Pascale Goupy

Enzymatic browning reactions in apple and apple products

This review examines the parameters of enzymatic browning in apple and apple products that is, phenolic compounds, polyphenoloxidases, and other factors (ascorbic acid and peroxidases), both qualitatively and quantitatively. Then the relationships between intensity of browning and the browning parameters are discussed, including a paragraph on the methods used for browning evaluation. Finally, the different methods for the control of browning are presented.

Antioxidant composition and activity of barley (Hordeum vulgare) and malt extracts and of isolated phenolic compounds

Phenolics have been identified and quantified in nine varieties of barley and their corresponding malts as flavan-3-ols, flavonols, phenolic acids and apolar esters. Flavan-3-ols are monomers, (+)-catechin and (−)-epicatechin, and polymers constituted mainly by units of (+)-catechin and (+)-gallocatechin. The most abundant compounds were the dimers procyanidin B3 and prodelphinidin B3. The main trimeric procyanidin was procyanidin C2. After malting, the phenolic content decreased for all varieties. Catechin monomers were the most affected. Beside polyphenols, barley and malt extracts contained other antioxidants: carotenoids (lutein and zeaxanthin) and tocopherols (α, δ and γ). The antioxidant activity was measured using three methods: capacity to react with DPPH. (ARP), inhibition of lipoxygenase activity (LoxI) and inhibition of cooxidation of β-carotene in a linoleate model system (AOP). The inhibition of cooxidation of β-carotene in a linoleate model system did not allow varieties to be discriminated. They all have high antioxidative properties. Using this assay, tocopherols were the best antioxidants. The ARP (antiradical power) was correlated positively with the amount of total flavan-3-ols (ru2009=u20090.89) and increased with the degree of polymerisation. The LoxI assay allowed discrimination of the nine varieties of barley and their corresponding malts but was not correlated with any compound, although flavan-3-ols were good inhibitors of lipoxygenase activity. n n© 1999 Society of Chemical Industry

Intestinal scavenger receptors are involved in vitamin K1 absorption

Background: Vitamin K intestinal absorption is poorly understood. Results: Modulating SR-BI and CD36 expression or function in cells and in mice led to control of vitamin K1 intestinal transport. Conclusion: Intestinal scavenger receptors impact vitamin K1 transport through the enterocyte. Significance: Understanding vitamin K1 intestinal absorption is crucial to maintain an optimal vitamin K status. Vitamin K1 (phylloquinone) intestinal absorption is thought to be mediated by a carrier protein that still remains to be identified. Apical transport of vitamin K1 was examined using Caco-2 TC-7 cell monolayers as a model of human intestinal epithelium and in transfected HEK cells. Phylloquinone uptake was then measured ex vivo using mouse intestinal explants. Finally, vitamin K1 absorption was compared between wild-type mice and mice overexpressing scavenger receptor class B type I (SR-BI) in the intestine and mice deficient in cluster determinant 36 (CD36). Phylloquinone uptake by Caco-2 cells was saturable and was significantly impaired by co-incubation with α-tocopherol (and vice versa). Anti-human SR-BI antibodies and BLT1 (a chemical inhibitor of lipid transport via SR-BI) blocked up to 85% of vitamin K1 uptake. BLT1 also decreased phylloquinone apical efflux by ∼80%. Transfection of HEK cells with SR-BI and CD36 significantly enhanced vitamin K1 uptake, which was subsequently decreased by the addition of BLT1 or sulfo-N-succinimidyl oleate (CD36 inhibitor), respectively. Similar results were obtained in mouse intestinal explants. In vivo, the phylloquinone postprandial response was significantly higher, and the proximal intestine mucosa phylloquinone content 4 h after gavage was increased in mice overexpressing SR-BI compared with controls. Phylloquinone postprandial response was also significantly increased in CD36-deficient mice compared with wild-type mice, but their vitamin K1 intestinal content remained unchanged. Overall, the present data demonstrate for the first time that intestinal scavenger receptors participate in the absorption of dietary phylloquinone.

New approaches for separating and purifying apple polyphenol oxidase isoenzymes: Hydrophobic, metal chelate and affinity chromatography

Abstract Apple polyphenol oxidase (PPO) was subjected to hydrophobic, metal chelate and affinity chromatography. Among numerous hydrophobic supports, Phenyl-Sepharose CL4B appeared to be the most appropriate matrix for the purification of highly hydrophobic proteins such as PPO. With immobilized copper affinity chromatography, four fractions were obtained. Using electrophoresis experiments, it was shown that these fractions differed from the isoenzymes separated by ion exchange on DEAE-Sepharose CL6B. Apple PPO was also adsorbed on synthesized affinity resins with competitive inhibitors ( p -coumaric and p -hydroxybenzoic acid) coupled via an azo linkage to hexamethylenediamineagarose. These affinity matrices were also used to evaluate the inhibition constants of the ligands. Lastly, apple PPO was successively chromatographed on hydrophobic, metal chelate and affinity columns. This protocol led to a 280-fold purified fraction representing 25% of the crude extract activity.

Increased diffusivity of lycopene in hot break vs. cold break purees may be due to bioconversion of associated phospholipids rather than differential destruction of fruit tissues or cell structures

Lycopene bioaccessibility is enhanced by processing, as explained by the destructuration of plant tissues, making diffusion easier. However, in tomato, the relationship between grinding intensity and lycopene release from purees suffers from uncertainty. In particular, hot break puree exhibited twice as much diffusible lycopene as compared to cold break, while both were processed with the same grinding intensity. To explain the difference, we systematically studied the diffusivity of particles according to their size and integrity, and used microscopic and physical analyses to reveal structural differences. Neither particle size distribution, nor cell destruction, nor plastid transformation exhibited any correlation to the differences in diffusivity. However, Raman microspectroscopy combined with a chemometric analysis revealed significant changes in lycopene spectra and a putative linkage to phospholipid transformation. Phospholipid profiling of five pairs of contrasted purees revealed that, during the cold break, a transition from complex phospholipids to more simple phosphatidic acid molecules systematically occurred.

Opposite Effects of the Spinach Food Matrix on Lutein Bioaccessibility and Intestinal Uptake Lead to Unchanged Bioavailability Compared to Pure Lutein

SCOPEnFood matrix is generally believed to alter carotenoid bioavailability, but its effect on xanthophylls is usually limited. This study thus aims to decipher the digestion-absorption process of lutein in the presence or not of a food matrix.nnnMETHODSnLutein transfer to gastric-like lipid droplets or artificial mixed micelles was assessed when lutein was added to test meals either as a pure molecule ((all-E)-lutein) or in canned spinach ((Z) + (all-E)-lutein). The obtained mixed micelles were delivered to Caco-2 cells to evaluate lutein uptake. Finally postprandial plasma lutein responses were compared in minipigs after the two test meals.nnnRESULTSnLutein transfer to gastric-like lipid droplets and to mixed micelles was higher when lutein was added in spinach than when it was added as pure lutein (+614% and +147%, respectively, pxa0<xa00.05). Conversely, lutein uptake was less effective when micellar lutein was from a meal containing spinach than from a meal containing its pure form (-55%, pxa0<xa00.05). In minipigs, postprandial lutein response was delayed with spinach but not significantly different after the two test meals.nnnCONCLUSIONnOpposite effects at the micellarization and intestinal cell uptake steps explain the lack of effect of spinach matrix on lutein bioavailability.

Carotenoids: Experimental Ionization Energies and Capacity at Inhibiting Lipid Peroxidation in a Chemical Model of Dietary Oxidative Stress

Carotenoids are important natural pigments and micronutrients contributing to health prevention by several mechanisms, including their electron-donating (antioxidant) activity. In this work, a large series of carotenoids, including 11 carotenes and 14 xanthophylls, have been investigated by wavelength-resolved atmospheric pressure photoionization mass spectrometry (DISCO line of SOLEIL synchrotron), thus allowing the experimental determination of their ionization energy (IE) for the first time. On the other hand, the carotenoids have been also investigated for their ability to inhibit the heme iron-induced peroxidation of linoleic acid in mildly acidic micelles, a simple but relevant chemical model of oxidative stress in the gastric compartment. Thus, the carotenoids can be easily classified from IC50 concentrations deduced from the time dependence of the lipid hydroperoxide concentration. With a selection of two carotenes and three xanthophylls a quantitative analysis is also provided to extract stoichio-kinetic parameters. The influence of the carotenoid structure (number of conjugated carbon-carbon double bonds, presence of terminal six-membered rings, hydroxyl, keto, and/or epoxy groups) on the IE, IC50, and stoichio-kinetic parameters is discussed in details. The data show that the antioxidant activity of carotenes is well correlated to their electron-donating capacity, which itself largely depends on the length of the conjugated polyene chain. By contrast, the IE of xanthophylls is poorly correlated to the polyene chain length because of the strong, and sometimes unexpected, electronic effects of the O-atoms. Although IE remains an approximate predictor of the antioxidant activity of xanthophylls, other factors (interaction with the aqueous phase, competing radical-scavenging mechanisms, the residual activity of the antioxidants oxidation products) probably play a significant role.