Proton-Detected Solid-State NMR of the Cell-Free Synthesized α-Helical Transmembrane Protein NS4B from Hepatitis C Virus

Proton-detected 100 kHz magic-angle-spinning (MAS) solid-state NMR is an emerging analysis method for proteins with only hundreds of microgram quantities, and thus allows structural investigation of eukaryotic membrane proteins. This is the case for the cell-free synthesized hepatitis C virus (HCV) nonstructural membrane protein 4B (NS4B). We demonstrate NS4B sample optimization using fast reconstitution schemes that enable lipid-environment screening directly by NMR. 2D spectra and relaxation properties guide the choice of the best sample preparation to record 2D ¹H-detected ¹H,¹⁵N and 3D ¹H,¹³C,¹⁵N correlation experiments with linewidths and sensitivity suitable to initiate sequential assignments. Amino-acid-selectively labeled NS4B can be readily obtained using cell-free synthesis, opening the door to combinatorial labeling approaches which should enable structural studies.     

Cultivation and downstream processing of microalgae and cyanobacteria to generate protein-based technofunctional food ingredients

Abstract

Microalgae are unicellular microorganisms that can be rich in proteins and are therefore a valuable ingredient in different foods. So far microalgae are mainly utilized in foods in low concentrations as a whole-cell ingredient even though it is known that proteins extracted from microalgae can possibly posess various technofunctional properties, such as high protein solubility, emulsification, foaming, and gelation properties. The widespread usage of protein-rich ingredients obtained from microalgae is for the most part prevented by the high price of the biomass, the lack of efficient downstream processes, and the adverse taste. The aim of this review is to give insights into the fundamental properties of the growth and processing of microalgae, highlight the advantages of microalgae ingredients and show potential applications based on the technofunctional, nutritional and sensory properties that were reported. Moreover, the existing challenges and knowledge gaps that hinder the application of microalgal proteins in foods are discussed.     

Malic acid production from renewables: a review

Malic acid derived from fossil resources is currently applied in the food and beverage industries with a medium global production capacity. However, in the transition from a fossil-based to a bio-based economy, biotechnologically produced l -malic acid may become an important platform chemical with many new applications, especially in the field of biopolymers. In this review, currently used petrochemical production routes to dl -malic acid are outlined and insights into possible bio-based alternatives for microbial l -malic acid production are provided. Besides ecological reasons, the possibility to produce enantiopure l -malic acid by microbial fermentation is the biggest advantage over chemical synthesis. State-of-the-art and open challenges concerning production host engineering, substrate choice and downstream processing are addressed. With regard to production hosts, a literature overview is given covering the leading natural production strains of Aspergillus, Ustilago and Aureobasidium, as well as Escherichia coli as the most important engineered recombinant host. The utilization of renewable substrates as an alternative to glucose is emphasized in particular as a key aspect for a competitive bio-based production. Out of the alternative substrates discussed in this review, the industrial side-streams crude glycerol and molasses seem to be most promising for large-scale l -malic acid production. © 2019 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Atomistic simulations of grain boundary migration in copper

While the motion of twist boundaries can be readily studied by atomistic simulations with molecular dynamics (MD) under the action of an elastic driving force, the approach fails for tilt boundaries. This is due to the interaction of the elastic stress with the grain boundary (GB) structure, which causes plastic strain by GB sliding. A novel concept, the orientation correlated driving force, is introduced to circumvent this problem. It is shown that this concept can be successfully applied to the study of the migration of tilt boundaries. The migration behavior of several twist and tilt GBs was investigated. The transition from low-to high-angle boundaries can be captured, and a structural transition of tilt boundaries was found at high temperatures, which also affected the migration behavior. The results compare well with experimental results of the motion high-angle boundaries, but for low-angle boundaries, the agreement is poor. This article is based on a presentation made in the “Hillert Symposium on Thermodynamics & Kinetics of Migrating Interfaces in Steels and Other Complex Alloys,” December 2–3, 2004, organized by The Royal Institute of Technology in Stockholm, Sweden.     

Efficient Low Dissipative High Order Schemes for Multiscale MHD Flows, II: Minimization of ∇·B Numerical Error

An adaptive numerical dissipation control in a class of high order filter methods for compressible MHD equations is systematically discussed. The filter schemes consist of a divergence-free preserving high order spatial base scheme with a filter approach which can be divergence-free preserving depending on the type of filter operator being used, the method of applying the filter step, and the type of flow problem to be considered. Some of these filter variants provide a natural and efficient way for the minimization of the divergence of the magnetic field (∇·B) numerical error in the sense that commonly used divergence cleaning is not required. Numerical experiments presented emphasize the performance of the ∇·B numerical error. Many levels of grid refinement and detailed comparison of the filter methods with several commonly used compressible MHD shock-capturing schemes will be illustratedA condensed version appears in the Proceedings of the International Conference on High Performance Scientific Computing, March 10-14, 2003, Hanoi, Vietnam. This is a revised version of a longer internal report, Feb. 19, 2004. The longer internal report was published as a RIACS Technical Report TR03.10, July 2003, NASA Ames Research Center     

Sensor guided handling and assembly of active micro-systems

Positioning accuracies in the range of a few micrometers and below are necessary for the assembly of active micro-systems. In order to reach these accuracies, an assembly system for sensor guided micro-assembly is developed in the Collaborative Research Centre 516 “Design and manufacturing of active micro-systems”. The combination of a parallel robot with an integrated 3D vision sensor and a micro-gripper enables to reach relative positioning accuracies below 1 μm.     

Handover and Uplink Power Control Performance in the 3.84 Mcps TDD mode of UTRA Network

In this article we consider the performance of the 3.84 Mcpstime-division duplex (TDD) mode of UTRA (Universal TerrestrialRadio Access) network. We emphasize two of the radio resourcemanagement algorithms, handover and uplink power control, whoserole in the overall system performance is studied extensively.First, a handover algorithm used in WCDMA (Wideband Code DivisionMultiple Access) standard is considered in a TDD-mode operation.This gives rise to a careful setting of different handoverparameters, and the evaluation of the effects to the systemperformance. Secondly, the specified uplink power controlalgorithm is considered. Since it is based on several user-mademeasurements which may involve both random and systematic errors acareful study about the suitability of the power control scheme iscarried out.     

Untersuchung der Verteilung und Intensität filamentierter Barrierenplasmen für die Behandlung von Oberflächen

The application of barrier discharges at atmospheric pressure in air expands on the market of plasma technology, because it is an ecological and cost‐effective alternative to other processes of surface treatment. These plasmas usually consist of a multitude of spatially and temporally localized filaments, whose distribution should be as even as possible for homogeneous treatment. This holds especially for the plasma treatment of sensitive goods such as wool or other textiles. In equipment for continuous pass of material the barrier arrangements often consist of a system cylinder – cylinder or cylinder – plane, whereby the gap width changes locally. Space distribution and intensity of filaments has been investigated by means of short‐time photography and spatially resolved measurement of current distribution and energy distribution derived from it. The local dependency found can be explained by means of a capacitive equivalent circuit.     

Occurrence and characterisation of the hydrogen-evolving enzyme in Frankia sp.

An increase in hydrogen evolution from the hydrogen-evolving enzyme in the actinomycete Frankia was recorded in the presence of nickel. Immunogold localisation analysis of the intracellular distribution of hydrogenase proteins indicated that they were evenly distributed in the membranes and cytosol of both hyphae and vesicles. In addition, molecular characterisation of the hydrogen-evolving enzyme at the proteomic level, using two-dimensional gel electrophoresis combined with mass spectrometry, confirmed that the Frankia hydrogen-evolving enzyme is similar to the cyanobacterial bidirectional hydrogenase of Anabena siamensis.