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Cupriavidus necator H16 gains increasing attention in microbial research and biotechnological application due to its diverse metabolic features. Here we present a tightly controlled gene expression system for C. necator including the pBBR1-vector that contains hybrid promoters originating from C. necator native tolC-promoter in combination with a synthetic tetO-operator. The expression of the reporter gene from these plasmids relies on the addition of the exogenous inducer doxycycline (dc). The novel expression system offers a combination of advantageous features as; (i) high and dose-dependent recombinant protein production, (ii) tight control with a high dynamic range (On/Off ratio), which makes it applicable for harmful pathways or for toxic protein production, (iii) comparable cheap inducer (doxycycline, dc), (iv) effective at low inducer concentration, that makes it useful for large scale application, (v) rapid, diffusion controlled induction, and (vi) the inducer does not interfere within the cell metabolism. As applications of the expression system in C. necator H16, the growth ability on glycerol was enhanced by constitutively expressing the E. coli glpk gene-encoding for glycerol kinase. Likewise, we used the system to overcome the expression toxicity of mevalonate pathway in C. necator H16. With this system, the mevalonate-genes were successfully introduced in the host and the recombinant strains could produce about 200 mg/l mevalonate.
Explorer CEOs: The effect of CEO career variety on large firms’ relative exploration orientation
(2018)
Prior studies demonstrate that firms need to make smart trade-off decisions between exploration and exploitation activities in order to increase performance. Chief executive officers (CEOs) are principal decision makers of a firm’s strategic posture. In this study, we theorize and empirically examine how relative exploration orientation of large publicly listed firms varies based on the career variety of their CEOs – that is, how diverse the professional experiences of executives were prior to them becoming CEOs. We further argue that the heterogeneity and structure of the top management team moderates the impact of CEO career variety on firms’ relative exploration orientation. Based on multisource secondary data for 318 S&P 500 firms from 2005 to 2015, we find that CEO career variety is positively associated with relative exploration orientation.
Interestingly, CEOs with high career varieties appear to be less effective in pursuing exploration, when they work with highly heterogeneous and structurally interdependent top management teams.
Against the background of growing data in everyday life, data processing tools become more powerful to deal with the increasing complexity in building design. The architectural planning process is offered a variety of new instruments to design, plan and communicate planning decisions. Ideally the access to information serves to secure and document the quality of the building and in the worst case, the increased data absorbs time by collection and processing without any benefit for the building and its user. Process models can illustrate the impact of information on the design- and planning process so that architect and planner can steer the process. This paper provides historic and contemporary models to visualize the architectural planning process and introduces means to describe today’s situation consisting of stakeholders, events and instruments. It explains conceptions during Renaissance in contrast to models used in the second half of the 20th century. Contemporary models are discussed regarding their value against the background of increasing computation in the building process.
Recent analysis of scientific data from Cassini and earth-based observations gave evidence for a global ocean under a surrounding solid ice shell on Saturn's moon Enceladus. Images of Enceladus' South Pole showed several fissures in the ice shell with plumes constantly exhausting frozen water particles, building up the E-Ring, one of the outer rings of Saturn. In this southern region of Enceladus, the ice shell is considered to be as thin as 2 km, about an order of magnitude thinner than on the rest of the moon. Under the ice shell, there is a global ocean consisting of liquid water. Scientists are discussing different approaches the possibilities of taking samples of water, i.e. by melting through the ice using a melting probe. FH Aachen UAS developed a prototype of maneuverable melting probe which can navigate through the ice that has already been tested successfully in a terrestrial environment. This means no atmosphere and or ambient pressure, low ice temperatures of around 100 to 150K (near the South Pole) and a very low gravity of 0,114 m/s^2 or 1100 μg. Two of these influencing measures are about to be investigated at FH Aachen UAS in 2017, low ice temperature and low ambient pressure below the triple point of water. Low gravity cannot be easily simulated inside a large experiment chamber, though. Numerical simulations of the melting process at RWTH Aachen however are showing a gravity dependence of melting behavior. Considering this aspect, VIPER provides a link between large-scale experimental simulations at FH Aachen UAS and numerical simulations at RWTH Aachen. To analyze the melting process, about 90 seconds of experiment time in reduced gravity and low ambient pressure is provided by the REXUS rocket. In this time frame, the melting speed and contact force between ice and probes are measured, as well as heating power and a two-dimensional array of ice temperatures. Additionally, visual and infrared cameras are used to observe the melting process.
Suspension depletion approach for exemption of infected Solanum jasminoides cells from pospiviroids
(2018)
Despite numerous studies, viroid elimination from infected plants remains a very challenging task. This study introduces for the first time a novel ‘suspension depletion’ approach for exemption of Solanum jasminoides plants from viroids. The proposed method implies initial establishment of suspension cultures of the infected plant cells. The suspended cells were then physically treated (mild thermotherapy, 33 °C), which presumably delayed the replication of the viroid. The viroid concentration in the treated biomass was monitored weekly using pospiviroid-specific PCR. After 10–12 weeks of continuous treatment, a sufficient decrease in viroid concentration was observed such that the infection became undetectable by PCR. The treated single cells then gave rise to microcolonies on a solid culture medium and the obtained viroid-negative clones were further promoted to regenerate into viroid-free plants. Three years of accumulated experimental data suggests feasibility, broad applicability, and good efficacy of the proposed approach.
Nanotubular tobacco mosaic virus (TMV) particles and RNA-free lower-order coat protein (CP) aggregates have been employed as enzyme carriers in different diagnostic layouts and compared for their influence on biosensor performance. In the following, we describe a label-free electrochemical biosensor for improved glucose detection by use of TMV adapters and the enzyme glucose oxidase (GOD). A specific and efficient immobilization of streptavidin-conjugated GOD ([SA]-GOD) complexes on biotinylated TMV nanotubes or CP aggregates was achieved via bioaffinity binding. Glucose sensors with adsorptively immobilized [SA]-GOD, and with [SA]-GOD cross-linked with glutardialdehyde, respectively, were tested in parallel on the same sensor chip. Comparison of these sensors revealed that TMV adapters enhanced the amperometric glucose detection remarkably, conveying highest sensitivity, an extended linear detection range and fastest response times. These results underline a great potential of an integration of virus/biomolecule hybrids with electronic transducers for applications in biosensorics and biochips. Here, we describe the fabrication and use of amperometric sensor chips combining an array of circular Pt electrodes, their loading with GOD-modified TMV nanotubes (and other GOD immobilization methods), and the subsequent investigations of the sensor performance.
The presentation of enzymes on viral scaffolds has beneficial effects such as an increased enzyme loading and a prolonged reusability in comparison to conventional immobilization platforms. Here, we used modified tobacco mosaic virus (TMV) nanorods as enzyme carriers in penicillin G detection for the first time. Penicillinase enzymes were conjugated with streptavidin and coupled to TMV rods by use of a bifunctional biotin-linker. Penicillinase-decorated TMV particles were characterized extensively in halochromic dye-based biosensing. Acidometric analyte detection was performed with bromcresol purple as pH indicator and spectrophotometry. The TMV-assisted sensors exhibited increased enzyme loading and strongly improved reusability, and higher analysis rates compared to layouts without viral adapters. They extended the half-life of the sensors from 4 - 6 days to 5 weeks and thus allowed an at least 8-fold longer use of the sensors. Using a commercial budget-priced penicillinase preparation, a detection limit of 100 µM penicillin was obtained. Initial experiments also indicate that the system may be transferred to label-free detection layouts.
Physical interaction with small solar system bodies (SSSB) is the next step in planetary science, planetary in-situ resource utilization (ISRU), and planetary defense (PD). It requires a broader understanding of the surface properties of the target objects, with particular interest focused on those near Earth. Knowledge of composition, multi-scale surface structure, thermal response, and interior structure is required to design, validate and operate missions addressing these three fields. The current level of understanding is occasionally simplified into the phrase, ”If you’ve seen one asteroid, you’ve seen one asteroid”, meaning that the in-situ characterization of SSSBs has yet to cross the threshold towards a robust and stable scheme of classification. This would enable generic features in spacecraft design, particularly for ISRU and science missions. Currently, it is necessary to characterize any potential target object sufficiently by a dedicated pre-cursor mission to design the mission which then interacts with the object in a complex fashion. To open up strategic approaches, much broader in-depth characterization of potential target objects would be highly desirable. In SSSB science missions, MASCOT-like nano-landers and instrument carriers which integrate at the instrument level to their mothership have met interest. By its size, MASCOT is compatible with small interplanetary missions. The DLR-ESTEC Gossamer Roadmap Science Working Groups‘ studies identified Multiple Near-Earth asteroid (NEA) Rendezvous (MNR) as one of the space science missions only feasible with solar sail propulsion. The Solar Polar Orbiter (SPO) study showed the ability to access any inclination, theDisplaced-L1 (DL1) mission operates close to Earth, where objects of interest to PD and for ISRU reside. Other studies outline the unique capability of solar sails to provide access to all SSSB, at least within the orbit of Jupiter, and significant progress has been made to explore the performance envelope of near-term solar sails for MNR. However, it is difficult for
sailcraft to interact physically with a SSSB. We expand and extend the philosophy of the recently qualified DLR Gossamer solar sail deployment technology using efficient multiple sub-spacecraft integration to also include landers for one-way in-situ investigations and sample-return missions by synergetic integration and operation of sail and lander. The MASCOT design concept and its characteristic features have created an ideal counterpart for thisand has already been adapted to the needs of the AIM spacecraft, former part of the NASA-ESA AIDA missionDesigning the 69th International Astronautical Congress (IAC), Bremen, Germany, 1-5 October 2018. IAC-18-F1.2.3 Page 2 of 17 combined spacecraft for piggy-back launch accommodation enables low-cost massively parallel access to the NEA population.
Physical interaction with small solar system bodies (SSSB) is key for in-situ resource utilization (ISRU). The design of mining missions requires good understanding of SSSB properties, including composition, surface and interior structure, and thermal environment. But as the saying goes "If you've seen one asteroid, you've seen one Asteroid": Although some patterns may begin to appear, a stable and reliable scheme of SSSB classification still has to be evolved. Identified commonalities would enable generic ISRU technology and spacecraft design approaches with a high degree of re-use. Strategic approaches require much broader in-depth characterization of the SSSB populations of interest to the ISRU community. The DLR-ESTEC GOSSAMER Roadmap Science Working Groups identified target-flexible Multiple Near-Earth asteroid (NEA) Rendezvous (MNR) as one of the missions only feasible with solar sail propulsion, showed the ability to access any inclination and a wide range of heliocentric distances as well as continuous operation close to Earth's orbit where low delta-v objects reside.
A field-effect biosensor employing tobacco mosaic virus (TMV) particles as scaffolds for enzyme immobilization is presented. Nanotubular TMV scaffolds allow a dense immobilization of precisely positioned enzymes with retained activity. To demonstrate feasibility of this new strategy, a penicillin sensor has been developed by coupling a penicillinase with virus particles as a model system. The developed field-effect penicillin biosensor consists of an Al-p-Si-SiO₂-Ta₂O₅-TMV structure and has been electrochemically characterized in buffer solutions containing different concentrations of penicillin G. In addition, the morphology of the biosensor surface with virus particles was characterized by scanning electron microscopy and atomic force microscopy methods. The sensors possessed a high penicillin sensitivity of ~ 92 mV/dec in a nearly-linear range from 0.1 mM to 10 mM, and a low detection limit of about 50 µM. The long-term stability of the penicillin biosensor was periodically tested over a time period of about one year without any significant loss of sensitivity. The biosensor has also been successfully applied for penicillin detection in bovine milk samples.