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The digestive characteristics of plant proteins are crucial for their nutritional value and utilization efficiency. In this study, an in vitro semi-dynamic digestion model was employed to investigate the gastric digestion process of soybean protein after treatment with phytase. The results found that phytase treatment reduced the phytate content in soybean proteins (22.83 ± 0.09 to 8.72 ± 0.07 mg/g), shifted its isoelectric point towards the alkaline range by 1 pH unit, and significantly improved its solubility at pH 4.0. Particularly for protein sample treated with phytase after acid precipitation, the formation of aggregates during digestion was weakened, resulting in a significantly higher digestion rate compared to untreated SPI, with digestion being at least 15 min faster than SPI. This study provides a strategy for preparing soybean protein with faster digestion and weaker clot-forming ability during digestion, which offers insights for the application of soybean protein in clinical nutrition products and specialized medical foods.

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The changing global climate brings a gradual yet constant and adverse shift in crop production. Grain crop plants, particularly cereals and legumes, respond varyingly to adverse climate, including reduction in grain yield and changes to their nutrient densities. An understanding of specific changes to crop systems under differing climatic conditions can help in planning diets to meet human nutrient sufficiency. Grain protein content is also affected by adverse environmental factors. Deficits in protein yield, linked to changes in grain or seed protein and antinutrient concentrations, have been reported in major food crops when exposed to elevated carbon dioxide, high temperature, drought, and humidity. These changes, in addition to affecting the quantity of indispensable or essential amino acids (IAA), also impact their bioavailability. Therefore, it is important to assess consequences of climate change on grain protein quality. An important tool to measure grain protein quality, is measuring its digestibility at the level of the ileum and its IAA concentration, linked to a metric called the Digestible IAA Score (DIAAS). A minimally invasive technique called the dual isotope tracer technique, which measures IAA digestibility after simultaneous administration of two different intrinsically labelled protein sources, one a test protein (2H/15N) and one a reference protein (13C) of predetermined digestibility, has been used in evaluation of grain protein IAA digestibility, and promises more in the evaluation of changes based on climate. This review discusses climate induced changes to grain protein quality through the prism of IAA digestibility, using the dual isotope tracer technique.

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With the global population projected to reach nine billion by 2050, the search for alternative protein sources has become critical. This study evaluated the digestibility of cricket protein powder compared with that of whey protein powder. Cricket protein powder had a slightly lower protein content but higher fat content than whey protein powder. Although both contained all essential amino acids, their quantities varied. The most abundant essential amino acid was leucine in both samples. The essential amino acid index (EAAI) for cricket protein powder reached 79% when utilising crude protein for calculation. When using the amino acid sum calculation method, it increased by nearly 13%. The EAAI for whey protein was then 94% when calculated based on crude protein, with a slight increase observed when using the amino acid sum calculation method. Cricket protein exhibited a gradual increase in digestibility during intestinal digestion, reaching nearly 80%, whereas whey protein digestibility surpassed 97%. Despite the lower digestibility of cricket protein compared with whey protein, it remains sufficiently high for consideration as a valuable protein source. This study highlights the potential of cricket proteins and underscores the importance of assessing their protein content and digestibility in evaluating their nutritional value.

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Sipunculus nudus (S. nudus), an edible marine invertebrate, is rich in myofibrillar proteins. However, its extremely low water solubility and relatively firm texture limit its practical applications. This study aimed to investigate the consequences of different ultrasound amplitude treatments on the structure, functional properties, and digestive characteristics of S. nudus salt soluble protein (SSP). The results showed that ultrasound treatment significantly reduced the particle size, surface tension, and the unordered structure of SSP, while having not impact the zeta potential. Additionally, the results of infrared spectroscopy and intrinsic fluorescence spectrum revealed that ultrasound treatment enhanced the hydrogen bonding and hydrophobic interaction within the components of SSP, leading to a more compact and uniformly distributed protein structure. These changes increased the solubility (increased from 12.07 % to 37.59 %) and optimized the functional properties of SSP (foamability and emulsifiability). Further, the results of in vitro digestion simulation revealed that the antioxidant proteopeptides of SSP were mainly produced in the small intestine, with the ABTS+ radical scavenging capacity ranging from 140 to 170 µg Trolox/mL. Additionally, the antioxidant activity of the digestive fluid was enhanced with increasing ultrasound amplitude. This work linked structural changes in denatured proteins to their functional properties and digestive characteristics. This study provided a new direction for developing easily digestible food ingredients.

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The purpose of this study was to assess the impact of food texture, oral processing, bolus characteristics, and in vitro digestive conditions on the starch and protein digestibility of al dente and soft-cooked commercial red lentil pasta. For that, samples were cooked as suggested by the provider and their texture properties were promptly analysed. Then, normal and deficient masticated pasta boluses were produced by four healthy subjects, characterised in terms of their oral processing, bolus granulometry, texture and viscoelastic properties, and finally subjected to static in vitro digestion, according to the INFOGEST consensus for both adults and the older adult population. Normal masticated boluses exhibited greater saliva impregnation and lower proportions of large particles, hardness, and stiffness than deficient masticated boluses. Likewise, insufficiently masticated al dente-cooked pasta boluses caused a delay in oral starch digestion owing to the larger particles attained during food oral processing, while reduced intestinal conditions in the elderly only interfere with the release of total soluble proteins in all samples. This work evidences the importance of considering the initial texture of products, oral capabilities, processing behaviour, and physical and mechanical properties of food boluses in digestion studies, opening new prospects in designing pulse-based foods that meet the nutritional requirements of the world's population.

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The study aims to assess the impact of oven-drying and decontamination on crude protein concentration and in vitro crude protein digestibility of yellow mealworms. Two kilograms of 12-wk-old mealworm larvae were subjected to freezing prior to the drying process. Approximately 1.5 kg of mealworm larvae were divided into 3 groups and exposed to oven-drying at temperatures of 50 °C for 36 h, 60 °C, and 70 °C for 24 h each. At intervals of 2 h, sets of 3 replicates were withdrawn to record water loss. Consistent weight stabilization was observed at 36 h for 50 °C (T50), 18 h for 60 °C (T60), and 14 h for 70 °C (T70). The remaining 0.5 kg of mealworm larvae was divided and dried under treatments T50, T60, and T70. Each treatment was then split into 2 portions, with one portion subjected to 90 °C for 15 min (denoted as T50-90, T60-90, T70-90) to eliminate microbial contamination. The 6 treatments were then used to determine concentrations of dry matter, crude ash, crude protein, pre-caecal protein digestibility, and dry matter residues after neutral detergent fiber, acid detergent fiber, and acid detergent lignin treatments. No interaction was observed between drying and decontamination treatments (P > 0.17). Pre-caecal crude protein digestibility increased with decreasing temperature (T50: 58% crude protein; T60: 51% crude protein; T70: 50% crude protein). Therefore, lower temperatures for longer times preserve crude protein digestibility. These findings are crucial for understanding how drying temperature and time impact protein bioavailability.

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Pigeon pea, a protein-rich legume with low protein digestibility (PD) due to its high polyphenol content and other antinutritional factors (ANFs). Consequently, processing methods are crucial to improve PD. We investigated the effects of thermal treatments (cooking, hydrothermal, autoclaving, infrared rays) treatments and germination on modulation of PD, its properties and association with ANFs in two distinct genotypes based on polyphenol content: high (Pusa Arhar 2018-4) and low (ICP-1452). Treatments improved in vitro PD and essential amino acid content, with autoclaving showing significantly higher PD (ICP-1452: 90.4%, Pusa-Arhar 2018-4: 84.32%) ascribed to disruption of tight protein matrices. Significant increase in ß-turn, reduction in protein: starch, protein: polyphenol interactions as well as breakdown of storage proteins revealed by the analysis of protein structural properties. This study suggests thermal treatments, particularly autoclaving, can enhance pigeon pea protein's nutritional quality for its utilization as a new ingredient in development of healthy foods.

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The objective of the review is to revisit the findings of the 2011 Food and Agriculture Organization of the United Nations (FAO) Expert Consultation on Dietary Protein Quality Evaluation in Human Nutrition, and to report on progress on uptake of the findings. It is evident that since 2011 there has been a concerted research effort to enhance an understanding of the protein quality of foods. The validity of the growing pig ileal protein digestibility assay has been confirmed and numerous studies reported using the growing pig as a model to give true ileal amino acid digestibility values for foods as consumed by humans. This has allowed for the determination of digestible indispensable amino acid scores (DIAAS) for a range of foods. A new non-invasive true ileal amino acid digestibility assay in humans which can be applied in different physiological states, called the dual-isotope assay, has been developed and applied to determine the DIAAS values of foods. It is concluded that DIAAS is currently the most accurate score for routinely assessing the protein quality rating of single source proteins. In the future, the accuracy of DIAAS can be enhanced by improved information on: the ideal dietary amino acid balance including the ideal dispensable to indispensable amino acid ratio; dietary indispensable amino acid requirements; effects of processing on ileal amino acid digestibility and lysine bioavailability. There is a need to develop rapid, inexpensive in vitro digestibility assays. Conceptual issues relating DIAAS to food regulatory claims, and to holistic indices of food nutritional and health status are discussed. The first recommendation of the 2011 Consultation regarding treating each indispensable amino acid as an individual nutrient has received little attention. Consideration should be given to providing food label information on the digestible contents of specific indispensable amino acids.

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Adding value to food industry by-products, like sunflower meal (SFM), through their utilization as ingredients in new food products can improve sustainability of food systems. This research investigated extrusion cooking to produce high-moisture meat analogues (HMMAs) made from blends of soy protein isolate and expeller-pressed SFM. The effects of feed moisture content [FMC] (60, 65, and 70%, wet basis) and SFM concentration (37.5, 50, and 62.5%, total blend weight basis) on physical and protein nutritional quality attributes of HMMAs were investigated. The processing temperatures (including cooling die), screw speed and feed rate were kept constant at 60-80-115-125-50-25 °C (from feeder to the die end), 200 rpm and 0.5 kg/h (dry basis), respectively. An increase in SFM concentration and FMC significantly (p < 0.05) reduced the mechanical energy requirements for extrusion. Cutting strength and texture profile analysis of HMMAs indicated softer texture with increases in SFM and FMC. X-ray microcomputed tomography analysis revealed that the microstructure of the HMMAs at the centre and towards the surface was different and affected by SFM concentration and FMC. The in vitro-protein digestibility corrected amino acid score of the HMMAs ranged between 85 and 91% and did not show significant (p < 0.05) changes as a function of FMC or SFM concentration. HMMAs produced from 37.5% SFM at 70% FMC showed no deficiency in essential amino acids for all age categories except for infants, suggesting the high potential of SFM and soy protein blends for creating nutritious meat alternative products. Overall, this work provided valuable insights regarding the effects of soy protein replacement by SFM on the textural, microstructural and nutritional quality of HMMA applications, paving the way for value-addition to this underutilized food industry by-product.

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The amount of chitin, a nitrogen-containing dietary fiber, in edible insects can mislead the exact nitrogen-to-protein conversion factor (NPF) and true protein content. We determined the amino acid score (AAS), protein digestibility-corrected AAS (PDCAAS), chitin content, and net NPF of five edible insects. Additionally, the effect of the amino acid composition of migratory locust on rat growth were investigated. The AAS of the insects were ranged from 63 to 94. The chitin contents were ranged from 1.6 g/100 g to 10.7 g/100 g. The PDCAAS, calculated by AAS and gut-intestinal digestibility, ranged from 44 to 81, which was lower than casein (97). The net NPF ranged from 4.93 to 5.76, which were lower than the conventional value. Dietary migratory locust, whose PDCAAS was the lowest, decreased growth and altered lipid metabolism. Therefore, a lower PDCAAS and overestimation of net NPF of insects can affect the true protein calculations and growth.

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Reducing the content of quickly absorbed carbohydrates and saturated fats in snack formulations while increasing the consumption of high-quality proteins are effective strategies to prevent obesity in childhood. Thus, the nutritional value, digestibility, and functionality of fava beans (Vicia faba L.) fermented with Pleurotus ostreatus were examined as potential ingredients for food design. Solid-state fermentation enhanced the protein content by 16% with a rise in essential (25%) and non-essential (15%) amino acids while decreasing total carbohydrate content and tannin levels. Moreover, fermentation modified the amino acid profile released during digestion, increasing amino acids such as valine, isoleucine, and threonine, which are vital for health and development in childhood. Furthermore, the bioaccessible fraction of the fermented bean showed a 60% of ACE inhibition and improved magnesium bioaccessibility. Consequently, fava beans fermented with Pleurotus ostreatus emerged as a new ingredient in the development of new protein-rich snacks tailored for children and adolescents.

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The escalating global population is anticipated to intensify the demand for high-quality proteins, necessitating the exploration of alternative protein sources. Edible insects are a promising solution, owing to their nutritional richness and sustainability. However, their digestibility and protein quality, particularly after culinary treatment, remains underexplored. In the present study, we investigated the effects of various culinary treatments on the protein digestibility of two insect species, Tenebrio molitor and Gryllus assimilis. Our findings revealed that culinary treatments such as boiling, roasting, drying, and microwave heating significantly influenced the digestibility of both insect species. Notably, drying emerged as the most effective method, leading to a substantial increase in digestibility. Furthermore, we assessed protein quality using the digestible indispensable amino acid score (DIAAS) and found that the choice of the calculation method significantly influenced the evaluation of protein quality. By including the sum of the anhydrous amino acids, we eliminated the potential overestimation of protein content and obtained a more reliable assessment of protein quality. Our results underscore the importance of culinary treatments and calculation methods in determining the suitability of insects as protein sources for human nutrition.

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Many improvements have been made to bring infant formula (IF) closer to human milk (HM) regarding its nutritional and biological properties. Nevertheless, the protein components of HM and IF are still different, which may affect their digestibility. This study aimed to evaluate and compare the protein digestibility of HM and IF using the infant INFOGEST digestion method. Pooled HM and a commercial IF were subjected to the infant INFOGEST method, which simulates the physiological digestion conditions of infants, with multiple directions, i.e. the curd state, gel images of SDS-PAGE, molecular weight distribution, free amino acid concentrations and in vitro protein digestion rate. HM underwent proteolysis before digestion and tended to have a higher protein digestion rate with finer curds during gastric digestion, than the IF. However, multifaceted analyses showed that the protein digestibility of HM and IF was not significantly different after gastrointestinal digestion. In conclusion, the infant INFOGEST method showed that the digestibility of HM and IF proteins differed to some extent before digestion and after gastric digestion, but not at the end of gastrointestinal digestion. The findings of this study will contribute to the refinement of IF with better protein digestibility in infant stomach.

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This review discusses different protein sources and their role in human nutrition, focusing on their structure, digestibility, and bioavailability. Plant-based proteins, such as those found in legumes, nuts, and seeds, may contain anti-nutritional factors that impact their bioavailability apart from structural and compositional differences from animal proteins. Animal proteins are generally highly digestible and nutritionally superior to plant proteins, with higher amino acid bioavailability. Alternative protein sources are also processed in different ways, which can alter their structure and nutritional value, which is also discussed.

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The reliance by North American regulatory authorities on in vivo rodent bioassays-Protein Correct-Amino Acid Score (PDCAAS) in the U.S. and Protein Efficiency Ratio (PER) in Canada-to measure the protein quality for protein content claim substantiation represents a major barrier for innovation in the development and marketing of protein foods. Although FAO in 2013 proposed a new method (Digestible Indispensable Amino Acid Score, DIAAS), it is still not used for protein content claim substantiation in any jurisdiction. Together with public health efforts to increase the consumption of plant-based foods, removing hurdles is key to incentivizing the food industry to measure protein digestibility in making food formulation decisions as well as in claiming protein content on product labels. To address this issue, a pathway has been proposed to position alternative methods for in vitro protein digestibility in collaborative studies to generate the data necessary for method approval by a certifying body. The latter is critical to the potential recognition of these methods by both Health Canada and the US FDA. The purpose of this article is to briefly summarize the state-of-the-art in the field, to inform the research community of next steps, and to describe the path engaging collaborative laboratories in a proficiency test as the first step in moving forward toward acceptance of in vitro digestibility methods. Throughout, a consultative and iterative process will be utilized to ensure the program goals are met. Success will be achieved when the proposed path results in the acceptance of an in vitro methods for protein digestibility used for PDCAAS determinations, which will enable increased protein analyses and improved nutrition labeling of protein foods.

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Protein is an essential macronutrient in our diet, source of nitrogen and essential amino acids, but the biological utilization of dietary protein depends on its digestibility and the absorption of amino acids and peptides in the gastrointestinal tract. The methods to define the amount and the quality of protein to meet human nutritional needs, such as the Digestible Indispensable Amino Acid Score (DIAAS), require the use of animal models or human studies. These in vivo methods are the reference in protein quality evaluation, but they are expensive and long-lasting procedures with significant ethical restrictions. Therefore, the development of rapid, reproducible and in vitro digestion methods validated with in vivo data is an old demand. This review describes the challenges of the in vitro digestion methods in the evaluation of the protein nutritional quality. In addition to the technical difficulties to simulate the complex and adaptable processes of digestion and absorption, these methods are affected by similar limitations as the in vivo procedures, i.e., analytical techniques to accurately determine bioavailable amino acids and the contribution of the endogenous nitrogen. The in vitro methods used for the evaluation of protein digestibility, with special attention on those showing comparative data, are revised, emphasizing their pros and cons. The internationally harmonized digestion protocol proposed by the INFOGEST network is being adapted to evaluate protein and amino acid digestibility. The inter-laboratory reproducibility of this protocol was demonstrated for dairy products. The in vivo/in vitro comparability results obtained to date with this protocol for several plant and animal sources are promising, but it requires an extensive validation with a wider range of foods and substrates with known in vivo digestibility. These in vitro methods will probably not be applicable to all foods, and therefore, it is important to identify their limitations, not to elude their use, but to apply them within the limits, by using the appropriate standards and references, and always as a complementary tool to in vivo tests to reduce their number.

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Soluble trehalose-conjugated quinoa proteins (T-QPs) were effectively prepared using the pH-shifting mechanism. The structural properties of the T-QPs were evaluated using a comparative evaluation, which included analyzing the amide I, surface charge and hydrophobicity, protein conformation, thermal stability, and protein structures. The results suggested that the development of the T-QPs was influenced mainly by no-covalent bonds. These interactions significantly influenced (P < 0.05) the quinoa proteins' conformation and higher-protein structure. T-QP had significant (P < 0.05) surface properties. Furthermore, the T-QPs exhibited improved solubility (79.7 to 88.4%) and digestibility (79.8 to 85.1%). Therefore, quinoa protein proved an excellent plant-based protein for conjugation with disaccharides. These findings provide significant insight into the potential development of modified proteins with enhanced solubility and digestibility by creating trehalose-conjugated plant-based proteins.

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This study evaluates the impact of high-intensity ultrasound (HIU) on the physicochemical properties and in-vitro digestibility of Atlantic cod (Gadus morhua). Various ultrasound durations (0-60 min) were applied to assess changes in color attributes, total antioxidant capacity (TAC), total flavonoid content (TFC), total phenolic content (TPC), total protein content, and in-vitro protein digestibility (IVPD). Results indicated HIU maximumly increased TAC, TFC, TPC, and peptide content before digestion by 7.28 % (US60), 3.00 % (US30), 32.43 % (US10), and 18.93 % (US60), respectively. While HIU reduced total protein content, it enhanced IVPD by up to 12.24 % (US30). Color attributes electron microscopy reflected structural changes in the cod samples, suggesting the effectiveness of HIU in altering protein structures. These findings highlight HIU's potential as a non-thermal technique for improving the sensory and nutritional quality of Atlantic cod, offering valuable insights for the seafood processing industry and consumers.

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This study aimed to investigate the impact of three cooking ways (sous vide (SV), frying (FR) and roasting (RO)) on pork protein digestion characteristics under conditions simulating healthy adult (control, C) and elderly individuals with achlorhydria (EA). Changes in degree of hydrolysis (DH), SDS-PAGE profiles, zeta potential, particle size and secondary structure during digestion were evaluated. Our results revealed the EA condition markedly affected the protein digestion process of pork with different cooking ways. The DH values of SV (25.62%), FR (21.38%) and RO (19.40%) under the EA condition were significantly lower than those of under the control condition (38.32%, 33.00% and 30.86%, respectively). Moreover, differences were also observed among three cooking ways under the EA condition. For a given cooking way, the differences between control and EA conditions gradually diminished from the gastric to the intestinal phase. Under a certain digestion condition, SV maintained the highest degree of digestion throughout the process, particularly under the EA condition. Therefore, we conclude that pork cooked by sous vide is more recommendable for the elderly considering protein digestibility.

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This review aims to provide an updated overview of the effects of protein extraction/recovery on antinutritional factors (ANFs) in plant protein ingredients, such as protein-rich fractions, protein concentrates, and isolates. ANFs mainly include lectins, trypsin inhibitors, phytic acid, phenolic compounds, oxalates, saponins, tannins, and cyanogenic glycosides. The current technologies used to recover proteins (e.g., wet extraction, dry fractionation) and novel technologies (e.g., membrane processing) are included in this review. The mechanisms involved during protein extraction/recovery that may enhance or decrease the ANF content in plant protein ingredients are discussed. However, studies on the effects of protein extraction/recovery on specific ANFs are still scarce, especially for novel technologies such as ultrasound- and microwave-assisted extraction and membrane processing. Although the negative effects of ANFs on protein digestibility and the overall absorption of plant proteins and other nutrients are a health concern, it is also important to highlight the potential positive effects of ANFs. This is particularly relevant given the rise of novel protein ingredients in the market and the potential presence or absence of these factors and their effects on consumers' health.

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