Jean-Paul Douliez

Long-chain fatty acids are insoluble in aqueous solution and form crystal precipitates. It is then of particular importance to determine the physicochemical parameters allowing their dispersion in water to improve their bioavailability and their utilization as surfactants. Herein, we report a study on salt-free catanionic systems in aqueous solution made of mixtures between palmitic or stearic fatty acids and alkylboladiamines (Abd's) differing by their alkyl chain length. Phase contrast microscopy, solid-state NMR, Fourier transform infrared spectroscopy, and small-angle neutron scattering were used to characterize the phase behavior of these systems at molar ratio of fatty acid to Abd of 1 and 2. Whatever the Abd and the molar ratio, fatty acids were embedded at low temperature in a bilayer gel phase which crystallizes after a period of rest. At an equimolar ratio, the gel phases transited upon raising the temperature to an isotropic phase made of wormlike micelles except in the case of the ethylenediamine chain for which a lamellar fluid phase was observed. At a molar ratio of 2 and high temperature, fatty acids were embedded in a lamellar fluid phase which self-orients with its stacking axis perpendicular to the magnetic field. However, for a long alkylboladiamine such as spermine, wormlike micelles formed. The phase behavior at high temperature is discussed in terms of molecular volume.

Cones are a very rare form of supramolecular self-assembly. It is shown that mixing of an unsymmetrical bola-fatty acid and a diamine (catanionic system) in dilute solutions can produce aggregates having a conical shape. These assemblies are made of frozen unsymmetrical monolayers in which bolas are arranged on a regular hexagonal lattice. Cones are stabilized by the introduction of pentagonal or n-gonal defects in the bola-lattice which yields the quantification of the cone apex angle according to the Euler theorems. The formation of cones represents a novel route for lipid and surfactant systems to minimize the edge energy of a flat membrane.

Glycerol derivatives of cutin and suberin monomers were synthesized by acid catalysis. Their dispersion in an aqueous solution was examined by phase contrast microscopy, neutron scattering, and solid state NMR. It is shown that the phase behavior strongly depends on the nature of the derivatives forming either lumps of aggregated membranes or well dispersed membranes.

The phase behavior of mixtures of palmitic acid (PA) and 1-monohexadecanoyl-rac-glycerol, palmitin, was studied by phase contrast microscopy and deuterium solid-state NMR. At pH 5, mixtures remained precipitated as lumps in solution. The NMR spectrum of the perdeuterated PA (PAd₃₁) at 300 K exhibited a shape and quadrupolar splittings, Δν, characteristic of lipids embedded in a gel phase. The alkyl chains remained in a trans conformation with their long molecular axis oriented at about 15° with respect to the bilayer normal. However, gauche defects were shown to occur at the end of the alkyl chain. At 330 K, the system underwent a phase transition to a hexagonal phase followed by an isotropic phase at 340 K. Upon cooling to 330 K, the spectrum in the hexagonal phase was oriented at 0° showing that the cylinders were oriented with their long axis parallel to the field. Up to 11 positions (from 15) of PAd₃₁ could be assigned. At pH 7 and 9 at room temperature, the mixtures were fully dispersed in a viscous solution of vesicles. The system underwent a phase transition at 320 K from a gel phase to a fluid phase with the bilayer normal oriented at 90° with respect to the field. Analogous experiments performed with PA selectively labeled on carbon C₂ allowed for the assignment of Δν for that position and suggested different conformations of the headgroup in the gel and fluid or hexagonal phases. The implications of these findings for the bio-availability of these fatty acids, in the understanding of the contribution of hydroxyl and carboxyl groups in the membrane formation, and for the production of simple self-oriented systems are discussed.

Study of the effect of protein chemical acylation on their functional properties or activity often brings valuable information regarding structure-function relationships. We performed such work on...

Recently, this laboratory has isolated from barley and beer extract an isoform of lipid transfer protein (LTP1), which was not fully sequenced (Jégou, S.; Douliez, J. P.; Mollé, D.; Boivin, P.; Marion, D. J. Agric. Food Chem.2000, 48, 5023−5029). It was named LTP1b and exhibited a molecular weight 294 Da higher than that of the known LTP1. This paper reports the finding of an LTP1 isoform in wheat that also exhibits an excess of 294 Da compared to the native protein. Amino acid sequencing, reduction and alkylation, and mass spectrometry showed that this new LTP1b possesses the same N-terminal sequence as the native LTP1, suggesting that the difference resides in the binding of an adduct which has a molecular weight of 294 Da. The aim of the present paper is to highlight various biophysical techniques that afford the identification of such an isoform-like LTP1 and to correlate this finding with other isoforms of LTP1 that were isolated from other plants but not fully sequenced. Keywords: Plant lipid transfer protein; wheat; isoform; adduct; post-translational modification