The potential for protein production from green plants

Data on protein yields show that forage crops, particularly alfalfa, produce several times more protein per acre than do seed crops. Amino acid analyses and estimations of biological values by enzymatic hydrolysis and feeding trials indicate that protein concentrates from green plants have high nutritive value. The protein concentrates from 10 plant species had a similar amino acid composition and biological value which indicates that good protein might be obtained from many plant species. It is suggested that the use of the fibrous residue as a feed for ruminants and the use of the protein concentrate as a high protein feed or base for processing into new protein foods may make it possible for the production of protein from green plants to compete with other sources of protein. This would markedly increase protein production per acre and allow the use of new plant species in our agriculture. The need for more research on protein production from different types of green plants and on ways to harvest, concentrate and process their proteins into edible forms is discussed.     

Apparent digestion coefficients for dry matter, protein and essential amino acids in terrestrial ingredients for Pacific shrimp Litopenaeus vannamei (Decapoda: Penaeidae)

Protein quality mainly depends on the essential amino acid (EAA) profile, but also on its bioavailability, because EAA digestibility is generally lower than the analyzed amounts. This information is needed in the aquaculture industry for aquafeed formulation. For this purpose, the apparent digestibility coefficients of dry matter, protein, and essential amino acids of eight feedstuffs of terrestrial origin were determined for the juvenile whiteleg shrimp Litopenaeus vannamei (15-19 g), using 1% chromic oxide as an inert marker. A reference diet was formulated and produced in the laboratory. Eight experimental diets were prepared each with 30% of one of the experimental ingredients added to the reference diet: casein, porcine byproduct meal poultry byproduct meal, corn meal, wheat gluten meal, soybean paste, sorghum meal, and wheat meal. The experiment consisted of a single-factor, completely randomized design with three replicates per treatment. Samples of ingredients, diets and feces were analyzed for nitrogen and amino acids. For amino acid assay, we used reverse-phase high performance liquid chromatography. To avoid partial loss of methionine and cystine, samples of ingredients, diets, and feces were oxidized with performic acid to methionine sulfone and cysteic acid prior to acid hydrolysis. The apparent dry matter and protein digestive utilization coefficients varied from 68% to 109% and from 70% to 103%, respectively. Apparent digestibility of protein for casein, soy paste, wheat meal and wheat gluten were very high (over 90%), corn gluten and poultry byproducts meal showed high protein digestibility (over 80%), but porcine byproducts meal and sorghum meal had low digestibility (76% and 70%, respectively). There was a reasonable, but not total, correspondence between apparent protein digestibility and average essential amino acid digestibility coefficients, except for arginine in corn gluten, phenylalanine and leucine in sorghum meal, phenylalanine in soy paste and lysine in wheat meal and poultry by-product meal. The most digestible feed ingredients for whiteleg shrimp were: wheat gluten, wheat meal and soy paste; poultry byproduct meal and corn gluten were less digestible and the lowest digestibility occurred in porcine byproduct meal and sorghum meal. Feedstuffs exhibited great variability in dry matter, protein and amino acid digestive utilization coefficients, which should be considered when formulating shrimp feeds.     

Molecular aspects of methionine biosynthesis

Methionine, lysine and threonine are essential amino acids required in the diets of non-ruminant animals. Major crops, such as corn, soybean and rice, are low in one or more of these amino acids. Currently, these amino acids are supplemented to animal feed to allow optimal growth--a costly process for farmers and consumer, therefore there is a great deal of interest in increasing essential amino acids in crops. The metabolism of methionine in plants is linked to the regulation of the aspartate pathway and is important for plant growth. In recent years, several key steps of this pathway have been identified at the molecular level, enabling us to initiate transgenic approaches to engineer the methionine content of plants.     

Fermentation of soybean hulls to ethanol while preserving protein value

Soybean hulls were evaluated as a resource for production of ethanol by the simultaneous saccharification and fermentation (SSF) process, and no pretreatment of the hulls was found to be needed to realize high ethanol yields with Saccharomyces cerevisiae D₅A. The impact of cellulase, β-glucosidase and pectinase dosages were determined at a 15% biomass loading, and ethanol concentrations of 25–30 g/L were routinely obtained, while under these conditions corn stover, wheat straw, and switchgrass produced 3–4 times lower ethanol yields. Removal of carbohydrates also concentrated the hull protein to over 25% w/w from the original roughly 10%. Analysis of the soybean hulls before and after fermentation showed similar amino acid profiles including an increase in the essential amino acids lysine and threonine in the residues. Thus, eliminating pretreatment should assure that the protein in the hulls is preserved, and conversion of the carbohydrates to ethanol with high yields produces a more concentrated and valuable co-product in addition to ethanol. The resulting upgraded feed product from soybean hulls would likely to be acceptable to monogastric as well as bovine livestock.     

Enrichment of cereal protein lysine content by altered tRNA(lys) coding during protein synthesis

The world's major crops are deficient in lysine and several other amino acids essential for human and animal nutrition. Increasing the content of these amino acids in cereals, our major source of dietary energy, can help feed a global population whose reliance upon dietary protein is growing faster than crop yields. Here we document the heritable expression in rice, the world's major cereal crop, of tRNA(lys) species that introduce lysine at alternative codons during protein synthesis, resulting in a significant enrichment of the lysine content of proteins in rice seeds without changing the types or quantities of the seed storage proteins.     

Ileal and total tract nutrient digestibility in blue foxes (Alopex lagopus) fed extruded diets containing different protein sources

This study compared apparent ileal and total tract digestibility of macronutrients and amino acids in blue foxes (Alopex lagopus) fed dry extruded diets. The control diet contained fish meal as the main protein source, while in the other diets 50% of the fish meal protein was replaced by not de-hulled solvent-extracted soybean meal, meat meal or bacterial protein meal. Ileal digestibility was obtained with animals modified with ileorectal anastomosis (IRA method). There were no significant differences in the average digestibility of CP, Lys, Trp, Val, Ala or Glu between IRA modified and intact animals. Average ileal digestibilities of His, Thr, Asp, Cys, Gly, Hyp, Pro, Ser, starch and total carbohydrates (CHO) were significantly lower compared with total tract values. Average ileal digestibility of Arg, Ile, Leu, Met, Phe, Tyr and crude fat was significantly higher than total tract digestibility. Digestibility measured in IRA modified animals showed less variability compared with values from intact animals. There were significant differences among diets in ileal as well as total tract digestibility of orude protein, most amino acids, starch and CHO, and in ileal digestibility of crude fat. Ileal crude protein digestibility of the diets ranged from 81.0-86.4% and total tract digestibility from 82.5-86.4%. Ileal amino acid digestibilities ranged from 70.1 (Asp) to 93.3% (Arg) and total tract values ranged from 72.8 (Cys) to 92.2% (Arg). Both ileal and total tract digestibilities of crude protein and most amino acids were significantly lower for diets containing meat meal or bacterial protein meal compared with the control fish meal-based diet and the diet with soybean meal. Although ileal digestibility may be more accurate than total tract digestibility in estimating the protein and amino acid supply in blue foxes, total tract digestibility may be acceptable because of numerically small differences between ileal and total tract digestibilities for protein and most amino acids. It was further concluded that ileal digestibilities of starch and CHO in the blue fox are lower than corresponding total tract digestibilities.     

Agricultural Seed Biomass for Migrating and Wintering Waterfowl in the Southeastern United States

ABSTRACT Waterfowl frequently acquire high-energy agricultural seeds in harvested and unharvested croplands during migration and winter. Estimates of agricultural seed biomass in harvested and unharvested corn, soybean, and grain sorghum fields do not exist or are outdated for the southeastern United States. Therefore, we estimated seed biomass in 105 harvested and 59 unharvested corn, soybean, and grain sorghum fields across 4 climate regions in Tennessee, USA, from September through January 2006 and 2007. We also used estimates of seed biomass to calculate duck-energy days (DEDs) in December and January when migratory waterfowl abundance peaks in the southeastern United States. Mean biomass of corn, soybean, and grain sorghum seed in harvested fields declined 239 kg/ha to 39 kg/ha, 118 kg/ha to 26 kg/ha, and 392 kg/ha to 19 kg/ha, respectively, from postharvest to January. Continuous monthly rates of decline were 64% for corn, 84% for soybean, and 74% for grain sorghum. Agricultural seed biomass in harvested corn and grain sorghum fields dropped below the waterfowl giving-up density (i.e., 50 kg/ha) in 3 months; soybean dropped below this threshold 1 month postharvest. Mean DEDs/ha in harvested corn, soybean, and grain sorghum fields were low (274, 90, and 27, respectively) in January, and DEDs were zero in >85% of fields. In unharvested corn, soybean, and grain sorghum fields, mean DEDs/ha in January were high (69,000, 18,000, and 26,000, respectively), and continuous rates of decline (3%, 7%, and 18%, respectively) were much lower than for harvested crops. Waterfowl biologists in the Southeast should use our estimates of agricultural seed biomass in DED calculations. We also recommend that biologists provide unharvested grain fields and natural wetlands for migrating and wintering waterfowl because seed resources are low in harvested agricultural fields.     

Protein quality of mixtures of animal protein meals

The protein quality of two-component mixtures of meat-and-bone meal, bone meal, blood meal and feather meal was characterized by rat experiments (net protein utilisation (NPU)) and by chemical indices calculated from amino acid composition (limiting essential amino acid, sum of essential amino acids, chemical score (CS), essential amino acid index (EAA-index) and predicted value (PV)).Substitution of meat-and-bone meal for bone meal on a nitrogen basis reduced the NPU value. This was also indicated by amino acid indices.Substitution of meat-and-bone meal for blood meal on a nitrogen basis also reduced the NPU value; however, this was not indicated by the chemical indices.In the mixtures examined, feather meal complemented meat-and-bone meal as indicated by both the NPU value and chemical indices. A complementary effect was also found with the mixtures of feather meal and blood meal.A significant correlation was found between NPU value and either predicted value (r=0.89), chemical score (0.83) or EAA-index (0.62), as calculated on cumulated data (n=20).     

Gelatinase Activity by Marine Fungi

Gelatinase activity was detectable in the culture filtrates of 13 of 14 species of marine fungi tested. The fungi were grown under conditions of submerged culture in a medium consisting of corn meal, soybean meal, and CaCO(3). The degree of enzymatic activity did not necessarily correlate with the amino nitrogen content of the filtrates. Eight of the filtrates contained measurable quantities of glutamic acid and alanine. Highest levels of gelatinase were obtained in the culture filtrates of Halosphaeria mediosetigera. Increasing the corn meal concentration of the medium resulted in even greater production of gelatinase by H. mediosetigera, whereas amino acid yields were enhanced by higher concentrations of soybean meal. Supplementation of the high corn meal medium with either a mineral mixture or distillers' solubles brought about a faster rate of production of gelatinase and, in the case of the distillers' solubles, a significantly higher level of enzyme, as well.     

Value-added subcritical water hydrolysate from rice bran and soybean meal

New value-added product was derived from agricultural by-products: rice bran and soybean meal by means of subcritical water (SW) hydrolysis. The effect of temperature (200-220 degrees C), reaction time (10-30 min), raw material-to-water weight ratio (1:5 and 2:5), was determined on the yields of protein, total amino acids, and reducing sugars in the soluble products. The suitable hydrolysis time was 30 min and the proper weight ratio of the raw material-to-water was 1:5. The reaction temperature suitable for the production of protein and amino acids was 220 degrees C for raw and deoiled rice bran, 210 degrees C for raw soybean meal, and 200 degrees C for deoiled soybean meal. The products were also found to have antioxidant activity as tested by ABTS(.)(+) scavenging assay. In addition, sensory evaluation of milk added with the hydrolysis product of deoiled rice bran indicated the potential use of the product as a nutritious drink.     

Amino acid requirement of growing pigs depending on amino acid efficiency and level of protein deposition. 2nd communication: threonine

This study was conducted to evaluate the variability of the efficiency of threonine in different feed proteins for growing pigs. This information is of importance for actual conclusions about threonine requirement within the exponential N-utilization model (Liebert and Gebhardt, 1986) used in our investigations. Wheat (as basal protein), high-protein soybean meal, low-protein soybean meal, rapeseed meal, field bean (Vicia faba), peas (Pisum sativum), corn gluten meal and soybean protein concentrate were used as protein sources. Fifty-six growing barrows (40-65 kg BW) of the genotype Piétrain x (Duroc x Landrace) were randomly allotted to eight N-balance experiments (n = 7). Diets were formulated with two main ingredients (wheat + one feed protein) with threonine as the first limiting amino acid in the mixture which was partly supplemented with crystalline amino acids. Based on N-balance data, the efficiency of threonine was determined in protein mixtures and individual feed proteins. Threonine requirement was calculated depending on efficiency of threonine and level of daily protein deposition. The results from the present studies indicate that the efficiency of threonine in different feed proteins varied in a wide range. Consequently, this factor has to be taken into account for requirement calculations. The threonine requirement depending on daily protein deposition (130, 145 and 160 g) and the efficiency of threonine according to different reference units (g/BW(kg)(-0.67)/d, g/d and % of threonine in the diet) were calculated. The threonine requirement of growing barrows (50 kg BW) corresponding to an average threonine efficiency was 8.52, 9.92 and 11.61 g/d for a daily protein deposition of 130, 145 and 160 g, respectively. The results for a daily protein deposition of 145 or 160 g are in agreement with actual studies and recommendations for threonine supply.     

The impact of protein quality on stable nitrogen isotope ratio discrimination and assimilated diet estimation

Accurately predicting isotopic discrimination is central to estimating assimilated diets of wild animals when using stable isotopes. Current mixing models assume that the stable N isotope ratio (δ¹⁵N) discrimination (?¹⁵N) for each food in a mixed diet is constant and independent of other foods being consumed. Thus, the discrimination value for the mixed diet is the combined, weighted average for each food when consumed as the sole diet. However, if protein quality is a major determinant of ?¹⁵N, discrimination values for mixed diets may be higher or lower than the weighted average and will reflect the protein quality of the entire diet and not that of the individual foods. This potential difference occurs because the protein quality of a mixed diet depends on whether, and to what extent, the profiles and amounts of essential amino acids in the individual foods are complementary or non-complementary to each other in meeting the animal’s requirement. We tested these ideas by determining the ?¹⁵N of several common foods (corn, wheat, alfalfa, soybean, and fish meal) with known amino acid profiles when fed singly and in combination to laboratory rats. Discrimination values for the mixed diets often differed from the weighted averages for the individual foods and depended on the degree of complementation. ?¹⁵N for mixed diets ranged from 1.1‰ lower than the weighted average for foods with complementary amino acid profiles to 0.4‰ higher for foods with non-complementary amino acid profiles. These differences led to underestimates as high as 44% and overestimates as high as 36% of the relative proportions of fish meal and soybean meal N, respectively, in the assimilated mixed diets. We conclude that using isotopes to estimate assimilated diets is more complex than often appreciated and will require developing more biologically based, time-sensitive models.     

Protein content and amino acid composition of cassava leaf

On the basis of leaf dry wt, the protein content of six varieties of cassava varied from 29.3 to 38.6% and the estimated leaf protein production ranged from 242 to 953 kg per ha. On the basis of fr. wt of leaf, the total amino acids ranged from 8.42 to 9.4% while the essential amino acids averaged 4.21% and the sulphur-containing amino acids only 0.25%. The amino acid composition profiles for the six varieties was similar.     

Genetic enhancement of Brassica napus seed quality

The ultimate value of the Brassica napus (canola) seed is derived from the oil fraction, which has long been recognized for its premium dietary attributes, including its low level of saturated fatty acids, high content of monounsaturated fatty acids, and favorable omega-3 fatty acid profile. However, the protein (meal) portion of the seed has also received favorable attention for its essential amino acids, including abundance of sulfur-containing amino acids, such that B. napus protein is being contemplated for large scale use in livestock and fish feed formulations. Efforts to optimize the composition of B. napus oil and protein fractions are well documented; therefore, this article will review research concerned with optimizing secondary metabolites that affect the quality of seed oil and meal, from undesirable anti-nutritional factors to highl value beneficial products. The biological, agronomic, and economic values attributed to secondary metabolites have brought much needed attention to those in Brassica oilseeds and other crops. This review focuses on increasing levels of beneficial endogenous secondary metabolites (such as carotenoids, choline and tochopherols) and decreasing undesirable antinutritional factors (glucosinolates, sinapine and phytate). Molecular genetic approaches are given emphasis relative to classical breeding.     

Review: Optimizing ruminant conversion of feed protein to human food protein

Ruminant livestock have the ability to produce high-quality human food from feedstuffs of little or no value for humans. Balanced essential amino acid composition of meat and milk from ruminants makes those protein sources valuable adjuncts to human diets. It is anticipated that there will be increasing demand for ruminant proteins in the future. Increasing productivity per animal dilutes out the nutritional and environmental costs of maintenance and rearing dairy animals up to production. A number of nutritional strategies improve production per animal such as ration balancing in smallholder operations and small grain supplements to ruminants fed high-forage diets. Greenhouse gas emission intensity is reduced by increased productivity per animal; recent research has developed at least one effective inhibitor of methane production in the rumen. There is widespread over-feeding of protein to dairy cattle; milk and component yields can be maintained, and sometimes even increased, at lower protein intake. Group feeding dairy cows according to production and feeding diets higher in rumen-undegraded protein can improve milk and protein yield. Supplementing rumen-protected essential amino acids will also improve N efficiency in some cases. Better N utilization reduces urinary N, which is the most environmentally unstable form of excretory N. Employing nutritional models to more accurately meet animal requirements improves nutrient efficiency. Although smallholder enterprises, which are concentrated in tropical and semi-tropical regions of developing countries, are subject to different economic pressures, nutritional biology is similar at all production levels. Rather than milk volume, nutritional strategies should maximize milk component yield, which is proportional to market value as well as food value when milk nutrients are consumed directly by farmers and their families. Moving away from Holsteins toward smaller breeds such as Jerseys, Holstein-Jersey crosses or locally adapted breeds (e.g. Vechur) would also reduce lactose production and improve metabolic, environmental and economic efficiencies. Forages containing condensed tannins or polyphenol oxidase enzymes have reduced rumen protein degradation; ruminants capture this protein more efficiently for meat and milk. Although these forages generally have lower yields and persistence, genetic modification would allow insertion of these traits into more widely cultivated forages. Ruminants will retain their niches because of their ability to produce valuable human food from low value feedstuffs. Employing these emerging strategies will allow improved productive efficiency of ruminants in both developing and developed countries.     

Dietary protein source affects lipid metabolism in the European seabass (Dicentrarchus labrax)

The study was undertaken to evaluate the effects of dietary protein sources on lipogenesis and fat deposition in a marine teleost, the European seabass (Dicentrarchus labrax). Four isonitrogenous (crude protein (CP, Nx6.25), 44% DM) and isoenergetic (22-23 kJ/g DM) diets were formulated to contain one of the following as the major protein source: fish meal (FM), one of two soy protein concentrates (SPC) and corn gluten meal (CGM). Apparent digestibility coefficients of the diets and raw ingredients, as well as soluble nitrogen (ammonia and urea) and phosphorus excretion were measured. Growth rates of seabass fed plant protein-based diets were significantly lower than those fed fish meal based diet. The protein utilisation was strongly correlated to the dietary essential amino acids index. Measurements of N excretion (ammonia and urea nitrogen) confirmed these data. Daily fat gain at the whole body level ranged between 1.1 to 1.7 g/kg BW, with the highest values being recorded in fish fed the fish meal based diet. Levels of plasma triglycerides and cholesterol were lower in fish fed soy protein diets than in those fed the diet solely based on fish meal. Soy protein rich diets decreased the activities of selected hepatic lipogenic enzymes (glucose 6-phosphate dehydrogenase, malic enzyme, ATP-citrate lysase, acetylcoenzyme A carboxylase and fatty acid synthetase). Highest lipogenic enzyme activities where found in fish fed the fish meal diet, except for fatty acid synthetase which was increased in seabass fed the corn-gluten meal based diets. Overall data suggest that dietary protein sources affects fat deposition and the lipogenic potential in European seabass.     

Pilot-scale purification of zeamatin, an antifungal protein from maize

Zeamatin is an antifungal protein isolated from the seeds of Zea mays. A practical process for isolation and purification was developed to increase recovery yields for future testing of zeamatin as a novel therapeutic drug. Zeamatin was extracted with buffer from corn meal milled to flour. Solids were removed from the extract using pressure filtration, and zeamatin was purified using two separate reverse-phase chromatography steps. Determination of the amino-terminal amino acid sequence, Western blotting using anti-zeamatin antibody, and activity against Candida albicans were used to confirm that the purified protein was zeamatin. From 5 kg of corn meal, approximately 110 mg of zeamatin was purified to apparent homogeneity.     

Effects of dietary soybean protein levels on energy budget of the southern catfish, Silurus meridionalis

Energy budgets were calculated for the southern catfish, Silurus meridionalis, fed diets replacing 0%, 13%, 26%, 39%, 52% and 65% fish meal protein with soybean meal (SBM) protein with or without methionine supplementation to apparent satiation at 27.5 degrees C. With increasing dietary soybean protein levels (SPL), the feed energy lost in feces, excretion and metabolism increased, while that available for growth decreased (P<0.05). When 0.12% or 0.26% methionine at 39% SPL was added to reach that in body carcass or the control group (0% SPL), no significant differences were found in each component of energy budgets. When 0.21% or 0.33% methionine at 52% SPL was added to reach the content of methionine in body carcass or the control group, energy spent on growth increased, but that on excretion and metabolism decreased (P<0.05). These results suggested that the differences in growth rate among the southern catfish fed the diets with different SPL were due to decreasing absorption rate, increasing excretion and metabolism with increasing dietary SPL. The most important factor limiting the use of soybean protein was the imbalance of essential amino acids, which resulted in more energy spent on metabolism and excretion, less energy on growth. Supplementation of methionine produced a relatively better amino acid profile and subsequently improved the utilization of soybean protein at high SPL, which resulted in less energy used for metabolism or lost in excretion and more energy available for growth.     

Enhancement of essential amino acid contents in crops by genetic engineering and protein design

The importance and urgency of providing humans and animals with quality proteins are reflected in the growing scientific and industrial interest in augmenting the nutritive value of the world's protein sources. Such nutritive value is determined by the protein content in 'essential amino acids', those that cannot be synthesized de novo and that must be supplied from the diet. It is the object of this review to discuss recent advances in the genetic modification of crops that aim to provide enhanced quantities of essential amino acids.