Interaction of a Rhodococcus sp. trehalose lipid biosurfactant with model proteins: thermodynamic and structural changes

One major application of surfactants is to prevent aggregation during various processes of protein manipulation. In this work, a bacterial trehalose lipid (TL) with biosurfactant activity, secreted by Rhodococcus sp., has been identified and purified. The interactions of this glycolipid with selected model proteins have been studied by using differential scanning calorimetry (DSC), Fourier-transform infrared (FTIR) spectroscopy, isothermal titration calorimetry (ITC), and fluorescence spectroscopy. Bovine serum albumin (BSA) and cytochrome c (Cyt-c) have been chosen because of their quite different secondary structures: BSA contains essentially no β-sheets and an average 66% α-helix, whereas Cyt-c possesses up to 25% β-sheets and up to 45% α-helical structure. Differential scanning calorimetry shows that addition of TL to BSA at concentrations below the critical micelle concentration (cmc) shifts the thermal unfolding temperature to higher values. FTIR indicates that TL does not alter the secondary structure of native BSA, but the presence of TL protects the protein toward thermal denaturation, mainly by avoiding formation of β-aggregates. Studies on the intrinsic Trp fluorescence of BSA show that addition of TL to the native protein results in conformational changes. BSA unfolding upon thermal denaturation in the absence of TL makes the Trp residues less accessible to the quencher, as shown by a decrease in the value of Stern-Volmer dynamic quenching constant, whereas denaturation in the presence of the biosurfactant prevents unfolding, in agreement with FTIR results. In the case of Cyt-c, interaction with TL gives rise to a new thermal denaturation transition, as observed by DSC, at temperatures below that of the native protein, therefore facilitating thermal unfolding. Binding of TL to native BSA and Cyt-c, as determined by ITC, suggests a rather nonspecific interaction of the biosurfactant with both proteins. FTIR indicates that TL slightly modifies the secondary structure of native Cyt-c, but protein denaturation in the presence of TL results in a higher proportion of β-aggregates than in its absence (20% vs 3.9%). The study of Trp fluorescence upon TL addition to Cyt-c results in a completely opposite scenario to that described above for BSA. In this case, addition of TL considerably increases the value of the dynamic quenching constant, both in native and denatured protein; that is, the interaction with the glycolipid induces conformational changes which facilitate the exposure of Trp residues to the quencher. Considering the structures of both proteins, it could be derived that the characteristics of TL interactions, either promoting or avoiding thermal unfolding, are highly dependent on the protein secondary structure. Our results also suggest the rather unspecific nature of these interactions. These might well involve protein hydrophobic domains which, being buried into the protein native structures, become exposed upon thermal unfolding.     

Development and Characterization of a Solid-Phase Biocatalyst Based on Cyclodextrin Glucantransferase Reversibly Immobilized onto Thiolsulfinate-Agarose

Reduction of disulfide bonds and introduction of ??de novo?? thiol groups in cyclodextrin glucantransferase from Thermoanaerobacter sp. were assessed in order to perform reversible covalent immobilization onto thiol-reactive supports (thiolsulfinate-agarose). Only the thiolation process dramatically improved the immobilization yield, from 0?% for the native and reduced enzyme, up to nearly 90?% for the thiolated enzyme. The mild conditions of the immobilization process (pH 6.8?C7.0 and 22?°C) allowed the achievement of 100?% coupling efficiencies when low loads were applied. Ionic strength was a critical parameter for the immobilization process; for high activity recoveries, 50?mM phosphate buffer supplemented with 0.15?M NaCl was required. The kinetic parameters, pH and thermal stabilities for the immobilized biocatalyst were similar to those for the native enzyme. For ??-cyclization activity, optimal pH range and temperature were 4.0?C5.4 and 85?°C. The possibility of reusing the support was demonstrated by the reversibility of enzyme?Csupport binding.     

Single-stage in situ synthesis of silver nanoparticles in antibacterial self-assembled overlays

In this work, a novel single-stage process for in situ synthesis of Ag nanoparticles (NPs) using the layer-by-layer (LbL) technique is presented. The Ag NPs were formed into nanotextured coatings based on sequentially adsorbed poly(allylamine hydrochloride) (PAH) and SiO₂ NPs. Such highly porous surfaces have been used in the fabrication of highly efficient ion release films for applications such as antibacterial coatings. In this approach, the amino groups of the PAH acted as reducing agent and made possible the in situ formation of the Ag NPs. This reduction reaction occurred during the LbL process as the coating was assembled, without any further step after the fabrication and stabilization of the multilayer film. Biamminesilver nitrate was used as the Ag⁺ ion source during the LbL process and it was successfully reduced to Ag NPs. All coatings were tested with gram-positive and gram-negative bacterial cultures of Escherichia coli, Staphylococcus aureus, and Lactobacillus delbrueckii showing an excellent antimicrobial behavior against these types of bacteria (more than 99.9% of killing efficiency in all cases).     

Liquid chromatographic/electrospray ionization tandem mass spectrometric study of polyphenolic composition of four cultivars of Fragaria vesca L. berries and their comparative evaluation

High-performance liquid chromatography coupled with ion spray mass spectrometry in the tandem mode with both negative and positive ionization was used for investigating a variety of polyphenolic compounds in four genotypes of Fragaria vesca berries. About 60 phenolic compounds belonging to the compound classes of phenolic acids, ellagitannins, ellagic acid derivatives, flavonols, monomeric and oligomeric flavanols, dihydrochalcones and anthocyanins were reported, providing for the first time a quite complete picture of polyphenolic composition of F. vesca berries. Some of the polyphenols herein investigated, such as a tris-galloyl-hexahydroxydiphenoyl-hexose, two castalagin/vescalagin-like isomers and peonidin-malonylglucoside, were described for the first time. Principal component analysis applied on original HPLC-MS/MS data, acquired in multiple reaction monitoring mode, successfully discriminated the four investigated cultivars on the basis of their polyphenolic composition, highlighting the fundamental role of mass spectrometry for food characterization. Copyright ? 2012 John Wiley & Sons, Ltd.     

Nanocrystalline CoSn2-carbon composite electrode prepared by using sonochemistry

A sonochemical method has been used to prepare negative electrode materials containing intermetallic nanoparticles and polyacrylonitrile (PAN). The ultrasound irradiation is applied to achieve small particle size. After annealing at 490 °C under Ar-flow, the polymer PAN is partially carbonized and the metallic nanoparticles are surrounded by a carbonaceous matrix. The main metallic phase is CoSn₂. The carbonaceous coating and the surface oxides have been explored by using XPS. The resulting CoSn₂-carbonaceous phase electrode (CoSn₂@C) shows improved electrochemical behavior (ca. 450 mAh/g after 50 cycles) in comparison with previous reports on pure crystalline CoSn₂. The reaction between CoSn₂@C and Li has been studied by using XRD and ¹¹⁹Sn Mössbauer spectroscopy. The formation of large grains of crystalline Li_xSn phases after the first discharge is discarded. The small particle size which is achieved by using ultrasonication and the carbonaceous matrix contribute to maintain the Co-Sn interactions during the electrochemical cycling. The aggregation of the nanosized metallic particles upon electrochemical cycling can be suppressed by the carbonaceous matrix (pyrolytic PAN).     

Determination of Pb,Zr, Ti,Sr, Cr,Nb and La in lead zirconate titanate ceramics by particle‐induced X‐ray emission

A particle‐induced X‐ray emission (PIXE) technique has been used in the determination of the principal components Pb, Zr and Ti and the substituting elements Sr, Cr, Nb and La in lead zirconate titanate ceramics. In general, precision of analysis was concentration dependent from each element under study. For Pb, precision varied between 0.13% and 0.16%, at higher concentration of 59.32–64.5%. It was around 6–9% for Sr, Cr and Nb at concentrations of 1% or lower. Particular attention was devoted to the estimation of the analysis trueness. With this purpose, three methods were applied: (1) comparison of PIXE and inductively coupled plasma optical emission spectrometry concentrations, (2) recovery study and (3) comparison with a laboratory standard. Trueness of analysis was around 100 ± 10% for the evaluated elements Pb, Zr, Ti, Sr and Cr. The expected stoichiometry and elemental composition homogeneity of a wide group of produced ceramics were confirmed by the PIXE technique. Copyright © 2012 John Wiley & Sons, Ltd.     

Trapping and injecting single domain walls in magnetic wire by local fields

A single domain wall (DW) moves at linearly increasing velocity under an increasing homogeneous drive magnetic field. Present experiments show that the DW is braked and finally trapped at a given position when an additional antiparallel local magnetic field is applied. That position and its velocity are further controlled by suitable tuning of the local field. In turn, the parallel local field of small amplitude does not significantly affect the effective wall speed at long distance, although it generates tail-to-tail and head-to-head pairs of walls moving along opposite directions when that field is strong enough.     

Extracting Past-Future Vacuum Correlations Using Circuit QED

We propose a realistic circuit QED experiment to test the extraction of past-future vacuum entanglement to a pair of superconducting qubits. The qubit P interacts with the quantum field along an open transmission line for an interval T_{on} and then, after a time-lapse T_{off}, the qubit F starts interacting for a time T_{on} in a symmetric fashion. After that, past-future quantum correlations will have transferred to the qubits, even if the qubits do not coexist at the same time. We show that this experiment can be realized with current technology and discuss its utility as a possible implementation of a quantum memory.     

Role of cellulose oxidation in the yellowing of ancient paper

The yellowing of paper on aging causes major aesthetic damages of cultural heritage. It is due to cellulose oxidation, a complex process with many possible products still to be clarified. By comparing ultraviolet-visible reflectance spectra of ancient and artificially aged modern papers with ab?initio time-dependent density functional theory calculations, we identify and estimate the abundance of oxidized functional groups acting as chromophores and responsible of paper yellowing. This knowledge can be used to set up strategies and selective chemical treatments preventing paper yellowing.     

Closed-form BER analysis of variable weight MPPM coding under gamma-gamma scintillation for atmospheric optical communications

In this paper, the performance of the variable weight multiple pulse-position modulation (MPPM) coding technique in an atmospheric optical communication environment under gamma-gamma optical scintillation is analyzed, proposing a closed-form expression for the average bit error rate (BER). This study is based on a hyperexponential fitting of the conditional BER in absence of turbulence fluctuations, leading to closed-form expressions that characterize the behavior of this nonlinear coding scheme. Finally, conditional and average BER expressions proposed here are corroborated with Monte Carlo simulations results.