Valorization of agro-industrial residues using Aspergillus heteromorphus URM0269 for protease production: Characterization and purification.

This study aimed to produce, characterize and purify a protease from Aspergillus heteromorphus URM0269. After production by solid fermentation of wheat bran performed according to a central composite design, protease was characterized in terms of biochemical, kinetic, and thermodynamic parameters for further purification by chromatography. Proteolytic activity achieved a maximum value of 57.43 U/mL using 7.8 g of wheat bran with 40 % moisture. Protease displayed high stability in the pH and temperature ranges of 5.0-10.0 and 20-30 °C, respectively, and acted optimally at pH 7.0 and 50 °C. The enzyme, characterized as a serine protease, followed Michaelis-Menten kinetics with a maximum reaction rate of 140.0 U/mL and Michaelis constant of 11.6 mg/mL. Thermodynamic activation parameters, namely activation Gibbs free energy (69.79 kJ/mol), enthalpy (5.86 kJ/mol), and entropy (-214.39 J/mol.K) of the hydrolysis reaction, corroborated with kinetic modeling showing high affinity for azocasein. However, thermodynamic parameters suggested a reversible mechanism of unfolding. Purification by chromatography yielded a protease purification factor of 7.2, and SDS-PAGE revealed one protein band with a molecular mass of 14.7 kDa. Circular dichroism demonstrated a secondary structure made up of 45.6 % α-helices. These results show the great potential of this protease for future use in the industrial area.

Exploration of Polyphenols Extracted from Cytisus Plants and Their Potential Applications: A Review.

The increasing world population means an increased demand for sustainable processes and products related to foods, particularly those with added health benefits. Plants can be an alternative source of nutritional and biofunctional ingredients. Cytisus plants are an underexploited bioresource, currently prevalent in the Mediterranean Basin and western Asia. This manuscript addresses the processing potential of Cytisus plants for the development of added-value products, including food formulations, food packaging, cosmetics, and therapeutic applications. Most research has reported that Cytisus spp. are a promising source of inexpensive bioactive polyphenol compounds. Cytisus flowers should be considered and exploited as raw materials for the development of new food ingredients (antioxidants, preservatives, additives, etc.), nutraceuticals, or even direct therapeutic agents (anticancer, antibacterial, etc.). In order to evaluate the socioeconomic effect of these underutilized plants, more research is needed to assess their valorization for therapeutic and dietary possibilities, as well as the economic impact.

Valorization of plant by-products in the biosynthesis of silver nanoparticles with antimicrobial and catalytic properties.

Biosynthesis based on natural compounds has emerged as a sustainable approach for the production of metallic nanoparticles (MNP). The main objective of this study was to biosynthesize stable and multifunctional silver nanoparticles (AgNP) using different plant by-products as reducers and capping agents. Extracts obtained from Eucalyptus globulus, Pinus pinaster, Citrus sinensis, Cedrus atlantica and Camellia sinensis by-products, were evaluated. From all plant by-products tested, aqueous extract of eucalyptus leaves (EL), green tea (GT) and black tea (BT) were selected due to their higher antioxidant phenolic content and were individually employed as reducers and capping agents to biosynthesize AgNP. The green AgNP showed zeta potential values of -31.8 to -36.3 mV, with a wide range of particle sizes (40.6 to 86.4 nm), depending on the plant extract used. Green AgNP exhibited an inhibitory effect against various pathogenic bacteria, including Gram-negative (P. putida, E. coli, Vibrio spp.) and Gram-positive (B. megaterium, S. aureus, S. equisimilis) bacteria with EL-AgNP being the nanostructure with the greatest antimicrobial action. EL-AgNP showed an excellent photodegradation of indigo carmine (IC) dye under direct sunlight, with a removal percentage of up to 100% after 75 min. A complete cost analysis revealed a competitive total cost range of 8.0-9.0 €/g for the biosynthesis of AgNP.

Recent advances in biorefineries based on lignin extraction using deep eutectic solvents: A review.

Considering the urgent need for alternative biorefinery schemes based on sustainable development, this review aims to summarize the state-of-the-art in the use of deep eutectic solvent pretreatment to fractionate lignocellulose, with a focus on lignin recovery. For that, the key parameters influencing the process are discussed, as well as various strategies to enhance this pretreatment efficiency are explored. Moreover, this review describes the challenges and opportunities associated with the valorization of extraction-derived streams and highlights recent advancements in solvent recovery techniques. Furthermore, the utilization of computational models for process design and optimization is introduced, as the initial attempts at the economic and environmental assessment of this lignocellulosic bioprocess based on deep eutectic solvents. Overall, this review offers a comprehensive perspective on the recent advances in this emerging field and serves as a foundation for further research on the potential integration of deep eutectic pretreatment in sustainable multi-product biorefinery schemes.

Polymeric biomaterials for wound healing.

Skin indicates a person's state of health and is so important that it influences a person's emotional and psychological behavior. In this context, the effective treatment of wounds is a major concern, since several conventional wound healing materials have not been able to provide adequate healing, often leading to scar formation. Hence, the development of innovative biomaterials for wound healing is essential. Natural and synthetic polymers are used extensively for wound dressings and scaffold production. Both natural and synthetic polymers have beneficial properties and limitations, so they are often used in combination to overcome overcome their individual limitations. The use of different polymers in the production of biomaterials has proven to be a promising alternative for the treatment of wounds, as their capacity to accelerate the healing process has been demonstrated in many studies. Thus, this work focuses on describing several currently commercially available solutions used for the management of skin wounds, such as polymeric biomaterials for skin substitutes. New directions, strategies, and innovative technologies for the design of polymeric biomaterials are also addressed, providing solutions for deep burns, personalized care and faster healing.

Microwave-assisted autohydrolysis of avocado seed for the recovery of antioxidant phenolics and glucose.

This study describes the valorization of avocado seed (AS) within a green biorefinery concept using microwave-assisted autohydrolysis. After the treatment at temperatures of 150-230 °C for 5 min, the resulting solid and liquor were characterized. The temperature of 220 °C led to the simultaneous optimal values of antioxidant phenolics/flavonoids (42.15 mg GAE/g AS, 31.89 RE/g AS, respectively) and glucose + glucooligosaccharides (38.82 g/L) in the liquor. Extraction with ethyl acetate allowed the recovery of the bioactive compounds while maintaining the polysaccharides in the liquor. The extract was rich in vanillin (99.02 mg/g AS) and contained several phenolic acids and flavonoids. The solid phase and the phenolic-free liquor were subjected to enzymatic hydrolysis to produce glucose, reaching values of 9.93 and 105 g glucose/L, respectively. This work demonstrates that microwave-assisted autohydrolysis is a promising technology to obtain fermentable sugars and antioxidant phenolic compounds from avocado seeds following a biorefinery scheme.

Evaluation of grape stems and grape stem extracts for sulfur dioxide replacement during grape wine production.

Sulfur dioxide (SO2), the main preservative in wine, may affect the sensory properties of the wines, as well as cause allergic reactions and headaches in sensitive people. The aim of this work was to evaluate the replacement of SO2 in Tempranillo wines with Mazuelo grape stem products. Five Tempranillo red wines were elaborated: positive control (60 mg/L SO2); negative control with no preservatives; Mazuelo extract (200 mg/L); Mazuelo extract combined with SO2 (100 mg/L + 20 mg/L); and Mazuelo stem (400 mg/L). The oenological parameters, antioxidant capacity, total phenolic (TP), total flavonoids (TF) and total anthocyanins (TA) contents were determined. Additionally, individual phenols were analyzed by HPLC-DAD-FLD. The spectrophotometric analyses showed that the wines had similar antioxidant capacities and concentrations of TP and TF. However, TA was more affected by the lack of SO2 as anthocyanins presented higher concentrations in positive control samples. The concentrations of individual phenols followed a similar path in all samples. Wines containing sulfites were more similar than the other treatments. However, these similarities were not reflected on the sensory analysis performed, as triangle test did not show differences between the wine with extract addition and the positive control wine. Therefore, Mazuelo stem extract could be a possible strategy for SO2 replacement. Nevertheless, further studies are necessary to confirm the potential of grape stem extracts as wine preservative.

Unveiling the techno-functional and bioactive properties of bee pollen as an added-value food ingredient.

Bee pollen is an imperative product for human use. Seven bee pollens were harvested from Morocco, and their chemical, biological and techno-functional properties were studied. All samples showed acceptable physicochemical and nutritional quality with a mean energy value of 239 kcal/100 g. FTIR spectra confirmed the presence of major constituents like carbohydrates, lipids, proteins and polyphenols. Moreover, pollens exhibited good techno-functional properties, like carbohydrate solubility (34.47-59.27 g/100 g), protein solubility (7.28-23.31 g/100 g), emulsifying stability (16.52-45.38 min), emulsifying activity (9.83-25.05 g/m3) water absorption capacity (1.06-2.19 g/g), oil absorption capacity (1.15-3.50 g/g) and water-oil absorption index (0.62-1.25). Bee pollen extracts revealed potent antioxidant capacity and digestive enzyme inhibitory activity associated with the presence of fifteen phenolic compounds belonging to flavons, flavonols, flavanones, flavan-3-ols, hydroxybenzoic and hydroxycinnamic acids, and stilbenes families. Present data indicate the possible application of bee pollen as a useful nutritional, bioactive and anti-foaming ingredient, replacing synthetic products in food industries.

Effect of Gastrointestinal Digestion on the Bioaccessibility of Phenolic Compounds and Antioxidant Activity of Fermented Aloe vera Juices.

Plant-based beverages are enriched by the fermentation process. However, their biocompounds are transformed during gastrointestinal digestion, improving their bioaccessibility, which is of primary importance when considering the associated health benefits. This study aimed to evaluate the effect of in vitro gastrointestinal digestion on phenolic compound bioaccessibility and antioxidant activity of novel Aloe vera juices fermented by probiotic Enterococcus faecium and Lactococcus lactis. Aloe vera juices were digested using the standardized static INFOGEST protocol. During digestion, phenolic compounds and antioxidant activity (DPPH, ABTS, and FRAP) were accessed. The digestion process was seen to significantly increase the total phenolic content of the fermented Aloe vera juices. The fermentation of Aloe vera increased the bioaccessibility of juice biocompounds, particularly for kaempferol, ellagic acid, resveratrol, hesperidin, ferulic acid, and aloin. The phenolics released during digestion were able to reduce the oxidative radicals assessed by ABTS and FRAP tests, increasing the antioxidant action in the intestine, where they are absorbed. The fermentation of Aloe vera by probiotics is an excellent process to increase the bioavailability of beverages, resulting in natural added-value functional products.

Deep Eutectic Solvents as a Green Tool for the Extraction of Bioactive Phenolic Compounds from Avocado Peels.

Avocado peels are the main agro-industrial residue generated during the avocado processing, being a rich source of bioactive compounds like phenolic compounds. The growing demand for more sustainable processes requires the development of new and effective methods for extracting bioactive compounds from industrial waste. Deep eutectic solvents (DESs) are a new sustainable alternative to toxic organic solvents due to their non-toxicity and biocompatibility. In this study, five selected DESs were applied for the extraction of bioactive phenolic compounds from avocado peels. The extraction efficiency was evaluated by measuring the total phenolics and flavonoids content. The best extraction results were obtained with choline chloride-acetic acid and -lactic acid (92.03 ± 2.11 mg GAE/g DAP in TPC and 186.01 ± 3.27 mg RE/g DAP); however, all tested DESs show better extraction efficiency than ethanol. All the obtained NADES extracts have high antioxidant activity (FRAP: 72.5-121.1 mg TE/g; TAC: 90.0-126.1 mg AAE/g). The synthesized DESs and avocado peels DES extracts had activity against all tested bacteria (Staphylococcus aureus, Streptococcus dysgalactiae, Escherichia coli and Pseudomonas putida), and the extracts prepared with choline chloride-acetic acid and -lactic acid have the highest antibacterial activity against all microorganisms. These results, coupled with the non-toxic, biodegradable, low-cost, and environmentally friendly characteristics of DESs, provide strong evidence that DESs represent an effective alternative to organic solvents for the recovery of phenolic bioactive compounds from agro-industrial wastes.

Exploring the Palynological, Chemical, and Bioactive Properties of Non-Studied Bee Pollen and Honey from Morocco.

Bee products are known for their beneficial properties widely used in complementary medicine. This study aims to unveil the physicochemical, nutritional value, and phenolic profile of bee pollen and honey collected from Boulemane-Morocco, and to evaluate their antioxidant and antihyperglycemic activity. The results indicate that Citrus aurantium pollen grains were the majority pollen in both samples. Bee pollen was richer in proteins than honey while the inverse was observed for carbohydrate content. Potassium and calcium were the predominant minerals in the studied samples. Seven similar phenolic compounds were found in honey and bee pollen. Three phenolic compounds were identified only in honey (catechin, caffeic acid, vanillic acid) and six phenolic compounds were identified only in bee pollen (hesperidin, cinnamic acid, apigenin, rutin, chlorogenic acid, kaempferol). Naringin is the predominant phenolic in honey while hesperidin is predominant in bee pollen. The results of bioactivities revealed that bee pollen exhibited stronger antioxidant activity and effective α-amylase and α-glycosidase inhibitory action. These bee products show interesting nutritional and bioactive capabilities due to their chemical constituents. These features may allow these bee products to be used in food formulation, as functional and bioactive ingredients, as well as the potential for the nutraceutical sector.

A Versatile Nanocarrier-Cubosomes, Characterization, and Applications.

The impact of nanotechnology on the exponential growth of several research areas, particularly nanomedicine, is undeniable. The ability to deliver active molecules to the desired site could significantly improve the efficiency of medical treatments. One of the nanocarriers developed which has drawn researchers' attention are cubosomes, which are nanosized dispersions of lipid bicontinuous cubic phases in water, consisting of a lipidic interior and aqueous domains folded in a cubic lattice. They stand out due to their ability to incorporate hydrophobic, hydrophilic, and amphiphilic compounds, their tortuous internal configuration that provides a sustained release, and the capacity to protect and safely deliver molecules. Several approaches can be taken to prepare this structure, as well as different lipids like monoolein or phytantriol. This review paper describes the different methods to prepare nanocarriers. As it is known, the physicochemical properties of nanocarriers are very important, as they influence their pharmacokinetics and their ability to incorporate and deliver active molecules. Therefore, an extensive characterization is essential to obtain the desired effect. As a result, we have extensively described the most common techniques to characterize cubosomes, particularly nanocarriers. The exceptional properties of the cubosomes make them suitable to be used in several applications in the biomedical field, from cancer therapeutics to imaging, which will be described. Taking in consideration the outstanding properties of cubosomes, their application in several research fields is envisaged.

Sambucus nigra flower and berry extracts for food and therapeutic applications: effect of gastrointestinal digestion on in vitro and in vivo bioactivity and toxicity.

The bioavailability of natural compounds should be assessed through different perspectives. Studying the behaviour of the extracts after digestion is often overlooked but is crucial for success in the development of active food ingredients. Thus, the bioaccessibility of S. nigra (flower and berry) extracts after in vitro gastrointestinal digestion and their effect on toxicity and bioactive potential were studied. The flower extract had a higher content of phenolic compounds, like rutin, chlorogenic acid and rosmarinic acid, while in the berry extract, rutin, resveratrol, ferulic acid and chlorogenic acid were the main phenolic compounds. The effect of the non-digested and digested extracts was significantly different on different cell lines. The IC50 of the normal cell line (L929) was the highest, indicating low toxicity. The IC50 of the cancerous cell lines (HeLa and HT29) was lower, particularly the extract obtained from the flower upon digestion. In the presence of an oxidant agent - tbHP, only the berry extract was able to significantly reduce the formation of ROS in the L929 cell line, while in the HeLa cells, all the extracts were able to reduce ROS formation. The in vivo Artemia salina lethality bioassay demonstrated a dose-dependent effect of extracts, and the berry digested extract induced the lowest mortality rate. The promising results obtained on the chemical and biological evaluation of the extracts indicate that the natural compounds isolated from S. nigra by-products can be used as potential ingredients for functional food formulations and/or as bio-therapeutic agents.

Unveiling the Antioxidant Therapeutic Functionality of Sustainable Olive Pomace Active Ingredients.

Olive pomace (OP) is the main residue that results from olive oil production. OP is rich in bioactive compounds, including polyphenols, so its use in the treatments of diseases related to oxidative stress, such as cancer, could be considered. The present work aimed to study the biological properties of different OP extracts, obtained by ohmic heating-assisted extraction and conventional heating, using water and 50% ethanol, in the treatment and prevention of colorectal cancer through Caco-2 cell models. Additionally, an in-silico analysis was performed to identify the phenolic intestinal absorption and Caco-2 permeability. The extracts were chemically characterized, and it was found that the Ohmic-hydroethanolic (OH-EtOH) extract had the highest antiproliferative effect, probably due to its higher content of phenolic compounds. The OH-EtOH induced potential modifications in the mitochondrial membrane and led to apoptosis by cell cycle arrest in the G1/S phases with activation of p53 and caspase 3 proteins. In addition, this extract protected the intestine against oxidative stress (ROS) caused by H2O2. Therefore, the bioactive compounds present in OP and recovered by applying a green technology such as ohmic-heating, show promising potential to be used in food, nutraceutical, and biomedical applications, reducing this waste and facilitating the circular economy.

Novel Bio-Functional Aloe vera Beverages Fermented by Probiotic Enterococcus faecium and Lactobacillus lactis.

Aloe vera has been medicinally used for centuries. Its bioactive compounds have been shown to be very effective in the treatment of numerous diseases. In this work, a novel functional beverage was developed and characterized to combine the health benefits of probiotic bacteria with the Aloe vera plant itself. Two Aloe vera juices were obtained by fermentation either by a novel isolated Enterococcus faecium or a commercial Lactococcus lactis. The extraction of Aloe vera biocompounds for further fermentation was optimized. Extraction with water plus cellulase enhanced the carbohydrates and phenolic compounds in the obtained extracts. The biotransformation of the bioactive compounds from the extracts during fermentation was assessed. Both probiotic bacteria were able to grow on the Aloe vera extract. Lactic acid and short-chain fatty acids (SCFA) together with fourteen individual phenolic compounds were quantified in the produced Aloe vera juice, mainly epicatechin, aloin, ellagic acid, and hesperidin. The amount of total phenolic compounds was maintained through fermentation. The antioxidant activity was significantly increased in the produced juice by the ABTS method. The novel produced Aloe vera juice showed great potential as a functional beverage containing probiotics, prebiotics, SCFA, and phenolic compounds in its final composition.

Correction: Silva et al. Chemical Profile and Bioactivities of Extracts from Edible Plants Readily Available in Portugal. Foods 2021, 10, 673.

The authors found a mistake in the original paper [...].

Nanocarriers as Active Ingredients Enhancers in the Cosmetic Industry-The European and North America Regulation Challenges.

"Flawless skin is the most universally desired human feature" is an iconic statement by Desmond Morris. Skin indicates one´s health and is so important that it affects a person's emotional and psychological behavior, these facts having propelled the development of the cosmetics industry. It is estimated that in 2023, this industry will achieve more than 800 billion dollars. This boost is due to the development of new cosmetic formulations based on nanotechnology. Nanocarriers have been able to solve problems related to active ingredients regarding their solubility, poor stability, and release. Even though nanocarriers have evident benefits, they also present some problems related to the high cost, low shelf life, and toxicity. Regulation and legislation are two controversial topics regarding the use of nanotechnology in the field of cosmetics. In this area, the U.S. FDA has taken the lead and recommended several biosafety studies and post-market safety evaluations. The lack of a global definition that identifies nanomaterials as a cosmetic ingredient is a hindrance to the development of global legislation. In the EU, the legislation regarding the biosafety of nanomaterials in cosmetics is stricter. "The cost is not the only important issue, safety and the application of alternative testing methods for toxicity are of crucial importance as well".

Protective Effect of Honey and Propolis against Gentamicin-Induced Oxidative Stress and Hepatorenal Damages.

Bee products are a promising source of phenolic compounds with strong antioxidant activity. The present study was designed to explore the protective effect of honey, propolis, and their combination on gentamicin-induced oxidative stress and hepatorenal dysfunction. This study was conducted on male Wistar rats by intraperitoneal injections of gentamicin (120 mg/kg BW/day, i.p.) or normal saline (1 ml/kg BW/day, i.p.) for 10 consecutive days. Honey (2 g/kg BW), propolis (100 mg/kg BW), or their combination were given daily by gavage to normal and gentamicin groups. Honey and propolis samples were evaluated for their phytochemical composition and antioxidant capacity. The in vitro investigations showed that the evaluated samples especially propolis extract have high antioxidant power associated with the presence of several phenolic compounds such as flavonoids, flavan-3-ols, hydroxybenzoic acids, hydroxycinnamic acids, and stilbenes, while honey contains only hydroxybenzoic acids and hydroxycinnamic acids. It was also shown that simultaneous treatment with honey or propolis extract alone or in association prevented changes caused by gentamicin administration and improved hepatic and renal functions. Changes caused by gentamicin administration, observed by in vivo experiments, include significant elevation of uric acid, urea, creatinine, and hepatic enzyme levels (ALT, AST, and ALP) and kidney biochemical changes (an increase of urea, uric acid, and creatinine and a decrease of albumin and total protein) as well as remarkable changes of renal and liver oxidative stress markers (CAT, GPx, and GSH) and elevation of MDA levels. Overall, it can be concluded that honey and propolis might be useful in the management of liver and renal diseases induced by xenobiotics.

Exploiting the Potential of Bioactive Molecules Extracted by Ultrasounds from Avocado Peels-Food and Nutraceutical Applications.

Natural bioactive compounds from food waste have fomented interest in food and pharmaceutical industries for the past decade. In this work, it purposed the recovery of bioactive avocado peel extract using an environmentally friendly technique: the ultrasound assisted extraction. The response surface methodology was applied in order to optimize the conditions of the extraction, ethanol-water mixtures and time. The optimized extracts (ethanol 38.46%, 44.06 min, and 50 °C) were chemically characterized by HPLC-ESI-MS and FTIR. Its antioxidant ability, as well as, its effect on cell metabolic activity of normal (L929) and cancer (Caco-2, A549 and HeLa) cell lines were assessed. Aqueous ethanol extracts presented a high content in bioactive compounds with high antioxidant potential. The most representative class of the phenolic compounds found in the avocado peel extract were phenolic acids, such as hydroxybenzoic and hydroxycinnamic acids. Another important chemical group detected were the flavonoids, such as flavanols, flavanonols, flavones, flavanones and chalcone, phenylethanoids and lignans. In terms of its influence on the metabolic activity of normal and cancer cell lines, the extract does not significantly affect normal cells. On the other hand, it can negatively affect cancer cells, particularly HeLa cells. These results clearly demonstrated that ultrasound is a sustainable extraction technique, resulting in extracts with low toxicity in normal cells and with potential application in food, pharmaceutical or nutraceutical sectors.

Chemical Characterization of Sambucus nigra L. Flowers Aqueous Extract and Its Biological Implications.

The main goal of this study was to chemically characterize an aqueous S. nigra flower extract and validate it as a bioactive agent. The elderflower aqueous extraction was performed at different temperatures (50, 70 and 90 °C). The extract obtained at 90 °C exhibited the highest phenolic content and antiradical activity. Therefore, this extract was analyzed by GC-MS and HPLC-MS, which allowed the identification of 46 compounds, being quercetin and chlorogenic acid derivatives representative of 86% of the total of phenolic compounds identified in hydrophilic fraction of the aqueous extract. Naringenin (27.2%) was the major compound present in the lipophilic fraction. The antiproliferative effects of the S. nigra extract were evaluated using the colon cancer cell lines RKO, HCT-116, Caco-2 and the extract's antigenotoxic potential was evaluated by the Comet assay in RKO cells. The RKO cells were the most susceptible to S. nigra flower extract (IC50 = 1250 µg mL-1). Moreover, the extract showed antimicrobial activity against Gram-positive bacteria, particularly Staphylococcus aureus and S. epidermidis. These results show that S. nigra-based extracts can be an important dietary source of bioactive phenolic compounds that contribute to health-span improving life quality, demonstrating their potential as nutraceutical, functional foods and/or cosmetic components for therapeutic purposes.