Heterologous Expression of Various PHA Synthase Genes in Rhodospirillum rubrum

The phototrophic non-sulfur purple bacterium Rhodospirillum rubrum is known for its metabolic versatility. Particularly, R. rubrum is able to synthesize PHA under heterotrophic or even autotrophic growth with carbon monoxide as carbon and energy source. R. rubrum has therefore become a promising candidate for future cheap PHA production. However, R. rubrum synthesizes lower amounts of PHAs in comparison to well-known PHA producers like Ralstonia eutropha H16 or recombinant Escherichia coli strains. Since the PHA synthase is the key enzyme of PHA biosynthesis, genes encoding for twelve different PHA synthases were heterologously expressed in two generated phaC deletion mutants of R. rubrum in this study. To clearly see the effect of the foreign PHA synthases, PHA-negative mutants were required. The single mutant R. rubrum ΔphaC2 showed a PHA-leaky phenotype (< 1 % PHA, wt/wt, of CDW), while the double mutant R. rubrum ΔphaC1ΔphaC2 accumulated no measurable PHA. Eight different PHA synthase genes of class I, and four of class IV were chosen for heterologous expression. All recombinant R. rubrum strains showed significant PHA synthesis and accumulation, although PHA contents in the recombinant strains of the single mutant R. rubrum ΔphaC2 were generally higher in comparison to those of the double mutant R. rubrum ΔphaC1ΔphaC2. Recombinant strains of the single mutant could be divided into two groups according to the accumulation of PHA in the cells. While recombinant strains dedicated to group one showed an increased PHA synthesis when compared to the wild type carrying an empty vector, strains of group two accumulated less PHA than the wild type. Finally, it was possible to increase the accumulation of PHA by up to 25 % due to heterologous expression of PHA synthase genes compared to the wild type

Diffusion based degradation mechanisms in giant magnetoresistive spin valves

Spin valve systems based on the giant magnetoresistive (GMR) effect as used for example in hard disks and automotive applications consist of several functional metallic thin film layers. We have identified by secondary ion mass spectrometry (SIMS) two main degradation mechanisms: One is related to oxygen diffusion through a protective cap layer, and the other one is interdiffusion directly at the functional layers of the GMR stack. By choosing a suitable material as cap layer (TaN), the oxidation effect can be suppressed.Comment: 3 pages, 3 figures. to be published in Appl. Phys. Let

Omnivory and grazer functional composition moderate cascading trophic effects in experimental Fucus vesiculosus habitats

We tested the relative strength of direct versus indirect effects of an aquatic omnivore depending on the functional composition of grazers by manipulating the presence of gastropod and amphipod grazers and omnivorous shrimp in outdoor mesocosms. By selectively preying upon amphipods and reducing their abundance by 70–80%, omnivorous shrimp favoured the dominance of gastropods. While gastropods were the main microalgal grazers, amphipods controlled macroalgal biomass in the experiment. However, strong predation on the amphipod by the shrimp had no significant indirect effects on macroalgal biomass, indicating that when amphipod abundances declined, complementary feeding by the omnivore on macroalgae may have suppressed a trophic cascade. Accordingly, in the absence of amphipods, the shrimp grazed significantly on green algae and thereby suppressed the diversity of the macroalgal community. Our experiment demonstrates direct consumer effects by an omnivore on both the grazer and producer trophic levels in an aquatic food web, regulated by prey availability. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00227-010-1602-6) contains supplementary material, which is available to authorized users

A Closer Look on the Polyhydroxybutyrate- (PHB-) Negative Phenotype of Ralstonia eutropha PHB-4

The undefined poly(3-hydroxybutyrate)- (PHB-) negative mutant R. eutropha PHB-4 was generated in 1970 by 1-nitroso-3-nitro-1-methylguanidine (NMG) treatment. Although being scientific relevant, its genotype remained unknown since its isolation except a recent first investigation. In this study, the mutation causing the PHA-negative phenotype of R. eutropha PHB-4 was confirmed independently: sequence analysis of the phaCAB operon identified a G320A mutation in phaC yielding a stop codon, leading to a massively truncated PhaC protein of 106 amino acids (AS) in R. eutropha PHB-4 instead of 589 AS in the wild type. No other mutations were observed within the phaCAB operon. As further mutations probably occurred in the genome of mutant PHB-4 potentially causing secondary effects on the cells' metabolism, the main focus of the study was to perform a 2D PAGE-based proteome analysis in order to identify differences in the proteomes of the wild type and mutant PHB-4. A total of 20 differentially expressed proteins were identified which provide valuable insights in the metabolomic changes of mutant PHB-4. Besides excretion of pyruvate, mutant PHB-4 encounters the accumulation of intermediates such as pyruvate and acetyl-CoA by enhanced expression of the observed protein species: (i) ThiJ supports biosynthesis of cofactor TPP and thereby reinforces the 2-oxoacid dehydrogenase complexes as PDHC, ADHC and OGDHC in order to convert pyruvate at a higher rate and the (ii) 3-isopropylmalate dehydrogenase LeuB3 apparently directs pyruvate to synthesis of several amino acids. Different (iii) acylCoA-transferases enable transfer reactions between organic acid intermediates, and (iv) citrate lyase CitE4 regenerates oxaloacetate from citrate for conversion with acetyl-CoA in the TCC in an anaplerotic reaction. Substantial amounts of reduction equivalents generated in the TCC are countered by (v) synthesis of more ubiquinones due to enhanced synthesis of MenG2 and MenG3, thereby improving the respiratory chain which accepts electrons from NADH and succinate

Quantitative AFM analysis of phase separated borosilicate glass surfaces

Phase separated borosilicate glass samples were prepared by applying various heat treatments. Using selective chemical etching we performed AFM measurement on the phase separated glass surfaces. A quantitative roughness analysis allowed us to measure precisely the dependence of the characteristic size of the phase domains on heating time and temperature. The experimental measurements are very well described by the theoretically expected scaling laws. Interdiffusion coefficients and activation energy are estimated from this analysis and are consistent with literature data

Controlled data storage for non-volatile memory cells embedded in nano magnetic logic

Among the beyond-CMOS technologies, perpendicular Nano Magnetic Logic (pNML) is a promising candidate due to its low power consumption, its non-volatility and its monolithic 3D integrability, which makes it possible to integrate memory and logic into the same device by exploiting the interaction of bi-stable nanomagnets with perpendicular magnetic anisotropy. Logic computation and signal synchronization are achieved by focus ion beam irradiation and by pinning domain walls in magnetic notches. However, in realistic circuits, the information storage and their read-out are crucial issues, often ignored in the exploration of beyond-CMOS devices. In this paper we address these issues by experimentally demonstrating a pNML memory element, whose read and write operations can be controlled by two independent pulsed currents. Our results prove the correct behavior of the proposed structure that enables high density memory embedded in the logic plane of 3D-integrated pNML circuits

Near-universal trends in brGDGT lipid distributions in nature

Bacterial brGDGT lipids are a prevalent tool in studies of terrestrial paleoclimate. Their distributions correlate empirically with environmental temperature and pH, and their ubiquity in terrestrial, freshwater, and marine environments gives them wide applicability. Whether correlations with temperature and pH emerge due to a physiological response of source organisms and/or a shift in bacterial community composition remains an open question with important implications for proxy development and application. We applied a newly described technique for grouping brGDGTs to a globally compiled dataset (n = 3129) consisting of all modern sample media known to host brGDGTs. We found strong resemblances in the relationships between brGDGT fractional abundances and both temperature and pH across nearly all sample types examined. We also found near-universal connections between the brGDGTs themselves. Given the markedly different bacterial communities expected to inhabit these settings, these widespread relationships may suggest physiological and/or biochemical bases for observed brGDGT distributions. &nbsp;</p

Intact Polar brGDGTs in Arctic Lake Catchments: Implications for Lipid Sources and Paleoclimate Applications

Paleotemperature histories derived from lake sediment archives provide valuable context for modern and future climate changes. Branched glycerol dialkyl glycerol tetraether (brGDGT) lipids are a valuable tool in such pursuits due to their empirical correlation with temperature and near ubiquity in nature. However, the relative contributions of terrestrial and lacustrine sources of brGDGTs to lake sediments is site-dependent and difficult to constrain. Here, we explored the potential for intact brGDGTs—the complete lipids with polar head groups (HGs) still attached—to provide insight into the sources of brGDGTs on the landscape and their contributions to the sedimentary record in a set of Arctic lakes. We measured core and intact brGDGTs in soils, surface and downcore sediments, water filtrates, and sediment traps across five lake catchments in the Eastern Canadian Arctic, with an emphasis on Lake Qaupat (QPT), Baffin Island. Soils were dominated by brGDGTs with a monoglycosyl (1G) HG, while lacustrine samples contained more phosphohexose (PH) brGDGTs, providing evidence for in situ brGDGT production in both settings. Core- and PH-brGDGT-IIIa were more abundant in sediments than in the soils or water column, implying an additional post-depositional source of brGDGTs. A hierarchical clustering analysis indicated that core brGDGTs in Lake QPT sediments were largely lacustrine in origin, while 1G-brGDGTs were primarily soil-derived. Additionally, we found evidence for preservation of intact brGDGTs—especially 1G-brGDGTs—downcore on thousand-year timespans, though in situ production deeper in the sediment column cannot be ruled out. Finally, we explored the possibility of reconstructing 1G-brGDGT-derived soil temperatures and core-brGDGT-derived lake temperatures in tandem from sedimentary archives

Crystalline surface structures induced by ion sputtering of Al-rich icosahedral quasicrystals

Low-energy electron diffraction patterns, produced from quasicrystal surfaces by ion sputtering and annealing to temperatures below ∼700 K, can be assigned to various terminations of the cubic CsCl structure. The assignments are based upon ratios of spot spacings, estimates of surface lattice constants, bulk phase diagrams vs surface compositions, and comparisons with previous work. The CsCl overlayers are deeper than about five atomic layers, because they obscure the diffraction spots from the underlying quasicrystalline substrate. These patterns transform irreversibly to quasicrystalline(like) patterns upon annealing to higher temperatures, indicating that the cubic overlayers are metastable. Based upon the data for three chemically identical, but symmetrically inequivalent surfaces, a model is developed for the relation between the cubic overlayers and the quasicrystalline substrate. The model is based upon the related symmetries of cubic close-packed and icosahedral-packed materials. The model explains not only the symmetries of the cubic surface terminations, but also the number and orientation of domains