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.

This study aimed to modify plant protein mixture to improve their functionality and digestibility by limited hydrolysis. Soy protein isolate and corn zein were mixed at the ratio of 5:1 (w/w), followed by limited hydrolysis using papain from 15 to 30 min. The structural characteristics, in vitro digestibility, and functional properties were evaluated. Also, DPPH radical scavenging activity was determined. The results indicated that the molecular weight of different modified samples was largely reduced by limited hydrolysis, and the proportion of random coil was significantly increased. Furthermore, the solubility, foaming, emulsifying and water-holding capacity of hydrolyzed protein mixture were significantly improved, which were close to those of whey protein isolate. In vitro digestibility after 30-min limited hydrolysis was remarkably elevated. In addition, the hydrolyzed protein mixture exhibited a higher antioxidant activity than those of untreated proteins. Overall, limited hydrolysis of protein mixture led to improved digestibility, functionality and antioxidant activity.

Plant-based proteins offer sustainable and nutritious alternatives to animal proteins with their techno-functional attributes influencing product quality and designer food development. Due to the inherent complexities of plant proteins, proper extraction and modifications are vital for their effective utilization. This review highlights the emerging sources of plant-based proteins, and the recent statistics of the techniques employed for pretreatment, extraction, and modifications. The pretreatment, extraction and modification approach to modify plant proteins have been classified, addressed, and the recent applications of such methodologies are duly indicated. Furthermore, this study furnishes novel perspectives regarding the potential impacts of emerging technologies on the intricate dynamics of plant proteins. A thorough review of 100 articles (2018-2024) shows the researchers\' keen interest in investigating novel plant proteins and how they can be used; seeds being the main source for protein extraction, followed by legumes. Use of by-products as a protein source is increasing rapidly, which is noteworthy. Protein studies still lack knowledge on protein fraction, antinutrients, and pretreatments. The use of physical methods and their combination with other techniques are increasing for effective and environmentally friendly extraction and modification of plant proteins. Several studies explore the effect of protein changes on their function and nutrition, especially with a goal of replacing ingredients with plant proteins that have improved or enhanced qualities. However, the next step is to investigate the sophisticated modification methods for deeper insights into food safety and toxicity.

Pea proteins are gaining increased interest from both the food industry as well as from consumers. Pea protein isolates (PPI) excel at forming meat-like textures upon heating while pea protein concentrates (PPC) are more challenging to transform into highly sought-after foods. PPCs are richer in dietary fibers (DF) and are more sustainable to produce than PPI. In this work, degradative enzymes were used to modify the functionality of PPC-water blends with a focus on texturization upon heating. Three enzyme solutions containing β-glucanases, hemicellulases, pectinases, xylanase, and cellulases were added to 65 wt% PPC blends. The effect of these enzymatic pretreatments was measured by monitoring the torque in a mixing reactor during blending, differential scanning calorimetry (DSC), high-pressure shear rheology (HPSR), and DF content and size analysis. Four endothermic peaks were detected in the DSC thermograms of PPC, namely at 63 °C, 77 °C, 105 °C and 123 °C. The first three peaks were attributed to phase transition and gelation temperatures of the starches and proteins constituting PPC. No endothermic peaks were measured for PPI blends. Enzyme solutions containing β-glucanases, hemicellulases, pectinases, and xylanases increased the endothermic energy of all peaks, hinting at an effect on the gelation properties of PPC. The same enzymes decreased the resistance to flow of PPC blends and induced a shift of the weight average molecular weight (Mw) distribution of soluble dietary fibers (SDF) towards smaller values while increasing the fraction of SDF by decreasing the insoluble dietary fiber (IDF) content. The solution containing cellulases did not change the DSC results or the viscosity of the PPC mixture, nor did it affect the IDF and SDF contents. On the other hand HPSR measurements of heated PPC samples up to 125 °C showed that all tested enzyme solutions decreased the complex viscosity of PPC-water blends to values similar to PPI-water blends. We demonstrated that degradative enzymes can enhance the functionality of less refined protein-rich ingredients based on pea and other vegetal sources. Using optimized enzyme blends for targeted applications can prove to be a key changer in the development and improvement of sustainable protein-rich foods.

The increasing world population requires the production of nutrient-rich foods. Protein is an essential macronutrient for healthy individuals. Interest in using plant proteins in foods has increased in recent years due to their sustainability and nutritional benefits. Dry and wet protein fractionation methods have been developed to increase protein yield, purity, and functional and nutritional qualities. This review explores the recent developments in pretreatments and fractionation processes used for producing pulse protein concentrates and isolates. Functionality differences between pulse proteins obtained from different fractionation methods and the use of fractionated pulse proteins in different food applications are also critically reviewed. Pretreatment methods improve the de-hulling efficiency of seeds prior to fractionation. Research on wet fractionation methods focuses on improving sustainability and functionality of proteins while studies on dry methods focus on increasing protein yield and purity. Hybrid methods produced fractionated proteins with higher yield and purity while also improving protein functionality and process sustainability. Dry and hybrid fractionated proteins have comparable or superior functionalities relative to wet fractionated proteins. Pulse protein ingredients are successfully incorporated into various food formulations with notable changes in their sensory properties. Future studies could focus on optimizing the fractionation process, improving protein concentrate palatability, and optimizing formulations using pulse proteins.

The fava bean protein isolate (FBPI) holds promise as a sustainable plant-based protein ingredient. However, native FBPIs exhibit limited functionality, including unsuitable emulsifying activities and a low solubility at a neutral pH, restricting their applications. This study is focused on the effect of ultrasonication (US) and pulsed electric fields (PEF) on modulating the techno-functional properties of FBPIs. Native FBPIs were treated with US at amplitudes of 60-90% for 30 min in 0.5 s on-and-off cycles and with PEF at an electric field intensity of 1.5 kV/cm with 1000-4000 pulses of 20 μs pulse widths. US caused a reduction in the size and charge of the FBPIs more prominently than the PEF. Protein characterization by means of SDS-PAGE illustrated that US and PEF caused severe-to-moderate changes in the molecular weight of the FBPIs. In addition, a spectroscopic analysis using Fourier-transform infrared (FTIR) and circular dichroism (CD) revealed that US and the PEF induced conformational changes through partial unfolding and secondary structure remodeling from an α-helix to a β-sheet. Crystallographic and calorimetric determinations indicated decreased crystallinity and lowered thermal transition temperatures of the US- and PEF-modified FBPIs. Overall, non-thermal processing provided an effective strategy for upgrading FBPIs\' functionality, with implications for developing competitive plant-based protein alternatives.

O-linked-β-N-acetylglucosamine (O-GlcNAcylation) is a distinctive posttranslational protein modification involving the coordinated action of O-GlcNAc transferase and O-GlcNAcase, primarily targeting serine or threonine residues in various proteins. This modification impacts protein functionality, influencing stability, protein-protein interactions, and localization. Its interaction with other modifications such as phosphorylation and ubiquitination is becoming increasingly evident. Dysregulation of O-GlcNAcylation is associated with numerous human diseases, including diabetes, nervous system degeneration, and cancers. This review extensively explores the regulatory mechanisms of O-GlcNAcylation, its effects on cellular physiology, and its role in the pathogenesis of diseases. It examines the implications of aberrant O-GlcNAcylation in diabetes and tumorigenesis, highlighting novel insights into its potential role in cardiovascular diseases. The review also discusses the interplay of O-GlcNAcylation with other protein modifications and its impact on cell growth and metabolism. By synthesizing current research, this review elucidates the multifaceted roles of O-GlcNAcylation, providing a comprehensive reference for future studies. It underscores the potential of targeting the O-GlcNAcylation cycle in developing novel therapeutic strategies for various pathologies.

Pulses have attracted much attention in the food industry due to their low cost, high yield, and high protein content, which promises to be excellent alternative protein sources. Recently, techniques for covalent and noncovalent binding of pulse proteins to polyphenols are expected to solve the problem of their poor protein functional properties. Additionally, these conjugates and complexes also show several health benefits. This review summarizes the formation of conjugates and complexes between pulse proteins and polyphenols through covalent and noncovalent binding and the impact of this structural change on protein functionalities and potential health benefits. Recent studies show that pulse protein functionalities can be influenced by polyphenol dose. This is mainly the case for adverse effects on solubility and enhancement in emulsifying capacity. Also, the conjugates/complexes exhibit antioxidant activity and can alter protein digestibility. The antioxidant activity of polyphenols could be retained after binding to proteins, while the effect on digestibility depends on the type or dosage of polyphenols. Considering the link between polyphenols and their potential health benefits, pulse polyphenols would be a good choice for producing the conjugates/complexes due to their low cost and proven potential benefits. Further studies on the structure-function-health benefits relationship of pulse protein-polyphenol conjugates and complexes are still required, as well as the validation of their application as functional foods in the food industry.

In this study, we investigated the biological profile of lectins isolated from raw and boiled Japanese red Kintoki beans (red kidney beans [RKB]; Phaseolus vulgaris). Lectins in beans showing agglutination activity were retained after heating. Raw and boiled RKB lectins were fractionated using carboxymethyl- and diethylaminoethyl-Sepharose, respectively. Boiled RKB lectins were evaluated for carbohydrate specificity as well as cytokine-inducing and antiproliferative activities against cancer cells and compared with raw RKB lectins. Raw RKB lectins showed specificity for thyroglobulin and fetuin, whereas boiled lectins showed specificity for N-acetylneuraminic acid. Raw RKB lectins showed low resistance to proteases and tolerated temperatures greater than 80°C for 5 min. Notably, lectins from raw and boiled beans showed antiproliferative activity against five types of cancer cells B16, LM8, HeLa, HepG2, and Colo 679. In particular, lectins from raw beans exhibited a significantly stronger activity than those from boiled beans. Anti-inflammatory effects were notably observed in crude extracts from raw and boiled beans. Specifically, lectins fractionated from boiled beans markedly inhibited the expression of tumor necrosis factor-α and interleukin-6. Overall, our results showed that RKB lectins from boiled beans exert anti-inflammatory and anticancer effects and could be developed as potential chemopreventive agents. PRACTICAL APPLICATION: Japanese red kidney beans (RKB) are cultivated in numerous parts of the temperate zone and consumed in many countries. Lectins from boiled beans exhibited anticancer activity, similar to lectins from raw beans. Additionally, crude and fractionated lectins from boiled beans showed anti-inflammatory activity. Thus, boiled RKB lectins have the potential to be used as a bioactive protein for medical research and could be developed as anticancer agents.

Different protein sources create distinct textures in plant-based meat due to differences in their hydration properties when exposed to different time, temperature, and shear regimes, which in turn depend upon their solubility, protein structure, and specific amino acids. This research aimed to identify these differences and manipulate them to reach a desired texture utilizing simple and reproducible analytical methods to characterize protein properties as either cold or heat swelling. Protein functionality was determined through least gelation concentration (LGC), water absorption index (WAI), and rapid visco analysis (RVA). Cold swelling or CS proteins (pea protein isolate, soy protein isolate, Arcon S soy protein concentrate) were characterized by an LGC < 14% and/ or WAI > 4.0 g/g, while LGC > 16% and/ or WAI < 4.0 g/g indicates proteins with heat swelling or HS properties (Arcon F soy protein concentrate, wheat gluten, and fava protein concentrate). An RVA peak time of around or less than 3 min (<75°C peak temperature) indicated CS properties while greater than 3.5 min (>80°C peak temperature) was considered HS. Protein mixes or treatments comprising mainly of different combinations and ratios of CS proteins were hypothesized to create a softer textured vegetable protein product or texturized vegetable protein (TVP) and those based on HS proteins a firmer TVP. Bulk density was higher for HS treatments (274-287 g/L) than for CS treatments (160-223 g/L). CS treatments exhibited a microstructure that was porous, while HS showed a dense, laminar microstructure. Texture profile analysis showed that CS treatments seemed to show a lower hardness (1154-1595 g) than the HS treatments (1893-2231 g). PRACTICAL APPLICATION: Controlling texture can be a valuable tool when producing a plant-based meat product. Different applications have various texture requirements. For example, a plant-based fish stick would require a softer texture than a hamburger or chicken nugget. By increasing the knowledge of how protein functionality affects meat analogue texture, the time needed to produce new products with novel textures can be reduced. Money could also be saved by being able to quickly replace ingredients with a more affordable or accessible protein with similar swelling abilities to preserve product quality.