Removal of anti-nutritional factors of rapeseed protein isolate (RPI) and toxicity assessment of RPI.

We prepared a detoxified rapeseed protein isolate (RPI) by phytase/ethanol treatment based on alkaline extraction and acidic precipitation. Contents of protein, fat, ash, moisture, crude fiber, glucosinolates, phytic acid, and phenolics and color were determined. To evaluate the safety of detoxified RPI, five groups of C57 mice (detoxified RPI [10 and 20 g kg-1]; commercial soybean protein isolate (SPI) [10 g kg-1]; non-detoxified RPI [10 g kg-1]; control) were used in the acute-toxicity test. Bodyweight and pathology parameters were recorded at different time points, followed by macroscopic examination, organ-weight measurement and microstructure examination. After pretreatment of rapeseed meals with phytase (enzyme : substrate ratio, 1 : 5 mg g-1) for 1.5 h and two-time ethanol extraction for precipitated protein, the chemical characteristics in RPI were protein (88.26%), fat (0.57%), ash (2.72%), moisture (1.90%), crude fiber (0.77%), glucosinolates (0 µmol g-1), phytic acid (0.17%), phenolics (0.36%) and whiteness (73.38). Treatment resulted in significant removal of anti-nutritional factors (ANFs) and increased whiteness in detoxified RPI compared with non-detoxified RPI, and lower than in cruciferin-rich canola protein isolate (Puratein®). Experimental-related effects on bodyweight, clinical observations, or clinicopathology, in mice treated with detoxified RPI were not observed except for a decreased thyroid gland/parathyroid gland index in mice treated with non-detoxified RPI. Furthermore, the no-observed-effect level (NOEL) was 10 g kg-1 of detoxified RPI, whereas the no-observed-adverse-effect-level (NOAEL) was the highest fed level of 20 g kg-1 of detoxified RPI. Overall, detoxified RPI prepared by the combined treatment of phytase and ethanol was considered safe under the conditions tested, in which the contents of the main ANFs were reduced significantly.

Assessment of physical, microstructural, thermal, techno-functional and rheological characteristics of apple (Malus domestica) seeds of Northern Himalayas.

In this research, two common apple seed cultivars Viz: 'Golden Delicious' (GD) and 'Red Delicious' (RD) of Northern Himalayan region were characterized for physical, techno-functional, microstructure, thermal, and rheological properties. Seeds showed a significant difference in width, arithmetic, and geometric mean diameters, volume, and surface area. Proximate analysis results revealed that seed flours have high oil content (> 20%) and are potentially rich sources of protein (> 40%). Color analysis of flours indicated their satisfactory whiter color with higher brightness values (L* ˃ 75), resulting from the reduced particle size which allows greater light penetration and relatively lower a* (< 1.5) and b* (< 11) values. Techno-functional attributes including water/oil absorption capacity, emulsifying capacity, and emulsion stability were significantly higher in RD than GD flour. There was also a significant difference in the average particle size of seed flours. Flour micrographs indicated the presence of oval/spherical-shaped starch granules embedded in dense protein matrix while, Differential Scanning calorimeter (DSC) revealed exothermic transition enthalpies for seed flours. Additionally, seed flours depicted high elastic modulus (G'), suggesting their suitability for modifying food texture. It was concluded that apple seeds exhibit significant potential for use in formulating protein-enriched foods while contributing to reducing industrial wastage.

Separating the Wheat from the Chaff: Nutritional Value of Plant Proteins and Their Potential Contribution to Human Health.

The quality and nutritional value of dietary proteins are determined by the quantity, digestibility and bioavailability of essential amino acids (EAA), which play a critical role in human growth, longevity and metabolic health. Plant-source protein is often deficient in one or more EAAs (e.g., branched-chain amino acids, lysine, methionine and/or tryptophan) and, in its natural form, is less digestible than animal-source protein. Nevertheless, dietary intake of plant-source protein has been promoted because of its potential health benefits, lower cost of production and lower environmental impact compared to animal-source protein. Implementation of dietary strategies that improve both human and planetary health are of critical importance and subject to growing interest from researchers and consumers. Therefore, in this review we analyse current plant protein intake patterns and discuss possible countermeasures that can enhance plant protein nutrition, examples include: (1) combining different plant proteins with complementary EAA profiles; (2) identification and commercial cultivation of new and novel high-quality plant proteins; (3) industrial and domestic processing methods; and (4) genome-editing techniques.

Risk assessment of heavy metal toxicity by sensitive biomarker δ-aminolevulinic acid dehydratase (ALA-D) for onion plants cultivated in polluted areas in Kosovo.

Biomarkers allow an integrated risk assessment of heavy metal pollution effects in living organisms. In this study, the biochemical effects of Cd, Cr, Ni, Pb and Zn pollution in agricultural soil and their accumulation in Alium cepa L. (onion) were evaluated with ALA-D enzyme response as a biomarker, along with δ-aminolevulinic acid (ALA) and total chlorophyll contents in leaves of this plant. Soil samples were randomly selected from agricultural areas in two regions, Mitrovica and Obiliqi, which are considered the most industrially polluted regions in Kosovo. Results show that Pb and Zn concentrations in soil samples from Mitrovica (1953-2576 mg kg -1) and Obiliqi regions (138-179 mg kg -1) and their bioaccumulation levels in onion were significantly higher in comparison with the control group. There was an adverse negative correlation between Pb or Zn concentration and ALA-D activity and total chlorophyll content, and a positive correlation with ALA content. This study indicates that ALA-D activity can be used as a very sensitive biomarker for evaluation of heavy metal pollution. The bioaccumulation of heavy metals from soil polluted areas poses a threat for food contamination and public health.

An automated flow system for in-line extraction of proteins and separation of jacalin using a low-cost stationary phase.

Lectins are carbohydrate binding proteins with many physiological and biotechnological applications. Isolation of proteins is normally time-consuming and encompasses multiple and, sometimes, complicated steps that hinder reproducibility and yield. Affinity chromatography is an efficient way to simplify and improve protein purification, however often requiring an expensive and fragile stationary phase. In this regard, automated flow-based systems minimize the time for extraction of species from solid samples without hindering the features of batch procedures. In this work, a new inexpensive affinity-based stationary phase was developed for in-line separation of jacalin, a galactose-binding lectin from jackfruit seeds. In the flow manifold, in-line extraction of proteins was also carried out with continuous monitoring using the spectrophotometric Biuret assay. For protein determination, linear response was observed from 3.0 to 15 g L-1. The results of the analysis of protein extracts from jackfruit seeds obtained with the herein described procedure and batch procedure agreed with 95% confidence level. Quantitative extraction of proteins from jackfruit seed powder required recirculation of extraction buffer for 15 min through a lab-made polymethylmethacrylate (PMMA) column containing 200 mg of the crude seed powder. In the chromatographic step, jacalin was isolated after 300 s. Therefore, three essential steps for jacalin isolation were performed in one manifold in a fast way, minimizing sample consumption and solution handling. Additionally, the versatile and multi-task developed flow manifold can be useful for routine analysis and preparative procedures, being adaptable for the extraction and separation of other species from solid matrixes with continuous monitoring of the processes.

Proteomic and physiological assessment of stress sensitive and tolerant variety of tomato treated with brassinosteroids and hydrogen peroxide under low-temperature stress.

The aim of current investigation was to perform proteomics and physio-chemical studies to dissect the changes in contrasting varieties (S-22 and PKM-1) of Lycopersicon esculentum under low-temperature stress. Plant grown under variable low-temperature stress were analysed for their growth biomarkers, antioxidant enzyme activities, and other physiological parameters, which headed toward the determination of protein species responding to low-temperature and 24-epibrassinolide (EBL) concentrations. The plants grown under temperatures, 20/14, 12/7, and 10/3 °C recorded significantly lower growth biomarkers, SPAD chlorophyll, net photosynthetic rate and carbonic anhydrase activity in S-22 and PKM-1. Moreover, the combined effect of EBL and hydrogen peroxide (H2O2) significantly improved the parameters mentioned above and consecutively upgraded the different antioxidant enzymes (CAT and SOD) with higher accumulation of proline under stress and stress-free environments. Furthermore, proteomics study revealed that the maximum number of differentially expressed proteins were detected in S-22 (EBL + H2O2); while treatment with EBL + H2O2 + low temperature lost expression of 20 proteins. Overall, three proteins (O80577, Q9FJQ8, and Q9SKL2) took a substantial part in the biosynthesis of citrate cycle pathway and enhanced the growth and photosynthetic efficiency of tomato plants under low-temperature stress.

¹H-NMR Profiling and Carbon Isotope Discrimination as Tools for the Comparative Assessment of Walnut (Juglans regia L.) Cultivars with Various Geographical and Genetic Origins-A Preliminary Study.

：The aim of the study was to investigate the differences between walnut genotypes of various geographical and genetic origins grown under the same or different environmental conditions. The biological material analyzed consisted in walnut kernels of 34 cultivars, nine advanced selections, and six hybrids harvested in 2015 and 2016, summing up to a total of 64 samples. The walnuts, walnut oil, and residue were characterized in respect to their chemical (proximate composition-fat, protein, nutritional value, fatty acids profile by ¹H-NMR) and carbon-13 isotopic composition. The data was used to statistically discriminate the cultivars according to composition, geographical area of origin, and year of harvest, comparing the Romanian cultivars, selections, and hybrids with the internationally available ones.

Assessment of peanut allergen Ara h1 in processed foods using a SWCNTs-based nanobiosensor.

The goals of this research were to develop a rapid single-walled carbon nanotube (SWCNT)-based biosensor and to employ it to commercial food products for Ara h1 detection. The SWCNT-based biosensor was fabricated with SWCNTs immobilized with antibody (pAb) through hybridization of 1-pyrenebutanoic acid succinimidyl ester (1-PBASE) as a linker. The resistance difference (ΔR) was calculated by measuring linear sweep voltammetry (LSV) using a potentiostat. Resistance values increased as the concentration of Ara h1 increased over the range of 1 to 105 ng/L. The specific binding of anti-Ara h1 pAb to antigen including Ara h1 was confirmed by both indirect ELISA kit and biosensor assay. The biosensor was exposed to extracts prepared from commercial processed food containing peanuts, or no peanuts, and could successfully distinguish the peanut containing foods. In addition, the application of present biosensor approach documented the precise detection of Ara h1 concentrations in commercially available peanut containing foods.

Isolation, Purity Assessment, and Proteomic Analysis of Nuclei.

The integrity of a subcellular proteomics is largely dependent on purity of the isolated compartment away from other contaminants. If high-purity nuclei is isolated, nuclear proteomics is a useful approach for investigating the mechanisms underlying plant physiological function. Although the isolation of high-purity nuclei from tissue or organ in plant is a difficult task, successful purification has been achieved through fractionation processes. For purification, there are five protocols such as (1) differential centrifugation, (2) discontinuous Percoll gradients, (3) continuous sucrose gradients, (4) combined continuous Percoll/sucrose gradients, and (5) continuous Percoll gradients. Furthermore, because purity assessment of purified nuclei is an important step, it is also described in this chapter.

Brazil nuts: Nutritional composition, health benefits and safety aspects.

Brazil nuts are among the richest selenium food sources, and studies have considered this Amazonian nut as an alternative for selenium supplementation. Besides selenium, Brazil nuts present relevant content of other micronutrients such as magnesium, copper, and zinc. The nutritional composition of nuts, also characterized by adequate fatty acid profile and high content of protein and bioactive compounds, has many health benefits. In the present review, we examine the nutritional composition of Brazil nuts, comparing it with other nuts, and describe the relevance of possible contaminants and metal toxicants observed in this nut for human health. Furthermore, we report different trials available in the literature, which demonstrate positive outcomes such as modulation of the lipid serum profile, enhancement of the antioxidant system and improvement of anti-inflammatory response. These effects have been assessed under different conditions, such as cognitive impairment, dyslipidemia, cancer, and renal failure.

Cowpea fresh pods - a new legume for the market: assessment of their quality and dietary characteristics of 37 cowpea accessions grown in southern Europe.

BACKGROUND: Cowpea is traditionally cultivated in some regions of southern Europe for its dried seeds; however, there is a scarcity of information on the quality and dietary characteristics of fresh pods, which are occasionally used in folk diets. This paper aims at covering this gap in knowledge, thereby contributing to the dissemination of fresh cowpea pods as a novel product for the market. The quality and dietary characteristics of pods from 37 accessions (Vigna unguiculata ssp. unguiculata and ssp. sesquipedalis) grown in southern Europe were assessed in an attempt to provide information on pod quality and nutritional properties and to identify relationships between quality traits and accession origin. RESULTS: Pods from the sesquipedalis accessions were heavier and larger, and reached commercial maturity 2 days later, than those from the unguiculata accessions. There were also large differences in the quality and dietary characteristics of the accessions. The pods of most accessions were rich in proteins, chlorophylls, carotenoids and phenolics, and showed high antioxidant activity and low concentrations of nitrates and raffinose-family oligosaccharides. Cluster analysis based on quality, dietary or antinutritional traits did not reveal any apparent grouping among the accessions. All the quality characteristics were independent of accession origin and subspecies. CONCLUSION: Most of the accessions produced fresh pods of good quality and high dietary value, suitable for introduction in the market and/or for use as valuable genetic material for the development of new improved varieties. © 2017 Society of Chemical Industry.

Assessment of Natural Variability of Maize Lipid Transfer Protein Using a Validated Sandwich ELISA.

Lipid transfer protein (LTP) is the main causative agent for rare food allergic reactions to maize. This paper describes a new, validated ELISA that accurately measures maize LTP concentrations from 0.2 to 6.4 ng/mL. The levels of LTP ranged from 171 to 865 µg/g of grain, a 5.1-fold difference, across a set of 49 samples of maize B73 hybrids derived from the Nested Association Mapping (NAM) founder lines and a diverse collection of landrace accessions from North and South America. A second set of 107 unique samples from 18 commercial hybrids grown over two years across 10 U.S. states showed a comparable range of LTP level (212-751 µg/g of grain). Statistical analysis showed that genetic and environmental factors contributed 63 and 6%, respectively, to the variance in LTP levels. Therefore, the natural variation of maize LTP is up to 5-fold different across a diverse collection of varieties that have a history of safe cultivation and consumption.

Quality assessment of dried okara as a source of production of gluten-free flour.

BACKGROUND: Okara is a by-product of soymilk and of tofu elaboration that is rich in protein, fiber and vegetable oils as a source of gluten-free flour. In order to take advantage of the nutritional characteristics of okara and to be able to determine an appropriate drying methodology, microwave, rotary dryer and freeze-drying were assessed. Furthermore, flour with an enzymatic treatment was characterized as well as its functional, physicochemical, and textural properties. RESULTS: The results showed that the physiochemical characteristics of the flour were affected by the drying process, reaching adequate water content, and high protein and fiber content. The freeze-drying process produced clearer flours with porous structure and high water absorption capacity, and with a higher protein denaturation. Okara dried by microwave and rotary dryer exhibited a denser structure with similar functional properties and improved textural characteristics such as firmness and consistency. The microwave-produced flour was darker due to the non-enzymatic browning reactions. The enzymatic treatment employed improved the consistency of the flour. CONCLUSION: It was possible to choose the drying process to be applied according to the feasible use of the flour, intended to preserve the favorable nutritional aspects of the okara flour. Based on the results, it can be affirmed that the physicochemical properties and attributes of okara are influenced by the drying process employed. Okara dried by freeze-drying resulted in a better product because it had a low final moisture content and the highest whiteness index. The flour presented a porous structure with high solubility, which is an indicator of potential applications in foods developments. © 2016 Society of Chemical Industry.

Antioxidant activity and nutritional assessment of under-utilized medicinal plants.

The present study investigates the nutritive and anti-nutritive composition of six species (Allium porrum, Amaranthus spinosis, Apium graveolens, Caralluma edulis, Chenopodium album, Urtica dioica). The studied species contained considerable amount of crude protein (4.53-11.41%), crude fat (1.25-3.74%), vitamin C (7.85-28.09mg 100-1 g) and ß-Carotene (18.29-169.33mg 100-1g). The mineral profile of Chenopodium album is considered as potential source of Zn (14.51±1.14mg 100-1g), Mn (67.71±0.85mg 100-1g), Se (8.45±0.49mg 100-1g) and Fe (182.08±1.52mg 100-1g) and Caralluma edulis as potential source of Fe (7.28± 0.03mg 100-1g). Ca and P content ranged 63-306mg 100-1 g and 12-392mg 100-1 g in all studied plants with exceptionally high level was found in Chenopodium album (1084mg 100-1g) and (3924mg 100-1g), respectively. Similarly, total phenol, tannin and phytic acid were found in the range of 105-354, 5-90 and 10-85mg 100-1g, respectively. Total oxalates and soluble oxalates contents were below 200mg 100-1g in five examined plants with the exception of Chenopodium album (413mg 100-1g). Antioxidant activity of Caralluma edulis was found maximum while the rest of the examined plants had moderate levels.

Chia (Salvia hispanica): A Review of Native Mexican Seed and its Nutritional and Functional Properties.

In this chapter, a review is made on various aspects of chia seed in order to provide an overall, yet comprehensive view, about this important commodity with the aim of updating the current state of knowledge on its composition, possible nutraceutical properties, and potential benefits for human health. Based on this approach, the discussion includes some comments on the main historical aspects, morphology of the seed, its importance in the diet of humans and stresses the main results issued from investigations on its three main components; lipid, protein, and fiber. The chapter closes with a discussion on the potential benefits for human health, highlighting the contradictions that still exist in this area and the need for continued research in this direction and considerations on the role of chia seed as a functional food.

Computational identification of protein kinases and kinase-specific substrates in plants.

The protein phosphorylation catalyzed by protein kinases (PKs) plays an essential role in almost all biological progresses in plants. Thus, the identification of PKs and kinase-specific substrates is fundamental for understanding the regulatory mechanisms of protein phosphorylation especially in controlling plant growth and development. In this chapter, we describe the computational methods and protocols for the identification of PKs and kinase-specific substrates in plants, by using Vitis vinifera as an example. First, the proteome sequences and experimentally identified phosphorylation sites (p-sites) in Vitis vinifera were downloaded. The potential PKs were computationally identified based on preconstructed Hidden Markov Model (HMM) profiles and ortholog searches, whereas the kinase-specific p-sites, or site-specific kinase-substrate relations (ssKSRs) were initially predicted by the software package of Group-based Prediction System (GPS) and further processed by the iGPS algorithm (in vivo GPS) to filter potentially false positive hits. All primary data sets and prediction results of Vitis vinifera are available at: http://ekpd.biocuckoo.org/protocol.php.

Effect of the addition of chia's by-product on the composition of fatty acids in hamburgers through chemometric methods.

BACKGROUND: Hamburger is a meat-based food that is easy to prepare and is widely consumed. It can be enriched using different ingredients, such as chia's by-product, which is rich in omega-3. Chemometrics is a very interesting tool to assess the influence of ingredients in the composition of foods. A complete factorial design 2(2) (two factors in two levels) with duplicate was performed to investigate the influence of the factors (1) concentration of textured soy proteins (TSP) and (2) concentration of chia flour partially defatted (CFPD) as a partial replacement for the bovine meat and porcine fat mix in hamburgers. RESULTS: The results of proximal composition, lipid oxidation, fatty acids sums, ratios, and nutritional indexes were used to propose statistical models. The factors TSP and CFPD were significant, and the increased values contributed to improve the composition in fatty acids, crude protein, and ash. Principal components analysis distinguished the samples with a higher content of chia. In desirability analysis, the highest level of TSP and CFPD was described as the optimal region, and it was not necessary to make another experimental point. CONCLUSION: The addition of chia's by-product is an alternative to increase the α-linolenic contents and to obtain nutritionally balanced food.

Effect of phenological stages on nutritional assessment of ten plant species in Tehsil Takht-e-Nasrati, District Karak, Pakistan.

The proximate analysis of ten plant species from Tehsil takht-e-Nasrati, district Karak was conducted at three phenological stages using standard techniques. The species showed variable results. In the test species, the moisture content varied from 2.11-14.12 %, 1.85-11.51 and 1.12-11.51%; the ash contents varied from 3.08-17.16%, 5.09-18.12% and 6.13-19.09%; the fibre contents varied from 2.14-11.09%, 2.43-12.31% and 2.78-13.01%; the carbohydrate contents varied from 54-85.1%, (50-84.1) and 47.1-81.1%; the fat contents varied from 1.06-5.46%, 1.09-5.98% and 1.56- 6.57 %; the protein contents varied from 2.11-12.41%, 3.14-15.87% and 4.12-18.54%; the energy value varied from 305.12-394.23 KCal/100g, 314.21-423.13 KCal /100g and 321.23-434.26 KCal /100g at vegetative stage, reproductive stage and post reproductive stage respectively. Proximate analysis of such plant species can help us to determine the health benefits achieved from their use in society. It is concluded that the poor livestock productivity in research area is partially due to insufficient amount of available poor quality forage that requires attention of the range mangers to improve the habitat and livestock breeds.

Proteomic analysis of the Actinidia deliciosa leaf apoplast during biotrophic colonization by Pseudomonas syringae pv. actinidiae.

For plant pathogenic bacteria, adaptation to the apoplast is considered as key in the establishment of the parasitic lifestyle. Pseudomonas syringae pv. actinidiae (Psa), the causal agent of the bacterial canker of kiwifruit, uses leaves as the entry site to colonize plants. Through a combined approach based on 2-DE, nanoLC-ESI-LIT-MS/MS and quantitative PCR, we investigated Psa colonization of the Actinidia deliciosa "Hayward" leaf apoplast during the bacterial biotrophic phase. A total of 58 differentially represented protein species were identified in artificially inoculated leaves. Although the pathogen increased its population density during the initial period of apoplast colonization, plant defense mechanisms were able to impede further disease development. We identified a concerted action of different proteins mainly belonging to the plant defense and metabolism category, which intervened at different times and participated in reducing the pathogen population. On the other hand, bacterial BamA was highly represented during the first week of leaf apoplast colonization, whereas OmpA and Cpn60 were induced later. In addition to presenting further proteomic information on the molecular factors actively participating in this pathosystem, our data characterize the early events of host colonization and will promote the eventual development of novel bioassays for pathogen detection in kiwiplants. BIOLOGICAL SIGNIFICANCE: This original study evaluates on a proteomic perspective the interaction occurring into the leaf apoplast between Actinidia deliciosa and its specific pathogen Pseudomonas syringae pv. actinidiae. Despite the initial bacterial multiplication, a concerted action of the plant defense mechanisms blocked the infection during 21days of apoplast colonization, as revealed by the number of differentially-represented proteins identified in artificially-inoculated and control leaves. Three bacterial proteins were also recognized among the over-represented molecules in infected plants. This study may contribute to improve breeding programs aimed at selecting resistant/tolerant kiwifruit cultivars toward P. syringae pv. actinidiae, which present a high representation of the plant proteins here shown to be involved in resistance mechanisms. In addition to present additional information on the molecular players actively participating in this pathosystem, our data will also facilitate the technological development of future bioassays for the detection of this pathogen in kiwiplants.

Nutritional characterisation and bioactive components of commercial carobs flours.

Food industry is interested in the utilisation of legume flours for the improvement the nutritional quality of cereal based foods. In this context, this research aimed at investigating the beneficial properties of different commercial carob seed flours -Ceratonia siliqua L.-. In particular, we determined chemical parameters (protein, fat, ash, soluble and insoluble fibre) by standard AOAC methods, lignans (secoisolariciresinol, lariciresinol, isolariciresinol, pinoresinol) by HPLC methods, the Total Polyphenol Content (TPC) by the Folin Ciocalteau method and the antioxidant properties by the FRAP assay. The carob germ flour and the raw carob seed flour reached the highest insoluble fibre, lignan and total polyphenols content and these results were matched by their antioxidant properties. Different carob flours showed a different distribution of the various lignans.