0000000001294469
AUTHOR
Camille Loupiac
IMAGINE: A Cold Neutron Imaging Station at the Laboratoire Léon Brillouin
International audience; A second cold neutron imaging station has been open to users at the Laboratoire Léon Brillouin. The station is designed for high resolution neutron imaging and tomography. The typical field of view is 100x100 mm2 with a spatial resolution of 100 μm. Betterspatial resolutions (∼50 μm) can be achieved when reducing the field of view down to 30x30mm2. The L/D ratio can be varied from 200 to1000with pinhole sizes ranging from 20 to7 mm. Future upgrades will provide capabilities for energy resolved measurements using either a velocity selector or a double crystal monochromator. The possibility to perform polarized neutron experiments will also be provided next year.
Effect of high pressure on the antimicrobial activity and secondary structure of the bacteriocin nisin
International audience; Effect of high pressure (HP) treatment on the antimicrobial properties and the structure of nisin was evaluated. Nisin solutions at pH 2.8 or 6.1 were treated by HP at 500 MPa – 10 min – 20 °C and their antimicrobial potency was determined. It appeared that HP clearly impacted the antimicrobial activity of nisin, with respective activity loss of 22.5% and 49.9% at pH 2.8 and 6.1. Structural analysis of nisin by circular dichroism and Fourier transform-infrared spectroscopies revealed that the decrease of nisin antimicrobial activity was likely due to the unfolding of the protein induced by HP. A loss of nisin β-turns structure, particularly significant at neutral pH,…
Effect of Ligands on HP-Induced Unfolding and Oligomerization of β-Lactoglobulin
ABSTRACTTo probe intermediate states during unfolding and oligomerization of proteins remains a major challenge. High pressure (HP) is a powerful tool for studying these problems, revealing subtle structural changes in proteins not accessible by other means of denaturation. Bovine β-lactoglobulin (BLG), the main whey protein, has a strong propensity to bind various bioactive molecules, such as retinol and resveratrol, two ligands with different affinity and binding sites. By combining in situ HP-small-angle neutron scattering (SANS) and HP-UV/visible absorption spectroscopy, we report the specific effects of these ligands on 3D conformational and local changes in BLG induced by HP. Dependin…
Effect of high hydrostatic pressure on extraction of B-phycoerythrin from Porphyridium cruentum: Use of confocal microscopy and image processing
International audience; The aim of the study was to extract B-phycoerythrin from Porphyridium cruentum while preserving its structure. The high hydrostatic pressure treatments were chosen as extraction technology. Different methods have been used to observe the effects of the treatment: spectrophotometry and confocal laser scanning microscopy followed by image processing analysis. Image processing led to the generation of masks used for the identification of three clusters: intra, extra and intercellular. All methods showed that high hydrostatic pressure treatments between 50 and 500 MPa failed to extract B-phycoerythrin from Porphyridium cruentum cells. The fluorescence emission was negati…
The impact of high hydrostatic pressure on structure and dynamics of beta-lactoglobulin
Abstract Methods Combining small-angle X-ray and neutron scattering measurements with inelastic neutron scattering experiments, we investigated the impact of high hydrostatic pressure on the structure and dynamics of β-lactoglobulin (βLG) in aqueous solution. Background βLG is a relatively small protein, which is predominantly dimeric in physiological conditions, but dissociates to monomer below about pH 3. Results High-pressure structural results show that the dimer–monomer equilibrium, as well as the protein–protein interactions, are only slightly perturbed by pressure, and βLG unfolding is observed above a threshold value of 3000 bar. In the same range of pressure, dynamical results put …
How neutron scattering experiments can target the structure and dynamics of milk proteins?
International audience; The powerful of neutron scattering techniques to characterize structure and dynamics of milk proteins is illustrated. Small angle neutron scattering and reflectivity are used to determine the structure and the interactions between milk proteins in solution, during gelation processes, or the protein structure at different interfaces. Experiments performed by inelastic and quasielastic neutron scattering allow one to observe the dynamics of water and proteins showing the major role of hydration on the dynamics of milk proteins.
Structure of calcium and zinc pectinate films investigated by FTIR spectroscopy
International audience; Calcium and zinc pectinate gels were prepared using a method which allowed calcium or zinc to diffuse from the cross-linking solution through a dialysis membrane to form a gel with amidated low-methoxyl pectin. The gel thus obtained was then dried, and the film structure was studied using FTIR spectroscopy as a function of the cation content (0%, 5%, 10%, and 15% w/v). Important consideration was given to the three functional groups (amide, carboxyl ester, and carboxylate groups) present in the pectin. When the zinc content was increased, the three wavenumber values corresponding to these three functional groups did not change significantly, while for calcium pectina…
Neutron scattering and imaging: a tool for archaeological studies
International audience; Neutron scattering and neutron imaging are powerful techniques for studying the structure of objects without damage, which is an essential prerequisite for investigations in Cultural Heritage domain, particularly in Archaeology. The deep penetration of neutrons in most materials allows for the study of relatively large objects. The contrast between similar materials, like metals in alloys, or that due to the presence of hydrogen atoms gives information about the internal structure of objects that have been modified or repaired in the past. Imaging and tomography give a 3-dimensional view of the whole object, permitting discrimination between different parts of the ob…
Neutron imaging and tomography: Applications in food science
International audience; Neutron imaging and tomography allow observing the structure of large objects (from few to hundred centimeters) at a resolution around 100 μm. This non-destructive method is based on the detection of the transmission of a neutron beam through an object in 2D (imaging) or 3D after rotating the sample (tomography). The components of the object attenuate the neutron beam differently, depending on their composition, and produce accurate pictures of objects which give information about their structure. Neutron imaging has been used for quality control purposes in industries (aircraft, motor engineering...), but also find application from materials sciences (fuel cells, li…
Impact of sucrose and water contents on macroscopic structure and stability of cereal based products : Thermodynamic versus dynamic
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Probing Protein−Membrane Interactions Using Solid Supported Membranes
International audience; Tethered bilayer lipid membranes have been used as a model system to mimic the interactions between the whey protein β-lactoglobulin and a lipid interface. The approach allowed for a detailed study of the lipid-protein interactions, the results being of possible importance in food and cosmetic applications. For such applications, lipid-protein interactions and the interfacial behavior are vital factors in controlling and manipulating process conditions such as emulsion stabilization and gelification. Lipid composition as well as the structural properties of the protein governed their interactions, which were probed by a combination of surface plasmon spectroscopy, ne…
The impact of cooking on meat microstructure studied by low field NMR and Neutron Tomography
International audience; We studied the impact of temperature of cooking on meat microstructure. The cooking temperature was verified by calorimetry, showing the disappearance of endothermic peaks when cooking temperature was increased. These observations correspond to the denaturation of different protein fractions at specific temperatures. 1H-low field NMR and neutron tomography were used to further understand the relationship between the observed protein denaturation and changes in meat microstructure after heating. Hahn’s echo and solid echo NMR sequences were applied to observe fast relaxation time corresponding to rigid protons. These protons were found to be associated with pools of p…
The cork viewed from the inside
International audience; Cork is the natural material stripped from the outer bark of cork oak. It is still the most used stopper to seal wine bottles and to preserve wine during storage. Cork stoppers are sorted in different classes according to apparent defects, named lenticels, which can be related to the cork macroporosity. The more lenticels there are, the worst cork quality is. The present work aims at investigating defects analysis of cork stoppers from two classes by comparing images recorded by digital photography and neutron imaging. Surface analysis of defects obtained from photography leads to more surface defects in class 4 (6.7%) than in class 0 (4.1%). Neutron radiography and …
Controlled loading and release of beta-lactoglobulin in calcium-polygalacturonate hydrogels
International audience; We show here how the structure of polygalacturonate (polyGalA) hydrogels cross-linked by Ca2+ cations via external gelation controls the loading and the release rate of betalactoglobulin (BLG), a globular protein. Hydrogels prepared from a polyGalA/BLG solution are found similar to those obtained from a polyGalA solution in our previous study (Maire du Poset, et al. Biomacromolecules 2019, 20(7), 2864-2872): they exhibit similar transparencies and gradients of mechanical properties and polyGalA concentration. The nominal BLG/polyGalA ratio of the mixtures is almost recovered within the whole mixed hydrogel despite such strong concentration gradients, except in the pa…
Structural studies of adsorbed protein (betalactoglobulin) on natural clay (montmorillonite)
International audience; In this work, the adsorption of a small globular protein (betalactoglobulin, BLG), on a natural montmorillonite clay (Mt) was investigated in acidic buffer (pH = 3). The combination of different characterization techniques such as zetametry, X-ray diffraction, transmission electronic microscopy, fluorescence and solid state nuclear magnetic resonance spectroscopies shed light on the interaction mechanism between the clay mineral and the proteins. For low BLG concentration, a slight increase of the interlayer spacing of the clay mineral was noticed as well as structural changes of the protein. In contrast, as the concentration of BLG increased, the adsorption led to a…
Protein-lipid interactions at the air-water interface.
International audience; Protein−lipid interactions play an important role in a variety of fields, for example in pharmaceutical research, biosensing, or food science. However, the underlying fundamental processes that govern the interplay of lipids and proteins are often very complex and are therefore studied using model systems. Here, Langmuir monolayers were used to probe the interaction of a model protein with lipid films at the air−water interface. The protein β-lactoglobulin (βlg) is the major component in bovine milk serum, where it coexists with the milk fat globular membrane. During homogenization of milk, βlg adsorbs to the interface of lipid fat globules and stabilizes the oil-in-…
How neutron scattering experiments can target the behavior of milk proteins
Any protein ingested intentionally or non-intentionally can be considered as a food protein. The scope of food protein chemistry covers agricultural production, processing, packaging, storage, distribution, retail and protein function as ingredients. Protein quality is an important consideration to the consumer as is allergy and other adverse reactions to some proteins. The major protein ingredients, material having 5 to 10% or more proteins are: dried non-fat milk, whey, caseinate, collagen, gelatin, myosin, blood, soybean flour, wheat gluten, egg albumin, yolk. Protein ingredients are used to achieve a wide range of functionalities and usages like adhesion, emulsification, gelation and fo…
SFN, French Neutron Community Meets at JDN23
JDN23 took place October 5–8, 2015 in Evian-Les-Bains (Haute Savoie, France), on the southern shore of the Lac Leman, facing Lausanne on the Swiss side. The first two days were devoted to a worksho...
Structural behaviour differences in low methoxy pectin solutions in the presence of divalent cations (Ca 2+ and Zn 2+ ): a process driven by the binding mechanism of the cation with the galacturonate unit
International audience; In this paper, we compare the interactions between low methoxy pectin (LMP) and either Ca2+ or Zn2+ in semi-dilute solutions. Intrinsic viscosity and turbidity measurements reveal that pectin-calcium solutions are more viscous, but yet less turbid, than pectin-zinc ones. To get a molecular understanding on the origin of this rather unexpected behavior, we further performed isothermal titration calorimetry, small angle neutron scattering experiments, as well as molecular dynamics simulations. Our results suggest that calcium cations induce the formation of a more homogeneous network of pectin than zinc cations do. The molecular dynamics simulations indicate that this …
Effect of iron chelates on oil–water interface, stabilized by milk proteins: The role of phosphate groups and pH. Prediction of iron transfer from aqueous phase toward fat globule surface by changes of interfacial properties
International audience; Iron incorporated into food systems induces oxidation and precipitation. The consequences are reduced bioavailability and a functional modification of other food components such as proteins. The iron-chelates such as ferrous bisglycinate represent a possibility to avoid side effects, since the iron is protected. The aim of this study is to investigate the effects of iron-chelates compounds on the properties of an oil/water interface stabilized by caseinate or β-lacotoglobulin, under environmental conditions at 20 °C. Analyses were performed using dynamic drop tensiometry during 5000 s. The aqueous bulk phase is an imidazole/acetate buffer (0.1 M), containing 0.4 × 10…
Myoglobin on Silica: A Case Study of the Impact of Adsorption on Protein Structure and Dynamics
International audience; If protein structure and function changes upon adsorption are well documented, modification of adsorbed protein dynamics remains a blind spot, despite its importance in biological processes. The adsorption of metmyoglobin on a silica surface was studied by isotherm measurements, microcalorimetry, circular dichroïsm, and UV-visible spectroscopy to determine the thermodynamic parameters of protein adsorption and consequent structure modifications. The mean square displacement and the vibrational densities of states of the adsorbed protein were measured by elastic and inelastic neutron scattering experiments. A decrease of protein flexibility and depletion in low freque…
In situ Phenotyping of Grapevine Root System Architecture by 2D or 3D Imaging: Advantages and Limits of Three Cultivation Methods
International audience; The root system plays an essential role in the development and physiology of the plant, as well as in its response to various stresses. However, it is often insufficiently studied, mainly because it is difficult to visualize. For grapevine, a plant of major economic interest, there is a growing need to study the root system, in particular to assess its resistance to biotic and abiotic stresses, understand the decline that may affect it, and identify new ecofriendly production systems. In this context, we have evaluated and compared three distinct growing methods (hydroponics, plane, and cylindric rhizotrons) in order to describe relevant architectural root traits of …
Protein Thermal Denaturation of Beef Muscle: Neutron Imaging and spectroscopies
International audience; Open Food System is an industrial innovation project having as goal the development of “smart” domestic appliances, which can pilot cooking process of meat and fish without any human intervention. My Ph.D research belongs to that huge project with the purpose to follow the evolution of cooking process at molecular scale, describing biochemical reactions occurring among muscle constituents and associating them to macroscopic characteristics (texture, colour, …). Thermal denaturation of muscle proteins was studied by spectroscopy technics, able to observe their conformational modifications due to the application of several temperature/time conditions. Spectroscopy sign…