Enzymatic fingerprinting reveals specific xyloglucan and pectin signatures in the cell wall purified with primary plasmodesmata.

Plasmodesmata (PD) pores connect neighbouring plant cells and enable direct transport across the cell wall. Understanding the molecular composition of these structures is essential to address their formation and later dynamic regulation. Here we provide a biochemical characterisation of the cell wall co-purified with primary PD of Arabidopsis thaliana cell cultures. To achieve this result we combined subcellular fractionation, polysaccharide analyses and enzymatic fingerprinting approaches. Relative to the rest of the cell wall, specific patterns were observed in the PD fraction. Most xyloglucans, although possibly not abundant as a group, were fucosylated. Homogalacturonans displayed short methylated stretches while rhamnogalacturonan I species were remarkably abundant. Full rhamnogalacturonan II forms, highly methyl-acetylated, were also present. We additionally showed that these domains, compared to the broad wall, are less affected by wall modifying activities during a time interval of days. Overall, the protocol and the data presented here open new opportunities for the study of wall polysaccharides associated with PD.

Comparative structural and vibrational investigations between cocoa butter (CB) and cocoa butter equivalent (CBE) by ESI/MALDI-HRMS, XRD, DSC, MIR and Raman spectroscopy.

A high quality chocolate requires not only a shiny surface, a crunchy and pleasant texture, but also a proper resistance to blooming. All these characteristics are influenced by the physical and chemical properties of the components, which are directly related to their crystalline structure. Some works found that the proportion of cocoa butter (CB), cocoa butter equivalent (CBE) and milk fatty acid (AMF) tend to strongly delay the blooming when mixing them. The goal of our research is to determine how the choice of adding CBE to the mixture delays chocolate blooming. ESI/MALDI-HRMS, X-ray, DSC, MIR and Raman investigations were used to analyze the structure features and the vibrational modes of CB and CBE. The comparison of these experimental results between CB and CBE made it possible to highlight markers of differentiation between CB and CBE which seems to explain the impact of CBE in the chocolate blooming. Part of these triglycerides remains in form IV instead. The presence of the latter seems to be a key parameter that favors the transformation deceleration to the form VI, which is responsible for the fat bloom development.

Physico-chemical studies of resveratrol, methyl-jasmonate and cyclodextrin interactions: an approach to resveratrol bioproduction optimization.

trans-Resveratrol (RSV) is a natural phenolic molecule of the stilbene family known for its anti-oxidant properties in the field of nutraceuticals and cosmetics. Its production by grapevine cell suspensions is induced by the addition to the culture medium of elicitor compounds, methyl jasmonate (MeJA) and cyclodextrins (CDs). Physico-chemical studies were performed to understand the mechanism of action of CDs on this bioproduction of RSV. Inclusion complexes of RSV in CDs were first observed and then interactions with MeJA were identified using various analytical techniques such as UV and nuclear magnetic resonance (NMR) spectroscopies, mass spectrometry (MS) and isothermal titration calorimetry (ITC).

Structural investigation of an exopolysaccharide substituted with a lactyl ether group produced by Raoultella terrigena Ez-555-6 isolated in the Chernobyl exclusion zone.

Raoultella terrigena strain Ez-555-6, isolated from a root nodule of Medicago sativa harvested in the Chernobyl exclusion zone, produces a non-referenced high-molecular-mass exopolysaccharide (EPS). The structure of this EPS was determined using a combination approach including monosaccharide composition (GLC-FID, HPAEC-PAD), determination of glycosylation sites (GLC-EIMS) and 1D/2D NMR ((1)H, (13)C) and ESIMS (HR, MS/MS) studies of oligosaccharides obtained from mild acid hydrolysis. The EPS was found to be a charged pentasaccharide with a repeating unit composed of D-galactose, D-glucose, D-mannose and D-glucuronic acid (1:2:1:1). Lactic acid and O-acetyl substituents were localized on galactose and glucose residues, respectively, as presented in the following structure:

Identification of an N-(hydroxysulfonyl)oxy metabolite using in vitro microorganism screening, high-resolution and tandem electrospray ionization mass spectrometry.

Preliminary metabolic profiling of a drug under pre-clinical development revealed the presence of a minor unknown metabolite with a positive ion electrospray ionization (ESI) mass spectrum identical to that of the unchanged compound. Since the low concentration of the compound did not allow any additional experiments, preparative bioconversion using fungi was used to obtain a substantial amount of the molecule. Negative ion ESI-MS and tandem mass spectrometry (MS/MS) in combination with accurate mass measurements obtained on a quadrupole/time-of-flight instrument (Q-TOF) led to the positive identification of a hydroxylamide sulfoconjugated metabolite.

Applications of liquid chromatography/dynamic liquid secondary ion mass spectrometry in structural elucidation studies.

The application of post-column splitting in liquid chromatography (LC) coupled to dynamic liquid secondary ion (or continuous-flow fast atom bombardment) mass spectrometry allows more flexibility with respect to the use of conventional LC methods. Gradient elution and the presence of involatile mobile phase modifiers do not therefore constitute insurmountable obstacles, especially in experiments where sensitivity is of secondary importance, such as in structural identification studies. The possibility of using conventional LC conditions presents a valuable gain in time by avoiding lengthy LC redevelopment procedures. This means a non-negligible asset in an environment where rapid answers are necessary.