This study investigates the optimization of extraction processes for bioactive compounds and proteins from the mushroom Craterellus cornucopioides by comparing Microwave-Assisted Extraction and Ultrasound-Assisted Extraction. Using Response Surface Methodology, the effects of temperature or amplitude, time, and solvent type were evaluated on total phenols, flavonoids, proteins, glutathione content, and antioxidant capacity measured by DPPH and FRAP assays. Additionally, the antimicrobial potential of the extracts was screened against various pathogens. Results demonstrated that water was the most effective solvent for nearly all parameters across both techniques, providing a unified optimum in the ultrasound system at six minutes and one hundred percent amplitude. However, a notable exception was observed for glutathione recovery in the microwave system, where ethanol proved superior to water. Ultrasound-assisted extraction consistently outperformed microwave extraction in protein yield and overall antioxidant potential, offering a more robust approach regarding process efficiency and bioactive yield. In conclusion, while both green techniques enhance recovery, ultrasound extraction with water establishes itself as the most consistent method for the simultaneous extraction of bioactives.

Mushrooms like Inonotus obliquus and Ganoderma lucidum show significant pharmacological promise. This review analyzes fungi as sources of natural inhibitors against Advanced Glycation End-products (AGEs)—key drivers of diabetes and neurodegeneration. We highlight that extracts from Lignosus rhinocerus and Auricularia auricula exhibit antiglycation potency (IC50 as low as 0.001 mg/mL) superior to aminoguanidine. Inhibitory effects are attributed to bioactive fractions including FYGL proteoglycans, uronic acid-rich polysaccharides, and fungal-specific metabolites like ergothioneine. These compounds act through multi-target mechanisms across the glycation cascade: competitive inhibition of Schiff base formation, trapping reactive dicarbonyls (e.g., methylglyoxal), transition metal chelation, and direct scavenging of reactive oxygen species (ROS). Furthermore, the review addresses the transition from in vitro potency to in vivo efficacy (RAGE pathway modulation), stability during food processing (UV-B irradiation), and critical safety issues regarding heavy metal bioaccumulation. Mushroom-derived inhibitors represent a sustainable therapeutic alternative to synthetic agents, offering broader protection against glycative stress. This synthesis provides a foundation for developing standardized mushroom-based nutraceuticals for managing AGE-related chronic disorders.

ABSTRACT This paper focuses on the application of non‐thermal Internet‐of‐Things (IoT) extraction of bioactive compounds from sugar beet leaves. High power ultrasound was used as a non‐thermal processing unit and was integrated with IoT hardware and software. In parallel, two other extraction methods were employed—thermal extraction and cold extraction. The bibliometric review showed that ultrasound extraction of sugar beet leaves has a research potential. Sugar beet leaves contain high levels of macronutrients, including sodium (9138.35 mg/kg), potassium (7275.87 mg/kg), calcium (6276.36 mg/kg), and magnesium (3612.79 mg/kg), as well as micronutrients such as zinc (39.63 mg/kg), manganese (51.85 mg/kg), copper (6.36 mg/kg), and boron (11.96 mg/kg). When it comes to extraction, the highest concentrations of amino acids were estimated for Glx b (193.00 and 134.41 μg/mL) in the ultrasound and thermal extracts. The ultrasound sample had a higher total polyphenol content (≈4.9 mg gallic acid equivalent/g d.m.) and total flavonoid content (≈6.1 mg quercetin eq./g d.m.) compared to the other two samples. The ultrasound method, applied within a non‐thermal IoT processing framework, resulted in higher concentrations of proteins. The highest concentration of the Ribulose bisphosphate carboxylase large chain peptides was obtained by the ultrasound and cold extraction procedures.

A real-time monitored Internet of Things (IoT)-integrated ultrasound-assisted extraction laboratory-scale prototype was developed and evaluated as a green and scalable processing platform for the recovery of high-value compounds from food-grade plant by-products. The key novelty of this study lies in the integration of IoT-based process monitoring and control with ultrasound-assisted extraction, enabling reproducible and energy-efficient recovery of proteins and bioactive compounds from plant matrices. Leaves of Vicia faba L. (broad bean) and Beta vulgaris L. (red beetroot), characterized by a protein content exceeding 30% of dry matter, were selected as representative raw materials. Ultrasound-assisted aqueous extraction yielded protein-rich extracts with mildly acidic pH values (5.94-6.81) and low electrical conductivity (0.043-4.355 mS/cm), indicating minimal matrix degradation and favourable processing conditions. The extracts contained water-soluble flavonoids, free amino acids, and proteins at concentrations suitable for further food and bioproduct applications. Proteomic profiling by nanoLC-MS/MS revealed the presence of ribulose-1,5-bisphosphate carboxylase/oxygenase as the dominant chloroplast protein, alongside hundreds of additional water-soluble proteins originating from multiple plant cell compartments, confirming the broad extraction capability of the ultrasound-assisted system. Continuous IoT-based monitoring ensured stable operating conditions and process reproducibility throughout extraction. The results demonstrate that the proposed IoTenabled ultrasound-assisted extraction prototype represents an effective green technology for the valorisation of plant-based by-products and shows strong potential for process optimization and scale-up toward industrial applications.

Dried leaf powders of red beetroot and broad bean, prepared with different particle sizes (<500 μm and ∼1000 μm), were investigated with respect to their thermal properties using differential scanning calorimetry and thermogravimetric analysis. Kinetic parameters of thermal degradation were derived from the thermogravimetric analysis data by applying the isoconversional Friedman method. Although the samples exhibited comparable thermal behavior, kinetic analysis revealed that powders with smaller particle sizes were less susceptible to thermal deterioration, as indicated by lower rate constants (kp). Further characterization of the red beetroot and broad bean leaf powders was performed using scanning electron microscopy, energy-dispersive X-ray spectroscopy, X-ray powder diffraction, and Fourier-transform infrared spectroscopy. The presence of a crystalline structure was detected in the samples of fava bean leaves powder. The findings provide useful structural and compositional insights for further usage of food industry waste material, which represents a valuable source of functional food ingredients.

Mushrooms have long been valued not only as food but also for their medicinal properties, especially in Eastern European traditional medicine. Species such as Inonotus obliquus, Fomitopsis officinalis, Piptoporus betulinus and Fomes fomentarius have been used to treat gastrointestinal problems, cancers, respiratory ailments and more. Modern research confirms their diverse pharmacological effects, including antitumor, immunomodulatory, antioxidant, antiviral, antibacterial and antidiabetic activities. In addition, mushrooms are widely incorporated into functional foods and nutraceuticals that promote health. Their sustainable cultivation, efficient use of agricultural residues, rapid growth cycles and resilience to environmental stressors make them an environmentally friendly source of food and pharmaceuticals. This review focuses on the potential of fungi to inhibit advanced glycation end products (AGEs)—harmful compounds formed through the non-enzymatic binding of sugars to proteins, lipids or nucleic acids. AGEs are strongly associated with the progression of chronic diseases such as diabetes, cardiovascular disorders, neurodegeneration and aging. Natural AGE inhibitors from mushrooms represent a promising therapeutic alternative to synthetic agents, as they may offer broader mechanisms of action with fewer adverse effects.

Sugar beet leaves, an underutilised by-product rich in proteins and polyphenols, were investigated for recovery of bioactive compounds using ultrasound-assisted extraction (UAE). Response Surface Methodology was applied to identify optimal conditions. The model predicted a maximum Bradford protein yield of 92.07 mg/g d.m. and a total flavonoid content (TFC) of 5.18 mg quercetin eq./g d.m. at 9 min, 100% amplitude, and a 1:50 solid-to-liquid ratio. Under the same conditions, the experimental values reached 94.6 mg/g d.m. (Bradford protein) and 4.71 mg quercetin eq./g d.m. (TFC), showing good agreement with predictions. Extraction time, amplitude, and ratio significantly affected Bradford protein yield, while amplitude was the only significant factor for TFC. In contrast, no significant effects were observed for total phenolic content or Lowry protein yield. These findings highlight the potential of UAE as a sustainable method for valorising sugar beet leaves into high-value natural products.

Knowledge about the composition (volatile and non-volatile) and functionality of natural extracts from Mediterranean plants serves as a basis for their further application. In this study, five selected plants were used for the extraction of plant metabolites. Leaves and flowers of Critmum maritimum, Rosmarinus officinalis, Olea europea, Phylliera latifolia and Mellisa officinalis were collected, and a total of 12 extracts were prepared. Extractions were performed under microwave-assisted conditions, with two solvent types: water (W) and a hydroalcoholic (ethanolic) solution (HA). Detailed extract analysis was conducted. Phenolics were analyzed by detecting individual bioactive compounds using high-performance liquid chromatography and by calculating total phenolic and total flavonoid content through spectrophotometric analysis. Higher concentrations of total phenolics and total flavonoids were obtained in the hydroalcoholic extracts, with the significantly highest total phenolic and flavonoid values in the rosemary hydroalcoholic extract (3321.21 mgGAE/L) and sea fennel flower extract (1794.63 mgQE/L), respectively; and the lowest phenolics in the water extract of olive leaves (204.55 mgGAE/L) and flavonoids in the water extracts of sea fennel leaves, rosemary, olive and mock privet (around 100 mgQE/L). Volatile organic compounds (VOC) were detected using HS-SPME/GC–MS (Headspace Solid-Phase Microextraction coupled with Gas Chromatography-Mass Spectrometry), and antioxidant capacity was estimated using DPPH (2,2-diphenyl-1-picrylhydrazyl assay) and FRAP (Ferric Reducing Antioxidant Power) methods. HS-SPME/GC–MS analysis of samples revealed that sea fennel had more versatile profile, with the presence of 66 and 36 VOCs in W and HA sea fennel leaf extracts, 52 and 25 in W and HA sea fennel flower extracts, 57 in rosemary W and 40 in HA, 20 in olive leaf W and 9 in HA, 27 in W mock privet and 11 in HA, and 35 in lemon balm W and 10 in HA extract. The lowest values of chlorophyll a were observed in sea fennel leaves (2.52 mg/L) and rosemary (2.21 mg/L), and chlorophyll b was lowest in sea fennel leaf and flower (2.47 and 2.25 mg/L, respectively), while the highest was determined in olive (6.62 mg/L). Highest values for antioxidant activity, determined via the FRAP method, were obtained in the HA plant extracts (up to 11216 mgAAE/L for lemon balm), excluding the sea fennel leaf (2758 mgAAE/L) and rosemary (2616 mgAAE/L). Considering the application of these plants for fresh fish preservation, antimicrobial activity of water extracts was assessed against Vibrio fischeri JCM 18803, Vibrio alginolyticus 3050, Aeromonas hydrophila JCM 1027, Moraxella lacunata JCM 20914 and Yersinia ruckeri JCM 15110. No activity was observed against Y. ruckeri and P. aeruginosa, while the sea fennel leaf showed inhibition against V. fisheri (inhibition zone of 24 mm); sea fennel flower was active against M. lacunata (inhibition zone of 14.5 mm) and A. hydrophila (inhibition zone of 20 mm); and rosemary and lemon balm showed inhibition only against V. fisheri (inhibition zone from 18 to 30 mm). This study supports the preparation of natural extracts from Mediterranean plants using green technology, resulting in extracts rich in polyphenolics with strong antioxidant potential, but with no clear significant antimicrobial efficiency at the tested concentrations.

Sustainable development integrates social, economic, technological, scientific, and environmental challenges through the framework of 17 Sustainable Development Goals. Industry 4.0, digitalisation, plant-based processing, environmental challenges, and alternative protein sources are vastly researched and present the foundation of innovative processing in the food processing industry. The exploitation of agroindustrial discards and plant-based by-products as an alternative source of valuable nutrients, including proteins, highlights the potential of upcycling (new economic value) while addressing sustainability challenges. Within the idea of nonthermal extraction of proteins and its process digitalisation, it is crucial to innovate, connect elements of Industry 4.0 and valorise the term sustainability. Innovative approaches in extractions such as ultrasound, plasma, pulsed electric fields, high-intensity pulses, and others, have an energy-saving effect with limited use of toxic chemicals and/or solvents. Usage of software for solvent selection, incorporating digitalisation, and development of protein and peptide databases can help in the prediction, optimisation, and efficiency of nonthermal extractions of proteins from agroindustrial discards. It is also important to select optimal delivery systems like electrospinning, electrospraying, or encapsulation as useful and efficient processes. The environmental, economic, and societal impact of innovative processing needs to be monitored and valorised using specific tools. Life cycle assessment methodology assesses the environmental impact throughout the life cycle of a commercial product, service, or process. The current review shows that extracting protein from agroindustrial discards and by-products of plant and animal origins is a complex matter. The need for a comprehensive approach that considers various viewpoints on extraction is suggested.

The incorporation of different ferrocene scaffolds into the peptide sequences induces the formation of hydrogen-bond-based secondary structural elements that are frequently observed in natural peptides and proteins. There are three simple ferrocene scaffolds for conjugation with amino acids and peptides that serve as templates for ferrocene peptidomimetics, namely ferrocene-1,1′-dicarboxylic acid (Fcd, I), 1′-aminoferrocene-1-carboxylic acid (Fca, III), and ferrocene-1,1′-diamine (Fcda, V). Here, we have investigated their ability to induce the turn structure upon conjugation with Val, Leu, and Phe. Furthermore, we also wanted to determine whether the branched side chains of Val, Leu, and Phe interfere with intramolecular hydrogen bonding (IHB). For these purposes, we performed a detailed spectroscopic analysis by measuring the concentration, temperature, and solvent dependence of the IR, NMR, and CD spectra. The effect of the different ferrocene scaffolds on the antioxidant activity of the prepared peptides was tested using the DPPH and ABTS methods, and was further rationalized using electrochemical measurements. It was found that the ferrocene scaffold has the greatest influence on the hydrogen bonding pattern, while the influence of the side branches of the amino acids is less relevant.

The chirality of the protein backbone influences both self-assembly and biological activity. In ferrocene-containing peptides, the sequence and chirality of the constitutive amino acids as well as the structure of the ferrocene scaffold strongly influence the conformational properties, whereas the biological activity is more strongly influenced by lipophilicity. A joint spectroscopic and computational study has shown that a relatively simple structural modification, such as changing the order of two amino acids in the dipeptide sequence from Pro–Ala (III) to Ala–Pro (IV), also alters the hydrogen bonding patterns from a mostly ten-membered (β-turn) to a seven-membered ring (γ-turn), which affects the antiproliferative activity of the ferrocene peptidomimetics studied. A systematic approach presented in this study allowed us to highlight the relevant structural variations that could lead to increased biological activity in similar ferrocene-based bioconjugates.

The development of turn-based inhibitors of protein–protein interactions has attracted considerable attention in medicinal chemistry. Our group has synthesized a series of peptides derived from an amino-functionalized ferrocene to investigate their potential to mimic protein turn structures. Detailed DFT and spectroscopic studies (IR, NMR, CD) have shown that, for peptides, the backbone chirality and bulkiness of the amino acid side chains determine the hydrogen-bond pattern, allowing tuning of the size of the preferred hydrogen-bonded ring in turn-folded structures. However, their biological potential is more dependent on their lipophilicity. In addition, our pioneering work on the chiroptical properties of aminoferrocene-containing peptides enables the correlation of their geometry with the sign of the CD signal in the absorption region of the ferrocene chromophore. These studies have opened up the possibility of using aminoferrocene and its derivatives as chirooptical probes for the determination of various chirality elements, such as the central chirality of amino acids and the helicity of peptide sequences.

In this research, bio-based films were developed using polyelectrolyte complexes derived from chitosan and gelatin for packaging fish oil. To further enhance the antioxidant functionality, the films were enriched with gallic acid and orange essential oils, either individually or in combination. Initially, the films were characterized for their physico-chemical, optical, surface, and barrier properties. Subsequently, the phenolic compounds and antioxidant capacity of the films were assessed. Finally, the films were tested as antioxidant cover lids for packaging fish oil, which was then stored at ambient temperature for 30 days, with periodical monitoring of oil oxidation parameters. This study revealed that the inclusion of gallic acid-induced possible crosslinking effects, as evidenced by changes in moisture content, solubility, and liquid absorption. Additionally, shifts in the FTIR spectral bands suggested the binding of gallic acid and/or phenols in orange essential oils to CSGEL polymer chains, with noticeable alterations in film coloration. Notably, films containing gallic acid exhibited enhanced UV barrier properties crucial for preserving UV-degradable food compounds. Moreover, formulations with gallic acid demonstrated decreased water vapor permeability, while samples containing orange essential oils had lower CO2 permeability levels. Importantly, formulations containing both gallic acid and essential oils showed a synergistic effect and a significant antioxidant capacity, with remarkable DPPH inhibition rates of up to 88%. During the 30-day storage period, fish oil experienced progressive oxidation, as indicated by an increase in the K232 value in control samples. However, films incorporating gallic acid or orange essential oils as active antioxidants, even used as indirect food contact, effectively delayed the oxidation, highlighting their protective benefits. This study underscores the potential of sustainable bio-based films as natural antioxidant packaging for edible fish oil or fresh fish, offering a promising tool for enhancing food preservation while reducing its waste.

Kombucha is a traditional beverage obtained by the microbial fermentation of tea using a symbiotic culture of bacteria and yeasts. In addition to several documented functional properties, such as anti-inflammatory activity and antioxidant activity, kombucha is often credited with high levels of vitamins, including riboflavin. To our knowledge, the vitamin B2 content in traditionally prepared kombucha has been determined in only two studies, in which the concentration measured by the HPLC technique ranged from 2.2 × 10−7 to 2.1 × 10−4 mol dm−3. These unexplained differences of three orders of magnitude in the vitamin B2 content prompted us to determine its concentration during the cultivation of kombucha under very similar conditions by spectrofluorimetry. The B2 concentrations during the 10-day fermentation of black tea ranged from 7.6 × 10−8 to 3.3 × 10−7 mol dm−3.

As a principal pigment in plants, chlorophyll a (Chl a) is widely used to evaluate quality changes and senescence process during storage of leafy vegetables. The determination of Chl a in plant extracts by spectrophotometric methods using various empirical equations is often unreliable. Considering the sensitivity of fluorescence detection, we report here a simple, inexpensive spectrofluorimetric method that can detect and quantify Chl a in plant extracts. The fluorescence standard used for the quantitative determination of Chl a was isolated in our laboratory from the extract of Anthrospira platensis (Spirulina). The method proved to be reliable, fast, and low cost in a study of the influence of the most commonly used domestic storage conditions on Chl a degradation in fresh-cut arugula.

Despite the large number of peptidomimetics with incorporated heteroannularly functionalized ferrocenes, few studies have investigated their bioactivity. Here, we report the biological evaluation and conformational analysis of enantiomeric dipeptides derived from 1′-aminoferrocene-1-carboxylic acid (Fca) and hydrophobic amino acids (AA = Val, Leu, Phe). The conformational properties of Y-AA-Fca-OMe (Y = Ac, Boc) were elucidated by experimental (IR, NMR, CD, and X-ray) and theoretical (DFT) methods. The prepared dipeptides were screened for their antimicrobial activity against selected Gram-positive and Gram-negative bacteria, lactic acid bacteria and yeasts, while their antioxidant activity was tested by DPPH and FRAP methods. Of all compounds tested, dipeptide d-2a showed the best antibacterial properties against S. aureus, B. subtilis, and P. aeruginosa at a concentration of 2 mM. The time–kill curves showed that antibacterial activity was concentration- and time-dependent. Chirality (d-) and a more polar-protecting group (Ac) were found to affect the biological activity, both antimicrobial and antioxidant. All investigated peptides are considered to be highly hydrophobic and chemically stable in both acidic and buffer media. Dipeptides d-1a–3a, which showed biological activity, were subjected to the determination of proteolytic activity, revealing very good resistance to proteolysis in the presence of chymotrypsin.

The effect of attaching the achiral, cyclic 1-aminocyclohexanecarboxylic acid (Ac6c) directly to the aminoferrocene unit (Ac6c−NH−Fc) appears to be a promising route for the development of a new chiroptical sensor based on a ferrocene chromophore. Three new compounds (Boc−AA−Ac6c−NH−Fc; AA = L-Ala, L-Val, L-Phe) were synthesized, spectroscopically characterized (IR, NMR, CD), and conformationally analyzed (DFT). The chiral information was transferred from the L-amino acid to the ferrocene chromophore by the predominant formation of P-helical structures with ten-membered hydrogen-bonded rings (β-turns). The perturbation of the ferrocene chromophore and the appearance of the negative CD signal near 470 nm originates from a relative orientation of the directly linked amide and cyclopentadienyl planes, described by the dihedral angle χ. The sterically demanding Ac6c amino acid makes trans-like configurations more favorable and thus restricts the dihedral angle χ, which then leads to the appearance of the negative peak near 470 nm in the CD curve.

The experimental (infrared, nuclear magnetic resonance, circular dichroism) and computational (density functional theory) methods allowed us to demonstrate the potential of the ferrocene chromophore to translate chiral information stored at N-terminally attached l-Ala, and transmitted through achiral (Aib)(n) sequence (n=1 to 3), into a characteristic signal in circular dichroism spectra near 470 nm. The sufficiently long tetrapeptide forms a robust and highly organized 3(10) helices capable of perturbing the environment of the highly symmetric ferrocene chromophore in an asymmetric manner. The origin of the sign in the circular dichroism spectra of the ferrocene chromophore (near the 470 nm) strongly correlates with the sign of the dihedral angle chi, accounting for a rotation of a substituent attached to the cyclopentadienyl ring that depends on the helicity of peptide sequence. These observations may help us in the design of future ferrocene-based probes for the assignment of the screw-sense preference of short peptides.

The synthesis and structural characterization of the ferrocene imide derivatives Fc−CO−NH−CO−Me (4), Fc−CO−NH−CO−Fc (7) and Fc−CO−NH−CO−Fn−CO−NH−CO−Fc (8) have been reported. The mononuclear, dinuclear and trinuclear ferrocene imides were prepared by the reaction of ferrocenecarboxamide (3), with acetyl chloride, ferrocenecarbonyl chloride (2) and ferrocene-1,1’-(dicarbonyl chloride) (6), respectively. IR spectroscopic analysis revealed the absence of intramolecular hydrogen bonds in solutions of imides 4, 7 and 8. The crystal packing of N-acetylferrocenecarboxamide (4) is characterized by N−H⋯O hydrogen bonds forming centrosymmetric dimers, while the molecules of its homologue N-methylferrocenecarboxamide (5) are self-assembled by intermolecular N−H⋯O bonds into infinite chains. A detailed conformational analysis (DFT study) suggests the cis-trans configuration of ferrocene imide derivative 7 in solution. The effect of different substituents attached to bridged imide nitrogen on conformational properties of bis-ferrocenyl imides was further investigated and results compared to the existing experimental data.