Stability and bioaccessibility of micronutrients and phytochemicals present in processed leek and Brussels sprouts during static in vitro digestion.

Vegetables are frequently processed before consumption. However, vegetable functionalization continues beyond ingestion as the human digestive tract exposes vegetable products to various conditions (e.g. elevated temperature, pH alterations, enzymes, electrolytes, mechanical disintegration) which can affect the stability of micronutrients and phytochemicals. Besides the extent to which these compounds withstand the challenges posed by digestive conditions, it is equally important to consider their accessibility for potential absorption by the body. Therefore, this study investigated the impact of static in vitro digestion on the stability (i.e. concentration) and bioaccessibility of vitamin C, vitamin K1, glucosinolates, S-alk(en)yl-l-cysteine sulfoxides (ACSOs) and carotenoids in Brussels sprouts (Brassica oleracea var. gemmifera) and leek (Allium ampeloprasum var. porrum). Water-soluble compounds, glucosinolates and ACSOs, remained stable during digestion while vitamin C decreased by >48%. However, all water-soluble compounds were completely bioaccessible. Lipid-soluble compounds were also stable during digestion but were only bioaccessible for 26-81%.

Impact of refrigerated storage on (bio)chemical conversions of health-related compounds in pretreated, pasteurized Brussels sprouts and leek.

Vegetable processing often consists of multiple processing steps. Research mostly focused on the impact of individual processing steps on individual health-related compounds. However, there is a need for more holistic approaches to understand the overall impact of the processing chain on the health potential of vegetables. Therefore, this work studied the impact of pretreatment (relatively intact versus pureed vegetable systems), pasteurization and subsequent refrigerated storage (kinetic evaluation) on multiple health-related compounds (vitamin C, vitamin K1, carotenoids, glucosinolates and S-alk(en)yl-L-cysteine sulfoxides (ACSOs)) in Brussels sprouts and leek. It could be shown that differences introduced by different types of pretreatment were not nullified during pasteurization and refrigerated storage. Clearly, enzymatic conversions controlled during pretreatment resulted in different health-related compound profiles still observable after pasteurization. Moreover, about -42% and -100% relative concentration differences of ACSOs and dehydroascorbic acid, respectively, were detected immediately after pasteurization, while glucosinolates concentrations decreased by about 47% during refrigerated storage. All other compounds were stable during pasteurization and refrigerated storage.

The volatile profile of pasteurized leek (Allium ampeloprasum var. porrum) and Brussels sprouts (Brassica oleracea var. gemmifera) (products), as a witness to (bio)chemical reactivity, influenced by pretreatment and successive refrigerated storage.

Processing can affect (bio)chemical conversions in vegetables and can act on their volatile properties accordingly. In this study, the integrated effect of pretreatment and pasteurization on the volatile profile of leek and Brussels sprouts and the change of this profile upon refrigerated storage were investigated. Pretreatments were specifically selected to steer biochemical reactivities to different extents. Volatile profiles were analyzed by headspace-solid phase microextraction-gas chromatography-mass spectrometry. For both vegetables, it was observed that different pretreatments prior to a pasteurization step led to diverse volatile profiles. The differences in volatile profiles observed in the different samples were presumably attributed to the different degrees of enzymatic conversions, further conversions of enzymatically formed products and thermally induced reactivities. Interestingly, the observed initial relative differences between volatile profiles of differently pretreated pasteurized samples were still observed after a refrigerated storage of 4 weeks at 4 °C. In conclusion, refrigerated storage only limitedly affected the resulting volatile profile.

Bean softening during hydrothermal processing is greatly limited by pectin solubilization rather than protein denaturation or starch gelatinization.

The rate liming step of bean softening during cooking was evaluated. Red kidney beans (fresh/non-aged and aged) were cooked at different temperatures (70-95 °C) and their texture evolution established. Softening of beans (loss of hard texture) with cooking and increasing cooking temperature was evident at ≥ 80 °C more so for non-aged than aged beans, evidencing hard-to-cook development during storage. Beans at each cooking time and temperature were subsequently classified into narrow texture ranges and bean cotyledons in the most frequent texture class evaluated for the extent of starch gelatinization, protein denaturation and pectin solubilization. During cooking, starch gelatinization was shown to precede pectin solubilization and protein denaturation, with these reactions progressing faster and to a greater extent with increasing cooking temperature. At 95 °C for instance (practical bean processing temperature), complete starch gelatinization and protein denaturation is attained earlier (∼10 and 60 min cooking, respectively and at comparable time moments for both non-aged and aged beans) than plateau bean texture (∼120 and 270 min for non-aged and aged beans)/plateau pectin solubilization. The extent of pectin solubilization in the cotyledons was consequently most correlated (negatively, r = 0.95) with and plays the most significant role (P < 0.0001) in directing the relative texture of beans during cooking. Ageing was shown to significantly retard bean softening. Protein denaturation plays a less significant role (P = 0.007) while the contribution of starch gelatinization is insignificant (P = 0.181). Thermo-solubilization of pectin in bean cotyledons is therefore the rate limiting step of bean softening towards attaining a palatable texture during cooking.

Phytate and mineral profile evolutions to explain the textural hardening of common beans (Phaseolus vulgaris L.) during postharvest storage and soaking: Insights obtained through a texture-based classification approach.

During adverse postharvest storage of Red haricot beans, the inositol phosphate content, particularly InsP6, decreased significantly, along with a significant increase in InsP5. Using a texture-based classification approach, the InsP6 content in cotyledons was shown an indicator for the extent of hard-to-cook (HTC) development during bean aging. This textural defect development was predominated by storage-induced InsP6 degradation, rather than phytate interconversions during soaking. Ca cations, released during storage, did not leach out significantly during subsequent soaking, suggesting that they were bound with the cell wall pectin in cotyledons, while Mg cations were mostly leached out into the soaking water due to their weak binding capacity to the pectin, and the cell membrane damages developed during HTC. Results obtained herein provide evidence for the pectin-cation-phytate mechanism in textural hardening (and its distribution after cooking) of common beans, and call for a more detailed Ca-relocation study during postharvest storage, soaking and cooking.

The role of mechanical collapse by cryogenic ball milling on the effect of high-pressure homogenization on the microstructural and texturizing properties of partially pectin-depleted tomato cell wall material.

In the current study, the effect of different particle size reduction techniques, namely high-pressure homogenization (HPH) and cryogenic ball milling (CBM), on the microstructural and texturizing properties of the tomato acid-unextractable fraction (AcUF) in suspension was studied. Partial pectin depletion was performed by nitric acid pectin extraction on the alcohol-insoluble residue. In the absence of the aforementioned mechanical treatments, the partially pectin-depleted material, i.e., the AcUF, showed a cellular morphology and a high texturizing potential. By short-time CBM in dry-state, the AcUF was extensively fractured and clumped, resulting in a collapsed structure with negligible texturizing potential and low water binding capacity. In contrast, HPH could disrupt the cell wall network (destroying the cellular morphology) resulting in a continuum of interacting material having very similar texturizing potential and a slightly higher water binding capacity than the AcUF before HPH. Furthermore, the potential of HPH to (re)functionalize the collapsed cryogenic ball milled AcUF by its shear-induced disruption was shown. Indeed, the debris-like cell wall remnants could to some extent be reopened by HPH, which resulted in a partial recovery of the original texturizing potential and an improved water binding capacity. However, the potential of HPH at 20 MPa to revert the detrimental effect of CBM decreased with increasing CBM treatment time.

Production and molecular characterization of tailored citrus pectin-derived compounds.

In this study, tailored-made citrus pectin-derived compounds were produced through controlled enzymatic and/or chemical modifications of commercial citrus pectin with different degrees of methylesterification (DM) and similar average molecular weight (MW). In the first treatment, degradation of the citrus pectin (CP) materials by endo-polygalacturonase (EPG) yielded pectins with average Mw's (between 2 and 60 kDa). Separation and identification of the oligosaccharide fraction present in these samples, revealed the presence of non-methylesterified galacturonic acid oligomers with degree of polymerization (DP) 1-5. In the second treatment, exploiting the combined effect of EPG and pectin lyase, compounds with MW between 2 and 21 kDa, containing methylesterified and non-methylesterified polygalacturonans (DP 1-6), were generated. Finally, CP was sequentially modified by chemical saponification and the action of EPG. A sample of DM 11% and MW 2.7 kDa, containing POS (DP 1-5), was produced. Diverse pectin-derived compounds were successfully generated for further studies exploring their functionality.

Thermal treatment of common beans (Phaseolus vulgaris L.): Factors determining cooking time and its consequences for sensory and nutritional quality.

Over the past years, the shift toward plant-based foods has largely increased the global awareness of the nutritional importance of legumes (common beans (Phaseolus vulgaris L.) in particular) and their potential role in sustainable food systems. Nevertheless, the many benefits of bean consumption may not be realized in large parts of the world, since long cooking time (lack of convenience) limits their utilization. This review focuses on the current insights in the cooking behavior (cookability) of common beans and the variables that have a direct and/or indirect impact on cooking time. The review includes the various methods to evaluate textural changes and the effect of cooking on sensory attributes and nutritional quality of beans. In this review, it is revealed that the factors involved in cooking time of beans are diverse and complex and thus necessitate a careful consideration of the choice of (pre)processing conditions to conveniently achieve palatability while ensuring maximum nutrient retention in beans. In order to harness the full potential of beans, there is a need for a multisectoral collaboration between breeders, processors, and nutritionists.

Microscopic evidence for pectin changes in hard-to-cook development of common beans during storage.

In this study, pectin changes during Red haricot bean storage under high temperature and high humidity conditions were investigated to understand the hard-to-cook (HTC) development from a microstructural point of view. First, to ensure repeatability of the microscopy results, a classification of the fresh and stored beans (aged at 35 °C and 83% relative humidity) into different hardening levels (the Non-aged, Aged and Very-hard aged sample) was performed based on the texture values of cooked half-cotyledons. Cell wall strength of the cotyledons was evaluated, showing that the aged samples (HTC seeds) exhibit stronger cell walls with more/stronger pectic cross-linkages than the Non-aged sample. After a sequential pectin extraction aiming at removing pectin fractions of different solubility, cell wall autofluorescence and immunolabeling of JIM7, LM9 and 2F4 epitopes in the residual materials were examined. Upon ageing, the samples exhibited an increased Ca2+-pectin and ferulic acid-pectin crosslinking, these pectic complexes being accumulated primarily at the intercellular spaces. The results suggest a contribution of both the pectin-cation-phytate hypothesis and the involvement of phenolic-pectin crosslinks in HTC development at the cotyledon during storage of common beans.

Arabinoxylan, ß-glucan and pectin in barley and malt endosperm cell walls: a microstructure study using CLSM and cryo-SEM.

The architecture of endosperm cell walls in Hordeum vulgare (barley) differs remarkably from that of other grass species and is affected by germination or malting. Here, the cell wall microstructure is investigated using (bio)chemical analyses, cryogenic scanning electron microscopy (cryo-SEM) and confocal laser scanning microscopy (CLSM) as the main techniques. The relative proportions of ß-glucan, arabinoxylan and pectin in cell walls were 61, 34 and 5%, respectively. The average thickness of a single endosperm cell wall was 0.30 µm, as estimated by the cryo-SEM analysis of barley seeds, which was reduced to 0.16 µm after malting. After fluorescent staining, 3D confocal multiphoton microscopy (multiphoton CLSM) imaging revealed the complex cell wall architecture. The endosperm cell wall is composed of a structure in which arabinoxylan and pectin are colocalized on the outside, with ß-glucan depositions on the inside. During germination, arabinoxylan and ß-glucan are hydrolysed, but unlike ß-glucan, arabinoxylan remains present in defined cell walls in malt. Integrating the results, an enhanced model for the endosperm cell walls in barley is proposed.

Cotyledon pectin molecular interconversions explain pectin solubilization during cooking of common beans (Phaseolus vulgaris).

Dynamics of pectin extractability in cotyledons and seed coats were explored for mechanistic insight into pectin changes due to aging and cooking of beans. In addition, changes in mineral distribution during cooking were determined in order to investigate their retention in the matrix. Pre-soaked fresh and aged beans were cooked in demineralized water for different times and the cotyledons, seed coats and cooking water were lyophilized. From cotyledon and seed coat powders, alcohol insoluble residue (AIR) was extracted and sequentially fractionated into water-, chelator- and sodium carbonate-extractable pectin (WEP, CEP and NEP, respectively). Characterization of pectin in AIR and pectin fractions revealed inherent structural differences between cotyledon and seed coat pectin with the latter exhibiting a lower degree of methylesterification (DM) and being more linear. Due to aging, WEP decreased whilst NEP substantially increased and the CEP fraction and DM of pectin in AIR did not change significantly, suggesting a more crucial role of increased covalent bonding than cation-mediated crosslinking in aging-induced hardening of beans. During cooking, some NEP was converted into WEP and no pectin depolymerization was observed from molar mass distribution profiles. Pectin changes due to aging and cooking of beans were more pronounced in the cotyledon compared to the seed coat. Whilst Ca2+, Fe2+ and Zn2+ were largely retained in the bean matrix during cooking, Mg2+ was largely leached from cotyledons into the cooking water. In conclusion, aging-induced hardening of beans and softening during cooking were found to be premised on interconversion of pectin fractions in cotyledons.

Lipid nanoparticles with fats or oils containing ß-carotene: Storage stability and in vitro digestibility kinetics.

The aim of this work was to study the formation of lipid nanoparticles (LNPs) with low (corn and olive oil) or high temperature melting lipids (cocoa butter and hydrogenated coconut oil). Moreover, their ß-carotene stability and in vitro digestibility kinetics was evaluated. Submicron LNPs (d43 ≈ 570-780 nm) were stabilized with Tween 80 at a surfactant-to-oil ratio (SOR) of 0.1. It was reduced below 200 nm at an SOR of 1. The ß-carotene retention at 25 °C was not related to the lipid type but rather to the particle size, being lower in samples with smaller particle sizes. Cocoa butter LNPs presented an equally complete digestion as corn oil LNPs and a high ß-carotene bioaccessibility, which was related to the high degree of micellarization of monoacylglycerols. This work evidences the potential of LNPs to protect lipophilic bioactive compounds with a high digestibility and bioaccessibility.

Impact of different sequences of mechanical and thermal processing on the rheological properties of Porphyridium cruentum and Chlorella vulgaris as functional food ingredients.

Microalgae are a promising and sustainable source for enhancing the nutritional value of food products. Moreover, incorporation of the total biomass might contribute to the structural properties of the enriched food product. Our previous study demonstrated the potential of Porphyridium cruentum and Chlorella vulgaris as multifunctional food ingredients, as they displayed interesting rheological properties after applying a specific combination of mechanical and thermal processing. The aim of the current study was to investigate the impact of a different sequence of high pressure homogenization (HPH) and thermal processing on the thickening and gelling potential of these microalgal biomasses in aqueous suspensions. Thermal processing largely increased the gel strength and viscosity of both microalgae, which was ascribed to larger and stronger aggregates as a result of partial solubilization of polymers, while subsequent HPH generally reduced the rheological properties. Interestingly, large amounts of intact cells were still observed for both microalgae when HPH was performed after a thermal treatment, irrespective of the applied homogenization pressure, implying that cell disruption was hindered by the preceding thermal treatment. Although thermal processing was regarded as the most effective processing technique to obtain increased rheological properties, the combination with a preceding HPH treatment should still be considered when cell disruption is desired, for instance to increase the bioavailability of intracellular components. Finally, biomass of P. cruentum showed the largest potential for use as a structuring agent, as the gel strength and viscosity in thermally treated suspensions of this microalga were about 10 times higher than for C. vulgaris.

Mechanistic insight into softening of Canadian wonder common beans (Phaseolus vulgaris) during cooking.

The relative contributions of cotyledons and seed coats towards hardening of common beans (Phaseolus vulgaris) were investigated and the rate-limiting process which controls bean softening during cooking was determined. Fresh or aged whole beans and cotyledons were soaked and cooked in demineralised water or 0.1 M NaHCO3 solution, and texture evolution, microstructure changes and thermal properties were studied. Fresh and aged whole beans cooked in demineralised water had significantly different softening rate constants and so did fresh and aged cotyledons. The comparable softening rate constants of aged whole beans and cotyledons indicated an insignificant role of the seed coat in hardening during storage. All samples cooked faster in 0.1 M NaHCO3 solution. Disintegration of cooked tissues followed by microscopic examination revealed a transition from cell breakage through a phase of cell breakage and separation to complete cell separation with increased cooking time wherefore texture decayed. Therefore, progressive solubilization of pectin in the middle lamella greatly promoted texture decay. While residual birefringence even after substantial cooking time suggested some molecular order of the starch, calorimetric analyses revealed complete starch gelatinisation before complete cell separation occurred. This implies an insignificant role of starch in texture decay during cooking but its hindered uncoiling into a viscous gel after gelatinisation due to the restricting cell wall could promote its retrogradation. Therefore, we suggest that the rate-determining process in bean softening relates to cell wall/middle lamella changes influencing pectin solubilization.

Structurally modified pectin for targeted lipid antioxidant capacity in linseed/sunflower oil-in-water emulsions.

The present work explored the lipid antioxidant capacity of citrus pectin addition to 5%(w/v) linseed/sunflower oil emulsions stabilized with 0.5%(w/v) Tween 80, as affected by pectin molecular characteristics. The peroxide formation in the emulsions, containing tailored pectin structures, was studied during two weeks of storage at 35°C. Low demethylesterified pectin (≤33%) exhibited a higher antioxidant capacity than high demethylesterified pectin (≥58%), probably due to its higher chelating capacity of pro-oxidative metal ions (Fe2+), whereas the distribution pattern of methylesters along the pectin chain only slightly affected the antioxidant capacity. Nevertheless, pectin addition to the emulsions caused emulsion destabilization probably due to depletion or bridging effect, independent of the pectin structural characteristics. These results evidence the potential of structurally modified citrus pectin as a natural antioxidant in emulsions. However, optimal conditions for emulsion stability should be carefully selected.

Shelf-life dating of shelf-stable strawberry juice based on survival analysis of consumer acceptance information.

BACKGROUND: Accurate shelf-life dating of food products is crucial for consumers and industries. Therefore, in this study we applied a science-based approach for shelf-life assessment, including accelerated shelf-life testing (ASLT), acceptability testing and the screening of analytical attributes for fast shelf-life predictions. Shelf-stable strawberry juice was selected as a case study. RESULTS: Ambient storage (20 °C) had no effect on the aroma-based acceptance of strawberry juice. The colour-based acceptability decreased during storage under ambient and accelerated (28-42 °C) conditions. The application of survival analysis showed that the colour-based shelf-life was reached in the early stages of storage (≤11 weeks) and that the shelf-life was shortened at higher temperatures. None of the selected attributes (a* and ΔE* value, anthocyanin and ascorbic acid content) is an ideal analytical marker for shelf-life predictions in the investigated temperature range (20-42 °C). Nevertheless, an overall analytical cut-off value over the whole temperature range can be selected. CONCLUSIONS: Colour changes of strawberry juice during storage are shelf-life limiting. Combining ASLT with acceptability testing allowed to gain faster insight into the change in colour-based acceptability and to perform shelf-life predictions relying on scientific data. An analytical marker is a convenient tool for shelf-life predictions in the context of ASLT. © 2017 Society of Chemical Industry.

Membrane fatty acid composition as a determinant of Listeria monocytogenes sensitivity to trans-cinnamaldehyde.

trans-Cinnamaldehyde, the major compound of cinnamon essential oil, is a potentially interesting natural antimicrobial food preservative. Although a number of studies have addressed its mode of action, the factors that determine bacterial sensitivity or tolerance to trans-cinnamaldehyde are poorly understood. We report the detailed characterization of a Listeria monocytogenes Scott A trans-cinnamaldehyde hypersensitive mutant defective in IlvE, which catalyzes the reversible transamination of branched-chain amino acids to the corresponding short-chain α-ketoacids. This mutant showed an 8.4 fold extended lag phase during growth in sublethal concentrations (4 mM), and faster inactivation in lethal concentrations of trans-cinnamaldehyde (6 mM). trans-Cinnamaldehyde hypersensitivity could be corrected by genetic complementation with the ilvE gene and supplementation with branched-chain α-ketoacids. Whole-cell fatty acid analyses revealed an almost complete loss of anteiso branched-chain fatty acids (BCFAs), which was compensated by elevated levels of unbranched saturated fatty acids and iso-BCFAs. Sub-inhibitory concentrations of trans-cinnamaldehyde induced membrane fatty acid adaptations predicted to reduce membrane fluidity, possibly as a response to counteract the membrane fluidizing effect of trans-cinnamaldehyde. These results demonstrate the role of IlvE in BCFA production and the role of membrane composition as an important determinant of trans-cinnamaldehyde sensitivity in L. monocytogenes.

Impact of processing on odour-active compounds of a mixed tomato-onion puree.

Gas chromatography-olfactometry revealed thirty-two odour-active compounds in a heat-processed tomato-onion puree, among which twenty-seven were identified by gas chromatography-olfactometry-mass spectrometry (GC-O-MS) and comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GC×GC-TOF MS). Based on the results of two olfactometric methods, i.e. detection frequency and aroma extract dilution analysis, the most potent aroma components include: dipropyl disulfide, S-propyl thioacetate, dimethyl trisulfide, 1-octen-3-one, methional, dipropyl trisulfide, 4,5-dimethylthiazole, 2-phenylacetaldehyde and sotolone. Processing of mixed vegetable systems can add complexity in their aroma profiles due to (bio)chemical interactions between the components. Therefore, the impact of different processing steps (i.e. thermal blanching, all-in-one and split-stream processes) on the volatile profile and aroma of a mixed tomato-onion puree has been investigated using a GC-MS fingerprinting approach. Results showed the potential to control the aroma in a mixed tomato-onion system through process-induced enzymatic modulations for producing tomato-onion food products with distinct aroma characteristics.

In vitro ß-Carotene Bioaccessibility and Lipid Digestion in Emulsions: Influence of Pectin Type and Degree of Methyl-Esterification.

Citrus pectin (CP) and sugar beet pectin (SBP) were demethoxylated and fully characterized in terms of pectin properties in order to investigate the influence of the pectin degree of methyl-esterification (DM) and the pectin type on the in vitro ß-carotene bioaccessibility and lipid digestion in emulsions. For the CP based emulsions containing ß-carotene enriched oil, water and pectin, the ß-carotene bioaccessibility, and lipid digestion were higher in the emulsions with pectin with a higher DM (57%; "CP57 emulsion") compared to the emulsions with pectin with a lower DM (30%; "CP30 emulsion") showing that the DM plays an important role. In contrast, in SBP-based emulsions, nor ß-carotene bioaccessibility nor lipid digestion were dependent on pectin DM. Probably here, other pectin properties are more important factors. It was observed that ß-carotene bioaccessibility and lipid digestion were lower in the CP30 emulsion in comparison with the CP57, SBP32, and SBP58 emulsions. However, the ß-carotene bioaccessibility of CP57 emulsion was similar to that of the SBP emulsions, whereas the lipid digestion was not. It seems that pectin type and pectin DM (in case of CP) are determining which components can be incorporated into micelles. Because carotenoids and lipids have different structures and polarities, their incorporation may be different. This knowledge can be used to engineer targeted (digestive) functionalities in food products. If both high ß-carotene bioaccessibility and high lipid digestion are targeted, SBP emulsions are the best options. The CP57 emulsion can be chosen if high ß-carotene bioaccessibility but lower lipid digestion is desired.

Process-Structure-Function Relations of Pectin in Food.

Pectin, a complex polysaccharide rich in galacturonic acid, has been identified as a critical structural component of plant cell walls. The functionality of this intricate macromolecule in fruit- and vegetable-based-derived products and ingredients is strongly determined by the nanostructure of its most abundant polymer, homogalacturonan. During food processing, pectic homogalacturonan is susceptible to various enzymatic as well as nonenzymatic conversion reactions modifying its structural and, hence, its functional properties. Consequently, a profound understanding of the various process-structure-function relations of pectin aids food scientists to tailor the functional properties of plant-based derived products and ingredients. This review describes the current knowledge on process-structure-function relations of pectin in foods with special focus on pectin's functionality with regard to textural attributes of solid plant-based foods and rheological properties of particulated fruit- and vegetable-derived products. In this context, both pectin research performed via traditional, ex situ physicochemical analyses of fractionated walls and isolated polymers and pectin investigation through in situ pectin localization are considered.