Fibrillization of lentil proteins is impacted by the protein extraction conditions and co-extracted phenolics.

Legume proteins can be induced to form amyloid-like fibrils upon heating at low pH, with the exact conditions greatly impacting the fibril characteristics. The protein extraction method may also impact the resulting fibrils, although this effect has not been carefully examined. Here, the fibrillization of lentil protein prepared using various extraction methods and the corresponding fibril morphology were characterized. It was found that an acidic, rather than alkaline, protein extraction method was better suited for producing homogeneous, long, and straight fibrils from lentil proteins. During alkaline extraction, co-extracted phenolic compounds bound proteins through covalent and non-covalent interactions, contributing to the formation of heterogeneous, curly, and tangled fibrils. Recombination of isolated phenolics and proteins (from acidic extracts) at alkaline pH resulted in a distinct morphology, implicating a role for polyphenol oxidase also in modifying proteins during alkaline extraction. These results help disentangle the complex factors affecting legume protein fibrillization.

Epigenetic regulation of N-hydroxypipecolic acid biosynthesis by the AIPP3-PHD2-CPL2 complex.

N-Hydroxypipecolic acid (NHP) is a signaling molecule crucial for systemic acquired resistance (SAR), a systemic immune response in plants that provides long-lasting and broad-spectrum protection against secondary pathogen infections. To identify negative regulators of NHP biosynthesis, we performed a forward genetic screen to search for mutants with elevated expression of the NHP biosynthesis gene FLAVIN-DEPENDENT MONOOXYGENASE 1 (FMO1). Analysis of two constitutive expression of FMO1 (cef) and one induced expression of FMO1 (ief) mutants revealed that the AIPP3-PHD2-CPL2 protein complex, which is involved in the recognition of the histone modification H3K27me3 and transcriptional repression, contributes to the negative regulation of FMO1 expression and NHP biosynthesis. Our study suggests that epigenetic regulation plays a crucial role in controlling FMO1 expression and NHP levels in plants.

Effect of UV Filters during the Application of Pulsed Light to Reduce Lactobacillus brevis Contamination and 3-Methylbut-2-ene-1-thiol Formation While Preserving the Physicochemical Attributes of Blonde Ale and Centennial Red Ale Beers.

Pulsed light (PL) is a novel, non-thermal technology being used to control the microbial spoilage of foods and beverages. Adverse sensory changes, commonly characterized as "lightstruck", can occur in beers when exposed to the UV portion of PL due to the formation of 3-methylbut-2-ene-1-thiol (3-MBT) upon the photodegradation of iso-α-acids. This study is the first to investigate the effect of different portions of the PL spectrum on UV-sensitive beers (light-colored blonde ale and dark-colored centennial red ale) using clear and bronze-tinted UV filters. PL treatments with its entire spectrum, including the ultraviolet portion of the spectrum, resulted in up to 4.2 and 2.4 log reductions of L. brevis in the blonde ale and centennial red ale beers, respectively, but also resulted in the formation of 3-MBT and small but significant changes in physicochemical properties including color, bitterness, pH, and total soluble solids. The application of UV filters effectively maintained 3-MBT below the limit of quantification but significantly reduced microbial deactivation to 1.2 and 1.0 log reductions of L. brevis at 8.9 J/cm2 fluence with a clear filter. Further optimization of the filter wavelengths is considered necessary to fully apply PL for beer processing and possibly other light-sensitive foods and beverages.

Phenolic profiles and their responses to pre- and post-harvest factors in small fruits: a review.

The consumption of small fruits has increased in recent years. Besides their appealing flavor, the commercial success of small fruits has been partially attributed to their high contents of phenolic compounds with multiple health benefits. The phenolic profiles and contents in small fruits vary based on the genetic background, climate, growing conditions, and post-harvest handling techniques. In this review, we critically compare the profiles and contents of phenolics such as anthocyanins, flavonols, flavan-3-ols, and phenolic acids that have been reported in bilberries, blackberries, blueberries, cranberries, black and red currants, raspberries, and strawberries during fruit development and post-harvest storage. This review offers researchers and breeders a general guideline for the improvement of phenolic composition in small fruits while considering the critical factors that affect berry phenolics from cultivation to harvest and to final consumption.

Determination of bound volatiles in blueberries, raspberries, and grapes with an optimized protocol and a validated SPME-GC/MS method.

Bound volatiles are odorless aroma reservoirs that modify flavor when released during food processing, and their determination is important to understand the aroma of fruit beverages. However, the generation of oxidation/degradation artifacts during analyses of glycosidically-bound volatiles has not been compared across fruit species and their dependence on diverse acidic and enzymatic hydrolytic conditions remains unclear. This work aimed to optimize and compare different hydrolytic conditions for the analysis of glycosidically-bound volatiles in blueberries, raspberries, and grapes with a solid-phase microextraction - gas chromatography/mass spectrometry (SPME-GC/MS) methodology. Enzymatic hydrolyses using AR2000® at 100 mg.mL-1 and Pectinex Ultra SPL® at 25-100 µL.mL-1 showed profiles characterized by the expected alcohols, while using AR2000® at 200-400 mg.mL-1 and citric acid at 50-100 mM resulted in profiles defined by artifacts (hydrocarbons, norisoprenoids, and aldehydes). (Z)-3-hexen-1-ol, 3-methyl-1-butanol, linalool, citronellol, and geraniol presented Odor Activity Values (OAV) > 1 for most small fruit genotypes.

Impact of hormone applications on ripening-related metabolites in Gewürztraminer grapes (Vitis vinifera L.): The key role of jasmonates in terpene modulation.

Terpenes play a formative role in grape and wine flavor, particularly for high-terpenic cultivars. Differences in terpene profiles influence grape varietal character and vintage quality. Little is known about the endogenous factors controlling terpene biosynthesis in grape. Through multiple experiments, six hormones (abscisic acid, ABA; ethylene, ETH; jasmonic acid, JA; methyl jasmonate, MeJA; indole-3-acetic acid, IAA; 1-naphthaleneacetic acid, NAA) that either promote or repress ripening were applied to Gewürztraminer clusters near veraison to gauge their effect on ripening and terpene biosynthesis. Jasmonates (JA, MeJA) increased terpene concentrations and the expression of terpene genes in grapes. Such increases were not associated to increases of other ripening-related metabolites such as sugars or anthocyanins. MeJA also affected the expression of several hormone related genes, increased IAA levels, and reduced sugar and anthocyanin concentration in grapes. This research provides novel insights into terpene regulation by ripening-related hormones and jasmonates in grapes.

Monolignol export by diffusion down a polymerization-induced concentration gradient.

Lignin, the second most abundant biopolymer, is a promising renewable energy source and chemical feedstock. A key element of lignin biosynthesis is unknown: how do lignin precursors (monolignols) get from inside the cell out to the cell wall where they are polymerized? Modeling indicates that monolignols can passively diffuse through lipid bilayers, but this has not been tested experimentally. We demonstrate significant monolignol diffusion occurs when laccases, which consume monolignols, are present on one side of the membrane. We hypothesize that lignin polymerization could deplete monomers in the wall, creating a concentration gradient driving monolignol diffusion. We developed a two-photon microscopy approach to visualize lignifying Arabidopsis thaliana root cells. Laccase mutants with reduced ability to form lignin polymer in the wall accumulated monolignols inside cells. In contrast, active transport inhibitors did not decrease lignin in the wall and scant intracellular phenolics were observed. Synthetic liposomes were engineered to encapsulate laccases, and monolignols crossed these pure lipid bilayers to form polymer within. A sink-driven diffusion mechanism explains why it has been difficult to identify genes encoding monolignol transporters and why the export of varied phenylpropanoids occurs without specificity. It also highlights an important role for cell wall oxidative enzymes in monolignol export.

Optimization and validation of a SPME-GC/MS method for the determination of volatile compounds, including enantiomeric analysis, in northern highbush blueberries (Vaccinium corymbosum L.).

Blueberry aroma is one of the most important quality traits that influences consumer purchasing decisions. This study aimed to optimize and validate a solid-phase microextraction-gas chromatography/mass spectrometry (SPME-GC/MS) method for the quantification of 73 volatile compounds in northern highbush blueberries. A SPME extraction of blueberries with water and specific proportions of sodium chloride, citric acid, and ascorbic acid, for 60 min at 50 °C using a divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS) fiber was optimal. The method was validated for sensitivity, reproducibility, linearity, and accuracy, and used to quantify volatile compounds through matrix-matched calibration curves in six blueberry cultivars ('Duke', 'Draper', 'Bluecrop', 'Calypso', 'Elliott', and 'Last Call'). Terpenes represented the most abundant volatile fraction, followed by aldehydes and alcohols. Linalool and 2-(E)-hexenal were key compounds that differentiated blueberry cultivars via Principal Component Analysis (PCA). Enantiomeric analyses revealed an excess of (-)-limonene, (-)-α-pinene, and (+)-linalool for all cultivars with potential impacts on the blueberry aroma.

Influence of Non-Saccharomyces on Wine Chemistry: A Focus on Aroma-Related Compounds.

Wine fermentation processes are driven by complex microbial systems, which comprise eukaryotic and prokaryotic microorganisms that participate in several biochemical interactions with the must and wine chemicals and modulate the organoleptic properties of wine. Among these, yeasts play a fundamental role, since they carry out the alcoholic fermentation (AF), converting sugars to ethanol and CO2 together with a wide range of volatile organic compounds. The contribution of Saccharomyces cerevisiae, the reference organism associated with AF, has been extensively studied. However, in the last decade, selected non-Saccharomyces strains received considerable commercial and oenological interest due to their specific pro-technological aptitudes and the positive influence on sensory quality. This review aims to highlight the inter-specific variability within the heterogeneous class of non-Saccharomyces in terms of synthesis and release of volatile organic compounds during controlled AF in wine. In particular, we reported findings on the presence of model non-Saccharomyces organisms, including Torulaspora delbrueckii, Hanseniaspora spp,Lachancea thermotolerans, Metschnikowia pulcherrima, Pichia spp. and Candida zemplinina, in combination with S. cerevisiae. The evidence is discussed from both basic and applicative scientific perspective. In particular, the oenological significance in different kind of wines has been underlined.

Development and assessment of GC/MS and HPAEC/PAD methodologies for the quantification of α-galacto-oligosaccharides (GOS) in dry beans (Phaseolus vulgaris).

The quantification of α-Galacto-oligosaccharides (GOS) in beans has been increasingly approached through different methodologies. However, reported GOS contents revealed up to 8-times disparity, which cannot be only attributed to the bean cultivar and underlines the need of using validated analytical methodologies. This study aimed to optimize and validate the extraction of the most abundant GOS found in beans, namely raffinose, stachyose and verbascose, and comparatively assess their determination by High-Performance Anion Exchange Chromatography/Pulsed Amperometric Detector (HPAEC/PAD) and Gas Chromatography/Mass Spectrometry (GC/MS). Hot sonication followed by shaking with 70% ethanol resulted in excellent GOS extraction efficiencies (92.54-107.94%). GC/MS determination was more reliable than HPAEC/PAD, with limits of quantification of 4.48-224.31 mg/kg and intra/inter-day repeatabilities <10%. The analysis of six bean varieties proved the feasibility of the GC/MS methodology, displaying total GOS contents from 1453.07 ±â€¯169.31 to 2814.34 ±â€¯95.28 mg/100 g. Stachyose was significantly (p < 0.05) the main GOS in all samples.

Systematic evaluation of the Folin-Ciocalteu and Fast Blue BB reactions during the analysis of total phenolics in legumes, nuts and plant seeds.

Folin-Ciocalteu and the more recent Fast Blue BB (FBBB) reactions are used for the quantification of total phenolics in food matrices. Despite its known interferences, Folin-Ciocalteu is still widely employed, considering Solid Phase Extraction (SPE) as clean-up step only in a few cases. Meanwhile, FBBB has shown no interferences for the determination of total phenolics in fruits and cereals, although its utilization in other popular matrices containing potential interferences, such as legumes, remains unexplored. In this study, the total phenolic content of 24 flours from legumes, cereals, fruits, nuts and plant seeds was evaluated by Folin-Ciocalteu and FBBB, with and without SPE. Folin-Ciocalteu showed interferences for 75% of the flours (attributed to reducing sugars and enediols), whereas FBBB only for legumes and nuts (attributed to the presence of tyrosine), both methods in those matrices requiring SPE. Although both SPE-FBBB and SPE-Folin-Ciocalteu presented excellent reproducibility, SPE-FBBB displayed 1.5 times higher sensitivity.

Real-Time Monitoring of Volatile Compounds Losses in the Oven during Baking and Toasting of Gluten-Free Bread Doughs: A PTR-MS Evidence.

Losses of volatile compounds during baking are expected due to their evaporation at the high temperatures of the oven, which can lead to a decrease in the aroma intensity of the final product, which is crucial for gluten-free breads that are known for their weak aroma. Volatiles from fermentation and lipids oxidation are transferred from crumb to crust, and they flow out to the air together with Maillard and caramelisation compounds from the crust. In this study, the release to the oven of volatile compounds from five gluten-free breads (quinoa, teff and rice flours, and corn and wheat starches) and wheat bread during baking and toasting was measured in real-time using proton transfer reaction-time of flight-mass spectrometry (PTR-ToF-MS). Baking showed different volatile release patterns that are described by bell-shaped curves, plateaus and exponential growths. Flour-based breads had the higher overall volatile release during baking, but also high ratios in the final bread, while starch-based breads showed high pyrazine releases due to moisture losses. Meanwhile, toasting promoted the release of volatile compounds from the bread matrix, but also the additional generation of volatiles from Maillard reaction and caramelisation. Interestingly, gluten-free breads presented higher losses of volatiles during baking than wheat bread, which could partially explain their weaker aroma.

The dynamics of starch hydrolysis and thickness perception during oral processing.

While it is known that salivary amylase changes perceived thickness in starch-based foods, the dynamics of oral breakdown of starch has not been related to that of sensory perception. This work examines the associations between in-mouth oral breakdown of starch by α-amylase and thickness of a semisolid product measured instrumentally and using a sensory panel. Pureed carrots made with added waxy maize (w/w) (0% (control), 0.4% (S0.4) and 0.8% (S0.8)), were tested for dilution and hydrolysis effects with the addition of water and saliva and measuring viscosity (shear of 10 s-1), viscoelasticity (10-0.01 Hz at 0.1% strain) and maltose release. Sensory testing was conducted using progressive profiling. Control and S0.8 pureed carrots showed similar viscosities at 10 s-1 while S0.4 was lower. Addition of saliva had the highest impact on viscosity drop for S0.8. Viscoelastic properties of pureed carrots decreased with starch addition which decreased further with water/saliva incorporation. Starch breakdown measured in terms of maltose release did not change with starch concentration and oral processing times. Unlike instrumental viscosity results, perceived thickness of purees increased with starch concentration. During oral processing, thickness decreased at a constant rate across all samples highlighting the dilution effect of saliva. A relation was observed only between oral viscosity and starch hydrolysis and not rheological measurements. This indicates that changes in starch hydrolysis during oral processing can be used as an indicator of rate of drop in perceived viscosity in starch-based semi-solid foods such as pureed carrots.

Okra seed and seedless pod: Comparative study of their phenolics and carbohydrate fractions and their impact on bread-making.

The outstanding amount of phenolics and pectins of okra seeds and seedless pods, respectively, is well-known. However, their impact on bread nutritional quality, and particularly on slowing down α-amylase activity during crumb digestion, has never been studied. In this work, the phenolic and carbohydrate fractions of developed fine and coarse flours from okra seeds (OS) and seedless pods (OP) were investigated as well as their impact on wheat bread physical and nutritional quality. The use of okra flours dramatically increased the amount of extractable (EPP) and non-extractable hydrolyzable phenolics (HPP) of wheat breads, attaining up to 210.8 and 2944.8 mg/100 g of EPP and HPP, respectively, with only a 5% replacement with OS. Interestingly, breads made with fine OS and OP exhibited a second digestion rate upon 50 min of digestion, indicating a time-dependence hypoglycemic effect of okra constituents whereby OS-breads presented the slowest digestion rate and extension among all breads.

Banana flour phenolics inhibit trans-epithelial glucose transport from wheat cakes in a coupled in vitro digestion/Caco-2 cell intestinal model.

Some fruit phenolics are reported to attenuate intestinal glucose transport through inhibitory action at the luminal brush border membrane. This effect may contribute, in part, to the ability of flavonoid-rich food to regulate glucose homeostasis of meals rich in available carbohydrates. For the first time, the potential of green banana flours to inhibit transepithelial glucose transport was investigated in the context of a model starchy meal (wheat cake) using a simulated digestion/Caco-2 human intestinal cell model. A 10% replacement of wheat flour with any of the four banana flours (native and extruded oven-dried and freeze-dried) resulted in cakes with significantly higher total phenolics (68-198 µg per 100 g, p < 0.05), especially using extruded banana flour (197-198 µg per 100 g), as measured by LC/MS. Banana cakes, especially those containing oven-dried and/or extruded banana flours, exhibited from 45.0 to 54.5% higher glucose transport inhibition than the control cake. Interestingly, the digesta of cakes made with freeze-dried and extruded banana flour presented a significantly higher phenolic content (1116 µM, p < 0.05) than the other digesta (745-791 µM), while the phenolic content in control digesta was only 548 µM. These results suggested that the amounts of quercetin and myricetin, even in traces, were critical determinants of glucose transport inhibition.

Unraveling the Inhibition of Intestinal Glucose Transport by Dietary Phenolics: A Review.

BACKGROUND: Glucose transport across the intestinal brush border membrane plays a key role in metabolic regulation. Depending on the luminal glucose concentration, glucose is mainly transported by the sodium- dependent glucose transporter (SGLT1) and the facilitated-transporter glucose transporter (GLUT2). SGLT1 is apical membrane-constitutive and it is active at a low luminal glucose concentration, while at concentrations higher than 50 mM, glucose is mainly transported by GLUT2 (recruited from the basolateral membrane). Dietary phenolic compounds can modulate glucose homeostasis by decreasing the postprandial glucose response through the inhibition of SGLT1 and GLUT2. METHODS: Phenolic inhibition of intestinal glucose transport has been examined using brush border membrane vesicles from rats, pigs or rabbits, Xenopus oocytes and more recently Caco-2 cells, which are the most promising for harmonizing in vitro experiments. RESULTS: Phenolic concentrations above 100 µM has been proved to successfully inhibit the glucose transport. Generally, the aglycones quercetin, myricetin, fisetin or apigenin have been reported to strongly inhibit GLUT2, while quercetin-3-O-glycoside has been demonstrated to be more effective in SGLT1. Additionally, epigallocatechin as well as epicatechin and epigallocatechin gallates were observed to be inhibited on both SGLT1 and GLUT2. CONCLUSION: Although, valuable information regarding the phenolic glucose transport inhibition is known, however, there are some disagreements about which flavonoid glycosides and aglycones exert significant inhibition, and also the inhibition of phenolic acids remains unclear. This review aims to collect, compare and discuss the available information and controversies about the phenolic inhibition of glucose transporters. A detailed discussion on the physicochemical mechanisms involved in phenolics-glucose transporters interactions is also included.

Manufacturing the ultimate green banana flour: Impact of drying and extrusion on phenolic profile and starch bioaccessibility.

Use of banana flours as functional ingredients is growing due to their nutritional benefits derived from phenolics and dietary fiber. However, the effect oven-drying, freeze-drying and extrusion on the phenolic compounds or starch digestibility is not understood. In this work, phenolic acids (gallic acid, caffeic acid), flavan-3-ols (epicatechin, catechin) and flavonols (quercetin-3-O-glucoside and myricetin) were quantified in banana flour processed by different methods. Epicatechin, the most abundant phenolic in all flours (up to 1.93 mg/100 g), was significantly reduced during thermal processing (oven-drying and extrusion). Meanwhile, phenolic acids and flavonols were found to be more thermally stable. Thus, oven-drying and extrusion generally improved the extractability of phenolic acids and flavonols. Freeze-drying resulted in native flours with significantly higher insoluble dietary fiber (up to 43.3%), although the digestible starch fraction was digested more rapidly than the oven-dried counterpart.

Analytical feasibility of a solvent-assisted flavour evaporation method for aroma analyses in bread crumb.

The analysis of bread aroma is essential in order to evaluate its quality as well as to improve it. The use of different methodologies for the analysis of volatile compounds lead to varying results. In the present study, the matrix effect, extraction efficiency, limits of detection and quantification as well as the precision of a proposed solvent-assisted flavour evaporation methodology were evaluated for the first time and compared with a reference method, both differing in the distillation step. The repeatability (<8%) and matrix effect (present in 15 of the 31 compounds) were improved with the proposed method but the extraction efficiencies (average of 52%) and the intermediate precision (>15%) were not as required. However, the applicability of the reference method was limited to breads with fat levels <2%. For breads higher in fat, the proposed method represents an alternative for aroma analysis.

Comparison of different extraction methodologies for the analysis of volatile compounds in gluten-free flours and corn starch by GC/QTOF.

Gluten-free bakery products usually exhibit weak aroma. Their main constituents are flours and starches, which contain aroma precursors but can also contribute additional volatiles in low concentrations. Static headspace (SHS), solid-phase microextraction (SPME) and solvent-assisted flavour evaporation (SAFE) coupled to GC/QTOF were compared for their efficacy in the analysis of volatiles in corn starch. SPME-GC/QTOF was selected as the most suitable methodology based on the number of detected compounds, LODs, repeatability as well as simplicity. It was successfully applied for the quantification of volatiles in corn starch and qualitative comparison of different gluten-free flours. Hexanal, 3-methyl-1-butanol, 1-pentanol, 1-octen-3-ol, acetic acid, furfural, benzaldehyde, (E)-2-nonenal, phenylethyl alcohol and short-medium chain acids were found in all the flours and corn starch. Quinoa flour and corn starch showed the highest contents of pyrazines, terpenes and esters, while teff, buckwheat and rice flours presented the highest contents of 3/2-methyl-1-butanol, acetoin and organic acids.