Plant-Based Fermented Beverages: Nutritional Composition, Sensory Properties, and Health Benefits.

Plant-based beverages have gained consumers' attention for being the main substitutes for dairy milk, especially for people with lactose intolerance, milk allergies, and a prevalence of hypercholesterolemia. Moreover, there is a growing demand for a more sustainable diet and plant-based lifestyle due to concerns related to animal wellbeing, environmental impacts linked to dairy production, and the rising cost of animal-derived foods. However, there are some factors that restrict plant-based beverage consumption, including their nutritional quality and poor sensory profile. In this context, fermentation processes can contribute to the improvement of their sensory properties, nutritional composition, and functional/bioactive profile. In particular, the fermentation process can enhance flavor compounds (e.g., acetoin and acetic acid) while decreasing off-flavor components (e.g., hexanal and hexanol) in the substrate. Furthermore, it enhances the digestibility and bioavailability of nutrients, leading to increased levels of vitamins (e.g., ascorbic acid and B complex), amino acids (e.g., methionine and tryptophan), and proteins, while simultaneously decreasing the presence of anti-nutritional factors (e.g., phytic acid and saponins). In contrast, plant-based fermented beverages have been demonstrated to possess diverse bioactive compounds (e.g., polyphenols and peptides) with different biological properties (e.g., antioxidant, anti-inflammatory, and antihypertensive). Therefore, this article provides an overview of plant-based fermented beverages including their production, technological aspects, and health benefits.

Application of Molecular Dynamics Simulations to Determine Interactions between Canary Seed (Phalaris canariensis L.) Bioactive Peptides and Skin-Aging Enzymes.

Food bioactive peptides are well recognized for their health benefits such as antimicrobial, antioxidant, and antihypertensive benefits, among others. Their drug-like behavior has led to their potential use in targeting skin-related aging factors like the inhibition of enzymes related with the skin-aging process. In this study, canary seed peptides (CSP) after simulated gastrointestinal digestion (<3 kDa) were fractioned by RP-HPLC and their enzyme-inhibition activity towards elastase and tyrosinase was evaluated in vitro. CSP inhibited elastase (IC50 = 6.2 mg/mL) and tyrosinase (IC50 = 6.1 mg/mL), while the hydrophobic fraction-VI (0.2 mg/mL) showed the highest inhibition towards elastase (93%) and tyrosinase (67%). The peptide fraction with the highest inhibition was further characterized by a multilevel in silico workflow, including physicochemical descriptor calculations, antioxidant activity predictions, and molecular dynamics-ensemble docking towards elastase and tyrosinase. To gain insights into the skin permeation process during molecular dynamics simulations, based on their docking scores, five peptides (GGWH, VPPH, EGLEPNHRVE, FLPH, and RPVNKYTPPQ) were identified to have favorable intermolecular interactions, such as hydrogen bonding of polar residues (W, H, and K) to lipid polar groups and 2-3 Å van der Waals close contact of hydrophobic aliphatic residues (P, V, and L). These interactions can play a critical role for the passive insertion of peptides into stratum corneum model skin-membranes, suggesting a promising application of CSP for skin-aging treatments.

Nutricosmetics: A new frontier in bioactive peptides' research toward skin aging.

Food derived bioactive peptides are small protein fragments (2-20 amino acids long) that can exhibit health benefits, beyond basic nutrition. For example, food bioactive peptides can act as physiological modulators with hormone or drug-like activities including anti-inflammatory, antimicrobial, antioxidant, and the ability to inhibit enzymes related to chronic disease metabolism. Recently, bioactive peptides have been studied for their potential role as nutricosmetics. For example, bioactive peptides can impart skin-aging protection toward extrinsic (i.e., environmental and sun UV-ray damage) and intrinsic (i.e., natural cell or chronological aging) factors. Specifically, bioactive peptides have demonstrated antioxidant and antimicrobial activates toward reactive oxygen species (ROS) and pathogenic bacteria associated with skin diseases, respectively. The anti-inflammatory properties of bioactive peptides using in vivo models has also been reported, where peptides have shown to decreased the expression of IL-6, TNF-α, IL-1ß, interferon-γ (INF-γ), and interleukin-17 (IL-17) in mice models. This chapter will discuss the main factors that trigger skin-aging processes, as well as provide examples of in vitro, in vivo, and in silico applications of bioactive peptides in relation to nutricosmetic applications.

In-depth peptidomic profile and molecular simulation studies on ACE-inhibitory peptides derived from probiotic fermented milk of different farm animals.

Investigations into ACE inhibitory properties of probiotic fermented bovine, camel, goat, and sheep milk were performed and studied for two weeks of refrigerated storage. Results from the degree of proteolysis suggested higher susceptibility of goat milk proteins, followed by sheep and camel milk proteins, to the probiotic-mediated proteolysis. ACE-inhibitory properties displayed continuous decline in ACE-IC50 values for two weeks of refrigerated storage. Overall, goat milk fermented with Pediococcus pentosaceus caused maximum ACE inhibition (IC50: 262.7 µg/mL protein equivalent), followed by camel milk (IC50: 290.9 µg/mL protein equivalent). Studies related to peptide identification and in silico analysis using HPEPDOCK score revealed presence of 11, 13, 9 and 9 peptides in fermented bovine, goat, sheep, and camel milk, respectively, with potent antihypertensive potential. The results obtained suggest that the goat and camel milk proteins demonstrated higher potential for generating antihypertensive peptides via fermentation when compared to bovine and sheep milk.

Canary Seed (Phalaris canariensis L.) Peptides Prevent Obesity and Glucose Intolerance in Mice Fed a Western Diet.

Previous research showed that canary seed (Phalaris canariensis L.) peptides (CSP) possess robust in vitro antiobesity properties via inhibition of pancreatic lipase (PL). Nevertheless, no studies have yet explored their antiobesity properties in vivo. Consequently, we investigated the effects of CSP in C57BL/6J mice under a Western diet (WD). Mice were assigned into groups and fed a normal diet (ND) or a WD accompanied by an oral dose of CSP (250 or 500 mg/kg/day), orlistat (40 mg/kg/day), or distilled water. The results showed that consuming CSP can provide metabolic benefits, including preventing weight gain by up to 20%, increasing glucose tolerance, and reducing insulin, leptin, and LDL/VLDL levels in plasma. Conversely, total ghrelin was unaffected by CSP-500, but decreased by CSP-250, and amplified by orlistat. Surprisingly, CSP-250 was more effective in preventing weight gain and promoting satiety than CSP-500. Parallel to this, protein absorption in CSP-500 was decreased, supported by a rise in fecal crude protein (+3.5%). Similarly, fecal fat was increased by orlistat (38%) and was unaffected by CSP-250 (3.0%) and CSP (3.0%), comparatively to WD (2.5%). Despite this, both CSP treatments were equally effective in decreasing hepatic steatosis and avoiding hyperlipidemia. Furthermore, the enzymatic analysis showed that CSP-PL complexes dissociated faster (15 min) than orlistat-PL complexes (41 min). Lastly, CSP did not affect expression of hepatic lipid oxidation genes ACO and PPAR-α, but reduced the expression of the hydrolase gene LPL, and lipogenesis related genes FAS and ACC. Taken together, these results suggest that CSP antiobesity mechanism relies on lipid metabolism retardation to increase fat transit time and subsequently suppress hunger.

Beyond Human Nutrition of Edible Insects: Health Benefits and Safety Aspects.

Nowadays, edible insects are considered an outstanding source of nutrients, primarily because they contain high-quality protein, amino acids, and vitamins. Insects are considered a promising alternative protein source towards alleviating future global food shortage problems due to their production considered as being more sustainable by using less agricultural land and water, as well as releasing a smaller amount of greenhouse gas emissions. However, other important aspects to consider about the consumption of edible insects include their health benefits and some safety aspects, which has been relatively overlooked. In this sense, edible insects contain bioactive compounds that can provide diverse bioactivities, such as antioxidant, antihypertensive, anti-inflammatory, antimicrobial, and immunomodulatory with a positive impact on human health. On the other hand, edible insects are a nutrient-rich food that can provide a perfect growth medium for diverse microorganisms, as well as possess some anti-nutritive factors. These two main aspects could represent food safety concerns for consumers. In this context, recent scientific evidence indicates that preservation methods, mainly thermal treatments, utilized in the cooking or processing of edible insects decreased the microbial levels and anti-nutritive factors, which suggests that edible insects do not represent a critical biological risk to humans. Besides, edible insects could have a positive effect on gut microbiota, either by their pre-biotic effect or their antimicrobial activity towards pathogens. Thus, this review is focused on studies related to the health benefits of edible insects and their isolated components, as well as discussion about potential issues related to their microbial content and anti-nutritive factors; this review will provide a synopsis on whether edible insects may be considered safe for human consumption.

Edible insects, a valuable protein source from ancient to modern times.

The current COVID-19 pandemic has demonstrated that we are not prepared to deal with food security amid unexpected situations; the FAO (Food and Agriculture Organization) has stipulated that the future of our food & agriculture looks challenging toward the year 2050; primarily in response to the fact that global population is expected to increase by 9 billion people by 2050. Although entomophagy has been practiced by humans for thousands of years, until recently, edible insects have gained special attention due to their high nutritional value (particularly their high protein and essential amino acid content) and lower environmental impact that could help alleviate the global food demand. Edible insects are classified into eight main orders belonging to Blattodea (cockroaches and termites), Coleoptera (beetles), Diptera (flies), Hemiptera (cicadas, stink bugs), Hymenoptera (bees, wasps, ants), Lepidoptera (butterflies, moths), Odonata (dragonflies), and Orthoptera (crickets, grasshoppers, locusts). Several traditional cooking (e.g., boiling, roasting, sun-drying) and processing technologies (e.g., pasteurization, enzymatic proteolysis, high pressure processing) have shown that it is feasible to prepare safe and nutritious insects and/or foods with insects. Nevertheless, challenges associated with consumers acceptance to eat insects, as well as potential presence of anti-nutritive factors and allergens, need to be carefully evaluated as the industry grows in the coming years. Foreseeing such food shortages during pandemics and future food security concerns, consumers, scientists, and the food industry need to consider the value of farming insects as promising protein sources.

Multifunctional Analysis of Chia Seed (Salvia hispanica L.) Bioactive Peptides Using Peptidomics and Molecular Dynamics Simulations Approaches.

Chia seed peptides (CSP) can be a source of multifunctional biopeptides to treat non-communicable diseases. However, interactions and binding affinity involved in targeting specific receptors remains unexplored. In this study, molecular simulation techniques were used as virtual screening of CSP to determine drug-like candidates using a multi-target-directed ligand approach. CSP fraction with the best bioactivities in vitro was sequenced. Then, a prediction model was built using physicochemical descriptors (hydrophobicity, hydrophilicity, intestinal stability, antiangiogenic, antihypertensive, and anti-inflammatory) to calculate potential scores and rank possible biopeptides. Furthermore, molecular dynamics simulations (MDS) and ensemble molecular docking analysis were carried out using four human protein targets (ACE, angiotensin converting enzyme; VEGF, vascular endothelial growth factor; GLUC, glucocorticoid and MINC, mineralocorticoid receptors). Five known-sequence peptides (NNVFYPF, FNIVFPG, SRPWPIDY, QLQRWFR, GSRFDWTR) and five de novo peptides (DFKF, DLRF, FKAF, FRSF, QFRF) had the lowest energy score and higher affinity for ACE and VEGF. The therapeutic effects of these selected peptides can be related to the inhibition of the enzymes involved in angiogenesis and hypertension, due to formation of stable complexes with VEGF and ACE binding sites, respectively. The application of MDS is a good resource for identifying bioactive peptides for future experimental validation.

Oxidative Stress Protection by Canary Seed (Phalaris canariensis L.) Peptides in Caco-2 Cells and Caenorhabditis elegans.

During oxidative stress, degenerative diseases such as atherosclerosis, Alzheimer's, and certain cancers are likely to develop. Recent research on canary seed (Phalaris canariensis) peptides has demonstrated the high in vitro antioxidant potential. Thus, this study aimed to assess the cellular and in vivo antioxidant capacity of a low-molecular-weight (<3 kDa) canary seed peptide fraction (CSPF) using Caco-2 cells and the Caenorhabditis elegans model. The results show that the CSPF had no cytotoxicity effect on Caco-2 cells at any tested concentration (0.3−2.5 mg/mL). Additionally, the cellular antioxidant activity (CAA) of the CSPF was concentration-dependent, and the highest activity achieved was 80% by the CSPF at 2.5 mg/mL. Similarly, incubation with the CSPF significantly mitigated the acute and chronic oxidative damage, extending the lifespan of the nematodes by 88 and 61%, respectively. Furthermore, it was demonstrated that the CSPF reduced the accumulation of reactive oxygen species (ROS) to safe levels after sub-lethal doses of pro-oxidant paraquat. Quantitative real-time PCR revealed that the CSPF increased the expression of oxidative-stress-response-related gene GST-4. Overall, these results show that the CSPFs relied on GST-4 upregulation and scavenging of free radicals to confer oxidative stress protection and suggest that a CSPF can be used as a natural antioxidant in foods for health applications.

Caenorhabditis elegans as an in vivo model for food bioactives: A review.

Caenorhabditis elegans (C. elegans) is being widely explored as an in vivo model to study the effects of food bioactives. These nematodes are largely advantageous over other in vivo models as they are relatively inexpensive, have a short generation time, and have a completely sequenced genome, among other advantages. C. elegans is a commonly used model to study diseases such as Alzheimer's and Parkinson's disease; however, researchers are finding they can also give insight into the health promoting effect of food-derived bioactive compounds. As consumers become more aware of the health benefits of the foods that they consume, the study of bioactive properties of foods and food constituents is becoming an important source of information. This review focuses on the advantages of using C. elegans as a model such as their short lifespans, high level of gene conservation relative to humans, and large number of progenies per reproductive cycle. They are also easily manipulated in order to perform controlled experiments on synchronous populations. Through review of recent literature, it is clear that C. elegans can be used to study a range of food derived compounds such as bioactive peptides, phenolic compounds, carbohydrates, and lipids. This review also provides information on potential challenges associated with working with this nematode. These challenges include the need for a sterile environment, potential inaccuracy when determining if the nematodes are dead, and the simplicity of the organism making it not suitable for all studies.

Integration of Molecular Docking Analysis and Molecular Dynamics Simulations for Studying Food Proteins and Bioactive Peptides.

In silico tools, such as molecular docking, are widely applied to study interactions and binding affinity of biological activity of proteins and peptides. However, restricted sampling of both ligand and receptor conformations and use of approximated scoring functions can produce results that do not correlate with actual experimental binding affinities. Molecular dynamics simulations (MDS) can provide valuable information in deciphering functional mechanisms of proteins/peptides and other biomolecules, overcoming the rigid sampling limitations in docking analysis. This review will discuss the information related to the traditional use of in silico models, such as molecular docking, and its application for studying food proteins and bioactive peptides, followed by an in-depth introduction to the theory of MDS and description of why these molecular simulation techniques are important in the theoretical prediction of structural and functional dynamics of food proteins and bioactive peptides. Applications, limitations, and future prospects of MDS will also be discussed.

Insects as an Alternative Protein Source.

The recent COVID-19 pandemic drastically affected food supply chains worldwide, showing the vulnerability of food security. Efforts to develop alternative protein sources that are sustainable and can help alleviate global food shortage problems should be prioritized. Insects have been part of our diet for thousands of years and still are today, and market trends show a global increase in the number of food-grade insect producers. The global market for edible insects has been forecasted to reach US$8 billion by the year 2030. Insects are highly nutritious and have bioactive peptides with potential therapeutic effects. This review provides an overview of the consumption of insects from ancient to modern times, discusses the rationale for using insects as alternative protein sources, and presents a summary of the major insects consumed worldwide as well as a brief description of the traditional and novel technologies currently used to process insects and/or extract their nutritional components.

Targeted Phenolic Characterization and Antioxidant Bioactivity of Extracts from Edible Acheta domesticus.

With entomophagy gaining popularity in the Western hemisphere as a solution for future food insecurity, research on alternative protein sources, such as edible insects, has become relevant. Most of the research performed on insects has been on their nutritional qualities; however, little is known regarding bioactive compounds, such as polyphenols, that, if present in the insect, could provide additional benefits when the insect is consumed. In this study, methanolic extracts of Acheta domesticus from two farms and their corresponding feeds were obtained using a microwave-assisted extraction. Targeted phenolic characterization was accomplished through LC-MS/MS leading to the identification of 4-hydroxybenzoic acid, p-coumaric acid, ferulic acid, and syringic acid as major phenolic compounds in both A. domesticus extracts. Furthermore, the in vitro antioxidant activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl radical cation (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical assays demonstrating the superior quenching activity of the A. domesticus extracts compared to the feeds. The discovery of phenolic compounds in A. domesticus implies the ability of this insect species to sequester and absorb dietary phenolics leading to possible added health benefits when consumed.

Processing insects for use in the food and feed industry.

Although insects are becoming more accepted as potential protein sources for food and feed, the appearance of the insect may be off-putting due to associations of disgust. Edible insects are more likely to be eaten if they are processed into non-recognizable forms. Thus, insects require the use of commercial processing methods that will render the protein suitable for food/feed formulation, while maintaining the safety, nutritional and sensory quality of the final product. Common methods that can be used include lipid extraction, enzymatic proteolysis, commercial thermal processing (e.g. blanching, pasteurization, and commercial sterilization), low-temperature processing (refrigeration and freezing), dehydration, and fermentation technology. Each method has advantages and disadvantages that need to be carefully considered as not all processing methods and/or conditions apply to all edible insects or insect flours. This article provides a brief overview of the most commonly used processing methods applicable for insects destined for food and feed.

Isolation and proteomic characterization of tropomyosin extracted from edible insect protein.

Edible insects are considered promising sustainable protein sources. Thermal treatments and proteolysis are commonly used to improve their safety and quality. However, their allergenicity remains mostly unexplored. Tropomyosin, a major insect pan-allergen, can be used to study processing effects on its immunoreactivity. In this study, selective precipitation was used to extract tropomyosin from heated and protease-treated crickets. Immunoinformatics predicted 31 epitope regions, while proteomic analysis suggested decreased amounts of intact epitope regions in microwave-heated/protease-treated crickets. Tropomyosin peptide sequences were identified in higher abundance in convection-heated samples. Finally, tropomyosin immunoreactivity by immunoblotting and ELISA, revealed that protease treatments under microwave heating had lower (p < 0.05) IgE and IgG reactivity. Based on results, processing insects using proteolysis and microwave-heating could be effective for generating hypoallergenic cricket protein ingredients. The use of proteomics and bioinformatics proved to be useful tools in understanding the impact of processing on allergenic reactivity of insect proteins.

Identification and Characterization of Edible Cricket Peptides on Hypertensive and Glycemic In Vitro Inhibition and Their Anti-Inflammatory Activity on RAW 264.7 Macrophage Cells.

Recent studies continue to demonstrate the potential of edible insects as a protein base to obtain bioactive peptides applicable for functional food development. This study aimed at identifying antihypertensive, anti-glycemic, and anti-inflammatory peptides derived from the in vitro gastrointestinal digests of cricket protein hydrolysates. After sequential fractionation, the protein digest subfraction containing the lowest molecular weight (<0.5 kDa), hydrophobic (C18) and cationic peptides (IEX) was found responsible for the most bioactivity. The cationic peptide fraction significantly reduced (p < 0.05) α-amylase, α-glucosidase, and angiotensin converting enzyme (ACE) activity in vitro, and also inhibited the expression of NF-κB in RAW 264.7 macrophage cells. A total of 28 peptides were identified with mass spectrometry (LC-MS/MS) and de novo sequencing from the potent fraction. Three novel peptides YKPRP, PHGAP, and VGPPQ were chosen for the molecular docking studies. PHGAP and VGPPQ exhibited a higher degree of non-covalent interactions with the enzyme active site residues and binding energies comparable to captopril. Results from this study demonstrate the bioactive potential of edible cricket peptides, especially as ACE inhibitors.

An insight to fermented edible insects: A global perspective and prospective.

The practice of eating insects is not a new phenomenon; however, the interest for their consumption has increased in recent years due to their recognized nutritional value (high content of micro- and macronutrient), potential health benefits (presence of bioactive substances), and low-environmental impact (use of less resources and reduced pollution levels). Currently, research on insects has focused on the promotion of various processing technologies for their use as either ingredients (in a non-recognizable form) to the development of innovative products, or as sources of novel bioactive compounds. In this context, evidence has suggested that alternative technologies, particularly fermentation, could be used the obtain diverse insect-based ingredients/products with unique properties. Therefore, the purpose of this narrative review was to provide an overview of the available literature on fermentation applied to obtain new insect-based products, to summarize the patents and patent-applications to protect fermented edible insect products and processes, as well as to enlist examples of current available products in the market.

Development of chia seed (Salvia hispanica) mucilage films plasticized with polyol mixtures: Mechanical and barrier properties.

Food packaging is one of the main contributors to the high rates of environmental contamination; therefore, interest has emerged on the use of biopolymers as alternative materials to replace conventional food packaging. Chia seed (Salvia hispanica) is recognized by having a high content of a polysaccharide called mucilage. The aim of this study was to evaluate the feasibility using of chia seed mucilage (CSM) and a polyol mixture containing glycerol and sorbitol for the development of films. CSM films with higher sorbitol content showed superior tensile strength (3.23 N/mm2) and lower water vapor permeability (1.3*10-9 g/m*s*Pa), but had poor flexibility compared to other treatments. Conversely, high glycerol content showed high elongation at break (67.55%) and solubility (22.75%), but poor water vapor permeability and tensile strength. Film formulations were optimized implementing a factorial design according to response surface methodology. Raman spectra analysis showed shifts from 854 to 872 cm-1 and 1061 to 1076 cm-1, ß (CCO) modes, indicating an increase in hydrogen bonding, responsible for the high tensile strength and decreased water vapor permeability observed in this study. The optimum conditions of polyol concentration were 1.3 g of glycerol and 2.0 g of sorbitol per g of CSM. Based on these results, chia seed mucilage can successfully be used to develop biofilms with potential to be used in drug delivery and edible food coating applications.

Purification and identification of an antioxidant peptide from perilla seed (Perilla frutescens) meal protein hydrolysate.

This study aimed to obtain antioxidant peptides from perilla seed meal (PSM), which is normally discarded as an industrial waste during seed oil extraction. PSM protein was hydrolyzed using trypsin and fractionated by ultrafiltration. Molecular weight fraction (<3 kDa) with the highest antioxidant activity was purified using prep-HPLC and analytical HPLC. The purification fold of the peptide (fraction V) obtained from PSM protein hydrolysate on DPPH radical scavenging activity, ABTS radical scavenging activity, and reducing power was 1.79-, 1.59-, and 1.81-fold, respectively, after the three-step purification procedure. The sequence of the purified peptide from fraction V that exhibited free radical scavenging activity and reducing power was identified as Ile-Ser-Pro-Arg-Ile-Leu-Ser-Tyr-Asn-Leu-Arg (1,330.77 Da). These results demonstrate that PSM protein, a by-product from the oil seed extraction, can be used as a source of natural antioxidant peptides for food and/or nutraceutical applications.