Raw and roasted pistachio nuts (Pistacia vera L.) are 'good' sources of protein based on their digestible indispensable amino acid score as determined in pigs.

BACKGROUND: Pistachio nuts may be consumed as raw nuts or as roasted nuts. However, there is limited information about the protein quality of the nuts, and amino acid (AA) digestibility and protein quality have not been reported. Therefore, the objective of this research was to test the hypothesis that raw and roasted pistachio nuts have a digestible indispensable AA score (DIAAS) and a protein digestibility corrected AA score (PDCAAS) greater than 75, thereby qualifying them as a good source of protein. RESULTS: The standardized ileal digestibility (SID) of all indispensable AAs, except arginine and phenylalanine, was less in roasted pistachio nuts than in raw pistachio nuts (P < 0.05). Raw pistachio nuts had a PDCAAS of 73, and roasted pistachio nuts had a PDCAAS of 81, calculated for children 2-5 years, and the limiting AA in the PDCAAS calculation was threonine. The DIAAS values calculated for children older than 3 years, adolescents, and adults was 86 and 83 for raw and roasted pistachio nuts respectively. The limiting AA in both raw and roasted pistachio nuts that determined the DIAAS for this age group was lysine. CONCLUSION: The results of this research illustrate that raw and roasted pistachio nuts can be considered a good quality protein source with DIAAS greater than 75; however, processing conditions associated with roasting may decrease the digestibility of AAs in pistachio nuts. © 2020 Society of Chemical Industry.

Digestible indispensable amino acid scores of nine cooked cereal grains.

True ileal digestibility (TID) values of amino acid (AA) obtained using growing rats are often used for the characterisation of protein quality in different foods and acquisition of digestible indispensable amino acid scores (DIAAS) in adult humans. Here, we conducted an experiment to determine the TID values of AA obtained from nine cooked cereal grains (brown rice, polished rice, buckwheat, oats, proso millet, foxtail millet, tartary buckwheat, adlay and whole wheat) fed to growing Sprague-Dawley male rats. All rats were fed a standard basal diet for 7 d and then received each diet for 7 d. Ileal contents were collected from the terminal 20 cm of ileum. Among the TID values obtained, whole wheat had the highest values (P<0·05), and polished rice, proso millet and tartary buckwheat had relatively low values. The TID indispensable AA concentrations in whole wheat were greater than those of brown rice or polished rice (P<0·05), and polished rice was the lowest total TID concentrations among the other cereal grains. The DIAAS was 68 for buckwheat, 47 for tartary buckwheat, 43 for oats, 42 for brown rice, 37 for polished rice, 20 for whole wheat, 13 for adlay, 10 for foxtail millet and 7 for proso millet. In this study, the TID values of the nine cooked cereal grains commonly consumed in China were used for the creation of a DIAAS database and thus gained public health outcomes.

Values for digestible indispensable amino acid scores (DIAAS) for some dairy and plant proteins may better describe protein quality than values calculated using the concept for protein digestibility-corrected amino acid scores (PDCAAS).

An experiment was conducted to compare values for digestible indispensable amino acid scores (DIAAS) for four animal proteins and four plant proteins with values calculated as recommended for protein digestibility-corrected amino acid scores (PDCAAS), but determined in pigs instead of in rats. Values for standardised total tract digestibility (STTD) of crude protein (CP) and standardised ileal digestibility (SID) of amino acids (AA) were calculated for whey protein isolate (WPI), whey protein concentrate (WPC), milk protein concentrate (MPC), skimmed milk powder (SMP), pea protein concentrate (PPC), soya protein isolate (SPI), soya flour and whole-grain wheat. The PDCAAS-like values were calculated using the STTD of CP to estimate AA digestibility and values for DIAAS were calculated from values for SID of AA. Results indicated that values for SID of most indispensable AA in WPI, WPC and MPC were greater (P<0·05) than for SMP, PPC, SPI, soya flour and wheat. With the exception of arginine and tryptophan, the SID of all indispensable AA in SPI was greater (P<0·05) than in soya flour, and with the exception of threonine, the SID of all indispensable AA in wheat was less (P<0·05) than in all other ingredients. If the same scoring pattern for children between 6 and 36 months was used to calculate PDCAAS-like values and DIAAS, PDCAAS-like values were greater (P<0·05) than DIAAS values for SMP, PPC, SPI, soya flour and wheat indicating that PDCAAS-like values estimated in pigs may overestimate the quality of these proteins.

Estimating In Vitro Protein Digestion and Protein Digestibility Corrected Amino Acid Score of Chicken Breasts Affected by White Striping and Wooden Breast Abnormalities.

An understanding regarding impacts of growth-related myopathies, i.e., white striping (WS) and wooden breast (WB), on the quality of dietary protein from cooked chicken breast is still limited. This study aimed at comparing protein content and in vitro protein digestion and estimating the in vitro protein digestibility corrected amino acid score (PDCAAS) of cooked chicken meat exhibiting different abnormality levels (i.e., normal, WS, and WS + WB). The results show that the WS + WB samples exhibited lower protein content, greater cooking loss, and greater lipid oxidation than those of normal samples (p < 0.05). No differences in protein carbonyls or the myofibril fragmentation index were found (p ≥ 0.05). Cooked samples were hydrolyzed in vitro using digestive enzyme mixtures that subsequently mimicked the enzymatic reactions in oral, gastric, and intestinal routes. The WS + WB samples exhibited greater values of free NH2 and degree of hydrolysis than the others at all digestion phases (p < 0.05), suggesting a greater proteolytic susceptibility. The in vitro PDCAAS of the WS + WB samples was greater than that of the other samples for pre-school children, school children, and adults (p < 0.05). Overall, the findings suggest that the cooked chicken breast with the WS + WB condition might provide greater protein digestibility and availability than WS and normal chicken breasts.

Options for substantiating protein content claims for conventional foods.

In Canada and the United States, front-of-package protein content claims require data to support the quality of the protein. In general, protein quality reflects the product of the amino acid composition of the food protein relative to human amino acid requirements and a measure of digestibility. The currently accepted method in both jurisdictions is the protein digestibility-corrected amino acid score (PDCAAS) that requires the measurement of true fecal protein (nitrogen) digestibility. The latter must be measured in vivo using a rat model. This requirement for animal testing is inconsistent with international efforts to reduce the usage of animals in testing for regulatory purposes. The current commentary positions four options to remove the need to use animal testing for determining protein quality, when considering protein content claim substantiation. These options include (i) a focus on protein quantity alone; (ii) the use of the amino acid score alone, with no correction for digestibility; (iii) the use of a fixed digestibility coefficient to estimate protein quality; and (iv) the use of in vitro methods to measure protein and/or amino acid digestibility. The relative merits and deficiencies of the options are positioned with the goal of encouraging dialogue within the regulatory agencies to move towards alternative approaches for substantiating protein content claims on foods, including those derived from plant-based sources.

Low protein digestibility-corrected amino acid score and net nitrogen-to-protein conversion factor value of edible insects.

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.

Digestible indispensable amino acid score (DIAAS): 10 years on.

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.

The in vivo and in vitro protein quality of three hemp protein sources.

In this work, the protein quality of defatted hemp hearts and protein-enriched hemp fractions was determined. Protein quality was assessed using a rodent bioassay to evaluate growth and protein digestibility, while amino acid composition was determined via HPLC. A method for determining in vitro protein digestibility was compared to in vivo methodology and used to generate an in vitro protein quality score. The true protein digestibility of hemp protein 2, a hemp protein concentrate, was significantly lower than that of either defatted hemp hearts or hemp protein 1, a hemp protein concentrate (p < .05). While there was no relationship between the in vivo and in vitro measurements of protein digestibility (R 2 = .293, p = .459), there was a significant correlation between the protein digestibility-corrected amino acid score (PDCAAS) determined in vivo and in vitro PDCAAS (R 2 = .989, p = .005). The protein efficiency ratio of hemp protein 1 was significantly lower than that of either defatted hemp hearts or hemp protein 2 (p < .05). These data highlight the nutritional capacity of hemp protein sources while also demonstrating the relationship between in vivo and in vitro methods for determining protein quality.

Impact of cooking on the protein quality of Russet potatoes.

Despite being low in crude protein, on a fresh weight basis, given their overall contribution to the North American diet, potatoes contribute approximately 2%-4% of the population's protein intake. However, the quality of the protein remains ill-defined. To that end, Russet potatoes were secured and subjected to various cooking conditions (raw [control], boiled, baked, microwaved, and fried [3, 6, and 9 min]) to determine the impact of cooking method on protein quality, as determined by amino acid score (AAS) and indices of in vivo true fecal protein digestibility (TFPD%; rodent bioassay) and in vitro protein digestibility (pH-drop, pH-Stat, and simulated gastrointestinal digestion both static and dynamic). The AAS of raw Russet potatoes was 0.67 ± 0.01, with histidine being the limiting AA. Frying led to a significant reduction in the AAS, however, other cooking methods yielded similar results to the raw control. The TFPD% of raw potato was low (40.5% ± 3.9%) and was significantly enhanced to over 80% with all cooking methods. Similar patterns were observed with all in vitro measures, however, all methods yielded higher values for the raw control samples. Final protein digestibility-corrected AAS (PDCAAS; product of AAS and TFPD%) values ranged from 0.27 (raw) to a high of 0.57 (boiled), with cooked values being comparable to other plant-based protein sources, including grains, and some nuts and pulses. In vitro PDCAAS values followed similar trends. This study defined the protein quality of cooked Russet potatoes and provides data for use in defining the quality of total protein consumed in the North American diet.

Protein quality of soy and the effect of processing: A quantitative review.

There is a growing demand for plant-based protein-rich products for human consumption. During the production of plant-based protein-rich products, ingredients such as soy generally undergo several processing methods. However, little is known on the effect of processing methods on protein nutritional quality. To gain a better understanding of the effect of processing on the protein quality of soy, we performed a quantitative review of in-vivo and in-vitro studies that assessed the indispensable amino acid (IAA) composition and digestibility of varying soy products, to obtain digestibility indispensable amino acids scores (DIAAS) and protein digestibility corrected amino acid scores (PDCAAS). For all soy products combined, mean DIAAS was 84.5 ± 11.4 and mean PDCAAS was 85.6 ± 18.2. Data analyses showed different protein quality scores between soy product groups. DIAAS increased from tofu, soy flakes, soy hulls, soy flour, soy protein isolate, soybean, soybean meal, soy protein concentrate to soymilk with the highest DIAAS. In addition, we observed broad variations in protein quality scores within soy product groups, indicating that differences and variations in protein quality scores may also be attributed to various forms of post-processing (such as additional heat-treatment or moisture conditions), as well as study conditions. After excluding post-processed data points, for all soy products combined, mean DIAAS was 86.0 ± 10.8 and mean PDCAAS was 92.4 ± 11.9. This study confirms that the majority of soy products have high protein quality scores and we demonstrated that processing and post-processing conditions can increase or decrease protein quality. Additional experimental studies are needed to quantify to which extent processing and post-processing impact protein quality of plant-based protein-rich products relevant for human consumption.

Effect of roasting pulse seeds at different tempering moisture on the flour functional properties and nutritional quality.

Knowledge on the functional and nutritional properties of wet roasted pulses can increase the utilization of processed pulses as ingredients in food products. This study investigated the effects of tempering different pulse [chickpea (CP), green lentil (GL), navy bean (NB) and yellow pea (YP)] seeds to 20 or 30% moisture prior to roasting (160℃ for 30 min) on the functional properties and nutritional quality of their resulting flours. The surface charge of each pulse remained the same (p > 0.05) after wet roasting and there were no significant (p > 0.05) differences between the different raw pulse flours. The oil holding capacity (OHC) of GL (~2 g/g) was not improved by wet roasting (p > 0.05) whereas the other pulses generally had better OHC for one or both of the tempering moistures used prior to roasting. Foaming properties of all pulses decreased after heat treatment with the exception of both foaming capacity (107%) and stability (~71%) for GL tempered to 20% moisture prior to roasting (p > 0.05). Raw GL had inferior foaming properties compared to the other raw pulse flours (p < 0.001). Emulsion properties of the wet roasted pulses were similar to those of the control (raw flour) for each pulse. Solubility decreased with roasting regardless of the tempering moisture (p < 0.05) whereas in general the in vitro protein digestibility increased. Small improvements (2.4-6.9% increase) in the in vitro protein digestibility-corrected amino acid score were found for GL and NB tempered to 20% moisture before roasting and roasted YP at either moisture content (p < 0.05). Wet roasting increased (p < 0.05) the rapidly digestible starch content, more so with a tempering moisture of 30%. Overall the results from this study will allow for the utilization of wet roasted pulses as ingredients based on their functional properties and protein quality.

A Rat Study to Evaluate the Protein Quality of Three Green Microalgal Species and the Impact of Mechanical Cell Wall Disruption.

The present study was conducted to evaluate the protein quality of microalgae species Chlorella vulgaris (CV), Chlorella sorokiniana (CS), and Acutodesmus obliquus (AO) and assess the impact of mechanical cell wall disruption. Male Sprague-Dawley rats, around 156 g after adaptation, were placed in metabolic cages and fed experimental diets that were either protein-free or contained 10% protein solely from one of the undisrupted or disrupted CV, CS, and AO. After 3 days, feces were collected for a period of 5 days and analyzed together with diet samples for crude protein contents. Apparent protein digestibility, true protein digestibility, amino acid score, and protein digestibility-corrected amino acid score were calculated. In vitro protein digestibility was measured using the pepsin-pancreatin method and the in vitro protein digestibility-corrected amino acid score was calculated. The crude protein contents of CV, CS, and AO were 53.5, 50.2, and 40.3%, respectively. The amino acid score of the first limiting amino acid was 1.10, 1.27, and 0.86, true protein digestibility was 64.7, 59.3, and 37.9% and protein digestibility-corrected amino acid score was 0.63, 0.64, and 0.29, respectively, for CV, CS, and AO. Mechanical cell disruption significantly improved protein digestibility without a substantial impact on the amino acid profile and score, resulting in the increase of protein digestibility-corrected amino acid score to 0.77, 0.81, and 0.46, respectively, for disrupted CV, CS, and AO. There was a strong correlation between in vitro protein digestibility and apparent protein digestibility (r = 0.986), and also between in vitro protein digestibility-corrected amino acid score and in vivo protein digestibility-corrected amino acid score (r = 0.994). The results suggest that the CV and CS are acceptable sources of protein for humans and animals and quality can be markedly improved by mechanical cell wall disruption. Additionally, in vitro protein digestibility measured using the pepsin-pancreatin method may be used to screen protein product candidates, save animals, reduce cost, and accelerate product development.

Thermal processing methods differentially affect the protein quality of Chickpea (Cicer arietinum).

Chickpea is a widely produced pulse crop, but requires processing prior to human consumption. Protein bioavailability and amino acid quantity of chickpea flour can be altered by multiple factors including processing method. For this reason, the protein quality of processed chickpea flour was determined using in vivo and in vitro analyses for processed chickpeas. Processing differentially affected the protein digestibility-corrected amino acid score (PDCAAS) of chickpeas with extruded chickpea (83.8) having a higher PDCAAS score than both cooked (75.2) and baked (80.03). Interestingly, the digestible indispensable amino acid score (DIAAS) value of baked chickpea (0.84) was higher compared to both extruded (0.82) and cooked (0.78). The protein efficiency ratio, another measure of protein quality, was significantly higher for extruded chickpea than baked chickpea (p < .01). In vivo and in vitro analysis of protein quality were well correlated (R 2 = .9339). These results demonstrated that under certain circumstances in vitro methods could replace the use of animals to determine protein quality.

Measuring Protein Content in Food: An Overview of Methods.

In order to determine the quantity of protein in food, it is important to have standardized analytical methods. Several methods exist that are used in different food industries to quantify protein content, including the Kjeldahl, Lowry, Bradford and total amino acid content methods. The correct determination of the protein content of foods is important as, often, as is the case with milk, it determines the economic value of the food product and it can impact the economic feasibility of new industries for alternative protein production. This editorial provides an overview of different protein determination methods and describes their advantages and disadvantages.

Determination of the protein quality of almonds (Prunus dulcis L.) as assessed by in vitro and in vivo methodologies.

Almonds (Prunus dulcis), such as all nuts, are positioned within the protein foods grouping within the current U.S. Dietary Guidelines. The ability to make claims related to the protein content of almonds, within the United States, requires substantiation via the use of the Protein Digestibility-Corrected Amino Acid Score (PDCAAS). The present study was designed to provide current estimates of PDCAAS, using both in vivo and in vitro assays, of key almond varietals from the 2017 California harvest. Additionally, historical protein and amino acid composition data on 73 separate analyses, performed from 2000 to 2014, were analyzed. Amino acid analysis confirmed lysine as the first-limiting amino acid, generating amino acid scores of 0.53, 0.52, 0.49, and 0.56 for Butte, Independence, Monterey, and Nonpareil varietals, respectively. True fecal protein digestibility coefficients ranged from 85.7% to 89.9% yielding PDCAAS values of 44.3-47.8, being highest for Nonpareil. Similar, albeit lower, results were obtained from the in vitro assessment protocol. Analysis of the historical data again positioned lysine as the limiting amino acid and yielded information on the natural variability present within the protein and amino acid profiles of almonds. Comparison of the 2017 AA profile, averaged across almond varietals, to the historical data provided strong evidence of persistence of amino acid composition and indices of protein quality over time.

Postprandial amino acid, glucose and insulin responses among healthy adults after a single intake of Lemna minor in comparison with green peas: a randomised trial.

A high protein content combined with its enormous growth capacity make duckweed an interesting alternative protein source, but information about postprandial responses in humans is lacking. The present study aimed to assess the postprandial serum amino acid profile of Lemna minor in healthy adults in comparison with green peas. A secondary objective was to obtain insights regarding human safety. A total of twelve healthy volunteers participated in a randomised, cross-over trial. Subjects received two protein sources in randomised order with a 1-week washout period. After an overnight fast, subjects consumed L. minor or peas (equivalent to 20 g of protein). After a baseline sample, blood samples were taken 15, 30, 45, 60, 75, 90, 120, 150 and 180 min after consumption to assess amino acid, glucose and insulin levels. Heart rate, blood pressure and aural temperature were measured before and after consumption, and subjects reported on gastrointestinal discomfort for four subsequent days. Compared with green peas, significantly lower blood concentrations of amino acids from L. minor were observed, indicating lower digestibility. L. minor consumption resulted in lower plasma glucose and insulin levels compared with peas, probably due to different glucose content. There were no significant differences concerning the assessed health parameters or the number of gastrointestinal complaints, indicating that a single bolus of L. minor - grown under controlled conditions - did not induce acute adverse effects in humans. Further studies need to investigate effects of repeated L. minor intake and whether proteins purified from L. minor can be digested more easily.

Research and regulatory gaps for the substantiation of protein content claims on foods.

Protein claims provide guidance to consumers seeking protein-rich foods. Protein claim regulations differ globally, and both Canada and the United States require protein quality assessments. A tripartite workshop identified the need to (i) harmonize, (ii) update existing amino acid composition and digestibility databases, (iii) develop non-animal bioassays, and (iv) evaluate the impact of protein claims on human health. The Protein-Digestibility Corrected Amino Acid Score method is recommended for current regulatory use in Canada.

The bioavailability of iron, zinc, protein and vitamin A is highly variable in French individual diets: Impact on nutrient inadequacy assessment and relation with the animal-to-plant ratio of diets.

Nutritional adequacy depends on nutrient intakes and bioavailability which strongly varies with the plant- or animal-origin of foods. The aim was to estimate iron, zinc, protein and vitamin A bioavailability from individual diets, and investigate its relation with the animal-to-plant ratio (A/P) of diets. Bioavailability was estimated in 1899 French diets using diet-based algorithms or food-group specific conversion factors. Nutrient inadequacy was estimated based on i) bioavailability calculated in each individual diet and ii) average bioavailability assumed for Western-diets. Mean iron absorption, zinc absorption, protein quality and ß-carotene conversion factor were 13%, 30%, 92%, and 17:1, respectively. Bioavailability displayed a high variability between individual diets, poorly explained by their A/P. Using individual bioavailability led to different inadequacy prevalence than with average factors assumed for Western-diets. In this population, the A/P does not seem sufficient to predict nutrient bioavailability and the corresponding recommended intakes. Nutritional adequacy should be assessed using bioavailability accounting for individual diets composition.

Impact of Processing on the Protein Quality of Pinto Bean (Phaseolus vulgaris) and Buckwheat (Fagopyrum esculentum Moench) Flours and Blends, As Determined by in Vitro and in Vivo Methodologies.

Blending of protein sources can increase protein quality by compensating for limiting amino acids present in individual sources, whereas processing grain flours by extrusion or baking can also alter protein quality. To determine the effect of baking and extrusion on the protein quality of blended flours from buckwheat and pinto beans, a rodent bioassay was performed and compared to an in vitro method of protein quality determination. Overall, extruded products had higher protein efficiency ratio values, increased digestibility, and greater protein digestibility corrected amino acid score (PDCAAS) values than baked products, with the extruded buckwheat/pinto blend having the greatest PDCAAS value of the experimental diets investigated. A correlation was found between both digestibility and PDCAAS values generated from in vitro and in vivo methods. The use of in vitro digestibility analysis should be investigated as a potential replacement for the current rodent assay for nutrient content claim purposes.

Determination of the protein quality of cooked Canadian pulses.

A study to determine the protein digestibility-corrected amino acid score and protein efficiency ratio of nine different cooked Canadian pulse classes was conducted in support of the establishment of protein quality claims in Canada and the United States. Split green and yellow pea, whole green lentil, split red lentil, Kabuli chickpea, navy bean, pinto bean, light red kidney bean, and black bean were investigated. Protein digestibility-corrected amino acid score (PDCAAS) and the protein efficiency ratio (PER) were determined using the appropriate rodent models. All pulses had high digestibility values, >70%, with PDCAAS values greater than 0.5, thereby qualifying as a quality protein in the United States, but only navy beans qualified as a good source of protein. All pulses except whole green lentils, split red lentils, and split green peas would qualify as sources of protein with protein ratings between 20 and 30.4 in Canada. These findings support the use of pulses as protein sources in the regulatory context of both the United States and Canada.