Tomato processing generates large amounts of by-products, with seeds representing an underutilized yet protein-rich fraction. This study investigated direct enzyme-assisted protein extraction from non-defatted tomato seeds. Various enzymes, enzyme/substrate ratios, pre-treatments, and incubation temperatures were evaluated and optimized. An enzyme/substrate ratio of 5% (w/w) was found to be optimal, with proteases alone outperforming cell wall-degrading enzymes and two-step extraction strategies. Bromelain, Protamex, and Trypsin, for the first time applied directly to non-defatted tomato seeds, achieved the highest protein recoveries (average 110.56 mg BSA eq/g DW). Among them, Trypsin also produced the highest reducing sugar content (25.07 mg GLU eq/g DW), indicating effective cell wall disruption. Digestates obtained from defatted and non-defatted tomato seeds showed comparable protein contents, demonstrating that defatting was unnecessary. Avoiding the defatting step improved process sustainability by reducing solvent use and energy consumption without significantly affecting protein extraction efficiency. Incubation at 37 °C was preferred over 60 °C, as similar yields were achieved under milder conditions while also reducing energy consumption by approximately three-fold (54,340 kJ vs 150,480 kJ for a 1000 L water-based scale-up simulation). These digestates showed significantly higher antioxidant and, for the first time in tomato seed extracts, anti-tyrosinase activities compared with controls. Protamex-derived samples exhibited the highest bioactivities (7.40 mg AA eq/g DW; 101.36 μg KA eq/g DW). Compared to conventional alkaline–acid extraction followed by enzymatic digestion, the direct enzymatic approach provided higher protein recovery. Overall, this method represents a sustainable strategy for producing bioactive peptide-rich extracts for food and non-food applications.

As the world's population grows and agricultural production increases, biorefineries are emerging to convert the resulting residues into bioproducts. However, most agri-residues remain unused, and expected new markets and independence from fossil- and crop-based equivalents have not materialised. In this work, a group of international experts has identified key gaps and interdependencies in integrated biorefinery systems, and proposed strategies to improve their efficiency, compatibility, and scalability. A particular attention is given to protein and polyester production to address the overlooked global protein deficit and persistent plastic pollution, respectively. First, early-stage guidance on embedding complex circularities is discussed to ensure safety and sustainability requirements are met. A dedicated Safe-and-Sustainable by-Design (SSbD) framework needs to be developed and integrated into a decision-support tool to, for example, prevent potential enrichment cycles of biological and chemical contaminants. Among the identified technical barriers, improving bioactive compound recovery through eco-friendly extraction processes, using specific enzymes, natural deep eutectic solvents and/or ionic liquids, is analysed. Simultaneously, valorisation of downstream residual flows should be ensured. Promising improvements of complementary biological conversion pathways have been identified at three levels to balance flexibility, cost, quality, and scalability: (i) upstream mild pre-treatment to convert resistant residues into carboxylate precursors (e.g. thermophilic hydrolysis); (ii) midstream high-performance microbial fermentation (e.g. continuous reactors, co-production of microbial proteins and polyhydroxyalkanoates (PHAs) of different chain lengths); and (iii)

Brewery spent grain (BSG), the primary by-product of beer production, accounts for 85

Wheat plays a key role in the EU economy, mainly in the food sector. However, the wheat value chain also generates large amounts of by-products, such as bran. From a circular economy perspective, it is essential to valorise wheat bran. Of particular interest is the possibility of recovering valuable compounds like ferulic acid (FA) which shows considerable promise. Different protocols have been tested for FA extraction, and alkaline hydrolysis is generally considered capable of achieving nearly 100

Nickel is economically important, but its mining is energy-intensive and environmentally damaging. Agromining, which employs hyperaccumulator plants, represents a sustainable alternative. Odontarrhena chalcidica (Brassicaceae) shows a Ni uptake potential promising for agromining. This study aimed at assessing morphological and physiological traits and Ni content of four O. chalcidica genotypes from Northern Greece. For the first time, a multidimensional approach was used to compare plant traits and Ni accumulation of different genotypes across various cultivation systems. Genotypes were phenotyped for morphological and physiological traits. Nickel accumulation in roots, stems, and leaves was quantified. Plants were grown in aeroponics, hydroponics and pot systems to assess trait consistency. Morphological traits were evaluated through biometric measurements and Environmental Scanning Electron Microscopy. Physiological photosynthetic parameters were analysed and chlorophyll content determined. Nickel content was quantified using monochromatic X-Ray Fluorescence spectroscopy. Significant phenotypic variability was observed among genotypes. Morphological parameters, such as stomatal and trichome densities, showed stability across cultivation conditions. Hydroponics and aeroponics proved to be suitable for phenotyping, as they did not significantly alter key morphological and physiological traits observed in pot. However, biomass yield and Ni uptake varied across cultivation system for both roots and leaves, with hydroponics being optimal for growth and Ni accumulation. The phenotypic diversity observed among genotypes proved to remain consistent across cultivation systems. Odontarrhena chalcidica traits observed in hydroponics and aeroponics are correlated with those observed in pot, thus could be the basis for future field agromining applications.

The wild plants Borago officinalis L. (Boraginaceae) and Hypericum perforatum L. (Hypericaceae) are known to have both food and medicinal uses. The aim of the work was to evaluate the bioaccessibility of potentially bioactive phytochemicals in organ-specific infusions and decoctions pre- and post- simulated in vitro oro-gastrointestinal human digestion aiming at both oral intake and topical applications. (Poly)phenols, reducing sugars, and proteins were analysed by spectroscopic and spectrometry techniques (NMR and HPLC-DAD) as well as their antioxidant and antimicrobial activities pre- and post-digestion. As expected, in vitro digestion altered the composition of the extracts. Specifically, the total amount of (poly)phenols as well as the largely correlated antioxidant activity decreased post-digestion, while reducing sugars and proteins increased. Catechins and rutin were detected in both plants. Rosmarinic acid was the most abundant compound in B. officinalis (on average 90% in infusions), while rutin and hyperoside in H. perforatum (up to 100% in stem digested extracts). Hypericum extracts exhibited similar activity against both Gram-positive and Gram-negative bacteria, while the Borago extracts showed higher activity against Gram-negative bacteria. Overall, the minimal inhibitory concentrations ranged from 0.47 to 0.94 mg/ml. In conclusion, the studied herbal preparations may provide significant amounts of bioactive compounds both pre- and post-digestion able to potentially mitigate oxidative stress-related disorders after ingestion and/or to treat skin infection diseases when applied topically.

This study investigated the valorization of rice industrial starch production fiber by-products, as source of valuable antioxidant phenolic compounds. Six enzymes and acid or alkaline solvents were tested. Total phenols yield and antioxidant activity were measured for best treatment selection. Specific phenols were identified by HPLC-DAD. Overall, 2% v/w Celluclast was identified as the best performing enzyme, leading to 436 mu g gallic acid eq/gFW phenol yield and increasing the recovery of ferulic and p-coumaric acids with respect to control and other enzymatic treatments. Among chemical treatments, NaOH was more efficient than HCl and H2SO4 and was more specific for ferulic acid release. All digestates showed antioxidant capacity, up to 243 and 502 mu g ascorbic acid eq/gFW for 2% v/w Celluclast and 1 M NaOH respectively. Both enzymatic and chemical treatments were successfully applied and, depending on extraction conditions, the release of specific phenolic compounds to obtain enriched digestates was achieved.

Cell-based assays are widely exploited for drug screening and biosensing, providing useful information about bioactivity of target analytes and complex biological samples. It is well recognized that 3D cell models are required to achieve highly valuable information, also from the perspective of replacing animal models. However, bioassays relying on 3D cell models are generally highly demanding in terms of facilities, equipment, and skilled personnel requirements. To reduce cost, increase sustainability, and provide a flexible 3D cell-based platform for bioassays, we here report a novel approach based on a 3D-printed microtissue device. To assess the suitability of this strategy for reporter gene technology, we selected to monitor two molecular pathways which were of interest in several applications, hypoxia signaling and the p53 pathway. The investigation of such pathways is highly relevant in fields spanning from drug screening to bioactivity monitoring for industrial by-product valorization. Microtissues of human hepatocarcinoma (HepG2) and human embryonic kidney (Hek293T) cell lines were obtained with a low-cost and sustainable chip platform and bioassays were developed to monitor the hypoxia-inducible factors (HIFs) and the p53 tumor suppressor pathway. HepG2 and Hek293T 3D cell models were genetically engineered to express the Luc2P from Photinus pyralis firefly either under the regulation of p53 or HIF response elements. The bioassays allowed quantitative assessment of hypoxia and tumoral activity with 1,10-phenanthroline for HIF and with doxorubicin for p53 pathway activation, respectively, showing good potential for applications of this sustainable and low-cost 3D-printed microfluidic platform for bioactivity analyses, drug screening, and precision medicine.

The effects of polyphenol-rich extract obtained from non-compliant defatted green coffee beans (dGCBs) on physicochemical and antioxidant properties, as well as on the sensory profile of vacuum-packed pork burgers stored at 4 °C for 14 days and after cooking were assessed. The dGCB extract obtained by means of supercritical water extraction was analyzed for its polyphenol profile, total phenolic content, radical scavenging, and ferric-reducing antioxidant activities (DPPH and FRAP), Fe2+-chelating capacity, and total iron. The most abundant polyphenol component observed in the dGCB extract was chlorogenic acid, and the alkaloid caffeine was also present. This extract showed antioxidant properties. Thereafter, five formulations of pork meat burgers with added NaCl (1%) were prepared; one without the antioxidant (negative control, C) and one with the use of a synthetic antioxidant (0.05% ascorbic acid = positive control, A), while the other three were supplemented with a different amount of dGCB extract (P15 = 0.15%; P30 = 0.30%; P60 = 0.60%). The addition of dGCB extract increased the antioxidant activity of the raw and cooked burgers and reduced the lipid oxidation of the cooked burgers (0.47, 0.21, and 0.20 vs. 1.28 and 0.55 mg MDA eq./Kg, for P15, P30, and P60 vs. C and A, respectively). No negative effects were observed on the meat’s color parameters and its stability during refrigerated storage and after cooking, nor on sensory attributes (color and aroma) for the lowest concentration of coffee extract. The results obtained indicate that 0.15% dGCB extract is a promising alternative to commercial synthetic antioxidants to improve the quality of refrigerated pork burgers.

EDITORIAL article Front. Plant Sci., 04 April 2023Sec. Technical Advances in Plant Science Volume 14 - 2023 | https://doi.org/10.3389/fpls.2023.1186180

In this work, the evaluation of the antioxidant activity of natural phenolic compounds is performed and compared to that of a conventional antioxidative agent. Phenolic molecules, extracted from industrial processing coffee residues, are added to a matrix of poly(1,4-butylene succinate) (PBS). The apparent activation energy (E-a) of the thermo-oxidative degradation is calculated by employing different methods like Kissinger-Akahira-Sunose, Flynn-Wall-Ozawa and Friedman. The results are compared with the antioxidant activity evaluation obtained through the ABTS radical scavenging assay. From the average activation energies, it is observed that the addition of the natural antioxidants led to an increase in the activation energy of the degradation process as a function of the phenolic compound content. This trend is confirmed by the results of the ABTS assay. Hence, this study proves that the active molecules extracted from agri-food waste could be employed to improve the antioxidant capacity of the biopolymer, even if the composition of the extract must be evaluated in order to mitigate the effects of other components.

In the last 20 years, an increased interest has been shown in the application of different types and combinations of enzymes to obtain phenolic extracts from grape pomace in order to maximize its valorization. Within this framework, the present study aims at improving the recovery of phenolic compounds from Merlot and Garganega pomace and at contributing to the scientific background of enzyme-assisted extraction. Five commercial cellulolytic enzymes were tested in different conditions. Phenolic compound extraction yields were analyzed via a Design of Experiments (DoE) methodology and a second extraction step with acetone was sequentially added. According to DoE, 2% w/w enzyme/substrate ratio was more effective than 1%, allowing a higher total phenol recovery, while the effect of incubation time (2 or 4 h) variation was more enzyme-dependent. Extracts were characterized via spectrophotometric and HPLC-DAD analyses. The results proved that enzymatic and acetone Merlot and Garganega pomace extracts were complex mixtures of compounds. The use of different cellulolytic enzymes led to different extract compositions, as demonstrated using PCA models. The enzyme effects were observed both in water enzymatic and in the subsequent acetone extracts, probably due to their specific grape cell wall degradation and leading to the recovery of different molecule arrays.

Wild food plants are widely consumed all over the world and many have both nutritional and therapeutic value due to the presence of biologically active compounds. The present research, for the first time, aims to compare primary and secondary metabolite levels among different plant organs (flower, leaf, stem, root, bark) of seven species (Borago officinalis L., Cynodon dactylon (L.) Pers., Foeniculum vulgare Mill., Hypericum perforatum L., Malva sylvestris L., Sambucus nigra L., Urtica dioica L.) collected in three different Italian regions (Liguria, Tuscany, Apulia). Plant organ samples were extracted with water or 95% (v/v) methanol and liquid fractions were analyzed using spectrophotometric assays. The best results were obtained for Hypericum perforatum L. samples, followed by Sambucus nigra L. and Borago officinalis L. As also confirmed via PCA analysis on normalized data, flower and leaf extracts of all species exhibited higher levels of polyphenols (up to 105.7 mg GA eq/gDW), reducing sugars (up to 389.2 mg GLUC eq/gDW), proteins (up to 675.7 mg BSA eq/gDW) and of antioxidant capacity (up to 263.5 mg AA eq/gDW). No differences among the regions of gathering were detected after spectrophotometric assays, which was confirmed via PCA analysis. These data contribute to further validate the traditionally reported healing effects of these species on human health.

Selenium (Se) hyperaccumulators are a unique group of plants that can accumulate this element in their aerial parts at concentrations exceeding 100 mg kgDW −1 . These plants actively search for Se in the soil, a phenomenon known as root foraging , reported to date only by few studies. In this study, the effect of localized Se enrichment, in the form of selenite and selenate, was investigated on the root architecture of two Se-hyperaccumulators ( Stanleya pinnata and Astragalus bisulcatus ) and two non-accumulators ( Brassica juncea and Medicago sativa ). Rhizoboxes were divided into two halves: one half was filled with control soil while the other with selenate or selenite (30 mg kgDW −1 ) spiked soil. Seedling were transferred into the interface of the two soils and allowed to grow for three weeks under controlled light and temperature conditions. Staneya pinnata exhibited equal root density in both halves of the rhizobox when grown in control/control and selenite/control soil treatments. However, in the presence of selenate, S. pinnata developed 76% of the roots towards the selenate-enriched half, indicating an active root foraging. In contrast, A. bisulcatus and the non-accumulators B. juncea and M. sativa did not show any preferential distribution of roots. This study revealed that only S. pinnata showed the ability to detect and forage for Se when provided as selenate. Non-accumulators did not show any morphological or Se-accumulation difference associated with the presence of Se in soil in either form.

BACKGROUND AND AIMS:Selenium hyperaccumulator species are of primary interest for studying the evolution of hyperaccumulation and for use in biofortification because selenium is an essential element in human nutrition. In this study, we aimed to determine whether the distributions of selenium in the three most studied hyperaccumulating taxa (Astragalus bisulcatus, Stanleya pinnata and Neptunia amplexicaulis) are similar or contrasting, in order to infer the underlying physiological mechanisms.METHODS:This study used synchrotron-based micro-X-ray fluorescence (µXRF) techniques to visualize the distribution of selenium and other elements in fresh hydrated plant tissues of A. racemosus, S. pinnata and N. amplexicaulis.KEY RESULTS:Selenium distribution differed widely in the three species: in the leaves of A. racemosus and N. amplexicaulis selenium was mainly concentrated in the pulvini, whereas in S. pinnata it was primarilylocalized in the leaf margins. In the roots and stems of all three species, selenium was absent in xylem cells, whereas it was particularly concentrated in the pith rays of S. pinnata and in the phloem cells of A. racemosus and N. amplexicaulis.CONCLUSIONS:This study shows that Astragalus, Stanleya and Neptunia have different selenium-handling physiologies, with different mechanisms for translocation and storage of excess selenium. Important dissimilarities among the three analysed species suggest that selenium hyperaccumulation has probably evolved multiple times over under similar environmental pressures in the US and Australia.

Aims The present study aimed at: (i) verifying the suitability of pure sewage sludge (SS) as growing medium for the hyperaccumulator species (Pteris vittata, Odontarrhena chalcidica, Astragalus bisulcatus and Noccaea caerulescens); (ii) evaluating the removal of As, Ni, Se and Zn operated by the chosen species; (iii) estimating the potential metal yields (bio-ore production) and connected monetary rewards in a small-scale field experiment. Methods Hyperaccumulator plants were first tested under controlled conditions, on three different SS (P1, P2, P3) characterized by the presence of one or more contaminants among As, Ni, Se and Zn. P1 sludge was then chosen for a small-scale field experiment. Hyperaccumulator seedlings were transferred on SS and cultivated for 16 weeks before harvesting. Results All hyperaccumulator species grew healthy on P1 SS, with A. bisulcatus and O. chalcidica reaching an average biomass of 40.2 and 21.5 g DW/plant. Trace metal concentrations in aerial parts were: As (P. vittata) 380 mg/kg DW, Ni (O. chalcidica) 683 mg/kg DW, Se (A. bisulcatus) 165 mg/kg DW, Zn (N. caerulescens) 461 mg/kg DW. The total removal of As, Ni, Se and Zn from SS due to phytoextraction was 5.8, 19, 18, 29% respectively. Conclusions This study demonstrated that phytoextraction can be applied to SS for the removal contaminants while recovering valuable metals. Se and As were identified as the most promising target element, while Ni and Zn removal was poorly efficient under the present experimental conditions.

Bioindicator plants are species that have the capacity to linearly uptake some elements (metal and metalloids) from the growing substrate, thus reflecting their concentration in the soil. Many factors can influence the uptake of these elements by plants, among which is the simultaneous presence of several metals, a common situation in contaminated or natural soils. A novel approach that can be used to validate the bioindication ability of a species growing on a polymetallic substrate is the design of experiment (DoE) approach. The aim of the present study was to apply the DoE in full factorial mode to model the Cu, Cd, Pb, Zn, and Cr bioindication capacity of Polygonum aviculare, used as the model plant. The results showed that P. aviculare has the ability to bioindicate Cd and Cr with a linear uptake (from 0.35 to 6.66, and 0.1 to 3.4 mg kg−1, respectively) unaffected by the presence of other metals. Conversely, the uptake of Pb, Cu, and Zn is strongly influenced by the presence of all the studied metals, making their concentration in the plant shoot not proportional to that of the soil. In conclusion, these preliminary results confirmed that the DoE can be used to predict the bioindicator abilities of a plant for several elements at the same time and to evaluate the interactions that can be established between variables in the growing medium and in the plant itself. However, more studies including other plant species are needed to confirm the effectiveness of this method.

One of the major challenges for the modern society, is the development of a sustainable economy also aiming at the valorization of agro-industrial by-products in conjunction with at a significant reduction of generated residues from farm to retail. In this context, the present study demonstrates a biotechnological approach to yield bioactive peptides from a protein fraction obtained as a by-product of the rice starch production. Enzymatic hydrolysis, with the commercial proteases Alcalase and Protamex, were optimized in bioreactor up to 2 L of volume. The two best digestates, selected with respect to peptide release and extract antioxidant capacity, were further fractionated (cut-offs of 10, 5, and 1 kDa) via cross-flow filtration. Amino acid composition indicated that most of the fractions showed positive nutritional characteristics, but a putative bitter taste. A fraction obtained with Alcalase enzyme (retentate 8 kDa) exerted anti-inflammatory potential, while the smaller molecular weight fractions (retentate 1–5 kDa and permeate < 1 kDa) were more active in tyrosinase inhibition. The latter were further sub-fractionated by size-exclusion chromatography. From the 15 most anti-tyrosinase sub-fractions, 365 peptide sequences were identified via liquid chromatography coupled with high resolution mass spectrometry. The present data support the possible exploitation of bioactive peptide from rice starch by-product as ingredients into food, nutraceutical, pharmaceutical, and cosmetic formulations.

Over the centuries, wild plants have constituted the main food ingredients and traditional medicine in rural communities. In the last decades, thousands of ethnobotanical studies have been conducted, with the aim of documenting the traditional knowledge on wild and cultivated plants both for food and therapeutic purposes. In the present work, 75 published papers related to Italian ethnobotanical knowledge on wild and cultivated plants traditionally used for medical purposes were analyzed and data on 1117 different species organized in the first dataset to target medicinal applications only. For each plant species, the Italian region of use, plant organs, mode of preparation, specific pathological group of application, citation index, and use index were listed. The different therapeutic applications were subdivided into nine main pathological groups according to the targeted human apparatus. Overall, the cited species with highest number of uses were related to the treatment of the digestive system and skin-ears-eyes-hair diseases, followed by diseases of the genito-urinary and respiratory systems. The 13 most relevant species were identified on the basis of their citation and use indexes. The present review on Italian medicinal flora aims to provide valuable information on wild and cultivated species, which are potential sources of plant-based therapeutic remedies, to preserve and reevaluate endangered traditional folk knowledge.

In the past, wild edible alimurgic plants became an important alternative food source when poverty, wars or drought made it difficult to access crops. These plants were considered rich in highly nutritional compounds and also frequently used as food-medicine given their health-promoting properties. With the aim of improving our knowledge on the content of beneficial or detrimental compounds in relation with past local dietary and curative traditions, 12 wild food plant species were collected from two study areas selected for their very different degree of industrialization, urbanization, and conservation of local past traditions among the population: the Bologna province (Northern Italy) and the Middle Agri Valley (Southern Italy). Protein, polyphenol flavonoid and biogenic amine (both free and conjugated) contents and antioxidant activity of raw and boiled wild food plant extracts, and of cooking water were analyzed by means of spectrophotometric and high-performance liquid chromatography methods. The results demonstrated that most of the phenolic compounds were released in the cooking water which also showed the highest antioxidant activity. Seventeen different phenolic compounds were identified, of which the health-related luteolin, luteolin-7-glucoside and rutin were the most abundant (e.g., S. pratensis L. and C. intybus L.). On the other hand, biogenic amines were absent or present at very low levels in cooking water of those very same species (e.g., S. pratensis L., T. officinalis Weber, C. vesicaria subsp. taraxacifolia and C. intybus L.) of which traditionally a decoction is used for therapeutic purposes. Free and conjugated spermidine and spermine were generally the most abundant biogenic amines, while none of the known detrimental monoamines (e.g., histamine) was detected. In conclusion, the present results seem to support past local popular traditions which indicated beneficial medical properties of some wild edible plant, as well as of their cooking water.