Relationship between amino acid scores and protein quality indices based on rat growth

Protein efficiency ratio (PER), relative PER (RPER), net protein ratio (NPR) and relative NPR (RNPR) values, and amino acid scores were calculated for 20 food products (casein, casein + Met, beef salami, skim milk, tuna, chicken frankfurters, sausage, heated skim milk, peanut butter, rolled oats, soy isolate, chick peas, pea concentrate, kidney beans, wheat cereal, pinto bean, lentils, rice-wheat gluten cereal, macaroni-cheese, and beef stew). In most cases, PER, RPER, NPR or RNPR ranked the products in the same order and positive correlations among the protein quality methods were highly significant (r=0.98–0.99). Amino acid scores (based on the first limiting amino acid, Lys-Met-Cys, Lys-Met-Cys-Trp or lys-Met-Cys-Trp-Thr) were positively correlated to the PER, RPER, NPR or RNPR data (r=0.61–0.75). Inclusion of the correction for true digestibility of protein improved the correlations between amino acid scores and the indices based on rat growth. The correlations were especially high between Lys-Met-Cys scores (corrected for true digestibility of protein) and PER, RPER, NPR or RNPR (r=0.86–0.91). Inclusion of the correction for true digestibility of individual amino acids did not result in further improvements of the correlations in most cases. It is concluded that adjusting amino acid scores for true digestibility of protein would be sufficient and further correction for digestibility of amino acids would be unnecessary in mixed diets.This paper was presented in part at the 71st annual FASAB meeting, Washington, DC, March 29–April 2, 1987.     

Influence of tryptophan supplementation of soy-based infant formulas on protein quality and on blood and brain tryptophan and brain serotonin in the rat model

A 2-wk feeding study with weanling rats was conducted to investigate theeffects of tryptophan supplementation of soy-based infant formulas on proteinquality and on the levels of blood and brain tryptophan and brain serotoninand 5-HIAA (5-hydroxyindole-3-acetic acid), a metabolite of serotonin. Thepowder and liquid concentrate forms of soy-based infant formulas obtainedfrom four manufacturers were tested. The tryptophan contents of the testsoy-based formulas (10.0–12.3 mg/g protein) were considerably lower comparedto those of human milk (17–19 mg/g protein). The infant formulas were fedas the sole source of protein in diets containing 8% protein, 20% fat, 5%cellulose and adequate amounts of minerals and vitamins. In general, supplementation of the diets containing test formulas with graded levels ofL-tryptophan (0.1 or 0.2%) had no effect on protein quality indices basedon rat growth such as PER (protein efficiency ratio), NPR (net protein ratio)and relative NPR (RNPR), but resulted in significant (p<;0.05) increases inthe concentrations of tryptophan in the plasma and brain, and serotonin and5-HIAA in brain of rats. The increased concentrations of brain tryptophan and5-hydroxyindoles (serotonin + 5-HIAA) in rats fed tryptophan-supplementedformulas compared to those fed unsupplemented formulas support the need forfurther research to investigate the influence of tryptophan supplementationof soy-based infant formulas on tryptophan metabolites and their potentialrelated effects on sleep latency and neurobehavioral developments in infants.     

Protein efficiency ratios and net protein ratios of selected protein foods

As a part of a cooperative study initiated to assess bothin vitro andin vivo protein quality methods, the protein efficiency ratio (PER) and net protein ratios (NPR) of 15 different protein sources were determined. Male weanling Sprague-Dawley rats were fed a 10% protein diet. Fourteen-day NPR and relative NPR (RNPR) values and 14- and 28-day PER and relative PER (RPER) values were calculated for each protein source. When protein quality values were expressed relative to ANRC casein, the 14- and 28-day PER data ranked the protein sources essentially in the same order. RPER values of nonfat dried skim milk (unheated) and tuna were more than 100% that of casein; nonfat dried skim milk (heated), chickpeas, and breakfast sausage were between 50 and 70% of that of casein; and pinto beans and rice-wheat gluten cereal did not support substantial growth of the rat. The NPR method did not always rank the protein sources in the same order as the PER method. For the poor quality proteins, RNPR values were much higher than the RPER values; however, the RNPR and RPER values agreed closely for high quality protein sources.A preliminary report of this work was given at the Annual Meeting of the Federation of American Societies for Experimental Biology, Washington, D.C., March 29–April 2, 1987. Fed. Proc. 1987; 46: 889.     

Digestibility of protein and amino acids in selected foods as determined by a rat balance method

Values (%) for true digestibility of crude protein and individual amino acids in 20 selected foods were determined by the rat balance (fecal) method. The products were fed as the sole source of protein in diets containing 8% crude protein (N × 6.25). Lowest true protein digestibility values (79–84) were obtained for pinto beans, kidney beans and lentils; intermediate values (89–92) were obtained for chick peas, beef stew, skim milk (over heated), rolled oats, whole wheat cereal, and pea protein concentrate; and highest values (94–100) were obtained for sausage, macaroni-cheese, rice-wheat gluten cereal, skim milk, tuna, soy isolate, peanut butter, chicken frankfurters, beef salami, casein and casein + methionine. In animal foods, peanut butter and soy isolate, the differences between true digestibility of crude protein and most individual amino acids were less than 5%. However, the values for true digestibility of methionine and cystine were up to 44% lower than those of crude protein in pinto beans, kidney beans, lentils, chick peas and pea concentrate. In these legumes, digestibility of crude protein was not a good predictor of digestibility of the limiting amino acids.     

Nutrient composition and biological evaluation of mesta (Hibiscus sabdariffa) seeds

Two varieties of mesta (Hibiscus sabdariffa) seeds were analysed for their proximate composition. Their protein (18.8–22.3%), fat (19.1–22.8%) and dietary fiber (39.5–42.6%) contents were found to be high. The seeds were found to be a good source of minerals like phosphorus, magnesium and calcium. Their lysine and tryptophan contents were also high. Sulphur containing amino acids were limiting in this seed protein and the chemical score of mesta seed protein was 40 and 57 for AMV-2 and Bhimili-1 varieties respectively. Mesta seed oil is rich in unsaturated fatty acids (70%), of which linoleic acid constituted 44%. Weanling rats were fed with 10% mesta seed protein before and after cooking for 4 weeks. Food intake of animals receiving raw mesta seed diets was significantly lower than those receiving cooked mesta seed diets as well as the casein control diet. Protein and dry matter digestibilities of raw and cooked mesta seed diets were lower than that of casein control diet. Cooking improved the food intake, gain in body weight, dry matter and protein digestibility of mesta seed diets. PER and NPU of cooked mesta seed diets were significantly higher than the corresponding raw diets. These results indicate that cooked mesta seed protein is of relatively good quality.     

Influence of sulfur-amino acid content variation in plant vs animal protein on serum and tissue lipids in rats

The objective of the study was to determine the effects of methionine and cysteine supplementation of soy protein isolate and casein on serum and tissue lipid levels in rats. Sixty male, weanling, Wistar-Kyoto rats were fed two sources of protein (casein or soy protein isolate) and three variations of sulfur-amino acid supplementation (none, methionine, or cysteine). At this level of protein intake (10% by weight), rats fed soy-based diets had similar serum lipid concentrations than rats fed casein-based diets. Choline was not added to the diet in order to be able to assess independent influences of methionine and cysteine on lipid metabolism. Overall, serum lipid values were greater in rats fed proteins supplemented with methionine while the addition of cysteine produced lower lipid levels. Liver lipid concentrations were increased tremendously upon cysteine supplementation of soy protein isolate. Protein quality, as determined by protein efficiency ratio, was improved by supplementation of either sulfur-amino acid; however, methionine had the greatest effect. Results indicate that the sulfur-amino acids influence lipid metabolism in the absence of dietary choline. The mechanism by which this occurs is not known.     

The protein quality of lupins (Lupinus mutabilis) alone and in combination with other protein sources

The chemical compositions, of raw and treated lupine flour were determined and compared with other plant protein sources. The protein content in the dry matter was 47.7% in untreated seeds ofLupinus mutabilis and about 56% in debittered seeds. The oil-cake contained 65.3% protein. The amino acid analysis showed that lupine protein is characterized by a low level of S-amino acids, the content of which amounts to only 50% of that of the FAO standard reference pattern. The protein quality was measured using the biological tests PER (Protein Efficiency Ratio), NPU (net protein utilization), and BV (biological value) in rats on diets with and without DL-methionine supplementation. PER determinations gave low values for the non-supplemented lupine proteins (1.34, semi-sweet variety; 1.53, water-extracted seeds; 1.19, oil-cake; 3.09, casein), but the PER's were improved by the addition of 0.2% DL-methionine to the diets (3.05, 2.69, 2.81, respectively). Raw as well as processed lupine protein showed an excellent apparent digestibility (80.0–85.8%; casein, 87.1%). The observed NPU and BV values confirmed the importance of methionine supplementation. The true digestibility of 92% was equivalent to that of casein. The complementation effects of mixing lupine protein with proteins from wheat, oat, barley, rice, maize, potato, quinua or fish were investigated by determination of the PER values of the respective mixtures. Feeding lupine protein with cereal proteins resulted in PER values that exceeded by far those of the proteins fed separately (true complementation). This result was not observed for the mixture of potato and lupine. High quality proteins like quinua and fish protein also showed no complementation effect with lupine protein but did improve the quality of the lupine protein to a great extent. The best results could be obtained with combinations of three different plant proteins, in which lupine protein always contributed one third of total protein. These mixtures resulted in PER values equal to those for casein or other animal proteins and may be applied as an economical way to prevent and combat malnutrition.     

Studies on protein quality of green gram (Phaseolus aureus)

Raw green gram contains trypsin inhibitor (5.6 × 10^–3 units/0.1 g) while cooking destroyed the trypsin inhibitor in green gram. No destruction was observed in the case of lysine, methionine, cystine, threonine and tryptophan. Sulphur containing amino acids have been found to be the only limiting amino acids in green gram from studies on blood levels of amino acids and PER experiments. Raw green gram, when fed as a sole source of protein promoted an extremely low growth rate, (13.2 g/4 weeks) and PER (0.77) in rats. Cooking slightly improved the nutritive value of the gram, as indicated by increase in gain in body weight (24 g/4 weeks) as well as PER (1.22). The presence of inhibitors in green gram did not appear to affect some of the essential amino acids as observed from the blood amino acid levels.     

Fertilizer effects on yield,mineral and amino acid composition,dietary fibre content and nutritive value of leeks

In pot experiments with greatly differing rates of N, P, S, K and Ca, dry matter (DM) yields of leek stems varied from 25 to 164 g/pot. Total-N and NO₃-N concentrations varied from 1.18 to 3.56% and from 10 to 1515 ppm in DM, respectively. Both N applications and P and K deficiency greatly increased total-N and NO₃-N content. S applications increased total-S content from 0.047 to 0.359% in DM, of which between approximately 100 to 25% were found in methionine+cystine. Total-N/total-S ratios decreased from 57 to 6 with the highest S level. P and K applications increased their respective content in DM two- and threefold. Severe Ca deficiency reduced Ca content from 0.495 to 0.045%. Iron, zinc, manganese and copper contents varied from 33–69, 14–26, 11–34 and 3.1–5.7 ppm in DM, respectively. Increasing N contents, whether due to N applications or P or K deficiency, decreased the content of all essential and some other amino acids in crude protein. Both S and severe P deficiency had a pronounced negative effect on amino acid composition and chemical score. Only glutamic acid (glutamine) and arginine were increased by increasing N contents. However, expressed as g/kg DM the concentrations of all amino acids were positively correlated with protein content. S and P deficiency reduced total dietary fibre (TDF) content of DM from 28.3 to 18.6% and 17.4%, respectively, of which between 53 and 60% were insoluble dietary fibre (IDF). Digestible energy (DE) was positively correlated with protein content (r=0.90**). In N-balance trials with rats, increasing protein concentrations (50% of total protein given as casein and supplemented with 1% methionine) raised the true digestibility (TD) of the protein from 44 to 72%. The biological value (BV) of protein was generally high, with a mean of 91.7 N deficiency tended to increase and S deficiency tended to decrease the BV.     

Seed protein fractions and amino acid composition in gram (Cicer arietinum)

Six chickpea strains were analysed for their protein content and various protein fractions. The protein content ranged from 20.9–25.27%. Albumin, globulin, prolamin and glutelin contents ranged from 8.39–12.31%; 53.44–60.29%; 3.12–6.89% and 19.38–24.40% respectively. Salt soluble proteins (albumin + globulin) and globulins resolved into 19–23 bands whereas albumin proteins resolved into 30–34 bands. The molecular weights of various polypeptides ranged from 10–91 kD. Amino acid analysis of total proteins revealed that glutamic acid was present in maximum concentration followed by aspartic acid and arginine. Just like other pulse proteins, chick pea proteins were also found deficient in sulphur containing amino acids.     

Estimation of optimum amino acid supplements to triticale

Based on the nitrogen balance (NB) data and the efficiency of amino acid utilization, optimum supplements of limiting amino acids (AA) to triticale were calculated and their effect on true N digestibility (TD), biological value of protein (BV) and net protein utilization (NPU) of a triticale-based diet was evaluated. To determine if AA balance must be considered in the calculation of the optimum supplements, the effect of a 20% of 40% excess of essential AA in lysine-, methionine-, threonine- or tryptophan-deficient diets was also studied. The AA excess had no significant effect on NB in diets deficient in lysine, methionine or threonine. However, NB in rats fed on the tryptophan-deficient diet increased as the AA excess increased. BV of the diet containing the optimized supplements of lysine, threonine, methionine and valine was comparable to that of lactalbumin or to the diet supplemented with all essential AA. The deletion of valine from the optimized supplement caused an insignificant decrease in BV. Due to the lower TD, NPU of diets containing the optimized AA supplements was lower than that of the diet containing all essential AA or of the lactalbumin-based diet.     

Effects of amylose and wheat bran on the levels of blood serum urea nitrogen (BUN), other blood parameters,growth and fecal characteristics in rats

A basal diet (containing 20% soybean protein isolate) was supplemented with limiting amino acids (AA, methionine, threonine and tryptophan); wheat bran (WB, 24.3%); amylose (AM, 10%) or a combination of AA and WB or AM to investigate their effects on the levels of blood serum urea nitrogen (BUN), other blood parameters, growth and fecal characteristics in young rats. After 3 weeks feeding, supplementation of the basal diet with AA or WB resulted in improved growth (gain/food ratio) while the addition of AM had no effect on rat growth. Addition of WB to the basal diet had no effect on BUN while the addition of AA and AM caused about 20 and 12% reduction in BUN values, respectively. Rats fed the AA-AM diet had the lowest BUN values (42% lower than those fed the basal diet). Fresh volume, fresh weight, moisture and dry weight of feces in rats fed the WB diets were about four to five fold higher than in those fed the AM diets. The results suggested that BUN may not be a good predictor of growth and protein quality in diets containing AM or a source of dietary fiber (WB).     

Rat bioassays for methionine availability in 16 food sources

Methionine availabilities of 16 test proteins were assessed by comparing ten day rat growth response to the test diets and reference (casein) diets. In a preliminary study, various concentrations of methionine and cystine were fed to determine methionine requirements and effect of excess cystine. Results indicated a methionine requirement of about 550 mg per 100 g diet. Cystine had a sparing affect of 50–55%, i.e., about 300 mg could be used to meet methionine requirements. Further additions of cystine (up to 2.6 times methionine) did not affect rat growth. Methionine availabilities were excellent (88–100%) for 15 of the 16 test foods; only pinto beans (58%) were low, but prior evidence indicates that the poor growth response was due to some factor other than availability.     

The nutritional quality of potato protein from intraspecific hybrids

The nutritional value of potato hybrids with Groups Andigena, Phureja, and Tuberosum parentage was measured by protein efficiency ratio (PER). Three diets at 6.0%, 7.75%, and 9.0% crude protein in potato flakes were fed to rats; 18 genotypes were analyzed and the hybrids had higher PER values than Group Tuberosum varieties, but increased tuber protein did not influence the PER values. The overall mean PER was 2.3, and 12 of the potato-flake diets were not different from the casein controls. Fortification with isoleucine did not improve the nutritional quality of the potato flakes. One diet was improved with the addition of both isoleucine and methionine. The influence of sulfur-containing amino acids in potato protein quality among these hybrids is not clear. It is suggested there may be two types of hybrids, one with dependence on these amino acids and a second type with a different protein factor(s) affecting the nutritional quality of the protein. Protein from these potato hybrids, as measured by PER, is among the highest in biological quality when compared with other proteins in plant foods.     

Nutritional and Antinutritional Characteristics of Seven South Indian Wild Legumes

Seeds from seven species of wild legumes of the South India were analyzed for proximate composition, mineral profiles, amino acid profiles of total seed protein, in vitro protein digestibility, and certain antinutritional factors to assess their potential as alternative sources of protein crops. The major findings of the study were as follows: crude protein ranged from 20.3 to 35.0%, crude lipid 3.1–9.6%, crude fiber 5.9–12.1%, ash 2.7–5.1%, and carbohydrates 49.2–61.8%. Minerals viz., sodium, potassium, calcium, magnesium, phosphorus, iron, copper, zinc, and manganese occurred in the range 42.9–135.9, 556.2–1639.5, 304.5–572.2, 174.9–686.7, 98.4–947.8, 3.6–16.4, 0.2–1.2, 2.0–30.0, and 1.0–3.9 mg/100 g seed flour, respectively. Profiles of amino acids of total seed proteins detected in the present study revealed that levels of valine, phenylalanine, tyrosine, isoleucine, and histidine of all the seven wild legume seed samples, threonine of Canavalia ensiformis and C. gladiata, leucine of Mucuna pruriens var. pruriens, and lysine of Cassia floribunda and C. obtusifolia were found to be higher than FAO/WHO (1991) requirement pattern. The in vitro protein digestibility of the legumes under study ranged from 63.39 to 83.32%. Antinutritional factors such as total free phenolics ranged from 0.41 to 5.96%, tannins from 0.04 to 0.60%, L-DOPA from 1.34 to 8.37% and trypsin inhibitor activity from 13.48 to 65.43 TIU/mg protein. The detected antinutritional factors probably have little nutritional significance if the seeds are properly processed.     

Safflower petals: A source of gamma linolenic acid

Safflower petals have been shown to have a lot of medicinal and therapeutic values. Indian safflower petal samples were analyzed for the red pigment carthamin, protein and oil contents. The petal oil (4.0–5.8%) was further analyzed for its fatty acids followed by alpha linolenic acid (15–19%) and palmitic acids (14–16%). Gamma linolenic acid, which has a lot of therapeutic value was present to an extent of 2–3%; decanoic and dodecanoic acids (2–5%) were also present.     

Influence of dietary soybean trypsin inhibitors and DL-ethionine on sulfur amino acid adequacy of diets for young rats

Weanling male Wistar rats were fed 20% protein diets based on casein or either of two combinations of soy protein isolate and ground raw soy providing three levels of soybean trypsin inhibitors (SBTI; 0,448 and 808 mg of trypsin inhibited per 100g of diet respectively). DL-ethionine was included at three levels (0,0.05% and 0.10%) with each level of SBTI. After 4, 8 and 12 weeks ofad libitum feeding, diets containing SBTI without DL-ethionine were associated with decreases in weight gain, feed efficiency, serum cholesterol and serum urea nitrogen. Higher levels of triglycerides, glutamate pyruvate transaminase (SGPT) and altered serum free amino acid levels were also found. Increased dietary levels of DL-ethionine also resulted in deficits in growth and feed efficiency, decreased serum cholesterol, increased SGPT and similar alterations in serum free amino acids. Combination of dietary SBTI with DL-ethionine resulted in even greater growth deficits and serum cholesterol decreases as well as increases in SGPT and serum triglycerides and changes in serum free amino acid levels. Methionine deficiency in the young rats fed SBTI and DL-ethionine was indicated by the changes in serum amino acids and growth deficits. Moderation of some effects over the 12 week test period suggested decreased methionine requirements in the older rats.     

The use ofEscherichia coli mutants to measure the bioavailability of essential amino acids in foods

The -galactosidase-based assay for lysine developed by Tuffnell & Payne was used to measure the bioavailabilities of cyst(e)ine, methionine, threonine and tryptophan in pronase digests of 17 foods. The digests were assayed by estimating the -galactosidase synthesis responses of fiveEscherichia coli mutants, each requiring one of the respective amino acids for protein synthesis. Deletion mutants were used whenever possible in order to ensure strain stability. Single digests of each food were assayed with 3 or 4 separate cultures of each mutant and the results were compared with those from the corresponding chemical assay. Omitting the anomalously low values for one food, the rank correlation coefficients of the bio- and chemo-assay values were 0.61 (cysteine), 0.91 (lysine), 0.95 (methionine), 0.64 (threonine) and 0.85 (tryptophan). Mean (± S.D.) relative amino acid bioavailabilities (casein = 100%) for the 17 foods were: cysteine, 53±23; lysine, 90±10; methionine, 95±18; threonine, 89±13; and tryptophan, 89±25. The cysteine mutant appeared to give unusually low bioavailability values except for the milk products. These amino acid mutants afford a rapid method for assaying the bioavailabilities of at least four of the five amino acids studied.     

Nutritional evaluation of spent grains from sorghum malts and maize grit

Potential use of the high protein by-product of beer production from 77% sorghum malt and 23% maize grit was investigated. Red sorghum spent grains (RSSG) and white sorghum spent grains (WSSG) contained 23.4 and 19.3% crude protein (CP), 54 and 43% dietary fiber (NDF), 1.44 and 0.78% ash, 4.5 and 3.2% hexane extract and tannin content of 7.5 and 1.0 mg/g catechin equivalent respectively. Magnesium was the most abundant mineral in both RSSG and WSSG — 185 and 140 mg/kg, respectively. Calcium, zinc, iron and copper were generally low. Both samples contained cadmium 1.12 (WSSG), 1.19 (RSSG) and lead at 1.38 mg/kg. Lysine was the limiting amino acid (chemical score 0.55) in both samples. Other essential amino acids were adequate or surplus. Stearic acid was the predominant fatty acid with varying levels of lauric, myristic, palmitic, and oleic acids in both samples. Feed intake and weight gain were highest in rats fed 26.3% WSSG (contributing 50% of the diet protein) but protein efficiency ratio (PER) and net protein retention (NPR) were highest in diets where spent grains contributed just 25% of the diet protein. True digestibility of diets decreased as dietary fiber content increased such that animals on diets containing 100% spent grain protein (above 20% NDF) lost weight.     

Changes in the carbohydrates and nitrogenous components during germination of proso millet,Panicum miliaceum

Since germination has been found to increase the nutritive qualities of grains, proso millet, a minor millet, was allowed to germinate for 1–7 days and analysed for protein fractions, free amino acids, lysine, tryptophan, methionine, non-protein nitrogen, starch, and sugars. Overnight soaking and germination up to 7 days significantly increased the free amino acids and total sugars while the content of dry weight and starch decreased. Increases in albumin and globulin and large decreases in prolamin accompanied sprouting. The percent protein in germinated grains was higher than in the initial grain as a result of dry matter loss during germination. Further, there was an increase in lysine, tryptophan and non-protein nitrogen contents during germination. No change was noticed in methionine content.