Generating Multi-Functional Pulse Ingredients for Processed Meat Products-Scientific Evaluation of Infrared-Treated Lentils.

In the last decade, various foods have been reformulated with plant protein ingredients to enhance plant-based food intake in our diet. Pulses are in the forefront as protein-rich sources to aid in providing sufficient daily protein intake and may be used as binders to reduce meat protein in product formulations. Pulses are seen as clean-label ingredients that bring benefits to meat products beyond protein content. Pulse flours may need pre-treatments because their endogenous bioactive components may not always be beneficial to meat products. Infrared (IR) treatment is a highly energy-efficient and environmentally friendly method of heating foods, creating diversity in plant-based ingredient functionality. This review discusses using IR-heating technology to modify the properties of pulses and their usefulness in comminuted meat products, with a major emphasis on lentils. IR heating enhances liquid-binding and emulsifying properties, inactivates oxidative enzymes, reduces antinutritional factors, and protects antioxidative properties of pulses. Meat products benefit from IR-treated pulse ingredients, showing improvements in product yields, oxidative stability, and nutrient availability while maintaining desired texture. IR-treated lentil-based ingredients, in particular, also enhance the raw color stability of beef burgers. Therefore, developing pulse-enriched meat products will be a viable approach toward the sustainable production of meat products.

Water-soluble phenolic compounds and their putative antioxidant activities in the seed coats from different lentil (Lens culinaris) genotypes.

The seed coat is a major byproduct of lentil processing with potential as a sustainable source of antioxidant polyphenols. Profiles of water-soluble phenolic compounds and antioxidant activities of seven genotypes of lentil which includes both normal-tannin and low-tannin seed coats were investigated. Antioxidant activities were assessed using four antioxidant assays, and phenolic compounds were quantified using liquid chromatography mass spectrometry (LC-MS). Total phenolic content (TPC) varied significantly among genotypes and ranged between 1519 ± 140 and 6502 ± 154 µg/g. Thirty phenolic compounds were identified with kaempferol tetraglycoside, catechin-3-glucoside and procyanidins being the dominant compounds in normal-tannin seed coats. Kaempferol tetraglycoside predominated (80-90%) in low-tannin seed coats. Antioxidant activities strongly correlated with TPC (r > 0.93) with a 6-9 times higher activity in normal-tannin than that of low-tannin lentils. Without flavan-3-ols and procyanidins, low-tannin seed coat may not exert strong antioxidant potential, whereas normal-tannin seed coat contains water-extractable natural phenolic compounds with promising antioxidant potential.

Sensory descriptors for pulses and pulse-derived ingredients: Toward a standardized lexicon and sensory wheel.

The organoleptic quality of pulses and their derived ingredients is fundamental in human utilization and evolution of food. However, the widespread use of pulses is hindered by their inherent sensorial aspects, which are regarded as atypical by the consumers who are unfamiliar to them. In most studies involving sensory assessment of pulses and pulse-ingredients using classical descriptive analysis methods, assessors establish their own lexica. This review is a synthesis of descriptive terms by which sensations emanating from pea, chickpea, lentil, faba bean, dry bean, bambara groundnut, lupin, pigeon pea and cowpea, and their derived ingredients have been described in the literature. Studies involving sensory assessment of processed whole seeds, slurries of raw flour, slurries of protein extracted from raw flour, and food products containing components of pulses were considered. The terms are categorized into those denoting basic taste, aroma, flavor, and trigeminal sensations. Bitterness is the most widely perceived basic taste. Beany, which is broad and complex with subcharacter notes, is predominantly used to describe aroma and flavor. The frequency of use of the collated terms in the reviewed studies was used to establish a sensory wheel. Inconsistency in the use of descriptive terms in the literature necessitates establishment of a standard lexicon that can be applied in both classical and increasingly popular rapid descriptive methods (e.g., check-all-that-apply) throughout the pulse value chain. This review is timely considering the dominance of pulses in plant-based foods and their increasing appeal to the food industry.

Subunit composition affects formation and stabilization of o/w emulsions by 11S seed storage protein cruciferin.

The 11S globulin cruciferin is the major storage protein in Brassicaceae/Cruciferae seeds and exists as a hexamer in its natural configuration. Arabidopsis thaliana cruciferin is composed of CRUA, CRUB and CRUC subunits. Wild type (WT) cruciferin and cruciferins composed only of identical CRUA, CRUB and CRUC subunits were examined for their ability to form and stabilize oil-in-water (o/w) emulsions. All proteins (0.9% at pH 7.4 and 2.0), except CRUC, formed stable canola oil or triolein emulsions with a dispersed phase volume fraction of 22-23%. A fine emulsion was formed by CRUB at pH 7.4 with droplet sizes of 6.8 and 8.6 µm for canola oil and triolein, respectively. The presence of 0.5 M NaCl reduced the level of adsorbed protein and protein load at the interface at pH 7.4, and resulted in emulsions that were less stable. Emulsions of CRUA and CRUB (pH 7.4, zero ionic strength, canola oil or triolein) had higher stability than emulsions with WT cruciferin up to 15 days after formation. CRUC formed a stable emulsion only at pH 2.0. The low solubility, low surface hydrophobicity and compact structure of the CRUC protein may contribute to its inferior emulsifying properties at neutral pH; however, acidic pH-induced dissociation of the hexameric assembly improved these properties. The abundance and exposure of hydrophobic residues in the hypervariable regions, extended loop regions, and solvent exposed surfaces of cruciferin are critical factors affecting o/w interface stabilization.

Structural Properties of Cruciferin and Napin of Brassica napus (Canola) Show Distinct Responses to Changes in pH and Temperature.

The two major storage proteins identified in Brassica napus (canola) were isolated and studied for their molecular composition, structural characteristics and the responses of structural features to the changes in pH and temperature. Cruciferin, a complex of six monomers, has a predominantly ß-sheet-containing secondary structure. This protein showed low pH unstable tertiary structure, and distinctly different solubility behaviour with pH when intact in the seed cellular matrix. Cruciferin structure unfolds at pH 3 even at ambient temperature. Temperature-induced structure unfolding was observed above the maximum denaturation temperature of cruciferin. Napin was soluble in a wider pH range than cruciferin and has α-helices dominating secondary structure. Structural features of napin showed less sensitivity to the changes in medium pH and temperature. The surface hydrophobicity (S0) and intrinsic fluorescence of tryptophan residue appear to be good indicators of cruciferin unfolding, however they were not the best to demonstrate structural changes of napin. These two storage proteins of B. napus have distinct molecular characteristics, therefore properties and functionalities they provide are contrasting rather than complementary.

Canola/Rapeseed Protein: Future Opportunities and Directions-Workshop Proceedings of IRC 2015.

At present, canola meal is primarily streamlined into the animal feed market where it is a competitive animal feed source owing to its high protein value. Beyond animal feed lies a potential game-changer with regards to the value of canola meal, and its opportunity as a high quality food protein source. An economic and sustainable source of protein with high bioavailability and digestibility is essential to human health and well-being. Population pressures, ecological considerations, and production efficiency underscore the importance of highly bioavailable plant proteins, both for the developed and developing world. Despite decades of research, several technologies being developed, and products being brought to large scale production, there are still no commercially available canola protein products. The workshop entitled "Canola/Rapeseed Protein-Future Opportunities and Directions" that was held on 8 July 2015 during the 14th International Rapeseed Congress (IRC 2015) addressed the current situation and issues surrounding canola meal protein from the technological, nutritional, regulatory and genomics/breeding perspective. Discussions with participants and experts in the field helped to identify economic barriers and research gaps that need to be addressed in both the short and long term for the benefit of canola industry.

Structural stability and Sin a 1 anti-epitope antibody binding ability of yellow mustard (Sinapis alba L.) napin during industrial-scale myrosinase inactivation process.

This study investigated the structural stability of yellow mustard (YM, Sinapis alba L.) napin and the changes of its Sin a 1 anti-epitope antibody-binding ability during myrosinase enzyme inactivation process. The food industry uses myrosinase-inactive non-pungent YM for uses beyond spice applications. Napin was isolated from seeds received from an industrial processor before (YM + M) and after (YM - M) myrosinase inactivation. Secondary and tertiary structural features and surface hydrophobicity parameters of napin were analyzed. The Sin a 1 content in YM seeds and the stability of Sin a 1-containing napin during simulated in vitro gastrointestinal (GI) digestion were determined by a non-competitive indirect enzyme-linked immunosorbent assay using the Sin a 1 anti-epitope antibody (AE-Ab) as the primary Ab. YM napin retained the dominant alpha-helical components of secondary and tertiary structure folds during this process. YM - M napin showed changes in hydrophobicity parameters of the molecules and binding ability of AE-Ab: 2.19 ± 0.48 g per 100 g of YM - M seeds vs. 1.49 ± 0.16 g per 100 g YM + M seeds. YM - M proteins were more susceptible for in vitro GI digestion and also showed a 30% reduction in AE-Ab binding ability upon digestion of napins. This suggests that the myrosinase inactivation process has induced the surface modification of napin, exposing Sin a 1 epitope, leading to an increase in AE-Ab binding. However, the epitope region of YM - M napin showed improved susceptibility for hydrolysis during GI digestion resulting in fewer available epitope regions, suggesting a possible reduction in napin immune reactivity.

Structural and physicochemical property relationships of cruciferin homohexamers.

Heteromeric cruciferin from wild type (WT) Arabidopsis thaliana and homomeric cruciferin CRUA, CRUB, and CRUC composed of identical subunits obtained from double-knockout mutant lines were investigated for their structural and physicochemical properties. A three-step chromatographic procedure allowed isolation of intact cruciferin hexamers with high purity (>95%). FT-IR and CD analysis of protein secondary structure composition revealed that all cruciferins were folded into higher order structures consisting of 44-50% ß-sheets and 7-9% α-helices. The structural and physicochemical properties of homohexameric CRUC deviated from that of CRUA and CRUB and exhibited a compact, thermostable, and less hydrophobic structure, confirming the predictions made using 3D homology structure models.

Characterization of Arabidopsis thaliana lines with altered seed storage protein profiles using synchrotron-powered FT-IR spectromicroscopy.

Arabidopsis thaliana lines expressing only one cruciferin subunit type (double-knockout; CRUAbc, CRUaBc, or CRUabC) or devoid of cruciferin (triple-knockout; CRU-) or napin (napin-RNAi) were generated using combined T-DNA insertions or RNA interference approaches. Seeds of double-knockout lines accumulated homohexameric cruciferin and contained similar protein levels as the wild type (WT). Chemical imaging of WT and double-knockout seeds using synchrotron FT-IR spectromicroscopy (amide I band, 1650 cm(-1), νC═O) showed that proteins were concentrated in the cell center and protein storage vacuoles. Protein secondary structure features of the homohexameric cruciferin lines showed predominant ß-sheet content. The napin-RNAi line had lower α-helix content than the WT. Lines entirely devoid of cruciferin had high α-helix and low ß-sheet levels, indicating that structurally different proteins compensate for the loss of cruciferin. Lines producing homohexameric CRUC showed minimal changes in protein secondary structure after pepsin treatment, indicating low enzyme accessibility. The Synchrotron FT-IR technique provides information on protein secondary structure and changes to the structure within the cell.

Release of angiotensin I-converting enzyme inhibitory peptides from flaxseed (Linum usitatissimum L.) protein under simulated gastrointestinal digestion.

The scope of this study was to determine the ability of flaxseed (Linum usitatissimum L.) proteins to release angiotensin I-converting enzyme inhibitory (ACEI) peptides during simulated gastrointestinal (GI) digestion using a static (SM; no absorption in the intestinal phase) and a dynamic model (DM; simultaneous absorption of digested products in the intestinal phase via passive diffusion). Gastric and gastric + small intestinal digests of flaxseed proteins of both models possessed ACEI activity. The ACEI activity of the gastric + small intestinal digest in the DM (IC(50) unabsorbed, 0.05 mg N/mL; IC(50) absorbed, 0.04 mg N/mL) was significantly higher (p < 0.05) than that of the SM (IC(50), 0.39 mg N/mL). Two peptides, a pentapeptide (Trp-Asn-Ile/Leu-Asn-Ala) and a hexapeptide (Asn-Ile/Leu-Asp-Thr-Asp-Ile/Leu), were identified in the most active ACEI fraction (0.5-1 kDa) of the absorbable flaxseed protein digest by de novo sequencing.

Proteins of Brassicaceae oilseeds and their potential as a plant protein source.

Among the commercially cultivated Brassicaceae (Cruciferae) plants, Brassica juncea, Brassica napus, Brassica rapa, and Sinapis alba store significant amounts of oil and protein in the seed. At present, Brassica seed proteins are primarily used for livestock feeding based on the nutritional value. The point of curiosity is whether the present knowledge on the protein structure, biochemical characteristics, nutritive value, and the recovery processes are inadequate to develop Brassica proteins into a usable plant protein source or these proteins are of substandard for uses beyond animal nutrition applications. Cruciferin (11S) and napin (2S) are the predominant storage proteins of Brassicaceae seeds that contribute to different properties and functions. A gamut of information is available on the chemistry, nutritional value, as well as the functionality in foods, and associated non-protein components of canola/rapeseed storage proteins. The intention of this article is to critically review what is known about the predominant storage proteins of commercially produced Brassicaceae seeds relative to the above aspects and identify the knowledge gaps.

Physicochemical, thermal and functional characterisation of protein isolates from Kabuli and Desi chickpea (Cicer arietinum L.): a comparative study with soy (Glycine max) and pea (Pisum sativum L.).

BACKGROUND: Chickpea (Cicer arietinum L.) seeds are a good source of protein that has potential applications in new product formulation and fortification. The main objectives of this study were to analyse the physicochemical, thermal and functional properties of chickpea protein isolates (CPIs) and compare them with those of soy (SPI) and pea (PPI) protein isolates. RESULTS: Extracted CPIs had mean protein contents of 728-853 g kg(-1) (dry weight basis). Analysis of their deconvoluted Fourier transform infrared spectra gave secondary structure estimates of 25.6-32.7% α-helices, 32.5-40.4% ß-sheets, 13.8-18.9% turns and 16.3-19.2% disordered structures. CPIs from CDC Xena, among Kabuli varieties, and Myles, among Desi varieties, as well as SPI had the highest water-holding and oil absorption capacities. The emulsifying properties of Kabuli CPIs were superior to those of PPI and Desi CPIs and as good as those of SPI. The heat-induced gelation properties of CPIs showed a minimum protein concentration required to form a gel structure ranging from 100 to 140 g L(-1) . Denaturation temperatures and enthalpies of CPIs ranged from 89.0 to 92.0 °C and from 2.4 to 4.0 J g(-1) respectively. CONCLUSION: The results suggest that most physicochemical, thermal and functional properties of CPIs compare favourably with those of SPI and are better than those of PPI. Hence CPI may be suitable as a high-quality substitute for SPI in food applications.

Quantitative detection of allergenic protein Sin a 1 from yellow mustard (Sinapis alba L.) seeds using enzyme-linked immunosorbent assay.

Allergy to yellow mustard (YM; Sinapis alba L.) seed proteins has been reported and is currently seen as a constraint that hampers expansion of YM protein utilization. The most predominant allergenic protein of YM seed has been recognized as Sin a 1. In this study, Sin a 1 was purified ( S. alba var. Andante), rabbit polyclonal antibodies (pAb) specific to Sin a 1 were generated, and a sandwich enzyme-linked immunosorbent assay (S-ELISA) was developed to detect and quantify Sin a 1 from YM. The S-ELISA method using Sin a 1-pAb and its horseradish peroxidase conjugate resulted in a detection limit of 0.3 microg/mL for purified Sin a 1. The Sin a 1 contents of six YM lines were in the range of 0.82-2.94 mg/g when assayed by the developed S-ELISA method. The results showed that S-ELISA could distinguish Sin a 1 in YM seed-derived extracts rapidly and could be applied in controlling and/or monitoring of YM allergenic proteins.