Techno-functional properties and antioxidant capacity of the concentrate and protein fractions of Leucaena spp. seeds / Propiedades tecno-funcionales y capacidad antioxidante del concentrado y fracciones proteicas de semillas de Leucaena spp

Introduction: The use of vegetable proteins as ingredients in food systems is based on their functional properties. The water and oil holding capacity, foaming, and emulsifying capacity/stability, and antioxidant assay of the protein fractions - albumins, globulins 7S/11S, glutelins and prolamins - isolated from Leucaena seed were evaluated. Objective: The objective of this study was to evaluate the functional properties and antioxidant capacity of the concentrate and protein fractions of ripe Leucaena spp. seeds. Materials and methods: Ripe Leucaena seeds were collected and evaluated in Oaxaca, Mexico (16°59'21''N 96°43'26''O) during the months of February-April 2021.The protein concentrate was isolated by isoelectric precipitation (pH=9, pH=4). The albumins, globulins, glutelins and prolamins were isolated based on their solubility properties in different extracting solutions. Results: Glutelins constituted the main protein fraction (75.88%). Prolamins were not found. The glutelins fractions showed the highest oil holding capacity (0.93±0.08 mL g-1). The albumins fraction had the highest water holding capacity (2.53±0.15 mL g-1), foaming capacity and foam stability (71.83±1.26 % and 70.00±0.00%, respectively) and antioxidant capacity (18.09±0.88%). The globulins exhibited the highest emulsifying capacity and emulsion stability (56.83±1.76% and 55.67±1.20%, respectively). Conclusions: The concentrate and protein fraction of Leucaena seeds showed different techno-functional and antioxidant properties of interest for the food industry, like those showed by other commercial vegetable proteins(AU)

Introducción: El uso de proteínas vegetales como ingredientes en sistemas alimentarios se basa en sus propiedades funcionales. Se evaluó la capacidad de retención de agua y aceite, la capacidad/estabilidad espumante y emulsionante y el ensayo antioxidante de las fracciones proteicas -albúminas, globulinas 7S/11S, glutelinas y prolaminas- aisladas de las semillas de Leucaena. Objetivo: El objetivo de este estudio fue evaluar las propiedades funcionales y la capacidad antioxidante del concentrado y las fracciones proteicas de las semillas maduras de Leucaena spp. Materiales y métodos: Las semillas maduras de Leucaena fueron recolectadas y evaluadas en Oaxaca, México (16°59'21''N 96°43'26''O) durante los meses de febrero-abril del año 2021. Se usó harina de Leucaena desgrasada para la preparación de las fracciones proteicas. El concentrado proteico se aisló por precipitación isoeléctrica (pH=9, pH=4). Las albúminas, globulinas, glutelinas y prolaminas se aislaron en función de sus propiedades de solubilidad en diferentes soluciones de extracción. Resultados: Las glutelinas constituyeron la principal fracción proteica (75,88%). No se encontraron prolaminas. La fracción de glutelinas mostró la mayor capacidad de retención de aceite (0.93±0,08 mL g-1). La fracción de albúminas presentó la mayor capacidad de retención de agua (2,53±0,15 mL g-1), capacidad espumante y estabilidad de la espuma (71,83±1,26% y 70,00±0,00%, respectivamente) y capacidad antioxidante (18,09±0,88%). Las globulinas mostraron la mayor capacidad emulsionante y estabilidad de la emulsión (56,83±1,76 y 55,67±1,20%, respectivamente). Conclusiones: El concentrado y las fracciones proteicas de las semillas de Leucaena mostraron diferentes propiedades tecno-funcionales y antioxidantes de interés para la industria alimentaria, similares a los reportados por diversas proteínas vegetales comerciales(AU)

Seaweed proteins are nutritionally valuable components in the human diet.

The global population is expected to reach 11 billion people by the year 2100 and will require sustainable sources of dietary protein. Most dietary protein originates from animal and terrestrial plant agriculture, which leads to deforestation, water pollution, and greenhouse gas emissions. Discovering alternative protein sources that are nutritionally adequate for the human diet without harmful environmental effects is imperative. Seaweeds are a promising option as they produce abundant protein with a low carbon footprint. Experimental evidence shows that seaweeds contain high concentrations of the essential amino acids (EAAs) necessary for human consumption, but seaweeds have yet to be evaluated with standardized metrics to compare their nutritional value to other protein sources. In this technical note, independent literature describing the EAA content and protein digestibility of 3 commonly consumed species of seaweeds was evaluated alongside traditional protein sources using a novel hybrid protein quality (HPQ) metric. HPQ is derived from the protein digestibility-corrected amino acid score and digestibility indispensable amino acid score but includes modifications to address the lack of in vivo digestibility data for seaweeds. Seaweed proteins are similar in quality to common plant protein sources such as peas, soy, and tree nuts. Furthermore, seaweed proteins from different species have complementary EAA profiles and can be mixed to form protein blends that are nutritionally on par with animal products such as milk and whey. Thus, seaweeds may be viable protein sources with a reduced footprint that provide beneficial ecosystem services.

Effect of heat treatment on the digestion behavior of pea and rice protein dispersions and their blends, studied using the semi-dynamic INFOGEST digestion method.

In the present study, the structuring and breakdown of a 5% protein dispersion prepared with commercial fractions of pea and rice isolates (PPI and RPI, respectively) were monitored by in vitro digestion. These proteins were blended in a 2 : 1 ratio of pea and rice, respectively, as this would deliver a high amino acid score. The effect of heating at 90 °C for 15 min on the digestion behavior was evaluated not only for the blend, but also for the respective protein isolate fractions, using the INFOGEST international consensus, semi-dynamic in vitro gastric model. Digesta were characterized by gel electrophoresis, light scattering, confocal laser scanning microscopy and size exclusion HPLC. Heating increased the solubility of PPI from 15.7% to 26.6% at pH 7.4. RPI showed low solubility (a maximum of 2.6% at pH 2), regardless of the treatment. Confocal microscopy observations evidenced major differences in the aggregates formed during digestion, with larger aggregates for heated PPI. While the unheated pea protein dispersions precipitated near the isoelectric pH, the heated counterpart formed macro-aggregates under the same conditions. In the case of RPI, there were no differences in structuring behaviour between unheated and heat treated reconstituted powder, due to their low solubility. Rice prolamins showed resistance to hydrolysis by pepsin and pancreatic enzymes. In the heated blend, macro-aggregates formed, but with a smaller size compared to heated pea protein alone, suggesting that pea protein aggregation was hindered by the presence of rice proteins. These results demonstrate how the composition of protein isolates can affect their in vitro digestion. However, pre-treatment of plant protein blends, such as heating, can modulate the rate and mechanism of digestion.

Impact of wheat globulin addition on dough rheological properties and quality of cooked noodles.

Impact of globulin addition on the functional and protein structural properties of dough and cooked noodles were investigated. The underlying mechanism was explored through analyzing the interaction between globulin and gluten by using SDS-PAGE, size exclusion chromatography, free sulfhydryl/disulfide bond analysis, laser scanning confocal microscopy and Fourier transform infrared spectroscopy. Results showed that the stiffness/hardness and maximum resistance of dough and cooked noodles were both increased when globulin addition was 1.5% or higher. Besides, extensibility of cooked noodles was also improved when the addition up to 3.0%. The addition of globulin facilitated weakening the S-S bonds in the gluten network and cross-linked with SDS-soluble gluten mainly through non-covalent interactions, especially hydrophobic interactions. Meanwhile, a more rigid protein network structure was observed. Additionally, following cooking, globulin addition accelerated the aggregation of protein molecules. When the addition reached 3%, the protein conformation was transformed from ß-sheets and random coils to ß-turns.

Characteristics of rice dreg protein isolate treated by high-pressure microfluidization with and without proteolysis.

The characteristics of rice dreg protein isolate (RDPI) treated by microfluidization (0, 40, 80, 120, and 160 MPa) with or without proteolysis were investigated. Alcalase, Neutrase, and the combination of the two (Alcalcase:Neutrase = 1:1 [w/w]) were adopted for hydrolysis. The surface hydrophobicity and solubility of RDPI were increased. As pressure increased, different structures of RDPI exhibited disaggregation (<120 MPa) and reaggregation (160 MPa), and the effect on proteolysis was significant. The solubility of Neutrase and combined enzyme hydrolysates was improved after microfluidization. Additionally, the optimum choice of microfluidization (40 MPa) and Neutrase was efficient for improving the DPPH radical scavenging activity. The results indicate that both pressure level and enzyme type synergistically determine the functionality and antioxidant activities of products. This work may provide an alternative methodology for improving the utilization of RDPI in the food industry through desirable modifications.

Protein structural properties and proteomic analysis of rice during storage at different temperatures.

Rice quality changes during storage, but there have been few studies of how rice proteins changes during aging. The present study characterized the structural properties of protein in stored rice and identified the mechanism of quality deterioration using proteomics. Compared with protein from newly harvested rice, the free sulfhydryl content of protein from stored rice was significantly reduced and the disulfide bond content and surface hydrophobicity was higher. Storage resulted in a loss of α-helix and ß-sheet structures and increase in ß-turn and random coil structures. High-molecular-weight protein subunits decomposed to produce low-molecular-weight subunits at 30 °C, while protein aggregation occurred at 70 °C. At 30 ℃ 157 differential proteins were found and 70 ℃ 395 such proteins occurred. Redox homeostasis, response to oxidative stress, glutathione metabolism, tricarboxylic acid cycle, glycolysis/gluconeogenesis, starch and sucrose metabolism, and fatty acid biosynthesis and degradation led to the different quality of stored rice.

Neuromodulatory peptides: Orally active anxiolytic-like and antidepressant-like peptides derived from dietary plant proteins.

Mental disorders are a severe health problem, and the number of patients is growing worldwide. Increased anxiety and decreased motivation due to excessive mental stress further accelerated the severity of the problem. Enzymatic digestion of food proteins produces bioactive peptides with various physiological functions, some of which exhibit neuromodulatory effects with oral administration. Recently, studies reported that some peptides produced from plant proteins such as soybeans, leaves, and grains exhibit emotional regulatory functions such as strong anxiolytic-like and antidepressant-like effects comparable to pharmaceuticals. Conventionally, researchers investigated bioactive peptides by fractionation of protein hydrolysates and structure-activity relationship. As a novel methodology for analyzing bioactive peptides, the information obtained by peptidomics simultaneous analysis of the digested fractions of proteins using mass spectrometry has been effectively utilized. Some small-sized peptides such as dipeptides and tripeptides released food-derived proteins show emotional regulating effects. Moreover, some middle-sized peptides produced after intestinal digestion may exhibit the emotional regulating effect via the vagus nerve, and the importance of the gut-brain axis is also focused. As the central mechanism of emotional regulation, it has been found that these plant-derived peptides regulated monoamine neurotransmitter signaling and hippocampal neurogenesis.

Physicochemical and functional properties of red lentil protein isolates from three origins at different pH.

Red lentils (Lens culinaris) present an attractive raw material for meat mimics due to its red-coloured proteins, abundance, high protein and low cost. However, data on its functional properties at various pH remain scarce. In this study, the physicochemical and functional properties of red lentil proteins (RLP) from three origins (USA, Nepal and Turkey), isolated by isoelectric precipitation, were evaluated. Amino acid profiles, water holding (ranging from 3.1 to 3.5 g/g) and oil absorption (ranging from 5.8 to 7.3 g/g) capacities of RLP samples were significantly different (p < 0.05). RLP consisted of legumin and vicilin, and comprised predominantly glutamine/glutamic acid (ranging from 8.72 to 10.55 g/100 g). Surface charge, protein solubility, foaming and emulsifying properties were the lowest and poorest at pH 5.2 (isoelectric point). Overall, good functional properties of RLP under high acidity and alkalinity conditions make it a promising protein for mimicking a wide range of meats.

Effect of Acetylation on Physicochemical and Functional Properties of Commercial Pumpkin Protein Concentrate.

The purpose of the present study was to determine the effects of acetylation with different doses of acetic anhydride on the chemical composition and chosen functional properties of commercial pumpkin protein concentrate (PPC). The total protein content decreased as compared to unmodified samples. Electrophoretic analysis revealed that in the acetylated pumpkin protein, the content of the heaviest protein (35 kDa) decreased in line with increasing concentrations of modifying reagent. Acetylation of PPC caused a significant increase in water-binding and oil-absorption capacity and for emulsifying properties even at the dose of 0.4 mL/g. Additionally, an increase in foaming capacity was demonstrated for preparations obtained with 2.0 mL/g of acetic anhydride, whereas acetylation with 0.4 and 1.0 mL/g caused a decrease in protein solubility as compared to native PPC.

Combined effect of pH treatment and the extraction pH on the physicochemical, functional and rheological characteristics of amaranth (Amaranthus hypochondriacus) seed protein isolates.

Results for the effect of extraction pH and pH treatment on the functional, physicochemical, rheological and thermal characteristics of amaranth protein isolates (APIs) are reported in this study. Four amaranth protein isolates (P1, P2, P3 and P4) were prepared by varying the extraction pH (9-11). These four protein isolate samples were further treated at pH values from 3 to 9. The total protein content and purity of protein isolates were found to be higher for P1 than P2, P3 and P4 samples. The particle size of P1 was significantly (p ≤ 0.05) higher (299.68 µm) than other samples. Solubility, emulsifying capacity and stability, foaming properties, water and oil binding capacities were higher for the P1 sample treated at pH 9. Gelation characteristics like storage modulus (G') and loss modulus (G") were higher for P1 samples. APIs obtained at extraction pH 9 (P1) also exhibited better thermal properties in comparison with other three samples.

Identification of dipeptidyl peptidase IV (DPP-IV) inhibitory peptides from vegetable protein sources.

Vegetable proteins are appearing as a sustainable source for human consumption. Food-derived peptides are an important field of research in terms of bioactive molecules. In this study, seven vegetable proteins were enzymatically hydrolysed following an optimised treatment (sequential hydrolysis with subtilisin-trypsin-flavourzyme) to obtain dipeptidyl peptidase IV (DPP-IV) inhibitory peptides. Hydrolysates were fractionated by size exclusion chromatography and, from the most bioactive fractions (corresponding to Glycine max, Chenopodium quinoa and Lupinus albus proteins); peptides responsible for this bioactivity were identified by mass spectrometry. Peptides with adequate molecular features and based on in silico analysis were proposed as DPP-IV inhibitors from soy (EPAAV) lupine (NPLL), and quinoa (APFTVV). These vegetable protein sources are adequate to obtain protein hydrolysates for functional food.

Drying methods affect physicochemical and functional properties of quinoa protein isolate.

Quinoa protein possesses great amino acid profiles and can be a potential food ingredient with broad applications. The objective of this study was to investigate the effect of different drying methods, namely freeze drying, spray drying, and vacuum drying on the functional and physicochemical properties of quinoa protein isolate, e.g., morphology, amino acid composition, SDS-PAGE profile, sulfhydryl/disulfide content, secondary structure, surface hydrophobicity, and thermal stability. The freeze-dried protein exhibited the highest emulsification capacity and stability and oil binding capacity, which was contributed to its higher surface hydrophobicity, while the spray-dried sample had the highest solubility and water absorption capacity at pH 7. Gels (8%) prepared with the freeze-dried protein had higher elastic and viscous modulus than that from others. The freeze-dried protein had the highest maximal denaturation temperature but lowest enthalpy, which may be attributed to its higher amount of random coil but lower percent of regular α-helix and ß-sheet structures. Overall, quinoa protein isolate from different processing methods demonstrated distinct functional properties. This information will be useful to optimize quinoa protein production and benefit its applications.

Optimization of extraction and deamidation of edible protein from evening primrose (Oenothera biennis L.) oil processing by-products and its effect on structural and techno-functional properties.

The optimization of ultrasound-assisted alkaline extraction and enzymatic deamidation by protein-glutaminase (PG) on evening primrose seed cake (EPSC) protein and its effect on structure (amino acid composition, secondary structure and electrophoresis pattern) and techno-functional properties (water-holding and oil-binding capacities, solubility, emulsifying and foaming properties) of EPSC protein were evaluated. The optimum conditions of the both processes were measured using response surface methodology (RSM). The maximum yield (26.4%) and protein content (86.1%) were reached at the optimized extraction conditions. Optimal conditions of PG deamidation based on reaching a high degree of deamidation (DD) with a simultaneously low degree of hydrolysis (DH). Under these conditions, DD and DH were 39.40 and 2.11%, respectively. Ultrasound-assisted alkaline extraction and enzymatic deamidation by PG have great potential to produce edible EPSC protein with modified techno-functional characteristics that can be used for several aims in the food and pharmaceutical applications.

The functionality of laccase- or peroxidase-treated potato flour: Role of interactions between protein and protein/starch.

Potato flour is used in bakery products, extruded products and snacks. However, it displays weaker gel strengths and thus the wholesome utility is curtailed significantly. To improve viscoelastic properties and stability of potato gels, herein potato flour was treated with laccase and peroxidase to create a protein network structure leading to stable gels. The results revealed that the secondary structure of potato proteins altered upon the enzyme treatment. The gels of peroxidase-treated potato flour (PPF) and laccase-treated potato flour (LPF) displayed larger anti-shear ability, thermal stability and stronger three-dimensional network structure compared to the native potato gel. The PPF and LPF gels exhibited stronger viscoelastic properties and structural stability compared to peroxidase-treated potato protein and laccase-treated potato protein gels. The outcome serves as a theoretical basis to improve the properties of potato gels and to promote the designing and the development of novel potato flour based functional food.

Natural edible materials made of protein-functionalized aerogel particles for postprandial hyperglycemia management.

α-Amylase inhibitors (α-AIs) delay digestion of dietary starch by inhibiting α-amylase in the gut, thereby reducing the postprandial glycemia, which is beneficial to the patients with obesity and diabetes. The proteinaceous α-AIs from wheat can effectively control starch digestion and regulate postprandial hyperglycemia. However, their gastric intolerance remains a challenge, which limits its commercial production and industrial application. In this study, sodium alginate/chitosan aerogels loaded with wheat protein α-AIs were prepared and evaluated as potential transportation and protection matrices for important components in food or pharmaceutical applications. Specifically, the biodegradable aerogel cross-linked with sodium alginate-chitosan-calcium chloride, has a large surface area and open porous structure, which can adsorb staple wheat proteins as an integrated edible material to block around 88,660 U/g of α-amylase activity. The aerogel particles were able to protect the activity of wheat α-AIs in the stomach, leading to the slow passage of the wheat α-AIs through the small intestine to inhibit starch digestion more effectively. Animal experiments further showed that the postprandial blood glucose levels in rats were effectively controlled through delayed increase, after administration of wheat protein-functionalized aerogel particles loaded with wheat α-AIs, which are natural biological macromolecules. This is a novel, safe, and economical method for the prevention and pretreatment of diabetes.

The health benefits, functional properties, modifications, and applications of pea (Pisum sativum L.) protein: Current status, challenges, and perspectives.

In recent years, the development and application of plant proteins have drawn increasing scientific and industrial interests. Pea (Pisum sativum L.) is an important source of high-quality vegetable protein in the human diet. Its protein components are generally considered hypoallergenic, and many studies have highlighted the health benefits associated with the consumption of pea protein. Pea protein and its hydrolysates (pea protein hydrolysates [PPH]) possess health benefits such as antioxidant, antihypertensive, and modulating intestinal bacteria activities, as well as various functional properties, including solubility, water- and oil-holding capacities, and emulsifying, foaming, and gelling properties. However, the application of pea protein in the food system is limited due to its poor functional performances. Several frequently applied modification methods, including physical, chemical, enzymatic, and combined treatments, have been used for pea protein to improve its functional properties and expand its food applications. To date, different applications of pea protein in the food system have been extensively studied, for example, encapsulation for bioactive ingredients, edible films, extruded products and substitution for cereal flours, fats, and animal proteins. This article reviews the current status of the knowledge regarding pea protein, focusing on its health benefits, functional properties, and structural modifications, and comprehensively summarizes its potential applications in the food industry.

Effects of daily 24-gram doses of rice or whey protein on resistance training adaptations in trained males.

BACKGROUND: Large (48-g), isonitrogenous doses of rice and whey protein have previously been shown to stimulate similar adaptations to resistance training, but the impact of consuming smaller doses has yet to be compared. We evaluated the ability of 24-g doses of rice or whey protein concentrate to augment adaptations following 8 weeks of resistance training. METHODS: Healthy resistance-trained males (n = 24, 32.8 ± 6.7 years, 179.3 ± 8.5 cm, 87.4 ± 8.5 kg, 27.2 ± 1.9 kg/m2, 27.8 ± 6.0% fat) were randomly assigned and matched according to fat-free mass to consume 24-g doses of rice (n = 12, Growing Naturals, LLC) or whey (n = 12, NutraBio Labs, Inc.) protein concentrate for 8 weeks while completing a standardized resistance training program. Body composition (DXA), muscular strength (one-repetition maximum [1RM]) and endurance (repetitions to fatigue [RTF] at 80% 1RM) using bench press (BP) and leg press (LP) exercises along with anaerobic capacity (Wingate) were assessed before and after the intervention. Subjects were asked to maintain regular dietary habits and record dietary intake every 2 weeks. Outcomes were assessed using 2 × 2 mixed (group x time) factorial ANOVA with repeated measures on time and independent samples t-tests using the change scores from baseline. A p-value of 0.05 and 95% confidence intervals on the changes between groups were used to determine outcomes. RESULTS: No baseline differences (p > 0.05) were found for key body composition and performance outcomes. No changes (p > 0.05) in dietary status occurred within or between groups (34 ± 4 kcal/kg/day, 3.7 ± 0.77 g/kg/day, 1.31 ± 0.28 g/kg/day, 1.87 ± 0.23 g/kg/day) throughout the study for daily relative energy (34 ± 4 kcals/kg/day), carbohydrate (3.7 ± 0.77 g/kg/day), fat (1.31 ± 0.28 g/kg/day), and protein (1.87 ± 0.23 g/kg/day) intake. Significant main effects for time were revealed for body mass (p = 0.02), total body water (p = 0.01), lean mass (p = 0.008), fat-free mass (p = 0.007), BP 1RM (p = 0.02), BP volume (p = 0.04), and LP 1RM (p = 0.01). Changes between groups were similar for body mass (- 0.88, 2.03 kg, p = 0.42), fat-free mass (- 0.68, 1.99 kg, p = 0.32), lean mass (- 0.73, 1.91 kg, p = 0.37), fat mass (- 0.48, 1.02 kg, p = 0.46), and % fat (- 0.63, 0.71%, p = 0.90). No significant between group differences were seen for BP 1RM (- 13.8, 7.1 kg, p = 0.51), LP 1RM (- 38.8, 49.6 kg, p = 0.80), BP RTF (- 2.02, 0.35 reps, p = 0.16), LP RTF (- 1.7, 3.3 reps, p = 0.50), and Wingate peak power (- 72.5, 53.4 watts, p = 0.76) following the eight-week supplementation period. CONCLUSIONS: Eight weeks of daily isonitrogenous 24-g doses of rice or whey protein in combination with an eight-week resistance training program led to similar changes in body composition and performance outcomes. Retroactively registered on as NCT04411173 .

Identification and Characterization of IgE-Reactive Proteins and a New Allergen (Cic a 1.01) from Chickpea (Cicer arietinum).

SCOPE: Chickpea (Cicer arietinum) allergy has frequently been reported particularly in Spain and India. Nevertheless, chickpea allergens are poorly characterized. The authors aim to identify and characterize potential allergens from chickpea. METHODS AND RESULTS: Candidate proteins are selected by an in silico approach or immunoglobuline E (IgE)-testing. Potential allergens are prepared as recombinant or natural proteins and characterized for structural integrity by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), circular dichroism (CD)-spectroscopy, and mass spectrometry (MS) analysis. IgE-sensitization pattern of Spanish chickpea allergic and German peanut and birch pollen sensitized patients are investigated using chickpea extracts and purified proteins. Chickpea allergic patients show individual and heterogeneous IgE-sensitization profiles with extracts from raw and boiled chickpeas. Chickpea proteins pathogenesis related protein family 10 (PR-10), a late embryogenesis abundant protein (LEA/DC-8), and a vicilin-containing fraction, but not 2S albumin, shows IgE reactivity with sera from chickpea, birch pollen, and peanut sensitized patients. Remarkably, allergenic vicilin, DC-8, and PR-10 are detected in the extract of boiled chickpeas. CONCLUSION: Several IgE-reactive chickpea allergens are identified. For the first time a yet not classified DC-8 protein is characterized as minor allergen (Cic a 1). Finally, the data suggest a potential risk for peanut allergic patients by IgE cross-reactivity with homologous chickpea proteins.

Improvement of PersiXyn2 activity and stability in presence of Trehalose and proline as a natural osmolyte.

Compatible solutes or osmolytes stabilize proteins against different stress conditions. In this study, the effect of trehalose and proline against pH and thermal stress is examined on PersiXyn2. Trehalose and proline lowered the optimum pH of PersiXyn2 and increased the optimum temperature of its activity, which is more desirable for its industrial application. These osmolytes improved PersiXyn2 storage stability for a long 24-day period. Also, both osmolytes retained the catalytic activity of the enzyme in the presence of different concentrations of metal ions but trehalose had the most effect in the presence of Mg2+ and proline had most significant effect in the presence of Mn2+. In addition, the effect of detergents on the enzyme in the presence of proline and trehalose increased the activity of PersiXyn2. Chemical stability studies using urea as a denaturant indicated that both of the osmolytes improved the conformational stability of PersiXyn2. According to molecular dynamic studies, a definite change in conformation of PersiXyn2 was detected in the presence of proline and trehalose. Based on findings in the current study, given the importance of xylanases activity and stability in extreme conditions for industrial applications, two natural osmolytes were able to activate and stabilize PersiXyn2.

Hydrolysis of plant proteins at the molecular and supra-molecular scales during in vitro digestion.

The digestion of plant protein is highly dependent on multiple factors, with two of the most important being the protein source and the food matrix. The present study investigated the effects of these two factors on the digestion of seitan (a wheat-based food), tofu, soya juice, and a homemade emulsion of soy oil and water that was stabilised with pea protein. The four plant matrices and their respective protein isolates/concentrates (wheat gluten, soya protein, pea protein) were subjected to in vitro static digestion following the INFOGEST consensus protocol. We monitored the release of α-amino groups during digestion. We found that food matrix had a strong influence on protein digestion: soya juice was more hydrolysed than fresh tofu (51.1% versus 33.1%; P = 0.0087), but fresh tofu was more hydrolysed than soya protein isolate (33.1% versus 17.9%; P < 0.0001). Likewise, the pea-protein emulsion was better hydrolysed than the pea-protein isolate (P = 0.0033). Differences were also detected between the two solid foods investigated here: a higher degree of hydrolysis was found for tofu compared to seitan (33.1% versus 11.8%), which was perhaps a function of the presence of numerous dense protein aggregates in the latter but not the former. Furthermore, freeze-drying more than doubled the final degree of hydrolysis of seitan (P < 0.0001), but had no effect on tofu (P = 1.0000). Confocal microscopy revealed that protein networks in freeze-dried seitan were strongly altered with respect to the fresh product; instead, protein networks in freeze-dried and fresh tofu were largely similar. Finally, we found that the protease:protein ratio had a strong effect on the kinetics of proteolysis: a 3.7-fold increase in the concentration of the soya protein isolate with respect to that of the soya juice decreased the final degree of hydrolysis from 50.3 to 17.9% (P = 0.0988).