The effects of conventional industrial processing steps on global phytochemical composition of broccoli, tomato and carrot purees were investigated by using a range of complementary targeted and untargeted metabolomics approaches including LC-PDA for vitamins, (1)H NMR for polar metabolites, accurate mass LC-QTOF MS for semi-polar metabolites, LC-MRM for oxylipins, and headspace GC-MS for volatile compounds. An initial exploratory experiment indicated that the order of blending and thermal treatments had the highest impact on the phytochemicals in the purees. This blending-heating order effect was investigated in more depth by performing alternate blending-heating sequences in triplicate on the same batches of broccoli, tomato and carrot. For each vegetable and particularly in broccoli, a large proportion of the metabolites detected in the purees was significantly influenced by the blending-heating order, amongst which were potential health-related phytochemicals and flavour compounds like vitamins C and E, carotenoids, flavonoids, glucosinolates and oxylipins. Our metabolomics data indicates that during processing the activity of a series of endogenous plant enzymes, such as lipoxygenases, peroxidases and glycosidases, including myrosinase in broccoli, is key to the final metabolite composition and related quality of the purees.

The relationship between small amplitude oscillatory rheological properties and microstructure of plant cell suspensions was studied. Carrot, broccoli and tomato were selected as model plant systems to generate particles with different microstructures: clusters of cells with smooth or rough edges and single cells. By analysing the compressive stress undergone by the plant cells under centrifugation, and comparing this to oscillatory rheometry, agreement was found between the compressive stress required to compress the dispersions to higher insoluble solids dry mass fractions, and the elastic shear modulus of the plant dispersions. This indicated that centrifugation is acting as a crude rheological measurement on the samples, rather than measuring any well-defined “particle phase volume”. We estimated the theoretical critical dry mass fraction above which smooth, roughly spherical, elastically interacting particles would acquire a non-zero G′, and compared this with the experimental values. Our results give evidence that for the three vegetable suspensions considered here, the elastic rheology observed is not coming simply from the packing of smooth particles, but is dominated in the dilute limit by attractive forces or interaction of asperities, and in the concentrated limit by deformation and buckling acting together. Improved understanding of the particles and their packing would help in the structuring of food products without adding other texturising or stabilising agents.

BACKGROUND:The food industry has shown an increased interest in the manufacture of healthier and more natural food products. By tailored processing fruit and vegetables can be used as structurants thus reducing artificial gums and stabilisers. The effect of different thermal and mechanical treatments, including high-pressure homogenisation, on the microstructural and rheological properties of carrot, broccoli and tomato dispersions was studied. As part of the rheological characterisation small oscillatory deformation as well as shear flow measurements were performed.RESULTS:Carrot and broccoli showed a different behaviour from tomato under the conditions studied. Changing the order of thermal and mechanical treatment led to microstructures with different flow properties. The resulting microstructures differed in the manner of cell wall separation: either breaking across the cell walls or through the middle lamella. High-pressure homogenisation decreased the viscosity of carrot and broccoli dispersions, while it increased the viscosity of tomato. Cryo-scanning electron microscopy showed that the cell walls of carrot and broccoli remained as compact structures after homogenisation whereas tomato cell walls were considerably swollen.CONCLUSIONS:Based on the type of vegetable, the different processes applied led to microstructures with different rheological properties. This study shows that particle size distribution, morphology and phase volume are important parameters to explain the complex relationship between rheology and microstructure for these types of systems.

To help understand how sugar interactions with proteins stabilise biomolecular structures, we compare the three main hypotheses for the phenomenon with the results of long molecular dynamics simulations on lysozyme in aqueous trehalose solution (0.75 M). We show that the water replacement and water entrapment hypotheses need not be mutually exclusive, because the trehalose molecules assemble in distinctive clusters on the surface of the protein. The flexibility of the protein backbone is reduced under the sugar patches supporting earlier findings that link reduced flexibility of the protein with its higher stability. The results explain the apparent contradiction between different experimental and theoretical results for trehalose effects on proteins.

Cryo In-SEM Raman has been used for the first time to localise carotene compounds in a food matrix. Raman spectra of lycopene and β-carotene have been obtained from sampling oil droplets and plant cell structures visualised with cryo-SEM in tomato and carrot based emulsions containing 5% oil. It was possible to identify the carotenoids in both the oil droplets and the cell walls. Furthermore our results gave some indication that the carotenoids were in the non-crystalline state. It has been suggested that a higher amount of carotenes solubilised into the oil phase of the food matrix would lead to a higher bioaccessibility, thus understanding the effect of processing conditions on micronutrients distribution in a food matrix might help the design of plant based food products with a better nutritional quality. This shows improved structural characterisation of the cryo-SEM with the molecular sensitivity of Raman spectroscopy as a promising approach for complex biological problems.

UNLABELLED The correlation between food microstructure and in vitro bioaccessibility of carotenes was evaluated for tomato and carrot emulsions (5% olive oil) subjected to high pressure homogenization (HPH) at varying degrees of intensity. The aim was to investigate whether additional mechanical disruption of the food matrix could be utilized to further increase the carotene bioaccessibility of an already pre-processed material. The carotene bioaccessibility of the samples was measured after simulated in vitro digestion, carotene release to the oil phase was estimated by Confocal Raman spectroscopy and, to measure active uptake of carotenes, Caco-2 cells were incubated with the digesta of selected samples. HPH did not notably affect the retention of carotenes or ascorbic acid but significantly increased both the release and micellar incorporation of α- and β-carotene in carrot emulsions 1.5- to 1.6-fold. On the other hand, in vitro bioaccessibility of lycopene from tomato was not increased by HPH under any of the conditions investigated. Instead, the results suggested that lycopene bioaccessibility was limited by a combination of the low solubility of lycopene in dietary lipids and entrapment in the cellular network. Carotene uptake by Caco-2 cells appeared to be mainly dependent upon the carotene concentration of the digesta, but cis-trans isomerization had a significant impact on the micellarization efficiency of carotenes. We therefore conclude that HPH is an interesting option for increasing the bioaccessibility of carotenes from fruits and vegetables while maintaining a high nutrient content, but that the results will depend on both food source and type of carotene. PRACTICAL APPLICATION A better understanding of the correlation between the processing of fruits and vegetables, microstructure and nutrient bioaccessibility can be directly applied in the production of food products with an increased nutritional value.

High-pressure homogenization, as a way to further mechanically disrupt plant cells and cell walls compared to conventional blending, has been applied to thermally treated and comminuted carrot and tomato material in the presence of 5% olive oil. Mixes of both vegetables in a 1:1 ratio were also included. Both the effect of homogenization pressure and the effect of multiple process cycles were studied. The different microstructures generated were linked to different rheological properties analyzed by oscillatory and steady state measurements. The results showed that while carrot tissue requires a high shear input to be disrupted into cells and cell fragments, tomato cells were broken across the cell walls already at moderate shear input, and the nature of the tomato particles changed to amorphous aggregates, probably composed of cell contents and cell wall polymers. All the plant stabilized emulsions generated were stable against creaming under centrifugation. While for tomato a low-pressure multiple cycle and a high-pressure single-cycle process led to comparable microstructures and rheological properties, carrot showed different rheological properties after these treatments linked to differences in particle morphology. Mixes of carrot and tomato showed similar rheological properties after homogenizing in a single or in a split-stream process. Practical Application: Following consumers' demand, the food industry has shown a growing interest in manufacturing products free of gums and stabilizers, which are often perceived as artificial. By tailored processing, fresh plant material could be used to structure food products in a more natural way while increasing their nutritional quality.

In this study we are trying to understand the effects of aqueous sodium halide solutions on conformational properties of trialanine. We performed long-scale MD simulations of a trialanine peptide in different sodium halide solutions with varying concentrations of the salt (0.20. 0.50, 1.0 and 2.0 M). By Clustering the conformational space of the tripeptide, we show that the molecular dynamics trajectories of this molecule form only a few major conformation clusters for each of the sixteen salt solutions under study. This gives its a way to compress the information contained in a terabyte of the simulation data to a small number of representative molecular geometries. These representative structures give insights into molecular mechanism of salt effects on peptide conformations. The results show that salts are able to make significant changes to the conformational landscape of trialanine. and that sodium chloride is particularly effective. (C) 2008 Elsevier B.V. All rights reserved.

A large-scale fully-atomistic molecular dynamics simulation of poly-l-glutamate demonstrates that a small amount of sodium chloride switches the preferred conformation from an extended conformation to a compact alpha-helix.

Molecular dynamics simulations demonstrate that differences in the interaction of sodium and potassium with the carboxylate side chains of alpha-Poly-L-glutamate (alpha-PGA) have a dramatic effect on the conformational properties of the polypeptide. Potassium ions cluster mainly in the second and third solvation shells of alpha-PGA because their low charge density makes the electrostatic interactions between them and alpha-PGA too weak for K* to compete with water for the first solvation shell of the alpha-PGA glutamic acid residuals. Unlike sodium ions, they do not switch the conformation of alpha-PGA from extended to alpha-helical. Potentials of mean force for pure water, sodium ion solutions, and potassium ion solutions show marked differences in ion association behavior. This supports the idea that Hofmeister effects depend upon direct ion-macromolecule interactions as well as interactions with water molecules in the first solvation shell rather than bulk water structuring.

In this work we are trying to gain insight into the mechanisms of ion-protein interactions in aqueous media at the molecular scale through fully atomistic molecular dynamics simulations. We present a systematic molecular simulation study of interactions of sodium and halide ions with a trialanine peptide in aqueous sodium halide solutions with different salts concentrations (0.20, 0.50, 1.0 and 2.0 M). Each simulation covers more than fifty nanoseconds to ensure the convergence of the results and to enable a proper determination of the tripeptide-ion interactions through the potentials of mean force. Changes in ion densities in the vicinity of different peptide groups are analysed and implications for the tripeptide conformations are discussed.

In this work we are trying to gain an insight on the molecular mechanisms of the salt effects on conformational stability of proteins with use of fully atomistic Molecular Dynamics simulations techniques. Such 'in silico' approach allows us to obtain quite realistic data on the time and scale resolutions that are unavailable for both 'in vitro' and 'in vivo' experimental techniques. We investigated a trialanine peptide which is the one of the simplest examples of biomolecules, bearing the essential features of proteins.

Fourier transform infrared (FTIR) spectroscopy, in combination with multivariate data analysis (multivariate curve resolution, MCR) is used to determine the phase compositions of bulk separated agarose/maltodextrin mixtures. Much higher concentrations of maltodextrin are found in the agarose rich phases than would be expected from the cloud point curve determined for this system. It is also shown that tie-line behaviour depends on the initial maltodextrin concentration. High performance size exclusion chromatography measurements on the separated phases suggest that this is due to a strong molar mass fractionation of maltodextrin during the phase separation.