://doi.org/10.1016/j.jbiotec.2014.08.028 SN - 1873-4863 (E-Journal); 0168-1656 (Print) IS - 192(A) SP - 204 EP - 214 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Wiegand, Sandra A1 - Dietrich, Sascha A1 - Hertel, Robert A1 - Bongaerts, Johannes A1 - Evers, Stefan A1 - Volland, Sonja A1 - Daniel, Rolf A1 - Liesegang, Heiko T1 - RNA-Seq of Bacillus licheniformis: active regulatory RNA features expressed within a productive fermentation JF - BMC genomics Y1 - 2013 SN - 1471-2164 VL - Vol. 14 SP - 667 PB - BioMed Central CY - London 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 - https://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 - JOUR A1 - Tippkötter, Nils A1 - Roth, Jasmine T1 - Purified Butanol from Lignocellulose – Solvent‐Impregnated Resins for an Integrated Selective Removal JF - Chemie Ingenieur Technik N2 - In traditional microbial biobutanol production, the solvent must be recovered during fermentation process for a sufficient space-time yield. Thermal separation is not feasible due to the boiling point of n-butanol. As an integrated and selective solid-liquid separation alternative, solvent impregnated resins (SIRs) were applied. Two polymeric resins were evaluated and an extractant screening was conducted. Vacuum application with vapor collection in fixed-bed column as bioreactor bypass was successfully implemented as butanol desorption step. In course of further increasing process economics, fermentation with renewable lignocellulosic substrates was conducted using Clostridium acetobutylicum. Utilization of SIR was shown to be a potential strategy for solvent removal from fermentation broth, while application of a bypass column allows for product removal and recovery at once. KW - Biofuel KW - Biorefinery KW - Butanol KW - Clostridium acetobutylicum KW - Lignocellulose Y1 - 2020 U6 - https://doi.org/10.1002/cite.202000200 SN - 1522-2640 N1 - Corresponding author: Nils Tippkötter VL - 92 IS - 11 SP - 1741 EP - 1751 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Unden, Gottfried A1 - Bongaerts, Johannes T1 - Alternative respiratory pathways of Escherichia coli: energetics and transcriptional regulation in response to electron acceptors JF - Biochimica et biophysica acta (BBA) - Bioenergetics Y1 - 1997 SN - 1879-2650 (E-Journal); 0005-2728 (Print) VL - Vol. 1320 IS - Iss. 3 SP - 217 EP - 234 ER - TY - CHAP A1 - Hering, T. A1 - Ulber, Roland A1 - Tippkötter, Nils T1 - Development of a screening system for antimicrobial surfaces T2 - New frontiers of biotech-processes (Himmelfahrtstagung) : 02-04 May 2016, Rhein-Mosel-Halle, Koblenz/Germany Y1 - 2016 SP - 129 PB - DECHEMA CY - Frankfurt am Main ER - TY - CHAP A1 - Capitain, C. A1 - Hering, T. A1 - Tippkötter, Nils A1 - Ulber, Roland T1 - Enzymatic polymerization of lignin model compounds and solubilized lignin in an aqueous ethanol extract T2 - New frontiers of biotech-processes (Himmelfahrtstagung) : 02-04 May 2016, Rhein-Mosel-Halle, Koblenz/Germany Y1 - 2016 SP - 151 EP - 152 PB - DECHEMA CY - Frankfurt am Main ER - TY - CHAP A1 - Artmann, Gerhard A1 - Meruvu, Haritha A1 - Kizildag, Sefa A1 - Temiz Artmann, Aysegül ED - Artmann, Gerhard ED - Temiz Artmann, Aysegül ED - Zhubanova, Azhar A. ED - Digel, Ilya T1 - Functional Toxicology and Pharmacology Test of Cell Induced Mechanical Tensile Stress in 2D and 3D Tissue Cultures T2 - Biological, Physical and Technical Basics of Cell Engineering N2 - Mechanical forces/tensile stresses are critical determinants of cellular growth, differentiation and migration patterns in health and disease. The innovative “CellDrum technology” was designed for measuring mechanical tensile stress of cultured cell monolayers/thin tissue constructs routinely. These are cultivated on very thin silicone membranes in the so-called CellDrum. The cell layers adhere firmly to the membrane and thus transmit the cell forces generated. A CellDrum consists of a cylinder which is sealed from below with a 4 μm thick, biocompatible, functionalized silicone membrane. The weight of cell culture medium bulbs the membrane out downwards. Membrane indentation is measured. When cells contract due to drug action, membrane, cells and medium are lifted upwards. The induced indentation changes allow for lateral drug induced mechanical tension quantification of the micro-tissues. With hiPS-induced (human) Cardiomyocytes (CM) the CellDrum opens new perspectives of individualized cardiac drug testing. Here, monolayers of self-beating hiPS-CMs were grown in CellDrums. Rhythmic contractions of the hiPS-cells induce membrane up-and-down deflections. The recorded cycles allow for single beat amplitude, single beat duration, integration of the single beat amplitude over the beat time and frequency analysis. Dose effects of agonists and antagonists acting on Ca2+ channels were sensitively and highly reproducibly observed. Data were consistent with published reference data as far as they were available. The combination of the CellDrum technology with hiPS-Cardiomyocytes offers a fast, facile and precise system for pharmacological and toxicological studies. It allows new preclinical basic as well as applied research in pharmacolgy and toxicology. Y1 - 2018 SN - 978-981-10-7904-7 U6 - https://doi.org/10.1007/978-981-10-7904-7_7 SP - 157 EP - 192 PB - Springer CY - Singapore ER - TY - JOUR A1 - Tippkötter, Nils A1 - Al-Kaidy, Huschyar A1 - Wollny, Steffen A1 - Ulber, Roland T1 - Functionalized magnetizable particles for downstream processing in single-use systems JF - Chemie Ingenieur Technik N2 - Biotechnological downstream processing is usually an elaborate procedure, requiring a multitude of unit operations to isolate the target component. Besides the disadvantageous space-time yield, the risks of cross-contaminations and product loss grow fast with the complexity of the isolation procedure. A significant reduction of unit operations can be achieved by application of magnetic particles, especially if these are functionalized with affinity ligands. As magnetic susceptible materials are highly uncommon in biotechnological processes, target binding and selective separation of such particles from fermentation or reactions broths can be done in a single step. Since the magnetizable particles can be produced from iron salts and low priced polymers, a single-use implementation of these systems is highly conceivable. In this article, the principles of magnetizable particles, their synthesis and functionalization are explained. Furthermore, applications in the area of reaction engineering, microfluidics and downstream processing are discussed focusing on established single-use technologies and development potential. Y1 - 2013 U6 - https://doi.org/10.1002/cite.201200130 VL - 85 IS - 1-2: Special Issue: Single-Use Technology SP - 76 EP - 86 PB - Wiley CY - Weinheim ER -