Draft genome sequence of the plant growth-promoting rhizobacterium Pseudomonas fluorescens Strain CREAC16 isolated from pea (Pisum sativum L.) rhizosphere

Herein, we report the draft genome sequence of Pseudomonas fluorescens strain CREA-C16, a plant growth-promoting rhizobacterium that was isolated from the rhizosphere of Pisum sativum L. plants. The genome sequence is ~6 Mb in size, with a G+C content of 60.1%, and includes 4,457 candidate protein-encoding genes

Wild-type and mutant bovine odorant-binding proteins to probe the role of the quaternary structure organization in the protein thermal stability

The exploration of events taking place at different timescales and affecting the structural and dynamics properties of proteins, such as the interactions of proteins with ligands and the subunits association/ dissociation, must necessarily be performed by using different methodologies, each of which specialized to highlight the different phenomena that occur when proteins are exposed to chemical or physical stress. In this work, we investigated the structure and dynamics of the wild-type bovine odorant-binding protein (wt-bOBP), which is a domain-swapped dimeric protein, and the triple mutant deswapped monomeric form of the protein (m-bOBP) to shed light on the role of the quaternary and tertiary structural organization in the protein thermal stability. Difference infrared spectra, 2D-IR correlation spectroscopy and molecular dynamics simulations were used to probe the effect of heating on protein structure and dynamics in microsecond and nanoseconds temporal ranges, respectively. The obtained results show that there is a heating-induced transition toward a less structured state in m-bOBP, that it is detectable around 70-80 degrees C. On the contrary, in wt-bOBP this transition is almost negligible, and changes are detectable in the protein spectra in the range of temperature between 75 and 85 degrees C. A detailed 3D inspection of the structure of the two proteins that takes into the account the spectroscopic results indicates that (a) ion pairs and hydrophobic interactions appear to be the major determinants responsible for the protein stability and (b) the protein intersubunit interactions confer an increased resistance toward the thermal stress

From untargeted metabolomics to the multiple reaction monitoring‐based quantification of polyphenols in chocolates from different geographical areas

Plants, including cocoa bean, are the main source of metabolites with multiple biological functions. Polyphenol extracts are widely used as a nutraceutical supplement for their well‐known health‐promoting role. In this paper, a preliminary untargeted metabolic screening was carried out by matrix‐assisted laser desorption/ionization (MALDI)‐time of flight (TOF)/TOF on a pool of chocolate samples made by cocoa beans of different geographical areas. Then, a targeted approach was developed for polyphenol quantification by an optimized Liquid chromatography (LC)–tandem mass spectrometry (MS/MS) method multiple reaction monitoring (MRM) ion mode. Detection limit of polyphenol standard ranged between 1 and 25 pg/μl with variation coefficient lower than 15%. External calibration curves were used for quantification of polyphenols in 18 samples. Fifty polyphenols were detected in a single LC–MRM/MS run and quantified by monitoring almost 90 transitions in a 5‐minute run. The polyphenols content of different cocoa beans from several countries was finally compared by principal component analysis (PCA) statistical analysis suggesting that the chocolate made by Ecuador cocoa beans showed the highest level of polyphenols

Molecular strategies for protein stabilization: the case of a trehalose/maltose-binding protein from Thermus thermophilus.

The trehalose/maltose-binding protein (MalE1) is one component of trehalose and maltose uptake system in the thermophilic organism Thermus thermophilus. MalE1 is a monomeric 48 kDa protein predominantly organized in alpha-helix conformation with a minor content of beta-structure. In this work, we used Fourier-infrared spectroscopy and in silico methodologies for investigating the structural stability properties of MalE1. The protein was studied in the absence and in the presence of maltose as well as in the absence and in the presence of SDS at different p(2)H values (neutral p(2)H and at p(2)H 9.8). In the absence of SDS, the results pointed out a high thermostability of the MalE1 alpha-helices, maintained also at basic p(2)H values. However, the obtained data also showed that at high temperatures the MalE1 beta-sheets underwent to structural rearrangements that were totally reversible when the temperature was lowered. At room temperature, the addition of SDS to the protein solution slightly modified the MalE1 secondary structure content by decreasing the protein thermostability. The infrared data, corroborated by molecular dynamics simulation experiments performed on the structure of MalE1, indicated that the protein hydrophobic interactions have an important role in the MalE1 high thermostability. Finally, the results obtained on MalE1 are also discussed in comparison with the data on similar thermostable proteins already studied in our laboratories

Nanobeads-based assays. the case of gluten detection

In order to verify if the use of nanobeads of poly[phenylacetylene-(co- acrylic acid)] (PPA/AA) in the ELISA test would affect the immune-activity of the antibodies (Ab) and/or the activity of the enzymes used to label the Ab anti-rabbit IGg, in this work we immobilized the horse liver peroxidase labelled Ab anti-rabbit IGg onto PPA/AA nanobeads. The gluten test was chosen as the model to demonstrate the usefulness of these nanobeads in immunoassays. The synthesis of PPA/AA nanobeads was performed by a modified emulsion polymerization. Self-assembly of nanospheres with mean diameter equal to 200nm was achieved by casting aqueous suspensions. The materials were characterized by traditional spectroscopic techniques, while the size and dispersion of the particles were analysed by scanning electron microscopy (SEM) measurements. The obtained results show that the immobilization process of the Abs onto PPA/AA did not affect either the immune-response of the Abs or the functional activity of the peroxidase suggesting the usefulness of PPA/AA for the design of advanced nanobeads-based assays for the simultaneous screening of several analytes in complex media. © 2008 IOP Publishing Ltd