A comparative characterisation of functional and structural properties of white and yellow lupin protein isolates

Hydrolyzed protein from Lupinus albus (INIA-Boroa variety) for the functional food industry: An example of partnership between farmers and academia

Proteins of White Lupin Seed, a Naturally Isoflavone-Poor Legume, Reduce Cholesterolemia in Rats and Increase LDL Receptor Activity in HepG2 Cells

Since recent literature has indicated that white lupin (Lupinus albus) may be a useful source of hypocholesterolemic proteins to be used in functional food formulation, our final goal is the development of a fast and automated high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC/ESI-MS/MS) method for the detection and the label-free semi-quantitation of the main lupin globulins in lupin foods and food ingredients. We present here some preliminary results in this direction. As a first step a total protein extract (TPE-WF) from lupin flakes was pre-fractionated by anion-exchange chromatography and each fraction was digested with trypsin and analyzed by HPLC/ESI-MS/MS. Subsequently, the tryptic digest of TPE-WF was directly analyzed by HPLC/ESI-MS/MS without any pre-fractionation. Eventually, in order to test the applicability of the method to real samples, a lupin beverage and two lupin protein isolates were analyzed. Both Mascot and Spectrum Mill MS Proteomics Workbench software were used to identify the protein composition in these samples and Spectrum Mill was used also to test the possibility of developing a label-free semi-quantitation method based on peptide hits. Encouraging results were obtained especially in the detection of the hypocholesterolemic component beta-conglutin.

Lupin protein isolate was treated using the combination of enzymatic hydrolysis (Papain, Alcalase 2.4 L and Pepsin) and lactic acid fermentation (Lactobacillus sakei ssp. carnosus, Lactobacillus amylolyticus and Lactobacillus helveticus) to investigate the effect on functional properties, sensory profile and protein integrity. The results showed increased foaming activity (2466–3481%) and solubility at pH 4.0 (19.7–36.7%) of all fermented hydrolysates compared to the untreated lupin protein isolate with 1613% of foaming activity and a solubility of 7.3 (pH 4.0). Results of the SDS-PAGE and Bead-Assay showed that the combination of enzymatic hydrolysis and fermentation of LPI was effective in reducing L. angustifolius major allergen Lup an 1 to a residual level of <0.5%. The combination of enzymatic hydrolysis and fermentation enables the production of food ingredients with good functional properties in terms of protein solubility and foam formation, with a balanced aroma and taste profile.

This study aimed to develop a cassava‐based rice analogue with improved nutritional value as an alternative to commercial white rice. The rice analogue formulations (RAFs) were developed by the substitution of modified cassava flour (MOCAF) with rice flour at different ratios as 1:0, 7:3, 5:5, 3:7, and 0:1, followed by the addition of cassava leaves (0, 10, and 20 wt%). The developed rice analogues were evaluated for physicochemical, functional, morphological, and antioxidant properties. The rice analogue containing a 5:5 ratio of rice flour to MOCAF (RAF 2) added with 20% cassava leaves presented the most desirable nutritional composition and functional properties. However, RAF 4 (100% MOCAF with 20% cassava leaves) showed the highest total polyphenol content (TPC) (198.8 mg gallic acid equivalent (GAE)/100 g), total chlorophyll content (TCC) (198 mg/ml), 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) inhibition (79%), and ferric reducing antioxidant power (FRAP) inhibition (85%). Hence, the addition of MOCAF and cassava leaves improved the nutritional value of cassava‐based rice analogues, which could be a healthy alternative to commercial rice in the daily diet.

Sweet lupin (Lupinus albus), a protein-rich legume devoid of anti-nutritional factors, is considered to have a high potential for protein nutrition in man. Results concerning the nutritional value of lupin protein are, however, conflicting in animals and very scarce in human subjects. Furthermore, where fibre-rich protein sources are concerned, the long-term nutritional results are often obscured, particularly since fibre-promoted colonic fermentation may bias the energy supply and redistribute N flux. We therefore studied, during the postprandial phase, the bioavailability and utilisation of lupin-flour protein in nine healthy men who had ingested a mixed meal containing intrinsically15N-labelled lupin flour as the protein source (Expt 1). The real ileal digestibility (RID) and ileal endogenous N losses (IENL) were assessed using a perfusion technique at the terminal ileum, and the N content and15N enrichment of ileal samples. Lupin flour exhibited a high RID of 91 (SD 3) % AND LOW IENL (5·4 (sd 1·3) mmol N/h). Postprandial dietary deamination was also assessed from body dietary urea and urinary dietary N excretion, and compared with results in nine healthy men following an isoenergetic meal containing a15N-soyabean-protein isolate with a similar RID, as a control (Expt 2). Postprandial dietary deamination was similar after lupin and soyabean meals (17 (sd 2) and 18 (sd 4) % ingested N respectively). We therefore conclude that lupin protein is highly bioavailable, even if included in fibre-rich flour, and that it can be used with the same efficiency as soyabean protein to achieve postprandial protein gain in healthy human subjects.

The present work takes an overarching look on how particles can alter the functional and structural properties of shape memory alloys (SMAs). We consider precipitates which form in binary and ternary NiTi-based alloys, including the high-temperature SMAs NiTiHf and NiTiZr. We also take a look at precipitates which form in Cu–Al–Zn, Cu–Al–Ni, and Co–Ni–Ga–SMAs. We take these alloy systems as examples to review different effects of particles on local alloy chemistry and local stress and strain states at particle/matrix interfaces. Their influence on the nucleation and growth of martensite and on the propagation of a martensitic transformation front is discussed, and the consequences for macroscopic functional and structural properties like phase transformation temperatures and widths of thermal hysteresis are highlighted. Emphasis is also placed on particles which form because elements like C and O are picked up during alloy production and affect functional and structural properties. Finally, it is suggested to take a look at additive manufactured SMAs with added inert nano/micro-particles, which would facilitate the investigation of the mechanical constraints, and which may lead to SMAs with improved structural and functional properties.

Simple SummaryIn the past, lupin seeds were not used in poultry feeding because of their alkaloid content, which affected the growth performance, producing negative results. Nowadays, new cultivars of lupins have reduced the anti-nutrition content and have been characterized by a high level of protein, similar to the level in soybean meal. Goose production is not popular and, generally, should be done by an extensive or semi-intensive system, which is corelated with small costs of feeds. Lupins can be used for crops where it is not possible to produce soybean (environmental conditions). Our study indicated that it is possible to produce goose meat with a good quality, where the feeding was based on various cultivars of lupins. Yellow, as well as white, lupin-rich feed had a positive effect on the meat traits and meat quality. Narrow-leaved lupin worsens the growth performance.The aim of the study was to analyze the quality of geese meat receiving feed with soybean meal (group 1), yellow lupin (group 2), narrow-leaved lupin (group 3), or white lupin (group 4). In total, 400 male White Kołuda® geese were randomly assigned to four groups, with 10 replicates and 10 birds each, during the 77-day rearing period. After the end of the rearing period, 10 birds from each group were slaughtered and dissected. Meat quality traits were measured. Based on the production results, it can be concluded that geese use fodder with yellow and white lupin to the same degree as in the case of the control group and higher body weight gain was recorded in the first rearing period. In contrast, the use of narrow-leaved lupin in mixtures for geese worsened the feed used. Meat traits were similar in all groups, including the content of muscles and fat in the carcass (p > 0.05), excluding abdominal fat. The weight of abdominal fat and its proportion in the carcass were higher (p < 0.05) in geese from group 4. A higher (p < 0.05) pH was found in group 1. The protein and intramuscular fat content in breast muscles was highest (p < 0.05) in geese from group 4, and a higher water content was found in group 1. The protein content in leg muscles was higher in group 3, and the fat content was higher in group 4 (p < 0.05). The color and water-holding capacity of meat were comparable in all groups (p > 0.05). The analysis revealed a positive effect of replacing soybean meal with alternative protein sources, especially yellow and white lupin, on the growth performance and quality of goose meat.

Comparative study of the emulsifying properties of blue lupin, white lupin, and soybean protein isolates

Starch is the main integral components of the millets and it is widely used in the food processing sectors for development of functional foods. The extraction and isolation methods of starch from millet grains are greatly affected their functional and structural properties. Therefore, in current study, the effects of different wet milling extraction methods such as water, acidic, and alkaline are investigated on the physical, structural, thermal, and functional properties of barnyard millet starch. The results show that the starch extracted from barnyard millets using alkaline method shows higher yield of starch (64.76%) with good amylose content (34.52%). In addition, the functional properties such as water binding capacity (211.39%), oil binding capacity (223.84%), higher color attributes (whiteness), thermal stability (91.62 °C), textural properties (74.70 g), and lower particle size (19.44 µm) are recorded in barnyard millet starch extracted using alkaline steeping method. The results of the field emission scanning electron microscopes (FESEM) are showed polygonal, spherical, and rounded shapes of all types of extracted starch granules. Whereas, the acidic method is more fused structures of starch granules as compared to water and alkaline methods. The results of the current study reveal that the application of alkaline method is the most potential to extract starch from barnyard millet with good quality and functional properties for further food applications.

Lupin-based food, due to the high content of functional proteins and phenolic compounds, are widely used in human nutrition. Unfortunately, proteins and phenolic compounds can easily interact with each other which results in formation of complexes that affect properties of both components. Therefore, in this study, composition of the seeds storage proteins isolated from Lupinus albus and L. angustifolius and their interactions with native flavonoids were investigated. Based on the chromatographic separations, six proteins fractions of lupin seeds storage proteins were identified. The results indicate that two dominant fractions, α-conglutin and β-conglutin, constitute up to 80% of all proteins present in the seeds. Three flavonoids interacting with the proteins were identified as apigenin C-glycosides. The lowest flavonoids content was noted in the main storage proteins while in both lupin seeds species over 90% of flavonoids interacted with the proteins present in late-embryogenesis abundant (LEA) protein fraction. Protein-phenolic compound complexes can affect the digestibility of proteins and bioavailability of phenolic compounds, and thus the functional and nutritional properties of products derived from lupin seeds can be changed. Therefore, a better understanding of factors affecting the nutritional value of lupin seeds proteins and flavonoids is necessary to optimize the biological use of this plant for human nutrition. © 2019 Society of Chemical Industry.

Several studies support the health-promoting benefits of lupins, particularly lupin proteins. It has been demonstrated that Lupinus albus gamma conglutin (Cγ) protein lowered blood glucose levels; thus, Cγ showed promise as a new anti-diabetic compound for type 2 diabetes (T2D) treatment. The aim of this study was to evaluate the effect of Cγ on Ins-1 gene expression and on pancreatic insulin content in streptozotocin-mediated diabetic rats. Cγ was isolated from Lupinus albus seeds. Its identification was confirmed with polyacrylamide gel electrophoresis under native and denaturing conditions. We used streptozotocin (STZ) to induce T2D on the 5th day of life of newborn male Wistar rats (n5-STZ). After 20 weeks post-induction, these animals (glycemia > 200 mg/dL) were randomly assigned to three groups that received the following one-week treatments: vehicle, 0.90% w/v NaCl (n5 STZ-Ctrl); glibenclamide, 10 mg/kg (n5 STZ-Glib); or Cγ, 120 mg/kg (n5 STZ-Cγ). Glucose and insulin levels were measured before and after treatment. Ins-1 gene expression was quantified using real time polymerase chain reaction and the pancreatic insulin content was evaluated with immunohistochemistry. Post-treatment, the n5 STZ-Cγ and n5 STZ-Glib groups showed reductions in glucose, increments in serum insulin, and increases in Ins-1 gene expression and beta cell insulin content compared to the n5 STZ-Ctrl group. The results showed that Cγ had beneficial effects on Ins-1 gene expression and pancreatic insulin content. These biological effects of Cγ strengthen its promising potential as a nutraceutical and/or new agent for controlling hyperglycemia.

BackgroundWhite lupin (Lupinus albus) is increasingly recognized as a sustainable, highprotein functional food ingredient, yet its potential allergenicity remains a significant concern for food safety. This study aimed to investigate the impact of thermal and biotechnological processing methods on the bioaccessibility and allergenic peptide profiles of white lupin seeds.MethodsLocal lupin seeds were subjected to four distinct processing techniques: boiling, microwave cooking, fermentation, and enzymatic treatment. To evaluate protein stability and allergen release under physiological conditions, the processed samples underwent simulated in vitro gastrointestinal digestion as well as standard protease hydrolysis. The resulting peptide sequences were characterized using liquid chromatography with tandem mass spectrometry (LC-MS/MS) and analyzed via in silico bioinformatics tools to predict potential allergenicity.ResultsThe results indicated that biotechnological processes, specifically enzymatic treatment and fermentation, generated a higher diversity of detectable peptides and distinct allergenic matches following digestion compared to thermal treatments alone. Furthermore, samples subjected to the simulated gastrointestinal digestion model revealed a broader spectrum of conserved epitope matches in allergen databases compared to standard hydrolysis, suggesting that this model provides a more accurate representation of the allergenic peptides likely to persist in the human digestive tract.DiscussionThese findings demonstrate that processing methods significantly modulate the structural integrity and predicted allergenic profile of lupin proteins, providing a molecular basis for selecting appropriate processing strategies in the development of safer lupin-based functional foods.

Background: The ingestion of dietary products containing sweet lupin (such as Lupinus albus or Lupinus angustifolius) has been reported to cause IgE-mediated allergic reactions. Recent studies have indicated lupin globulins as important IgE binding proteins. The aim of the present study was to generate and characterize monoclonal antibodies (mAbs) against lupin seed proteins. Methods: Mice were immunized with a protein isolate from L. albus and mAbs were obtained by hybridoma techniques. Albumins and globulins were extracted, and the globulin fraction was separated further into conglutins by anion exchange chromatography. Specificities, binding patterns and applications of the mAbs were investigated by immunochemical methods. Results: Five mAbs were produced: Lu11 (an IgG2b antibody), Lu8, Lu18, Lu34 and Lu35 (all IgM antibodies). The mAbs reacted strongly with protein isolates from both L. albus and L. angustifolius. All mAbs are directed towards the lupin globulin fraction; Lu11 and Lu18 recognize α-conglutin, while Lu8, Lu34 and Lu35 recognize β-conglutin. In addition, Lu11 inhibited the binding of IgE from patients with positive skin prick tests to lupin proteins in a competitive ELISA by approximately 30%. Furthermore, preliminary results show that Lu11 can be used to develop a sensitive method for the detection of α-conglutin in foods. Conclusions: Lupin globulins are immunogenic and α-conglutin is a potential allergen. This is the first study describing mAbs against the candidate lupin allergens, emphasizing the importance of additional studies on conglutins in lupin allergy.

Complex service-oriented networked applications can be built in a flexible way by appropriate composition of individual services. Services are self-contained pieces of functionality that reside in remote nodes in the network and that communicate only via the exchange of messages. Service composition must be done in compliance with the application-level requirements i.e., quality of service (QoS) constraints. In this way, not only data dependencies must be satisfied but also other non-functional execution requirements must be considered (i.e., resource limitations and timing properties). This paper describes the structure of service-based applications, and it deepens in the characterization of the QoS properties of individual services. It explores the implications of application-level QoS properties in order to select particular services. The proposed characterization presents two dimensions of QoS properties, separating data-related information (functional properties) from physical resources information (non-functional properties). This scheme is proposed and used in the iLAND project where target applications are networked systems for which QoS informaiton is relevant; the execution in these systems must comply with functional and non-functional requirements, especially timing requirements. In this project, the communication middleware performs QoS-based service composition in a time-deterministic manner. To validate the proposed characterization, a service based video application demonstrator is implemented following this scheme, and it shows to be fully compliant with the requirements of the application. It has also proved to be a simple and easy-to-use scheme.