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Alternative protein sources have been required to meet the significant plant protein demand. Agro-industrial by-products such as leaves have considerable potential as a source of macromolecules once they are mostly discarded as waste. The current study evaluated dried cassava leaves as a protein source. First, alkaline extraction parameters (solid-liquid ratio, pH, and temperature) were optimized and the run that result in the highest protein yield were acidified at pH 2.5 or 4. The influence of carbohydrate solubilized on protein precipitation was also evaluated by removing it via alcoholic extraction prior to precipitation. The experimental design showed that high pH and temperature conditions associated with a low solid-liquid ratio led to increased protein yields. The presence of carbohydrates in the supernatant significantly influenced protein precipitation. The protein concentrate had around 17.51% protein when it was obtained from a supernatant with carbohydrates, while protein content increased to 26.88% when it was obtained from carbohydrate-free supernatant. The precipitation pH also influenced protein content, whereas protein content significantly decreased when pH increased from 2.5 to 4. The natural interaction between carbohydrates and proteins from cassava leaves positively influenced the emulsion stability index and the foaming capacity and stability. Thus, the presented results bring insights into challenges in extracting and precipitation proteins from agro-industrial by-products.

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Plant proteins with improved solubility, foaming, and emulsifying properties are required to meet the demand for plant-based foods. This study evaluated the influence of alkaline extraction combined with enzyme- and ultrasound-assisted extraction on lentil protein structure and functionality. Enzyme- and ultrasound-assisted extractions were not capable of increasing the protein yield compared to alkaline extraction alone. However, the purity of isolated protein was dependent on the extraction process, ranging from 82.7% to 90%. Although the molecular mass, zeta potential profiles, and denaturation temperature were not dependent on the extraction method, the enthalpy of denaturation for protein obtained solely by alkaline extraction was significantly lower than that for assisted processes, indicating that protein denaturation is caused by an alkaline process. Changes in protein structure were also suggested by solubility analyses that showed that lentil proteins obtained by enzyme-assisted and ultrasound-assisted extraction have better solubility at pH 7 when compared to alkaline extraction alone. The surface activity of lentil protein was evidenced by interfacial and surface analysis, and it was influenced by the extraction process applied. We demonstrated that combining alkaline extraction with assisted processes, especially ultrasound technology, results in concentrates/isolates with higher solubility as compared to ones obtained solely by the traditional alkaline method, even though the choice of extraction method should depend on the desired functionality.

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RESEARCH BACKGROUND: Extracted from the pulp of an Amazonian fruit, buriti oil is rich in micronutrients with antioxidant properties and high biological value. The few studies available indicate that this oil could be used in a wide range of applications; however, there are no studies that work on the improvement in the characteristics of this oil for commercial application. The enzymatic interesterification is one of the tools available to improve the properties of oils and fats and our recent studies have demonstrated that the lipase could specifically act on buriti oil to produce structured lipids rich in oleic acid, while preserving most of the minor compounds present in this oil. Still looking for ways to expand the applicability of this raw oil, in this work, we are interested in studying the behaviour of this structured oil in nanostructured lipid carriers (NLCs). EXPERIMENTAL APPROACH: The NLCs were produced with interesterified buriti oil and the stability, droplet size, electrical charge, microstructure, polymorphism and antioxidant activity of the samples were evaluated by ORAC and FRAP methods. RESULTS AND CONCLUSIONS: The results showed that the interesterification formed more unsaturated triacylglycerols (TAGs), and NLCs prepared with interesterified buriti oil had smaller droplets than NLCs with crude buriti oil. Particles remained stable throughout the storage period and NLCs exhibited complex polymorphism with the presence of three crystalline forms. The oxygen radical absorbance capacity (ORAC) value was approx. 23% higher in nanolipid carries with structured lipids than in the nanolipid carriers with crude buriti oil, and the ferric reducing antioxidant power (FRAP) value 16% higher, demonstrating the influence of interesterification on the antioxidant activity of nanocarriers. Thus, NLCs prepared with interesterified buriti oil had small droplets, high stability and antioxidant capacity, and have a potential for nutritional and biological applications. NOVELTY AND SCIENTIFIC CONTRIBUTION: This research showed that interesterification positively influenced the physicochemical properties of NLCs, producing the oil rich in oleic acid, high stability and antioxidant capacity. Therefore, it may be interesting to use these nanocarriers to obtain efficient carrier systems for future applications.

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This research aimed to evaluate the pretreatment of soybean with the carbohydrase multi-enzyme complex "Viscozyme L", during the extraction process; in order to improve the recovery of proteins and isoflavones in soybase, and reduce the loss of these compounds through the okara residue. Three concentrations of enzyme were studied at 50 °C, along with an experiment carried out without enzyme addition (control experiment). The results were also evaluated in relation to standard soybase processing. In comparison to the standard and control processes, the enzymatic pretreatment reduced up to 85% the total amount of okara residue. Due to the action of the multi-enzyme complex, protein and total isoflavone recovery increased from 42% to 83% and from 59% to 93%, respectively. The application of Viscozyme L also favored the conversion of conjugated forms of isoflavone to aglycone in the soybase, representing up to 50% of the total isoflavones. The enzymatic pretreatment of soybean with carbohydrase improved the nutritional quality of the soybase, while at the same time reducing residue generation; showing that the proposed food process can be considered environmentally friendly method.

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Enzymatic interesterification is used to manipulate oil and fat in order to obtain improved restructured lipids with desired technological properties. However, with raw materials containing significant amounts of bioactive compounds, the influence of this enzymatic process on the bioactivity of the final product is still not clear. Thus, the aim of this study is to evaluate the antioxidant potential and modulatory effects of two raw materials from the Amazonian area, buriti oil and murumuru fat, before and after lipase interesterification, on human hepatoma cells (HepG2). The results indicate that minor bioactive compounds naturally found in the raw materials and their antioxidant capacity are preserved after enzymatic interesterification, and that the restructured lipids modulate HepG2 endogenous antioxidant enzyme.