Biological and chemical evaluation of chick pea seed proteins as affected by germination,extraction and α-amylase treatment

The effects of germination, extraction (double extraction with 70% ethanol and water at isoelectric point) and -amylase treatments of chick pea seed flours on crude protein, total carbohydrate, protein efficiency ratio (PER), biological value (BV), true digestibility (TD), net protein utilization (NPU), essential amino acid composition, in-vitro protein digestibility (IVPD) and actual amino acid indices (essential amino acid index or amino acid score) were evaluated. Crude protein content was increased (8–149%), while total carbohydrate was decreased (11–62%) by germination, extraction and -amylase treatments. Alpha-amylase treatment was more efficient in reducing total carbohydrate and increasing the protein content than that of extraction treatment. The protein quality of chick pea flours as measured by PER, BV, TD, NPU, IVPD and corrected amino acid indices (actual amino acid indices×IVPD) was significantly improved by these treatments. The protein quality of germinated--amylase treatment was comparble with casein, while germinated--amylase treaded seeds appeared nutritionally superior to casein. The results indicate that the germinated--amylase and germinated--amylase-extracted treatments could be used successfully as a source of concentrated high quality protein for baby food production. The corrected amino acid indices gave better prediction of PER, BV, TD and NPU (r=93 to 97) than actual amino acid indices (r=45 to 71). PER was highly correlated with corrected amino acid score (r=0.93). The PER could be predicted from the following simple regression equation: PER=–1.827+0.0561×corrected amino acid score.     

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.     

Changes in carbohydrate and nitrogenous components and amylase activities during germination of grain amaranth

Grain amaranth (Amaranthus hypochondriacus), Yercaud local variety, was soaked overnight and germinated for 192 h taking the soaked grains as the zero time (0 h) sample. The changes in the activities of - and -amylases, starch, sugar, protein and lysine contents during germination are reported. Activity of -amylase was high in the 0 h soaked grains, while -amylase activity was high in 72 h germinated grains. The joint action of the amylases resulted in a decrease of starch content from 0 to 192 h in germinated grains and an increase in total sugars during the initial period of germination.Protein nitrogen was found to decrease from 48 h to 192 h accompanied by an increase in free amino acid and non-protein nitrogen contents. Total lysine content was found to be increased by 31% in 24 h germinated grain amaranth. Protein fractionation of raw, soaked and 24 h germinated grain amaranth showed that the distribution of different types of proteins varied during germination of the grains. An increase of water soluble protein content was noticed in 24 h germinated grains.     

Nutritional quality of germinated cowpea flour (Vigna unguiculata) and its application in home prepared powdered weaning foods

Amino acid profiles, protein digestibility corrected aminoacid scores (PDCAAS), chemical scores, essential amino acid indexes, andcalculated biological values of control-cowpea flour (CCF), germinated cowpeaflour (GCF) prepared from cowpeas germinated at 25 ^°C for either24 h or 48 h and weaning foods prepared from cowpea flours weredetermined. Locally available rice, cowpea flour, banana-pumpkin slurry,and skim milk powder and sucrose in the ratio 35:35:15:15:5 were used to formulateweaning food containing not less than 15% protein. The ingredients werecooked into a slurry and oven-dried to produce flakes. The nutritional andsensory qualities of the weaning products were evaluated. Germination hadlittle effect on the amino acid profile of cowpeas. In vitro proteinquality and starch digestibility were improved in germinated cowpea flour. The PDCAAS of 24 h germinated cowpea flour (GCF) weaning food washigher (55.49%) than CCF-weaning food (46.74%). Vitamin A activityin 24 h GCF weaning food was higher than in CCF-weaning food. Invitro starch digestibilities of 24 h GCF and 48 h GCF-weaning foods werehigher than that of CCF weaning food. The 24 h GCF-weaning food which hada higher overall acceptability score by sensory panelist than 48 h GCFand CCF-weaning food is recommended for household consumption.     

Effects of germination and cooking on polyphenols and in vitro protein digestibility of horse gram and moth bean

Changes in polyphenols and in vitro protein digestibility due to germination and cooking were studied in horse gram and moth bean. Horse gram had higher amounts of polyphenols than moth bean. Polyphenol content in horse gram decreased from 1.6% to 1.1% at 48h germination, but in moth bean there was a reduction in polyphenol content from 1.3% to 1.0% at 24 h germination with a significant increase from 1.0% to 1.7% at 36 h germination. Seeds germinated for 48 h also exhibited higher amounts of polyphenols than in raw moth bean. The germination had no effect on total protein content, but a progressive increase in free amino acid content was observed in both legumes. In the ungerminated form, horse gram protein was more digestible than moth bean protein. The in vitro protein digestibility of both the legumes increased during germination. Cooking decreased the protein digestibility of horse gram either in ungerminated or germinated form, but such a decrease was noticed in moth bean at only 36 h germination. Moth bean either in raw form or germinated for 24 h did not exhibit a decrease in in vitro protein digestibility after cooking. Cooking decreased polyphenol content by 20%–50%. The apparent decrease in polyphenols in moth bean and horse gram due to cooking has been attributed to a decrease in assayable polyphenols.     

Changes in chemical and selected biochemical components,protein quality,and digestibility of mung bean (Vigna radiata) during gemination and cooking

The effect of germination on the nutritive value of mung beans (Vigna radiata) was studied in raw and cooked samples after 0, 2 and 4 days of germination. Water content increased, but crude protein, lipids, crude fiber, ash and carbohydrates decreased with germination. Trypsin inhibitor activity did not change significantly while hemagglutinins were absent. Essential amino acids (methionine, tyrptophan and lysine) decreased with germination.Net protein ratio (NPR) and protein efficiency ratio (PER) in rats decreased with germination especially in cooked sprouts. True digestibility did not change consistently due to germination or cooking. In conclusion, loss of protein quality in mung beans occurs upon germination and cooking.Publication INCAP/UNU-4.     

Nutritive value of malted millet flours

The changes in proximate composition, phytate phosphorus, thiamine and ascorbic acid content of finger millet, pearl millet and foxtail millet during progressive germination were studied. Germination resulted in a slight decrease in total protein and minerals, a marked fall in phytate-phosphorus and a significant increase in the ascorbic acid content of the millets. An increase in lysine and tryptophan but no appreciable changes in threonine and sulfur amino acid content of the millets were observed as a result of germination. However, the protein efficiency ratio values of ungerminated control seeds, 48 h germinated green malt and kilned malt were not significantly different.     

Preparation and properties of flours and protein concentrates from raw, fermented and germinated fluted pumpkin (Telfairia occidentalis Hook) seeds

In vitro protein digestibility, chemical composition and selected functional properties of flours and protein concentrates prepared from raw, fermented and germinated fluted pumpkin (Telfairia occidentalis Hook) seeds were studied. Protein concentrates prepared by an alkaline extraction process had increased crude protein contents (61.5–70.8%) compared to flour samples (46.4–52.7%). The yields of protein concentrates ranged from 24.5% to 29.4% while values for protein recoveries varied between 64.8% and 65.2%. Protein concentrates also had increased foam volume and decreased foam stability (100% decrease over a 2 h period), compared to flour samples. Fermentation and germination were observed to significantly (p<0.05) lower polyphenol and phytic acid contents, but increased protein digestibility of fluted pumpkin seed flours and concentrates. Both raw flour and concentrate were significantly (p<0.05) higher in water absorption capacity than germinated or fermented flours and concentrates. Protein concentrates had comparatively better fat absorption properties than the flour samples. Hence protein concentrates may prove to have useful applications in ground meat formulations.     

Changes in oil,sugars and nitrogenous components during germination of sunflower seeds,Helianthus annuus

Since germination has been found to improve the nutritional quality of grains, sunflower seeds Helianthus annuus were germinated for up to five days, and the cotyledons were analyzed for oil, sugar, free amino acid, non-protein nitrogen, lysine, tryptophan and methionine contents. Protein was fractionated. Soaking and germination increased the non-protein nitrogen, total free amino acid, lysine and tryptophan contents. Protein content and dry weight decreased. The oil content decreased significantly after 72 hours of germination. The reducing sugars increased gradually until day five of germination.The saline soluble albumin and globulin fractions decreased while the glutelin content increased during germination. No changes were noticed in the prolamin and methionine contents.     

Amino acid composition protein quality and water-soluble vitamin content of germinated cowpeas (Vigna unguiculata)

Amino acid composition, protein digestibility, calculated protein efficiency ratio (C-PER and DC-PER), chemical scores and water-soluble vitamin content of cowpea seeds germinated at 25°C or 30°C for 24h were determined. Also, the effect of processing steps (heated-air drying, decortication and cooking) on these parameters were examined. Germination had little effect on amino acid profile of cowpeas. In vitro protein digestibility was not improved significantly by germination nor by decortication but was improved by cooking. C-PER and DC-PER ranged from 1.95 to 2.21 and from 1.63 to 1.82, respectively. DC-PER compared well withreported rat PER of cowpea products and seemed more sensitive than C-PER. Based on whole egg values, chemical scores ranged from 37.7 to 45.8% (mean±SD; 42.2±2.4%). Germination increased the contents of niacin, thiamin and riboflavin significantly. Decortication resulted in up to 30% loss in niacin while thiamin content was reduced 41% by cooking.     

Protein evaluation of four oat (Avena sativa L.) cultivars adapted for cultivation in the south of Brazil

Four oat cultivars adapted for soil and climate conditions in the southern region of Brazil were evaluated for protein nutritive value. Evaluations were done both in vitro and in vivo. In vitro evaluation was done by essential amino acid profile, available lysine, amino acid scoring, and protein digestibility corrected amino acid-scoring (PDCAAS). Nitrogen balance indices and PER were determined in vivo with rats. In all four cultivars (UFP-15, UFP-16, CTC-03, UFRGS-14), lysine was the most limiting amino acid. Available lysine, amino acid score and PDCAAS were highest for cultivar UFRGS-14 and lowest for CTC-03. When compared to casein, only nitrogen retention for UFRGS-14 did not differ statistically (p>0.05); all other indices of protein quality were inferior to casein for the oat cultivars. The oat cultivars tended to be identical among themselves, except for apparent protein digestibility which was significantly higher in the UFRGS-14 and CTC-03 cultivars. On average, the PER values of the oat cultivars were 82% of casein; the net protein utilization was 88% of casein as determined in vivo and 49% by the estimation in vitro (PDCAAS).     

Studies on weaning diets in Nigeria: 2 protein sources

Protein isolates were extracted from melon, groundnut, and soybean cakes in an aqueous system that could potentially yield both oil and protein from oilseeds quantitatively. Protein isolates obtained were 91.2, 78.6 and 79.5% of the total protein content of melon, groundnut and soybean cakes respectively. Both cakes and isolates were assessed in a rat bioassay while breadfruit, cassava, and corn starch were also investigated as sources of carbohydrate. Results showed that there were no significant differences between feed intake, weight gain and PER of rats fed the cake and the protein isolates except in soybean. Weight gain, PER and NPR were highest in cassava-soybean protein isolate diet followed by cassavacasein diet (control) while performance was only moderate with melon- and groundnutcassava diets. With melon protein isolate, cassava and breadfruit were better carbohydrate sources than corn starch. In simulated weaning diets, cassava-sucrose-melon-soybean protein isolate based diet performed better (weight gain 33.4 g, PER 1.63, NPR 2.23) than a commercial weaning food (weight gain 30.6 g PER 1.57, NPR 2.18) while replacement of sucrose and cassava at 30% by breadfruit yielded a cost effective diet with good performance (weight gain 29.4 g, PER 1.51, NPR 2.12). Diets where soybean protein served as the sole source of protein or breadfruit, white and yellow maize ogi served as the main source of carbohydrate performed only moderately.     

Influence of spices on utilization of sorghum and chickpea protein

Influence of eight common Indian spices on the protein quality of sorghum and chickpea was studied. Spices used include red chillies (Capsicum annum), black pepper (piper nigrum), coriander seeds (Coriandrum sativum) cumin seeds (Cuminum cyminum), garlic (Allium sativum), asafoetida (Ferula foetida), dry ginger (Zingiber officinale) and ajowan (Carum copticum). Addition of spices did not affect protein digestibility (TD) of sorghum. The BV of all sorghum diets with spices was higher than that of control diet. However, it was significant only in case of diets combined with red chilli + coriander (11) mix, black pepper + cumin (11) mix, coriander and cumin. Addition of spices did not have any effect on TD or BV of chickpea diets.     

Effect of processing on the in vitro digestibility of proteins and carbohydrates in some Indian legumes

The effect of different types of processing such as boiling, pressure cooking, puffing, frying, germination, and germination followed by cooking on the protein and carbohydrate digestibility of chickpea, horsegram and cowpea were studied in in vitro systems. In the case of chickpea, the protein digestibility was not significantly improved by any of the treatments. However, for horsegram and cowpea, improvement in protein digestibility was observed after some of the different processing treatments. Frying decreased the protein digestibility in all of the pulses. All of the treatments, except germination, caused a marked increase in in vitro carbohydrate digestibility.     

Nutritional Evaluation and Shelf Life Studies of Papads Prepared from Wheat–Legume Composite Flours

The present study was carried out to develop papads from wheat and legume blends and to analyze them for organoleptic acceptability, nutritional quality, and keeping quality. Papad is a dehydrated product prepared from dhals or rice. A firm but pliable dough is made from the flours of dhals or rice with addition of suitable seasoning. It is shaped into balls, rolled out thin, dried and toasted over open fire so as to give a light, crisp product. Mung flour papads were kept as control and wheat flour, chickpea flour, and pea flour were used to supplement mung flour papads. Three different proportions (10, 20, 30%) of each flour were used to supplement Mung flour. Papads with wheat flour (10%), chickpea flour (20%), and pea flour (10%) level of supplementation were found to be most acceptable and these papads were subjected to nutritional evaluation. Protein content significantly increased on supplementation with legume flours at all levels. Fat content was significantly higher in chickpea flour supplemented papads. Ash content varied significantly from 10.17 to 10.78% in papads. Total carbohydrates decreased significantly on supplementation with chickpea flour. Copper content increased significantly on supplementation. A significant decrease in phytic acid and trypsin inhibitor of supplemented papads was found. In vitro protein digestibility significantly increased on supplementation but a significant decrease in in vitro starch digestibility was found in supplemented papads. Storage studies showed that chickpea flour and pea flour supplemented papads can be stored safely for 60 days and wheat flour supplemented papads for 30 days both at room and refrigeration temperatures.     

Production of African bread-fruit (Treculia africana) and soybean (Glycine max) seed based food formulations, 1: Effects of germination and fermentation on nutritional and organoleptic quality

Germination and fermentation were investigated as methods of improving the nutritional and organoleptic properties ofsoybean and African breadfruit seed based food formulations.Four products consisting of germinated-fermented soy-breadfruitseeds (GFSB), nongerminated-fermented soy-bread-fruit seeds(NGFSB), germinated-nonfermented soy-breadfruit seeds (GNFSB)and nongerminated-nonfermented soy-breadfruit seeds (NGNFSB)were prepared. Phytic acid contents, in vitro protein digestibilities, protein efficiency ratios (PER), net proteinratios (NPR), flavor, appearance and overall acceptabilitywere evaluated. Germination followed by natural lactic fermentation significantly (p < 0.05) reduced the phytic acid by a factor of 11.6 in NGNFSB compared to reductionfactors of 2.1 and 1.5 in GNFSB and NGFSB, respectively.The in vitro protein digestibility (%), PER and NPR valuesof 73.4, 2.46 and 3.62 for GFSB; 71.1, 2.35 and 3.46 for NGFSB;68.7, 2.16 and 3.41 for GNFSB were significantly (p < 0.05)higher than the 64.7, 1.82 and 2.11 for NGNFSB. The meansensory scores were 5.26–5.67 for GNFSB, 4.66–4.94 forNGNFSB, 4.33–4.80 for GFSB and 4.27–4.34 for NGFSB on a7-point rating scale.     

Changes in nitrogenous and other chemical constituents,protein fractions and in vitro protein digestibility of germinating fluted pumpkin ( Telfairia occidentalis Hook) seed

The effect of 7 days of germination on levels of nitrogenous and other nutrition related parameters, protein fractions and in vitro protein digestibility of fluted pumpkin (Telfairia occidentalis) seed was studied. The non-protein nitrogen gradually increased and the protein nitrogen content decreased during germination. Albumin and globulin fractions were found to be the major seed proteins of fluted pumpkin seeds, constituting about 58.6% of the total protein of the ungerminated (raw) seeds. The protein fractions, albumin and glutelin, were observed to increase by 61.5% and 57.0%, respectively, while a 54.6% decrease was noted in the prolamine fraction. The globulin fraction increased at the beginning of germination but decreased at the end. Germination significantly (p 0.05) increased the crude protein, nitrogen solubility and in vitro protein digestibility but decreased the fat, phytic acid and polyphenol contents of the seeds.     

Contribution of low molecular weight antioxidants to the antioxidant screen of germinated soybean seeds

Soybean seed of the Mazovia variety (Glycine max) were germinated in the dark at a constant temperature of 25°C up to 168 h (24 h × 7). Changes in content of low molecular weight antioxidants (LMWA) in the course of germination were studied. The analyzed compounds were: total phenolics, soluble proteins, tocopherols (-T, -T, -T, -T), reduced and oxidized glutathione, L-ascorbic acid and L-dehydroascorbic acid. The changes in total antioxidant status (TAS) and the peroxyl radical-trapping capacity (PRTC) of the processed seeds were measured and compared to those of the raw seeds. On the basis of data obtained, the contribution of LMWA to the TAS of raw and germinated soybean seeds was calculated. The results showed a very high molar percentage contribution of phenolic compounds (49.7%) in opposition to glutathione (7.0%), tocopherols (4.0%) and soluble protein (5.7%) to the total TAS of raw soybean seeds. The germination process led to the increased contribution of phenolic compounds (67.7%), soluble proteins (9.3%) and ascorbic acid (4.4%) to the total TAS of germinated seeds and as a result, a decrease in the antioxidant gap was observed compared to the total TAS of raw soybean seeds. This study showed that the contribution of experimentally obtained total PRTC to the total TAS of raw and germinated soybean seeds after seven days ofthe process was 48% and 25.2%, respectively. The PRTC results were comparable to those obtained in TAS only when calculation was done by multiple mean values for content of investigated compounds and their relative potency with respect to Trolox. The PRTC values obtained in this manner were higher than those obtained experimentally indicating that high molecular weight antioxidants as well as are able to quench peroxyl radicals as well as low molecular weight antioxidants. Therefore, calculation of PRTC is a more accurate method that eliminates the puzzling question of unidentified antioxidants shown as the antioxidant gap in the TAS assay.     

The degradation of lectins,phaseolin and trypsin inhibitors during germination of white kidney beans,Phaseolus vulgaris L.

White kidney beans (Phaseolus vulgaris), cv Processor, contain a relatively high content of phaseolin (storage protein), lectins and a special group of glycoproteins as well as a considerable amount of protein-type trypsin inhibitors. Protein digestion of raw Processor beans in monogastrics, for example pigs, is disturbed by poorly digested, phaseolin lectins, which can bind to carbohydrates in brush border membranes of the small intestinal epithelium, and trypsin inhibitors. The effect of the germination of white kidney beans on lectins, phaseolin and trypsin inhibitors was studied in order to achieve a degradation of lectins, phaseolin and trypsin inhibitors and an increase ofin vitro enzymatic hydrolysis of the protein of bean flour. Therefore, whole bean extracts were examined throughout a germination period of up to seven days for their lectin and phaseolin pattern, lectin content, binding capacities of functional lectins towards brush border membranes and trypsin inhibitor content. In addition thein vitro enzymatic hydrolysis by pepsin and pancreatin of the protein from flours of (un)germinated white kidney beans was studied. SDS-PAGE demonstrated a degradation of E-lectins and a disappearance of L-lectins and phaseolin during germination. Results indicated a decrease of the lectin content by 85%, a loss of binding capacities of functional lectins towards brush border membranes by 91%, and a decrease of trypsin inhibitors by 76%, in bean flour after germination for seven days. A maximum inin vitro enzymatic hydrolysis of protein from bean flour was already established after germination for half a day.     

Specific role of endopeptidases in modulating the nature of protein digestion products

The impact of various endopeptidases on the nature of protein digestion products was measured with the digestion cell technique. After a 30 min pepsin pre-digestion, casein and rapeseed concentrates were hydrolyzed with various amounts of pancreatin, trypsin and/or chymotrypsin. This hydrolysis was performed in a dialysis tube (molecular weight cut-off 1000) with continuous collection of the digested material. The addition of pure trypsin or chymotrypsin to pancreatin (Enzyme:Substrate 125) did not change the digestibility of casein. Only a higher pancreatin level (Enzyme: Substrate 112.5) increased the total protein digestibility without affecting the amino acid spectra. Rapeseed digestibility was markedly increased by the addition of pure trypsin to pancreatin. Lysine and arginine, target amino acids of trypsin, were favored at the expense of chymotrypsin and elastase target amino acids. Supplementation of pancreatin with, chymotrypsin enhanced rapeseed digestibility without affecting the relative amino acid digestibility. The impact of a higher pancreatin ratio (Enzyme: Substrate 112.5) was similar to that of enriched pancreatin but the rate of amino acid release was modified. The differences between protein sources were mainly attributed to protein structure.