Varying precipitation conditions allow directing the composition and physical properties of soy protein concentrates.

Soy protein concentrates (SPCs) are common food ingredients. They typically contain 65% (w/w) protein and ∼30% (w/w) carbohydrate. SPCs can be obtained with various protein precipitation conditions. A systematic study of the impact of these different protein precipitation protocols on the SPC protein composition and physical properties is still lacking. Here, SPCs were prepared via three different protocols, that is, isoelectric (pH 3.5-5.5), aqueous ethanol (50%-70% [v/v]), and Ca2+ ion (5-50 mM) based precipitations, and analyzed for (protein) composition, protein thermal properties, dispersibility, and water-holding capacity. SPCs precipitated at pH 5.5 or by adding 15 mM Ca2+ ions had a lower 7S/11S globulin ratio (∼0.40) than that (∼0.50) of all other SPC samples. Protein in SPCs obtained by isoelectric precipitation denatured at a significantly higher temperature than those in ethanol- or Ca2+ -precipitated SPCs. Precipitation with 50%-60% (v/v) ethanol resulted in pronounced denaturation of 2S albumin and 7S globulin fractions in SPCs. Additionally, increasing the precipitation pH from 3.5 to 5.5 and increasing the Ca2+ ion concentration from 15 to 50 mM caused a strong decrease of both the dispersibility of the protein in SPC and its water-holding capacity at pH 7.0. In conclusion, this study demonstrates that the SPC production process can be directed to obtain ingredients with versatile protein physicochemical properties toward potential food applications. PRACTICAL APPLICATION: This study demonstrates that applying different protein precipitation protocols allows obtaining SPCs that vary widely in (protein) composition and physical properties (such as protein dispersibility and water-holding capacity). These varying traits can greatly influence the suitability of SPCs as functional ingredients for specific applications, such as the production of food foams, emulsions, gels, and plant-based meat alternatives. The generated knowledge may allow targeted production of SPCs for specific applications.

Hydrothermal treatments of starch impact reaction patterns during subsequent chemical derivatization.

Differences in derivatization patterns (using a fluorescent reagent, fluorescein isothiocyanate) of wheat, pea, and potato starches between native granular (NAT) starches and their respective annealed (ANN) and heat-moisture treated (HMT) starches were investigated to reveal structural changes associated with starch hydrothermal treatments. Size-exclusion chromatography with fluorescence and refractive index detection assessed the reactivity of amylose (AM), intermediate chains (IM1 and IM2), and amylopectin branch chains (AP1, AP2, and AP3) within the different starches. Shifts in X-ray diffraction patterns of HMT starches and in the gelatinization properties of both ANN and HMT starches confirmed molecular rearrangement. The reaction homogeneity (wheat and pea) and the overall extent of reaction (pea and potato) increased for HMT starches compared to other starches. The lower reactivities of IM2 chains (HMT starch) and AP3 chains (ANN starch) relative to NAT starches, indicated their involvement in molecular rearrangements and improved double helical order. IM2 and AP branch chains in ANN pea starch also were less reacted than NAT starch chains, suggesting their co-crystallization. Molecular rearrangements in ANN and HMT starches led to altered swelling and pasting viscosities. Thus, changes in the relative crystallinity of individual starch branch chains induced by hydrothermal processing impact the final physical properties.

Sucrose substitution in cake systems is not a piece of cake.

Successful sucrose replacement in cake systems requires thorough understanding of its functionality. Time-domain 1H NMR showed that water in the viscous aqueous phase isolated from cake batter by ultracentrifugation [i.e. the batter liquor (BL)] exhibits low mobility by its low T2 relaxation time (T2,D RT). This is due to its interactions with sucrose or sucrose replacers. The T2,D RT itself is positively related with the effective volumetric hydrogen bond density of sucrose or sucrose replacers. Sucrose additionally co-determines the quantity and viscosity of cake BL and thereby how much air the batter contains at the end of mixing. Like sucrose, maltitol and oligofructose provide adequate volumes of BL with low water mobility and thus sufficient air in the batter, while the rather insoluble mannitol and inulin do not. Differential scanning calorimetry and rapid viscosity analysis revealed, however, that, in contrast to sucrose and maltitol, oligofructose fails to provide appropriate timings of starch gelatinisation and protein denaturation, resulting in poor cake texture. The shortcomings of mannitol and oligofructose in terms of respectively ensuring appropriate gas content in batter and biopolymer transitions during baking can be overcome by using mixtures thereof. This work shows that successful sucrose substitutes or substitute mixtures must provide sufficient BL with low water mobility and ensure appropriate timings of starch and protein biopolymer transitions during baking.

Promotion of starch retrogradation by enzymatic elongation of amylopectin chains does not reduce glycemic responses: a randomized cross-over clinical trial.

The molecular structure of amylopectin (AP) governs the propensity of its chains to re-associate into crystalline arrangements after starch gelatinization. Amylose (AM) crystallization and AP re-crystallization (i.e. retrogradation) decrease starch digestibility. The aim of this work was to enzymatically elongate AP chains using an amylomaltase (AMM, i.e. 4-α-glucanotransferase) from Thermus thermophilus to promote AP retrogradation and to investigate the impact thereof on in vivo glycemic responses in healthy subjects. Participants (n = 32) consumed two oatmeal porridges (containing 22.5 g available carbohydrates) prepared with or without the enzymatic modification and stored at 4 °C for 24 h. Finger-prick blood samples were taken fasting and at intervals during 3 h following test-meal consumption. The incremental area under the curve (iAUC0-180) was determined. The AMM was very effective at elongating the AP chains at the expense of AM, resulting in increased retrogradation capacity upon storage at low temperature. Nevertheless, postprandial glycemic responses were not different after consumption of either the AMM modified oatmeal porridge or its unmodified counterpart (iAUC0-180 = 73 ± 30 vs. 82 ± 43 mmol min L-1, respectively; p = 0.17). Unexpectedly, promoting starch retrogradation by selectively modifying its molecular structure did not result in reduced glycemic responses, challenging the notion that starch retrogradation negatively impacts glycemic responses in vivo.

Olive-Derived Antioxidant Dietary Fiber Modulates Gut Microbiota Composition and Attenuates Atopic Dermatitis Like Inflammation in Mice.

SCOPE: Epidemiological data suggest that altered gut microbiota contributes to the development of atopic dermatitis (AD). The effect of an olive-derived antioxidant dietary fiber (OADF) in relieving AD symptoms in a murine model of 2,4-dinitrofluorobenzene (DNFB)-induced AD is examined and the effect of OADF in modulating host gut microbiota is explored. METHODS AND RESULTS: Mice are fed with either standard diet or standard diet + OADF for 3 weeks prior to induction of AD and maintained on the same diet throughout the DNFB application period. Dietary OADF causes significant improvement of AD-like symptoms with reduced serum levels of immunoglobulin (Ig)E, interleukin (IL)-1ß, IL-6, C-X-C motif ligand (CXCL)1, and increased serum levels of IL-10. OADF supplementation restore gut microbiota composition that are altered in AD mice. Specifically, OADF increases the proportion of intestinal bacteria (Ruminococcaceae UCG014, GCA900066575, UBA1819) associated with enhanced butyrate production, along with inhibiting Clostridiales vadin BB60 which are more prevalent in AD mice. CONCLUSION: OADF modulates gut microbiota composition, improves cytokine profile and butyrate production influencing AD-associated immune response. Results highlight the importance of the gut-skin axis for the AD dietary therapeutic agents.

Incubation tests mimicking fermentation reveal that phytate breakdown is key to lower the cadmium concentrations in cacao nibs.

Earlier studies revealed that cadmium (Cd) concentrations in cacao nibs can decrease by a factor up to 1.3 during fermentation. Here, fermentation was mimicked by incubating beans at different temperatures, and acetic acid and ethanol concentrations in the incubation media. Nib Cd concentrations decreased during incubation by mobilisation in the nibs and subsequent outward migration to the testa and the incubation solution. This was most pronounced when high concentrations of acetic acid were combined with high temperature, while ethanol had no statistically significant effect. Incubation under typical fermentation conditions (45 °C and 20.0 g acetic acid L-1) reduced the nib Cd concentration by a factor 1.3. This factor increased to 1.6 under more extreme conditions, i.e. 65 °C and 40 g acetic acid L-1. The final nib Cd concentrations correlated well to nib phytate concentrations (R2 = 0.56), suggesting hydrolysis of phytate and mobilisation of the associated Cd2+.

The influence of varying levels of molecular oxygen on the functionality of azodicarbonamide and ascorbic acid during wheat bread making.

Air, and thus also molecular oxygen (O2), is incorporated in wheat flour dough during mixing. O2 participates in several (enzymatic) reactions, including those resulting in the oxidation of free sulfhydryl groups, thereby increasing dough strength and bread volume. We here incorporated different O2 levels in dough by mixing dough samples for a fixed time under different modified atmospheres which led to significant changes in dough free sulfhydryl contents and bread volumes. Although altering the mixing time not only impacted how much O2 was incorporated in dough but also the mechanical input, the changes in dough and bread properties when using different mixing times, largely depended on differences in O2 uptake. When used in bread recipes, redox agents such as azodicarbonamide (ADA) and ascorbic acid (AH2) impact the dough sulfhydryl contents and bread volumes. The effect of different levels of O2 incorporation on dough samples which contained ADA or AH2 was studied by altering the mixing time or the O2 content in the mixing atmosphere. Lower ADA levels were needed when dough was mixed under an atmosphere enriched in O2. As AH2 requires O2 to be converted to dehydroascorbic acid (DHA) to exert its improver effect, it came as a surprise that when it was included in a dough which was prepared under O2 enriched conditions, no additional impact was obtained and that, even under reduced O2 conditions, its use still resulted in an increased bread volume. These findings suggest that AH2 oxidase very effectively uses O2 to form DHA.

The use of time domain 1 H NMR to study proton dynamics in starch-rich foods: A review.

Starch is a major contributor to the carbohydrate portion of our diet. When it is present with water, it undergoes several transformations during heating and/or cooling making it an essential structure-forming component in starch-rich food systems (e.g., bread and cake). Time domain proton nuclear magnetic resonance (TD 1 H NMR) is a useful technique to study starch-water interactions by evaluation of molecular mobility and water distribution. The data obtained correspond to changes in starch structure and the state of water during or resulting from processing. When this technique was first applied to starch(-rich) foods, significant challenges were encountered during data interpretation of complex food systems (e.g., cake or biscuit) due to the presence of multiple constituents (proteins, carbohydrates, lipids, etc.). This article discusses the principles of TD 1 H NMR and the tools applied that improved characterization and interpretation of TD NMR data. More in particular, the major differences in proton distribution of various dough and cooked/baked food systems are examined. The application of variable-temperature TD 1 H NMR is also discussed as it demonstrates exceptional ability to elucidate the molecular dynamics of starch transitions (e.g., gelatinization, gelation) in dough/batter systems during heating/cooling. In conclusion, TD NMR is considered a valuable tool to understand the behavior of starch and water that relate to the characteristics and/or quality of starchy food products. Such insights are crucial for food product optimization and development in response to the needs of the food industry.

Hydration of Wheat Flour Water-Unextractable Cell Wall Material Enables Structural Analysis of Its Arabinoxylan by High-Resolution Solid-State 13C MAS NMR Spectroscopy.

To enable its structural characterization by nuclear magnetic resonance (NMR) spectroscopy, the native structure of cereal water-unextractable arabinoxylan (WU-AX) is typically disrupted by alkali or enzymatic treatments. Here, WU-AX in the wheat flour unextractable cell wall material (UCWM) containing 40.9% ± 1.5 arabinoxylan with an arabinose-to-xylose ratio of 0.62 ± 0.04 was characterized by high-resolution solid-state NMR without disrupting its native structure. Hydration of the UCWM (1.7 mg H2O/mg UCWM) in combination with specific optimizations in the NMR methodology enabled analysis by solid-state 13C NMR with magic angle spinning and 1H high-power decoupling (13C HPDEC MAS NMR) which provided sufficiently high resolution to allow for carbon atom assignments. Spectral resonances of C-1 from arabinose and xylose residues of WU-AX were here assigned to the solid state. The proportions of un-, mono-, and di-substituted xyloses were 59.2, 19.5, and 21.2%, respectively. 13C HPDEC MAS NMR showed the presence of solid-state fractions with different mobilities in the UCWM. This study presents the first solid-state NMR spectrum of wheat WU-AX with sufficient resolution to enable assignment without prior WU-AX solubilization.

Investigating the Sequence Determinants of the Curling of Amyloid Fibrils Using Ovalbumin as a Case Study.

Highly ordered, straight amyloid fibrils readily lend themselves to structure determination techniques and have therefore been extensively characterized. However, the less ordered curly fibrils remain relatively understudied, and the structural organization underlying their specific characteristics remains poorly understood. We found that the exemplary curly fibril-forming protein ovalbumin contains multiple aggregation prone regions (APRs) that form straight fibrils when isolated as peptides or when excised from the full-length protein through hydrolysis. In the context of the intact full-length protein, however, the regions separating the APRs facilitate curly fibril formation. In fact, a meta-analysis of previously reported curly fibril-forming proteins shows that their inter-APRs are significantly longer and more hydrophobic when compared to straight fibril-forming proteins, suggesting that they may cause strain in the amyloid state. Hence, inter-APRs driving curly fibril formation may not only apply to our model protein but rather constitute a more general mechanism.

Induction of Maize Starch Gelatinization and Dissolution at Low Temperature by the Hydrotrope Sodium Salicylate.

Complete aqueous dissolution of starch requires the use of temperatures exceeding 100 °C. During and after cooling of the resultant aqueous solutions, starch polymers precipitate by aggregation and crystallization. Low-temperature gelatinization and dissolution of maize starch (MS) is induced, and the stability of the resultant solutions is enhanced when they contain the hydrotrope sodium salicylate (NaSal). Differential scanning calorimetry and optical microscopy evidence showed that MS gelatinization shifts to lower temperatures and that the associated enthalpy decreases with increasing NaSal concentrations. The enhanced gelatinization and dissolution are probably brought about by the association of NaSal with the more hydrophobic MS structural moieties. The minimum NaSal content of aqueous mixtures allowing full gelatinization of MS at room temperature, that is, about 11 wt %, was found to be independent of MS content (in the range 10-66.7 wt % MS).

Bioavailability and Health Impact of Ingested Amyloid-like Protein Fibrils and their Link with Inflammatory Status: A Need for More Research?

The use of amyloid-like protein fibrils (ALFs) in food formulations looks very promising in terms of improving techno-functional properties, but raises some concerns in terms of food safety, because of their structural resemblance to disease-related endogenous amyloids. This review focuses on the biological fate and potential health implications of ingested ALF structures in both healthy and predisposed individuals. A comprehensive overview of ALF gastrointestinal digestion, intestinal absorption, and systemic dissemination is provided, in addition to a thorough assessment of potential ALF cross-seeding of endogenous precursor proteins linked to (non)neurodegenerative amyloidosis. In general, this study concludes that the health impact of ALF consumption remains widely understudied and merits additional research efforts to determine the exact extent to which ALF ingestion may influence the general health status.

Heat-induced denaturation and aggregation of protein in quinoa (Chenopodium quinoa Willd.) seeds and whole meal.

Physical barriers hinder about 20-25% of the protein from being extracted from whole meal. Heat-induced denaturation and aggregation of protein in quinoa seeds and in whole meal was investigated. Maximally 37% of the protein in seeds covalently aggregate when boiling for 15 min. Although embryonic cell walls surrounding protein bodies remain intact during boiling of seeds, protein aggregation is not hindered. 11S Globulin monomers first dissociate into their acidic and basic subunits which further assemble into large (> 500 kDa) mainly disulfide-linked aggregates. 2S Albumins are not involved in covalent aggregation but partially leach during seed boiling. The presence of disrupted food matrix constituents in whole meal delays denaturation and causes less aggregation of protein in whole meal than in seeds. Globulins still dissociate into their subunits but less and mainly small covalent aggregates (ca. 100-500 kDa) are formed. These novel insights allow developing new quinoa-based food products.

Structural factors governing starch digestion and glycemic responses and how they can be modified by enzymatic approaches: A review and a guide.

Starch is the most abundant glycemic carbohydrate in the human diet. Consumption of starch-rich food products that elicit high glycemic responses has been linked to the occurrence of noncommunicable diseases such as cardiovascular disease and diabetes mellitus type II. Understanding the structural features that govern starch digestibility is a prerequisite for developing strategies to mitigate any negative health implications it may have. Here, we review the aspects of the fine molecular structure that in native, gelatinized, and gelled/retrograded starch directly impact its digestibility and thus human health. We next provide an informed guidance for lowering its digestibility by using specific enzymes tailoring its molecular and three-dimensional supramolecular structure. We finally discuss in vivo studies of the glycemic responses to enzymatically modified starches and relevant food applications. Overall, structure-digestibility relationships provide opportunities for targeted modification of starch during food production and improving the nutritional profile of starchy foods.

Investigation of starch functionality and digestibility in white wheat bread produced from a recipe containing added maltogenic amylase or amylomaltase.

In the crumb of fresh white wheat bread, starch is fully gelatinized. Its molecular and three-dimensional structure are major factors limiting the rate of its digestion. The aim of this study was to in situ modify starch during bread making with starch-modifying enzymes (maltogenic amylase and amylomaltase) and to investigate the impact thereof on bread characteristics, starch retrogradation and digestibility. Maltogenic amylase treatment increased the relative content of short amylopectin chains (degree of polymerization ≤ 8). This resulted in lower starch retrogradation and crumb firmness upon storage, and reduced extent (up to 18%) of in vitro starch digestion for fresh and stored breads. Amylomaltase only modestly shortened amylose chains and had no measurable impact on amylopectin structure. Modification with this enzyme led to slower bread crumb firming but did not influence starch digestibility.

Gas cell stabilization by aqueous-phase constituents during bread production from wheat and rye dough and oat batter: Dough or batter liquor as model system.

Proper gas cell stability during fermentation and baking is essential to obtain high-quality bread. Gas cells in wheat dough are stabilized by the gluten network formed during kneading and, from the moment this network locally ruptures, by liquid films containing nonstarch polysaccharides (NSPs) and surface-active proteins and lipids. Dough liquor (DL), the supernatant after ultracentrifugation of dough, is a model system for these liquid films and has been extensively studied mostly in the context of wheat bread making. Nonwheat breads are often of lower quality (loaf volume and crumb structure) than wheat breads because their doughs/batters lack a viscoelastic wheat gluten network. Therefore, gas cell stabilization by liquid film constituents may be more important in nonwheat than in wheat bread making. This manuscript aims to review the knowledge on DL/batter liquor (BL) and its relevance for studying gas cell stabilization in wheat and nonwheat (rye and oat) bread making. To this end, the unit operations in wheat, rye, and oat bread making are described with emphasis on gas incorporation and gas cell (de)stabilization. A discussion of the knowledge on the recoveries and chemical structures of proteins, lipids, and NSPs in DLs/BLs is provided and key findings of studies dealing with foaming and air-water interfacial properties of DL/BL are discussed. Next, the extent to which DL/BL functionality can be related to bread properties is addressed. Finally, the extent to which DL/BL is a representative model system for the aqueous phase of dough/batter is discussed and related to knowledge gaps and further research opportunities.

Free wheat flour lipids decrease air-liquid interface stability in sponge cake batter.

The impact of free wheat flour lipids on air-liquid interface stability during sponge cake making was investigated. Therefore, the molecular population at the air-liquid interface in batters prepared with flour of which part of the lipids had been either relocated or removed prior to batter preparation was determined. Surface-active molecules were isolated from batter using a foam separation protocol. Diluted batter was whipped and the resulting foam was used as model system for the air-liquid interface in sponge cake batter. Relocating flour lipids prior to batter making enabled them to adsorb at the air-liquid interface in the foam. This limited the degree of protein adsorption at the air-liquid interface, but it did not impact the composition of the adsorbed protein population. Removing flour lipids prior to batter making resulted in foam containing relatively higher levels of lipids mainly originating from egg yolk. Prior removal of flour lipids impacted neither foam protein content nor foam protein composition. The resultant molecular population improved air-liquid interface stability in sponge cake batter. Thus, free wheat flour lipids and wheat flour lipids set free by solvent treatment decrease air-liquid interface stability in sponge cake batter mainly because they limit protein adsorption and, as such, interfere with the protein-dominated interface.

Normal-Phase HPLC-ELSD to Compare Lipid Profiles of Different Wheat Flours.

Normal-phase high-performance liquid chromatography (HPLC) is widely used in combination with evaporative light scattering detection (ELSD) for separating and detecting lipids in various food samples. ELSD responses of different lipids were evaluated to elucidate the possibilities and challenges associated with quantification by means of HPLC-ELSD. Not only the number and type of polar functional groups but also the chain length and degree of unsaturation of (free or esterified) fatty acids (FAs) had a significant effect on ELSD responses. Tripalmitin and trilinolein yielded notably different ELSD responses, even if their constituting free FAs produced identical responses. How FA structure impacts ELSD responses of free FAs is thus not predictive for those of triacylglycerols and presumably other lipids containing esterified FAs. Because ELSD responses of lipids depend on the identity of the (esterified) FA(s) which they contain, fully accurate lipid quantification with HPLC-ELSD is challenging and time-consuming. Nonetheless, HPLC-ELSD is a good and fast technique to semi-quantitatively compare the levels of different lipid classes between samples of comparable FA composition. In this way, lipid profiles of different flours from near-isogenic wheat lines could be compared.

1H Diffusion-Ordered Nuclear Magnetic Resonance Spectroscopic Analysis of Water-Extractable Arabinoxylan in Wheat (Triticum aestivum L.) Flour.

The structural heterogeneity of water-extractable arabinoxylan (WE-AX) impacts wheat flour functionality. 1H diffusion-ordered (DOSY) nuclear magnetic resonance (NMR) spectroscopy revealed structural heterogeneity within WE-AX fractions obtained via graded ethanol precipitation. Combination with high-resolution 1H-1H correlation NMR spectroscopy (COSY) allowed identifying the relationship between the xylose substitution patterns and diffusion properties of the subpopulations. WE-AX fractions contained distinct subpopulations with different diffusion rates. WE-AX subpopulations with a high self-diffusivity contained high levels of monosubstituted xylose. In contrast, those with a low self-diffusivity were rich in disubstituted xylose, suggesting that disubstitution mainly occurs in WE-AX molecules with large hydrodynamic volumes. In general, WE-AX fractions precipitating at higher and lower ethanol concentrations had higher and lower self-diffusivity and more and less complex substitution patterns. Although 1H DOSY NMR, as performed in this study, was valuable for elucidating WE-AX structural heterogeneity, physical limitations arose when studying WE-AX populations with high molecular weight dispersions.

Use of Amylomaltase to Steer the Functional and Nutritional Properties of Wheat Starch.

The fine molecular structure of starch governs its functionality and digestibility, and enzymatic approaches can be utilized to tailor its properties. The aim of this study was to investigate the in situ modification of starch by amylomaltase (AMM) from Thermus thermophilus in model starch systems subjected to hydrothermal treatments under standardized conditions and the relationship between molecular structure, rheological properties and in vitro digestibility. When low dosages of AMM were added to a wheat starch suspension prior to submitting it to a temperature-time profile in a Rapid Visco Analyzer, the increased peak viscosity observed was attributed to partial depolymerization of amylose, which facilitated starch swelling and viscosity development. At higher dosages, the effect was smaller. The low cold paste viscosity as a result of the activity of AMM reflected substantial amylose depolymerization. At the same time, amylopectin chains were substantially elongated. The longer amylopectin chains were positively correlated (R2 = 0.96) with the melting enthalpies of retrograded starches, which, in turn, were negatively correlated with the extent (R2 = 0.92) and rate (R2 = 0.79) of in vitro digestion. It was concluded that AMM has the potential to be used to deliver novel starch functionalities and enhance its nutritional properties.