Search Results (11)

Nervonic acid (NA), a very-long-chain monounsaturated fatty acid, is known for its benefits in treating neurological diseases and promoting brain health. In this study, we utilized two different receptors, Brassica juncea (B. juncea, rich in erucic acid, C22:1) and Brassica napus (B. napus, high in oleic acid, C18:1), to overproduce NA through systematic metabolic engineering. Two multi-gene vector constructs, Napin-3 and Napin-5 (CgKCS::SLC1-1::DGAT1; CgKCS::SLC1-1::BnFAE1::LdLPAAT::DGAT1), are driven by seed-specific napin promoters. In B. juncea, Napin-3 and Napin-5 expression elevated NA levels to 48.7% and 46.3% in seed oil, respectively, compared to 2.5% in wild types. In B. napus, Napin-3 and Napin-5 expression achieved NA levels of 45% and 39.6%, respectively, while NA is absent in wild types. To our knowledge, this represents the highest NA production in plants to date, with stable oil content and yield, enabling cost-effective NA production. In B. juncea, a significant increase in NA is observed alongside a decrease in C18:1, C20:1, and C22:1 levels; in B. napus, the rise in NA is accompanied by a decrease in C18:1, and an increase in C20:1 and C22:1. These patterns reflect the dynamic equilibrium of fatty acids following NA conversion, influenced by the Dynamic Substrate Tugging (DST) Mechanism, in the form of either an EA-tugging mode or C18:1-tugging mode mechanism, depending on the cellular context. NA is an elongation product derived from C18:1, catalyzed by CgKCS with broad substrate specificity, indicating that plants with high levels of C18:1, similarly to those rich in C22:1, serve as excellent candidates for NA production. This “green factory” for NA production provides strong support for its pharmaceutical, nutraceutical, and industrial applications. The exogenous and the endogenous enzymes coordinate function remodeling of the intra-seed fatty acid elongation flux through the DST strategy, thereby systematically enhancing the synthesis and accumulation efficiency of the target fatty acid. Full article

Canola is the second-largest cultivated oilseed crop in the world and produces meal consisting of about 35–40% proteins. Despite this, less than 1% of the global plant-based protein market is taken up by canola protein. The reason behind such underutilization of canola protein and its rapeseed counterpart could be the harsh conditions of the industrial oil extraction process, the dark colour of the meal, the presence of various antinutrients, the variability in the protein composition based on the source, and the different properties of the two major protein components. Although academic research has shown immense potential for the use of canola protein and its rapeseed counterpart in emulsion development and stabilization, there is still a vast knowledge gap in efficiently utilizing canola proteins as an effective emulsifier in the development of various emulsion-based foods and beverages. In this context, this review paper summarizes the last 15 years of research on canola and rapeseed proteins as food emulsifiers. It discusses the protein extraction methods, modifications made to improve emulsification, emulsion composition, preparation protocols, and emulsion stability results. The need for further improvement in the scope of the research and reducing the knowledge gap is also highlighted, which could be useful for the food industry to rationally select canola proteins and optimize the processing parameters to obtain products with desirable attributes. Full article

During the Advanced Plant Habitat experiment 2, radish plants were grown in two successive grow-outs on the International Space Station (ISS) for 27 days each. On days 10, 18, and 24, leaf punch (LP) samples were collected and frozen. At harvest, bulb tissue was sampled with oligo-dT functionalized Solid Phase Gene Extraction (SPGE) probes. The space samples were compared with samples from ground controls (GC) grown at the Kennedy Space Center (KSC) under the same conditions as on the ISS, with notably elevated CO₂ (about 2500 ppm), and from lab plants grown under atmospheric CO₂ but with light and temperature conditions similar to the KSC control. Genes corresponding to peroxidase (RPP), glucosinolate biosynthesis (GIS), protein binding (CBP), myrosinase (RMA), napin (RSN), and ubiquitin (UBQ) were measured by qPCR. LP from day 24 and bulb samples collected at harvest were compared with RNA-seq data from material that was harvested, frozen, and analyzed after return to Earth. The results showed stable transcription in LP samples in GC but decreasing values in ISS samples during both grow-outs, possibly indicative of stress. SPGE results were similar between GC and ISS samples. However, the RNA-seq analyses showed different transcription profiles than SPGE or LP results, possibly related to localized sampling. RNA-seq of leaf samples showed greater variety than LP data, possibly because of different sampling times. RSN and RPP showed the lowest transcription regardless of method. Temporal analyses showed relatively small changes during plant development in space and in ground controls. This is the first study that compares developmental changes in space-grown plants with ground controls based on a comparison between RNA-seq and qPCR analyses. Full article

Preventing oxidation and microbial spoilage are both major concerns in food industries. In this context, this study aimed to valorize the total rapeseed meal proteins with controlled enzymatic proteolysis to generate potent mineral-chelating peptides from cruciferins while keeping intact the antimicrobial napins. Implementation of proteolysis of total rapeseed protein isolate with the Prolyve^® enzyme highlighted an interesting selective hydrolysis of the cruciferins. Hence, the mechanism of this particular hydrolysis was investigated through a Design of Experiments method to obtain a model for the prediction of kinetics (cruciferin degradation and napin purity) according to the operating conditions applied. Then, multicriteria optimization was implemented to maximize the napin purity and yield while minimizing both enzymatic cost and reaction time. Antioxidant assays of the peptide fraction obtained under the optimal conditions proved the high metal-chelating activity preservation (EC₅₀ = 247 ± 27 µg) for more than three times faster production. This fraction might counteract lipid oxidation or serve as preventing agents for micronutrient deficiencies, and the resulting purified napins may have applications in food safety against microbial contamination. These results can greatly help the development of rapeseed meal applications in food industries. Full article

The aim of the study presented here was to determine if there is a correlation between the presence of specific protein domains within tree nut allergens or tree nut allergen epitopes and the frequency of bioactive fragments and the predicted susceptibility to enzymatic digestion in allergenic proteins from tree nuts of cashew (Anacardium occidentale), pecan (Carya illinoinensis), English walnut (Juglans regia) and pistachio (Pistacia vera) plants. These bioactive peptides are distributed along the length of the protein and are not enriched in IgE epitope sequences. Classification of proteins as bioactive peptide precursors based on the presence of specific protein domains may be a promising approach. Proteins possessing a vicilin, N-terminal family domain, or napin domain contain a relatively low occurrence of bioactive fragments. In contrast, proteins possessing the cupin 1 domain without the vicilin N-terminal family domain contain a relatively high total frequency of bioactive fragments and predicted release of bioactive fragments by the joint action of pepsin, trypsin, and chymotrypsin. This approach could be utilized in food science to simplify the selection of protein domains enriched for bioactive peptides. Full article

The study aimed to investigate the antibacterial activity of Mustard (Brassica juncea) and Moringa (Moringa oleifera) leaf extracts and coagulant protein for their potential application in water treatment. Bacterial cell aggregation and growth kinetics studies were employed for thirteen bacterial strains with different concentrations of leaf extracts and coagulant protein. Moringa oleifera leaf extract (MOS) and coagulant protein showed cell aggregation against ten bacterial strains, whereas leaf extract alone showed growth inhibition of five bacterial strains for up to 6 h and five bacterial strains for up to 3 h. Brassica juncea leaf extract (BJS) showed growth inhibition for up to 6 h, and three bacterial strains showed inhibition for up to 3 h. The highest inhibition concentration with 2.5 mg/mL was 19 mm, and furthermore, the minimum inhibitory concentration (MIC) (0.5 mg/mL) and MBC (1.5 mg/mL) were determined to have a higher antibacterial effect for <3 KDa peptides. Based on LCMS analysis, napin was identified in both MOS and BJS; furthermore, the mode of action of napin peptide was determined on lipoprotein X complex (LpxC) and four-chained structured binding protein of bacterial type II topoisomerase (4PLB). The docking analysis has exhibited moderate to potent inhibition with a range of dock score −912.9 Kcal/mol. Thus, it possesses antibacterial-coagulant potential bioactive peptides present in the Moringa oleifera purified protein (MOP) and Brassica juncea purified protein (BJP) that could act as an effective antimicrobial agent to replace currently available antibiotics. The result implies that MOP and Brassica juncea purified coagulant (BJP) proteins may perform a wide degree of antibacterial functions against different pathogens. Full article

Rapeseed oil-extracted expeller cake mostly contains protein. Various approaches have been used to isolate, detect and measure proteins in rapeseeds, with a particular focus on seed storage proteins (SSPs). To maximize the protein yield and minimize hazardous chemical use, isolation costs and the loss of seed material, optimization of the extraction method is pivotal. For some studies, it is also necessary to minimize or avoid seed-to-seed cross-contamination for phenotyping and single-tissue type analysis to know the exact amount of any bioactive component in a single seed, rather than a mixture of multiple seeds. However, a simple and robust method for single rapeseed seed protein extraction (SRPE) is unavailable. To establish a strategy for optimizing SRPE for downstream gel-based protein analysis, yielding the highest amount of SSPs in the most economical and rapid way, a variety of different approaches were tested, including variations to the seed pulverization steps, changes to the compositions of solvents and reagents and adjustments to the protein recovery steps. Following SRPE, 1D-SDS-PAGE was used to assess the quality and amount of proteins extracted. A standardized SRPE procedure was developed and then tested for yield and reproducibility. The highest protein yield and quality were obtained using a ball grinder with stainless steel beads in Safe-Lock microcentrifuge tubes with methanol as the solvent, providing a highly efficient, economic and effective method. The usefulness of this SRPE was validated by applying the procedure to extract protein from different Brassica oilseeds and for screening an ethyl methane sulfonate (EMS) mutant population of Brassica rapa R-0-18. The outcomes provide useful methodology for identifying and characterizing the SSPs in the SRPE. Full article

The aim of the study was to investigate the antibacterial and anticoagulant activity of Moringa (Moringa oleifera) seed extracts and coagulant protein for their potential application in water treatment. Pathogenic microorganisms were obtained from Ramachandra Hospital, Chennai, India. Bacterial cell aggregation and growth kinetics studies were employed for six bacterial strains with different concentrations of seed extracts and coagulant protein. Moringa seed extract and coagulant protein showed cell aggregation against six bacterial strains, whereas seed extract alone showed growth inhibition of all six bacterial strains for up to 6 h, compared to that of control. Escherichia coli and Salmonella para typhi B did not develop resistance against coagulant protein. The results imply that Moringa oleifera is likely an efficient low-molecular bioactive peptide (with <7.5 kDa plant-based coagulant and antimicrobial peptides, confirmed by applying amino acid sequences), using liquid chromatography–mass spectrometry and HPLC, with the corresponding sequences from Napin-1A peptide posing different degrees of antibacterial activity against different pathogenic organisms. Full article

To improve the protein nutritional quality of canola (Brassica napus L.) meal, further investigation of the effects of processing conditions and post-production treatments is desirable. The impact of barrel dry heat temperature (20 °C (cold press) and 100 °C (expeller)) and moist heat pressure (MHP) duration time on general nutritional properties, Maillard reaction product (MRP) formation, in vitro protein degradability, and molecular and microscopic structural characteristics of canola meals were investigated. Increased MHP duration reduced (p < 0.05) dry matter, soluble protein, rapidly degradable protein, yellowness (early MRP), whiteness (late MRPs), absorbance at 294 nm (intermediate MRPs), and amide I; and increased (p < 0.05) non-protein N, neutral detergent fibre, neutral detergent insoluble crude protein (CP), intermediately and slowly degradable protein, in vitro effective CP degradability, redness, degree of colour change, and browning. Increased dry heat temperature reduced (p < 0.01) CP and rapidly degradable protein, constricted amide II, reduced (p < 0.05) protein solubility in 0.5% KOH and increased (p < 0.05) acid-detergent fibre and intermediate MRPs. Browning index and redness exhibited potential as rapid indicators of effective CP degradability and soluble protein, respectively. Dry heat and MHP altered (p < 0.05) lipid-related functional groups. Dry heat affected napin solubility, and MHP altered cruciferin and napin solubility. Application of MHP induced the formation of proteolysis-resistant protein aggregates with crevices containing oil bodies. Induced changes may impact the supply of proteins and amino acids and subsequently the yield and composition (protein and lipid) of milk produced by dairy cows. Full article

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 β-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. Full article

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. Full article