Alginate Beads with Encapsulated Bioactive Substances from Mangifera indica Peels as Promising Peroral Delivery Systems.

Since various bioactive substances are unstable and can degrade in the gastrointestinal tract, their stabilization is crucial. This study aimed to encapsulate mango peel extract (MPE) into edible alginate beads using the ionotropic gelation method for the potential oral delivery of bioactive substances. Mango peels, generally discarded and environmentally harmful, are rich in health-promoting bioactive substances. The alginate beads were examined for entrapment efficiency, particle size, morphology, thermal stability, physiochemical interactions, release profile under gastrointestinal conditions, and antibacterial efficacy. The study demonstrated the successful encapsulation of MPE with an efficiency of 63.1%. The in vitro release study showed the stability of the alginate beads in simulated gastric fluid with a maximum release of 45.0%, and sustained, almost complete release (99.4%) in simulated intestinal fluid, indicating successful absorption into the human body. In both fluids, the MPE release followed first-order kinetics. Encapsulation successfully maintained the antibacterial properties of MPE, with significant inhibitory activity against pathogenic intestinal bacteria. This is the first study on MPE encapsulation in alginate beads, presenting a promising oral delivery system for high-added-value applications in the food industry for dietary supplements, functional foods, or food additives. Their production is sustainable and economical, utilizing waste material and reducing environmental pollution.

Multifunctional Iron Oxide Nanoparticles as Promising Magnetic Biomaterials in Drug Delivery: A Review.

A wide range of applications using functionalized magnetic nanoparticles (MNPs) in biomedical applications, such as in biomedicine as well as in biotechnology, have been extensively expanding over the last years. Their potential is tremendous in delivery and targeting systems due to their advantages in biosubstance binding. By applying magnetic materials-based biomaterials to different organic polymers, highly advanced multifunctional bio-composites with high specificity, efficiency, and optimal bioavailability are designed and implemented in various bio-applications. In modern drug delivery, the importance of a successful therapy depends on the proper targeting of loaded bioactive components to specific sites in the body. MNPs are nanocarrier-based systems that are magnetically guided to specific regions using an external magnetic field. Therefore, MNPs are an excellent tool for different biomedical applications, in the form of imaging agents, sensors, drug delivery targets/vehicles, and diagnostic tools in managing disease therapy. A great contribution was made to improve engineering skills in surgical diagnosis, therapy, and treatment, while the advantages and applicability of MNPs have opened up a large scope of studies. This review highlights MNPs and their synthesis strategies, followed by surface functionalization techniques, which makes them promising magnetic biomaterials in biomedicine, with special emphasis on drug delivery. Mechanism of the delivery system with key factors affecting the drug delivery efficiency using MNPs are discussed, considering their toxicity and limitations as well.

Mango Peels as an Industrial By-Product: A Sustainable Source of Compounds with Antioxidant, Enzymatic, and Antimicrobial Activity.

Plant waste materials are important sources of bioactive compounds with remarkable health-promoting benefits. In particular, industrial by-products such as mango peels are sustainable sources of bioactive substances, with antioxidant, enzymatic, and antimicrobial activity. Appropriate processing is essential to obtain highly bioactive compounds for further use in generating value-added products for the food industry. The objective of the study was to investigate and compare the biological activity of compounds from fresh and dried mango peels obtained by different conventional methods and unconventional extraction methods using supercritical fluids (SFE). The highest total phenolic content (25.0 mg GAE/g DW) and the total content of eight phenolic compounds (829.92 µg/g DW) determined by LC-MS/MS were detected in dried mango peel extract obtained by the Soxhlet process (SE). SFE gave the highest content of proanthocyanidins (0.4 mg PAC/g DW). The ethanolic ultrasonic process (UAE) provided the highest antioxidant activity of the product (82.4%) using DPPH radical scavenging activity and total protein content (2.95 mg protein/g DW). Overall, the dried mango peels were richer in bioactive compounds (caffeic acid, chlorogenic acid, gallic acid, catechin, and hesperidin/neohesperidin), indicating successful preservation during air drying. Furthermore, outstanding polyphenol oxidase, superoxide dismutase (SOD), and lipase activities were detected in mango peel extracts. This is the first study in which remarkable antibacterial activities against the growth of Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) and Gram-positive bacteria (Bacillus cereus and Staphylococcus aureus) were evaluated by determining the microbial growth inhibition rate after 12 and 24 h incubation periods for mango peel extracts obtained by different methods. Ethanolic SE and UAE extracts from dried mango peels resulted in the lowest minimum inhibitory concentrations (MIC90) for all bacterial species tested. Mango peels are remarkable waste products that could contribute to the sustainable development of exceptional products with high-added value for various applications, especially as dietary supplements.

Influence of Supercritical Carbon Dioxide on the Activity and Conformational Changes of α-Amylase, Lipase, and Peroxidase in the Solid State Using White Wheat Flour as an Example.

Green technologies using renewable and alternative sources, including supercritical carbon dioxide (sc-CO2), are becoming a priority for researchers in a variety of fields, including the control of enzyme activity which, among other applications, is extremely important in the food industry. Namely, extending shelf life of e.g., flour could be reached by tuning the present enzymes activity. In this study, the effect of different sc-CO2 conditions such as temperature (35-50 °C), pressure (200 bar and 300 bar), and exposure time (1-6 h) on the inactivation and structural changes of α-amylase, lipase, and horseradish peroxidase (POD) from white wheat flour and native enzymes was investigated. The total protein (TPC) content and residual activities of the enzymes were determined by standard spectrophotometric methods, while the changes in the secondary structures of the enzymes were determined by circular dichroism spectrometry (CD). The present work is therefore concerned for the first time with the study of the stability and structural changes of the enzyme molecules dominant in white wheat flour under sc-CO2 conditions at different pressures and temperatures. In addition, the changes in aggregation or dissociation of the enzyme molecules were investigated based on the changes in particle size distribution and ζ-potential. The results of the activity assays showed a decrease in the activity of native POD and lipase under optimal exposure conditions (6 h and 50 °C; and 1 h and 50 °C) by 22% and 16%, respectively. In contrast, no significant changes were observed in α-amylase activity. Consequently, analysis of the CD spectra of POD and lipase confirmed a significant effect on secondary structure damage (changes in α-helix, ß-sheet, and ß-turn content), whereas the secondary structure of α-amylase retained its original configuration. Moreover, the changes in particle size distribution and ζ-potential showed a significant effect of sc-CO2 treatment on the aggregation and dissociation of the selected enzymes. The results of this study confirm that sc-CO2 technology can be effectively used as an environmentally friendly technology to control the activity of major flour enzymes by altering their structures.

Transglutaminase in Foods and Biotechnology.

Stabilization and reusability of enzyme transglutaminase (TGM) are important goals for the enzymatic process since immobilizing TGM plays an important role in different technologies and industries. TGM can be used in many applications. In the food industry, it plays a role as a protein-modifying enzyme, while, in biotechnology and pharmaceutical applications, it is used in mediated bioconjugation due to its extraordinary crosslinking ability. TGMs (EC 2.3.2.13) are enzymes that catalyze the formation of a covalent bond between a free amino group of protein-bound or peptide-bound lysine, which acts as an acyl acceptor, and the γ-carboxamide group of protein-bound or peptide-bound glutamine, which acts as an acyl donor. This results in the modification of proteins through either intramolecular or intermolecular crosslinking, which improves the use of the respective proteins significantly.

Enzymatic, Antioxidant, and Antimicrobial Activities of Bioactive Compounds from Avocado (Persea americana L.) Seeds.

The aim of this research was to identify and quantify biologically active compounds from avocado (Persea americana L.) seeds (AS) utilizing different techniques with the use of ultrasound (US), ethanol (EtOH), and supercritical carbon dioxide (scCO2) for possible applications in (bio)medicine, pharmaceutical, cosmetic, or other relevant industries. Initially, a study of the process efficiency (η) was carried out, which revealed yields in the range of 2.96-12.11 wt%. The sample obtained using scCO2 was found to be the richest in total phenols (TPC) and total proteins (PC), while the sample obtained with the use of EtOH resulted in the highest content of proanthocyanidins (PAC). Phytochemical screening of AS samples, quantified by the HPLC method, indicated the presence of 14 specific phenolic compounds. In addition, the activity of the selected enzymes (cellulase, lipase, peroxidase, polyphenol oxidase, protease, transglutaminase, and superoxide dismutase) was quantified for the first time in the samples from AS. Using DPPH radical scavenging activity, the highest antioxidant potential (67.49%) was detected in the sample obtained with EtOH. The antimicrobial activity was studied using disc diffusion method against 15 microorganisms. Additionally, for the first time, the antimicrobial effectiveness of AS extract was quantified by determination of microbial growth-inhibition rates (MGIRs) at different concentrations of AS extract against three strains of Gram-negative (Escherichia coli, Pseudomonas aeruginosa, and Pseudomonas fluorescens) bacteria, three strains of Gram-positive (Bacillus cereus, Staphylococcus aureus, and Streptococcus pyogenes) bacteria, and fungi (Candida albicans). MGIRs and minimal inhibitory concentration (MIC90) values were determined after 8 and 24 h of incubation, thus enabling the screening of antimicrobial efficacy for possible further applications of AS extracts as antimicrobial agents in (bio)medicine, pharmaceutical, cosmetic, or other industries. For example, the lowest MIC90 value was determined for B. cereus after 8 h of incubation in the case of UE and SFE extracts (70 µg/mL), indicating an outstanding result and the potential of AS extracts, as the MIC values for B. cereus have not been investigated so far.

Sustainable Biodegradable Biopolymer-Based Nanoparticles for Healthcare Applications.

Biopolymeric nanoparticles are gaining importance as nanocarriers for various biomedical applications, enabling long-term and controlled release at the target site. Since they are promising delivery systems for various therapeutic agents and offer advantageous properties such as biodegradability, biocompatibility, non-toxicity, and stability compared to various toxic metal nanoparticles, we decided to provide an overview on this topic. Therefore, the review focuses on the use of biopolymeric nanoparticles of animal, plant, algal, fungal, and bacterial origin as a sustainable material for potential use as drug delivery systems. A particular focus is on the encapsulation of many different therapeutic agents categorized as bioactive compounds, drugs, antibiotics, and other antimicrobial agents, extracts, and essential oils into protein- and polysaccharide-based nanocarriers. These show promising benefits for human health, especially for successful antimicrobial and anticancer activity. The review article, divided into protein-based and polysaccharide-based biopolymeric nanoparticles and further according to the origin of the biopolymer, enables the reader to select the appropriate biopolymeric nanoparticles more easily for the incorporation of the desired component. The latest research results from the last five years in the field of the successful production of biopolymeric nanoparticles loaded with various therapeutic agents for healthcare applications are included in this review.

(Magnetic) Cross-Linked Enzyme Aggregates of Cellulase from T. reesei: A Stable and Efficient Biocatalyst.

Cross-linked enzyme aggregates (CLEAs) represent an effective tool for carrier-free immobilization of enzymes. The present study promotes a successful application of functionalized magnetic nanoparticles (MNPs) for stabilization of cellulase CLEAs. Catalytically active CLEAs and magnetic cross-linked enzyme aggregates (mCLEAs) of cellulase from Trichoderma reesei were prepared using glutaraldehyde (GA) as a cross-linking agent and the catalytic activity and stability of the CLEAs/mCLEAs were investigated. The influence of precipitation agents, cross-linker concentration, concentration of enzyme, addition of bovine serum albumin (BSA), and addition of sodium cyanoborohydride (NaBH3CN) on expressed activity and immobilization yield of CLEAs/mCLEAs was studied. Particularly, reducing the unsaturated Schiff's base to form irreversible linkages is important and improved the activity of CLEAs (86%) and mCLEAs (91%). For increased applicability of CLEAs/mCLEAs, we enhanced the activity and stability at mild biochemical process conditions. The reusability after 10 cycles of both CLEAs and mCLEAs was investigated, which retained 72% and 65% of the initial activity, respectively. The thermal stability of CLEAs and mCLEAs in comparison with the non-immobilized enzyme was obtained at 30 °C (145.65% and 188.7%, respectively) and 50 °C (185.1% and 141.4%, respectively). Kinetic parameters were determined for CLEAs and mCLEAs, and the KM constant was found at 0.055 ± 0.0102 mM and 0.037 ± 0.0012 mM, respectively. The maximum velocity rate (Vmax) was calculated as 1.12 ± 0.0012 µmol/min for CLEA and 1.17 ± 0.0023 µmol/min for mCLEA. Structural characterization was studied using XRD, SEM, and FT-IR. Catalytical properties of immobilized enzyme were improved with the addition of reducent NaBH3CN by enhancing the activity of CLEAs and with addition of functionalized aminosilane MNPs by enhancing the activity of mCLEAs.

Effect of Green Food Processing Technology on the Enzyme Activity in Spelt Flour.

In this research, a new approach to enzyme inactivation in flour was presented by supercritical technology, considered a sustainable technology with lower energy consumption compared to other technologies that use ultra-high temperature processing. Total protein concentration and the activity of enzymes α-amylase, lipase, peroxidase, polyphenol oxidase, and protease were determined in flour pre-treated with scCO2. During the study, it was observed that the activity of enzymes such as lipase and polyphenol oxidase, was significantly reduced under certain conditions of scCO2 treatment, while the enzymes α-amylase and protease show better stability. In particular, polyphenol oxidase was effectively inactivated below the 60% of preserved activity at 200 bar and 3 h, whereas α-amylase under the same conditions retained its activity. Additionally, the moisture content of the scCO2-treated spelt flour was reduced by 5%, and the fat content was reduced by 58%, while the quality of scCO2-treated flour was maintained. In this regard, the sustainable scCO2 process could be a valuable tool for controlling the enzymatic activity of spelt flour since the use of scCO2 technology has a positive effect on the quality of flour, which was verified by the baking performance of spelt flour with the baked spelt bread as an indicator of quality.

Synergistic Effect of Supercritical and Ultrasound-Assisted Ginger (Zingiber officinale Roscoe) Extracts.

Proper processing of natural material is crucial to obtain an extract with high content of biologically active components. Dried, grinded ginger roots were extracted by ultrasonic method and supercritical extraction with CO2. The aim of the study was to determine if a mixture of the two types of extracts attained by different methods and solvents exhibits better bioavailability than each extract itself. Therefore, both extracts were analytically evaluated and then mixed in a ratio of 1:1. The supercritical extract (SCG extract) and the mixed extract (mixG extract) had high antioxidant activity (78% and 73%) and total phenols (827 mg/g ext. and 1455 mg/g ext.), which is also consistent with the levels of gingerol (303 mg/g ext. and 271 g/g ext.) and shogaol (111 mg/g ext. and 100 g/g ext.) in the extracts. In comparison to both pure extracts higher levels of total phenols were found in the extract mixG. This could be the reason for the significant inhibition of melanoma cells and antimicrobial potential (against Staphylococcus aureus, Escherichia coli, and Candida albicans). The combination of the extracts resulted in a significant increase in the inhibition of selected microbial and melanoma cells WM-266-4 compared to the control. Cell viability decreased below 60% when mixG extract was applied. Antimicrobial activity has been confirmed.