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International audience; A novel and simple method for the measurement of cloud point temperatures of solutions is presented. Cloud point determination , which is currently used to establish the phase diagrams of protein solutions, is indicative of proteins interactions and constitutes a useful tool for food products engineering. We describe a novel experimental setup that allows screening of a large number of physical-chemical conditions in one measurement and the determination of cloud point temperatures both above and below ambient temperature. We use a simple method to avoid solvent evaporation and condensation, so that the setup can be used for solutions prepared with a volatile solvent. We present the operating parameter range and the precision of the measurement. The optical properties of the system are calibrated with solutions of known transmittance, and the determination of cloud point temperatures is validated on a standard non-ionic surfactant solution. Finally, we demonstrate the efficiency of the method by determining the phase diagram of a wheat protein extract, soluble in a water/ethanol mixture. Complemented with differential scanning calorimetry measurements, the liquid-liquid phase transition can be determined up to a protein concentration of 250 g/L, a range inaccessible with conventional methods for this protein extract.

International audience; The self-assembling properties of glycerol esters in water are well known. Still, few data on glycerol monoesters of undecylenic acid are available. The aim of this study was to highlight the behavior of glycerol monoundecenoate (GM-C11:1) in different diluted and concentrated states. Its self-assembling properties in water and upon solid inorganic surfaces were investigated in the diluted state using surface tension experiments, atomic force microscopy, and cryogenic transmission electron microscopy studies. In the concentrated state, the gelling properties in the presence of water were investigated using polarized light microscopy, differential scanning calorimetry (DSC), and small-angle X-ray scattering (SAXS) experiments. GM-C11:1 at 100 mg/L self-assembles at the liquid/air interfaces as aggregates of approximately 20 nm in diameter, organized into concentric forms. These aggregates are spherical globules composed of several molecules of GM-C11:1. At higher concentrations (1000 and 104 mg/L), GM-C11:1 is able to uniformly coat liquid/air and liquid/solid interfaces. In bulk, GM-C11:1 forms spontaneously aggregates and vesicles. In a more concentrated state, GM-C11:1 assembles into lamellar Lβ and Lα forms in water. By cross-referencing SAXS and DSC findings, we were able to distinguish between interlamellar water molecules strongly bound to GM-C11:1 and other molecules remaining unbound and considered to be “mobile” water. The percentage of water strongly bound was proportional to the percentage of GM-C11:1 in the system. In this case, GM-C11:1 appears to be an effective molecule for surface treatments for which water retention is important.

Innovative dry chemo- and chemo-mechanical pretreatments form an interesting approach for modifying the native physico-chemical composition of lignocellulose facilitating its microbial conversion to carboxylates. Here, the impact of four dry-pretreatment conditions on the microbial transformation of wheat straw was assessed: milling to 2 mm and 100 µm, and NaOH chemical impregnation at high substrate concentrations combined with milling at 2 mm and 100 µm. Pretreatment effect was assessed in the light of substrate structure and composition, its impact on the acidogenic potential and the major enzyme activities of a rumen-derived microbial consortium RWS. Chemo-mechanical pretreatment strongly modified the substrate macroporosity. The highest carboxylate production rate was reached after dry chemo-mechanical treatment with NaOH at 100 µm. A positive impact of the dry chemo-mechanical treatment on xylanase activity was observed also. These results underline that increasing substrate macroporosity by dry chemo-mechanical pretreatment had a positive impact on the microbial acidogenic potential.

International audience; An original method based on atomic force microscopy (AFM) in contact mode was developed to abrade progressively the surface of tablets made of starch or gluten polymers isolated from wheat. The volume of the material removed by the tip was estimated from the analysis of successive topographic images of the surface, and the shear force was measured by keeping a constant normal force. Our data together with a simple tribological model provide clear evidence for a higher hardness and shear strength of starch compared to gluten. Gluten appears to have mechanical properties close to soft materials, such as talc, whereas starch displays higher hardness close to calcite. Our results are in a better agreement with structural properties of gluten (complex protein network) and starch (granular and semi-cristalline structure) than earlier studies by micro-indentation. This work shows that the AFM scratching method is relevant for the characterization of any polymer surface, in particular in application to materials made of different polymers at the nano-scale.

International audience; A mixture of nine microorganisms (six bacteria and three yeasts) from the microflora of surface-ripened cheeses were subjected to in vitro digestive stress in a three-compartment "dynamic gastrointestinal digester" (DIDGI). We studied the microorganisms (i) grown separately in culture medium only (ii) grown separately in culture medium and then mixed, (iii) grown separately in culture medium and then included in a rennet gel and (iv) grown together in smear-ripened cheese. The yeasts Geotrichum candidum, Kluyveromyces lactis and Debaryomyces hansenii, were strongly resistant to the whole DIDGI process (with a drop in viable cell counts of less than < 1 log CFU mL(-1)) and there were no significant differences between lab cultures and cheese-grown cultures. Ripening bacteria such as Hafnia alvei survived gastric stress less well when grown in cheese (with no viable cells after 90 min of exposure of the cheese matrix, compared with 6 CFU mL(-1) in lab cultures). The ability of Corynebacterium casei and Staphylococcus equorum to withstand digestive stress was similar for cheese and pure culture conditions. When grow in a cheese matrix, Brevibacterium aurantiacum and Arthrobacter arilaitensis were clearly more sensitive to the overall digestive process than when grown in pure cultures. Lactococcus lactis displayed poorer survival in gastric and duodenal compartments when it had been grown in cheese. In vivo experiments in BALB/c mice agreed with the DIDGI experiments and confirmed the latter's reliability.

The cross-linking of plant cell walls by ferulate dehydrodimerization reactions is well established. Recently, a (5-5 ')/(8 '-O-4 '') dehydrotrimer of ferulic acid (A) was isolated from alkali extracts of maize bran. Its structure was elucidated on the basis of an extensive structural analysis (UV, MS, 1D and 2D NMR). This first identified ferulic acid dehydrotrimer has revealed that parietal polysaccharide chains can be more extensively cross-linked than has been previously recognized. To produce the (5-5 ')/(8 '-O-4 '') dehydrotrimer (A) in suf-ficient high-purity quantities and allow the development of analytical methodologies (e.g., LC/GC, immunohistochemistry) to identify and quantify this peculiar trimer in plant extracts or samples, its first total convergent synthesis has been achieved in 10 steps and in 18% yield starting from commercially available vanillin by horseradish peroxidase mediated enzymatic aryl-aryl coupling, aldolization/crotonization, and Wittig olefination reactions as the key steps.

International audience; Prolamins, the main storage proteins of wheat seeds, are synthesized and retained in the endoplasmic reticulum (ER) of the endosperm cells, where they accumulate in protein bodies (PBs) and are then exported to the storage vacuole. The mechanisms leading to these events are unresolved. To investigate this unconventional trafficking pathway, wheat gamma-gliadin and its isolated repeated N-terminal and cysteine-rich C-terminal domains were fused to fluorescent proteins and expressed in tobacco leaf epidermal cells. The results indicated that gamma-gliadin and both isolated domains were able to be retained and accumulated as protein body-like structures (PBLS) in the ER, suggesting that tandem repeats are not the only sequence involved in gamma-gliadin ER retention and PBLS formation. The high actin-dependent mobility of gamma-gliadin PBLS is also reported, and it is demonstrated that most of them do not co-localize with Golgi body or pre-vacuolar compartment markers. Both gamma-gliadin domains are found in the same PBLS when co-expressed, which is most probably due to their ability to interact with each other, as indicated by the yeast two-hybrid and FRET-FLIM experiments. Moreover, when stably expressed in BY-2 cells, green fluorescent protein (GFP) fusions to gamma-gliadin and its isolated domains were retained in the ER for several days before being exported to the vacuole in a Golgi-dependent manner, and degraded, leading to the release of the GFP 'core'. Taken together, the results show that tobacco cells are a convenient model to study the atypical wheat prolamin trafficking with fluorescent protein fusions.