Structure and chemical composition of layers adsorbed at interfaces with champagne.

The structure and the chemical composition of the layer adsorbed at interfaces involving champagne have been investigated using native champagne, as well as ultrafiltrate (UFch) and ultraconcentrate (UCch) obtained by ultrafiltration with a 10(4) nominal molar mass cutoff. The layer adsorbed at the air/liquid interface was examined by surface tension and ellipsometry kinetic measurements. Brewster angle microscopy demonstrated that the layer formed on polystyrene by adsorption or drop evaporation was heterogeneous, with a domain structure presenting similarities with the layer adsorbed at the air/liquid interface. The surface chemical composition of polystyrene with the adlayer was determined by X-ray photoelectron spectroscopy (XPS). The contribution of champagne constituents varied according to the liquid (native, UFch, and UCch) and to the procedure of adlayer formation (evaporation, adsorption, and adsorption + rinsing). However, their chemical composition was not significantly influenced either by ultrafiltration or by the procedure of deposition on polystyrene. Modeling this composition in terms of classes of model compounds gave approximately 35% (w/w) of proteins and 65% (w/w) of polysaccharides. In the adlayer, the carboxyl groups or esters represent about 18% of carbon due to nonpolypeptidic compounds, indicating the presence of either uronic acids in the complex structure of pectic polysaccharides or of polyphenolic esters. This structural and chemical information and its relationship with the experimental procedures indicate that proteins alone cannot be used as a realistic model for the macromolecules forming the adsorption layer of champagne. Polysaccharides, the other major macromolecular components of champagne wine, are assembled with proteins at the interfaces, in agreement with the heterogeneous character of the adsorbed layer at interfaces.

Polyphenol-beta-casein complexes at the air/water interface and in solution: effects of polyphenol structure.

The interactions between proteins and plant polyphenols are responsible for astringency and haze formation in beverages and may participate in foam stabilization. The effect of phenolic compounds with different structures, namely, catechin (C), epicatechin (Ec), epigallocatechin (Egc), epicatechin gallate (EcG), and epigallocatechin gallate (EgcG), on the surface properties at the air/liquid interface of beta-casein, chosen as model protein, were monitored by tensiometry and ellipsometry. The formation of complexes in the bulk phase was measured by electrospray ionization mass spectrometry (ESI-MS). Adsorption of polyphenols from pure solution was not observed. Surface pressure, surface concentration, and dilational modulus of the protein adsorption layer were greatly modified in the presence of galloylated flavanol monomers (EcG and EgcG) but not of lower molecular weight polyphenols (<306 g/mol). The formation of polyphenol-protein aggregates in the bulk, as evidenced by ESI-MS and light scattering experiments, was related to the slowdown of protein adsorption.

Characterization by optical measurements of the effects of some stages of champagne technology on the adsorption layer formed at the gas/wine interface.

This study analyzes the effects of some important factors of champagne technology on the ellipticity and Brewster angle microscopy (BAM) of the air/champagne interface in view of using the optical properties of the adsorption layer of base wine to forecast the stability of the champagne bubble collar. Using standard, ultrafiltered, and ultraconcentrated wines it was observed that champagne can lose amphiphilic macromolecules which adsorb on the inner glass wall of the bottle during storage, particles such as dead yeasts can adhere to the adsorption layer, a weak increase of the ethanol content during bottle fermentation can reduce significantly the ellipticity of the adsorption layer, and CO2 has no significant effect on the properties of that layer. Surprisingly, no visible differences of the adsorption layer were noticed between the experimental champagnes of the 2004 vintage of three vine varieties (Chardonnay, Pinot noir, and Pinot meunier). From analysis of all samples it is proposed that the mean value and standard deviation of the ellipticity measured during 30 min after pouring the wine in a Petri dish are physical quantities which satisfactorily characterize the adsorption layer of champagne. When needed, further characterization of the adsorption layer may be obtained by a detailed analysis of the kinetics of ellipticity during the same period and inspection of the BAM images of the interface.

Effect of sucrose on the properties of caffeine adsorption layers at the air/solution interface.

Sweet and bitter tastes are known to be mediated by G-protein-coupled receptors. The relationship between the chemical structure of gustable molecules and their molecular organization in saliva (aqueous solution) near the surface of the tongue provides a useful tool for elucidating the mechanism of chemoreception. The interactions between stimulus and membrane receptors occur in an anisotropic system. They might be influenced by the molecular packing of gustable molecules within an aqueous solvent (saliva) close to the receptor protein. To investigate the molecular organization of a sweet molecule (sucrose), a bitter molecule (caffeine), and their mixture in an aqueous phase near a "wall", a hydrophobic phase, we modeled this using an air/liquid interface as an anisotropic system. The experimental (tensiometry and ellipsometry) data unambiguously show that caffeine molecules form an adsorption layer, whereas sucrose induces a desorption layer at the air/water interface. The adsorption of caffeine molecules at the air/water interface gradually increases with the volume concentration and is delayed when sucrose is added to the solution. Spectroscopic ellipsometry data show that caffeine in the adsorption layer has optical properties practically identical to those of the molecule in solution. The results are interpreted in terms of molecular association of caffeine with itself at the interface with and without sucrose in the subphase, using the theory of ideal gases.

Phase separation in molecular layers of macromolecules at the champagne-air interface.

Bubble and foam stability, which are essential for the hallmark of champagne, rely on the concentration of amphiphilic macromolecules originating from the grape, which form molecular layers at the interface between champagne and gas. Ellipsometry and Brewster angle microscopy experiments were conducted at the air-champagne interface to analyse the lateral organization of the layers of macromolecules. Several kinds of phase separations - leading in some cases to two-dimensional foams - were identified. At the beginning of layer formation, condensed domains develop at the expense of dilute domains. Thereafter, phase separations occur within the condensed domains. These findings may allow advances in the implementation of methods predicting bubble and foam stability of champagnes.

Influence of lysozyme treatments on champagne base wine foaming properties.

The objective of this study was to estimate the effect of lysozyme on the foaming characteristics of Champagne base wine. Lysozyme additions were made to the musts and also to the wines before and after bentonite or charcoal treatments, which remove endogenous proteins. Treatments with bentonite diminished foamability and foam stability of wines, whatever the dose (30 or 80 g/hL) and variety [Chardonnay, -28%; Pinot noir, -20% (at 30 g/hL)]. An addition of lysozyme in must raised Pinot noir wine foamability by 21%, whereas the difference is hardly perceptible for Chardonnay wine (+3%). Pinot noir and Chardonnay wines, originating from lysozyme-treated musts, in addition to bentonite treatment on the wine, presented higher foamability than wines treated only with bentonite. Lysozyme was removed (91-100%) by the bentonite treatment. Then, it was not responsible for the increase in foamability but seemed to have a protective effect on the wine proteins. When wines were initially treated with bentonite (150 g/hL) and then enriched with 80 g/hL lysozyme, this enzyme was not able to restore foaming properties. Treatments with charcoal always diminished foamability. The average increase in foamability due to an addition of lysozyme after charcoal treatment (80 g/hL) was 23%. Results showed a real positive effect of lysozyme on foam stability when wines have to be treated with charcoal (+25% and +56% for the Pinot noir wine and the Chardonnay wine, respectively, at 30 g/hL).

Characterisation by drop tensiometry and by ellipsometry of the adsorption layer formed at the air/champagne wine interface.

A foam ring composed of small bubbles on the surface of a champagne glass is one of its hallmarks. The equilibrium state of that ring is linked with the rate of formation and of disappearance of bubbles. The stability of bubbles is usually ascribed to the occurrence and to the properties of an adsorption layer formed at the gas/liquid interface. Our goal is to characterise such an adsorption layer at the gas/wine interface in order to understand its role in bubble stability. Alcohol in wine lowers the surface tension to 49 mN/m. The adsorption of other molecules may cause a further decrease of 2 mN/m. Such a situation makes the study of adsorption by surface tension measurement inaccurate. To overcome this problem, we have diluted the wine four times with water before its surface tension measurement by pendant drop shape analysis. In these conditions, ethanol lowers the surface tension to 64 mN/m and the adsorption of other molecules of the wine can be monitored over 6-8 mN/m. The usual behaviour of such a diluted wine is a lowering of the surface tension during at least 20 min after drop formation. Since the role of macromolecules on the foaming properties of wine had been previously observed, we have chosen to evaluate the effect of this fraction of the wine molecules on its surface properties. Thus, wines were ultrafiltrated on a membrane with a 10000 molecular mass cut-off. The ultrafiltrate (UF) does not show any decrease of its surface tension over a 20-min period while the ultraconcentrate (UC) has a kinetics similar to that of unfiltered wine. Mixtures of UF and UC have behaviours intermediate between those of these products. A technological treatment of the wine with bentonite, believed to lower the content of macromolecules, yields a wine similar to UF. The effect of ultrafiltration was also analysed by spectroscopic ellipsometry. UF has a spectrum similar to that of a water/alcohol mixture with the same ethanol content and its ellipticity is stable during at least 20 min. On the contrary, wine or UC show spectra with the features of an adsorption layer and those characteristics increase during more than 20 min. Two varieties of vine were compared: 'Chardonnay' and 'Pinot noir'. The former is known to have better foaming properties than the latter. Its surface properties measured in this study are also more pronounced than those of Pinot noir. However, the representation of the dilational modulus against the surface pressure (which, in some instances, may be a mathematical transformation of the state equation) puts all the samples (wines, UF and UC of each) on the same master curve, a fact in favour of a common nature for all the adsorption layers. It can be concluded that surface properties of champagne wines are mostly determined by ethanol and by macromolecules with a molecular mass larger than 10000. Moreover, the adsorption layers seem to have the same nature, irrespective of the vine variety and of the concentration ratio of the wine.

Asymmetric Multiblock Copolymers at the Gas-Liquid Interface: Phase Diagram and Surface Pressure.

A theoretical model of copolymers made of N blocks is studied at the air-water interface. Each block is made of a sequence A of ZA hydrophobic and of a sequence B of ZB hydrophilic monomers. The A and B sequences cannot cross the interface. The conformation of an adsorbed polymer is determined as a random walk of N elements whose size is the Flory radius of a single sequence. The structure of the interfacial layer is determined as a function of alpha = ZA/ZB and of the surface concentration using scaling law arguments. Only three different regions are found in the phase diagram to describe the change of surface regime as a function of the total surface concentration. The energy of flower-like micelles of polymers is calculated and compared with the energy of adsorbed macromolecules in order to determine the surface concentration at saturation. The surface pressure is also calculated as a function of the surface concentration in the three different regions of the phase diagram. It is found that these surface pressure isotherms are not affected by the solvent quality except when the properties of the interfacial layer are dominated by a purely two-dimensional behavior (semidiluted regime of the whole polymer or of the A sequences on the air side of the interface). Finally the properties of this model are compared with experimental data obtained with protein adsorbed layers and encouraging agreement is found although proteins are much more complicated polymers than this crude model. Copyright 1999 Academic Press.

Effect of Ethanol on the Structure and Properties of beta-Casein Adsorption Layers at the Air/Buffer Interface.

The adsorption of beta-casein at the air-solution interface has been monitored in equilibrium conditions by neutron reflectivity. It was observed that for a bulk concentration of 100 mg/L, the amount of protein adsorbed per unit surface increases from 2.8 to 4.4 mg/m2 when the ethanol concentration in the bulk changes from 0 to 20% (v/v). Surface pressure measurements on aqueous solutions indicate that the surface pressure is higher when both protein and alcohol are added than when a single substance is in the solution. The addition of protein has an effect when the alcohol concentration is less than 20%. These results are consistent with the occurrence at the interface of a protein network leaving a surface fraction available for ethanol. A thermodynamic model has been developed using scaling law arguments to model the surface pressure and dilational modulus measurements. It introduces an exponent which is characteristic of the solvent "quality" and of the structure of the interfacial layer. The results are interpreted as showing that ethanol modifies the solvent properties, the interactions between the protein and the solvent, and the structure of the adsorption layer. The main transition seems to occurr at 6% ethanol. Copyright 1998 Academic Press.

A structural study of beta-casein adsorbed layers at the air-water interface using X-ray and neutron reflectivity.

New details on the structure of beta-casein adsorbed layers, at the air-water interface, have been obtained using X-ray and neutron reflectivity. The experimental data are fitted well by a power law model and the results discussed in terms of the distribution of amino-acid sequences between trains, loops and tails. This distribution seems to be consistent with statistical theories established for flexible polymers. The trains are present in close proximity to the surface as a dense layer 8-9 A thick. At low surface coverage, the tail effect is negligible and the adsorbed layer is composed of nearly 60% amino-acid sequences in trains and the remaining in loops. When the bulk concentration is increased, a substantial part of the amino-acid residues has to be accommodated in loops and long tails; the adsorbed layer becomes more extended (80-100 A). A striking feature is observed for a high bulk concentration (10(-1) wt.%): trains are forced to eject out of the interface.

Thermal denaturation and gelation of rubisco: effects of pH and ions.

In order to understand the mechanism of thermal gelation of rubisco, its native and heat denatured states were characterized by absorbance, fluorescence and circular dichroïsm spectroscopies as well as by differential scanning calorimetry in the presence of various salts. It appears that during the denaturation process, divalent anions are released while divalent cations are fixed by the protein, while it is disorganized and while the environment of its aromatic chromophores becomes more hydrophilic. The pH transition of gelation is shifted 1-2 pH units higher than the transition of denaturation temperature which occurs near the isoelectric point of the native molecule. This shift probably corresponds to the breaking of saline bridges within the protein molecule. Finally, a large effect of divalent cations on the phase diagram indicates that a particular denatured state is attained when these cations are in the denaturation medium.

Differential scanning calorimetric studies of the effects of ions and pH on ribulose 1,5-bisphosphate carboxylase/oxygenase.

Lucerne rubisco (ribulose 1,5 bisphosphate carboxylase/oxygenase, EC 4.1.1.39) was purified by ammonium sulfate fractionation and gel chromatography. Differential scanning calorimetry (d.s.c.) was used to study the effects of pH in the range 3.5 to 11.4, and the effects, at pH 7.5, of various salts at ionic strength lower than 0.3 (NaH2PO4, (NH4)2SO4, Na2SO4, NaCl, MgCl2, CaCl2) on the thermal denaturation of the protein. Van't Hoff and calorimetric enthalpies, and the change in heat capacity between the native and denatured states were calculated from the experimental data. The effects of salts on the thermal denaturation seem to follow the Hofmeister series. In some cases, the thermal denaturation may be interpreted as a two-step transition of the polypeptide chains. Moreover, comparison of the results with literature data suggests that the thermal denaturation parameters depend on the botanical origin of rubisco and are affected by the conditions of its purification.

[Lipoxygenase activity of wheat seeding chloroplasts]. / Activité lipoxygénasique de chloroplastes de plantules de Blé.

Chloroplasts have been isolated from Wheat shoots by density gradient centrifugation. When added to a linoleic acid and Tween 20 suspension these chloroplasts induce an oxygen consumption. The hydroperoxides synthesized from linoleic and oxygen have been characterized by spectrophotometry and thin layer chromatography.