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In the present work, methanolic extracts from thyme and dittany plants were prepared and characterized in terms of their polyphenolic content through analytical and spectrophotometric techniques. Rosmarinic acid, thymol and carvacrol were found to be the main components of the extracts, which were further biologically assessed for their antioxidant, anti-tyrosinase, anti-lipase and antibacterial activity against Gram-negative and Gram-positive bacteria. As found, thyme extracts exhibited superior antioxidant activity (SC50 at 33.9 μg mL−1), while dittany extracts inhibited the microbial growth to a great extent against Bacillus subtilis strain (MIC at 0.5 mg mL−1) and E. coli strain (MIC at 2 mg mL−1). Furthermore, the thyme extract was proven to strongly inhibit the activity of lipase from Candida rugosa (IC50 at 63.9 μg mL−1), comparable to the standard inhibitor orlistat, while its inhibitory effect against mushroom tyrosinase was weak. On the other hand, the dittany extract presented an inhibitory effect against the tested lipase (IC50 over 500 μg mL−1) and an activation effect against tyrosinase (at concentrations > 500 μg mL−1). Additionally, molecular docking studies of the main compounds of the extracts showed that rosmarinic acid plays a crucial role on the inhibitory activity of the extracts against lipase, while thymol has a stronger effect on inhibiting tyrosinase. Furthermore, both extracts were employed in the preparation of gelatin-deep eutectic solvent (DES) hydrogels that were further studied for their antioxidant and antibacterial activity. The results showed that the incorporation of the extracts offered antibacterial properties to the biopolymer-based hydrogels and enhanced the antioxidant activity of gelatin up to 85%. [ABSTRACT FROM AUTHOR]

This study aimed to explore the capacity of immobilized lipases on the acetylation of six aglycon flavonoids, namely myricetin, quercetin, luteolin, naringenin, fisetin and morin. For this purpose, lipase B from Candida antarctica (CaLB) and lipase from Thermomyces lanuginosus (TLL) were immobilized onto the surface of ZnOFe nanoparticles derived from an aqueous olive leaf extract. Various factors affecting the conversion of substrates and the formation of monoesterified and diesterified products, such as the amount of biocatalyst and the molar ratio of the substrates and reaction solvents were investigated. Both CaLB and TLL-ZnOFe achieved 100% conversion yield of naringenin to naringenin acetate after 72 h of reaction time, while TLL-ZnOFe achieved higher conversion yields of quercetin, morin and fisetin (73, 85 and 72% respectively). Notably, CaLB-ZnOFe displayed significantly lower conversion yields for morin compared with TLL-ZnOFe. Molecular docking analysis was used to elucidate this discrepancy, and it was revealed that the position of the hydroxyl groups of the B ring on morin introduced hindrances on the active site of CaLB. Finally, selected flavonoid esters showed significantly higher antimicrobial activity compared with the original compound. This work indicated that these lipase-based nanobiocatalysts can be successfully applied to produce lipophilic derivatives of aglycon flavonoids with improved antimicrobial activity. [ABSTRACT FROM AUTHOR]

Bioactive compounds that can be recovered by the solid wastes of the olive oil sector, such as polyphenols, are known for their significant antioxidant and antimicrobial activities with potential application in nutraceutical, cosmetic, and food industries. Given that industrial demands are growing, and the polyphenol market value is ever increasing, a systematic study on the recovery of natural antioxidant compounds from olive pomace using ultrasound-assisted extraction (UAE) was conducted. Single-factor parameters, i.e., the extraction solvent, time, and solid-to-liquid ratio, were investigated evaluating the total phenolic content (TPC) recovery and the antioxidant activity of the final extract. The acetone–water system (50% v/v, 20 min, 1:20 g mL−1) exhibited the highest total phenolic content recovery (168.8 ± 5.5 mg GAE per g of dry extract). The olive pomace extract (OPE) was further assessed for its antioxidant and antibacterial activities. In DPPH, ABTS, and CUPRAC, OPE exhibited an antioxidant capacity of 413.6 ± 1.9, 162.72 ± 3.36 and 384.9 ± 7.86 mg TE per g of dry extract, respectively. The antibacterial study showed that OPE attained a minimum inhibitory activity (MIC) of 2.5 mg mL−1 against E. coli and 10 mg mL−1 against B. subtilis. Hydroxytyrosol and tyrosol were identified as the major phenolic compounds of OPE. Furthermore, active chitosan–polyvinyl alcohol (CHT/PVA) films were prepared using different OPE loadings (0.01–0.1%, w/v). OPE-enriched films showed a dose-dependent antiradical scavenging activity reaching 85.7 ± 4.6% (ABTS) and inhibition growth up to 81% against B. subtilis compared to the control film. Increased UV light barrier ability was also observed for the films containing OPE. These results indicate that OPE is a valuable source of phenolic compounds with promising biological activities that can be exploited for developing multifunctional food packaging materials. [ABSTRACT FROM AUTHOR]

Light controls the developmental, physiological, morphological, and metabolic responses of many fungi. Most fungi respond primarily to blue, red, and green light through their respective photoreceptors. In this study, a screening of different light wavelengths' effects on submerged Pleurotus ostreatus cultivation in baffled flasks was conducted. P. ostreatus growth was not inhibited in all tested conditions, while an equal or higher protein content was observed in comparison with dark conditions. Red and green light favored exopolysaccharide (EPS) production while red and blue light favored intracellular polysaccharide (IPS) production. To focus on EPS production, the effect of red and green light wavelengths on the production of the polysaccharide via submerged cultivation of P. ostreatus LGAM 1123 was tested. Submerged cultivation using red light in baffled flasks resulted in EPS production of 4.1 ± 0.4 g/L and IPS content of 23.1 ± 1.4% of dry weight (dw), while green light resulted in EPS production of 4.1 ± 0.2 g/L and 44.8 ± 5.2% dw IPS content. Similar production levels were achieved in a 3.5 L bioreactor using red light. The EPS produced using red light revealed a polysaccharide with a higher antioxidant activity compared to the polysaccharides produced by green light. In addition, the analysis of the crude polysaccharides has shown differences in biochemical composition. The structural differences and β glucan's existence in the crude polysaccharides were confirmed by FT-IR analysis. Overall, these polysaccharides could be used in the food industry as they can enhance the functional health-promoting, physicochemical, and sensory properties of food products. [ABSTRACT FROM AUTHOR]

Nowadays, biosensors are gaining increasing interest in foods' and beverages' quality control, owing to their economic production, enhanced sensitivity, specificity, and faster analysis. In particular, colorimetric biosensors can be combined with color recognition applications on smartphones for the detection of analytes, rendering the whole procedure more applicable in everyday life. Herein, chitosan (CS) films were prepared with the deep eutectic solvent (DES) choline chloride/urea/glycerol (ChCl:U:Gly). Glucose oxidase (GOx), a widely utilized enzyme in quality control, was immobilized within CS films through glutaraldehyde (GA), leading to the formation of CS/GOx films. The optimized GOx concentration and DES content were determined for the films. Moreover, the effect of the pH and temperature of the glucose oxidation reaction on the enzymatic activity of GOx was studied. The structure, stability, and specificity of the CS/GOx films as well as the Km values of free and immobilized GOx were also determined. Finally, the analytical performance of the films was studied by using both a spectrophotometer and a color recognition application on a smartphone. The results demonstrated that the films were highly accurate, specific to glucose, and stable when stored at 4 °C for 4 weeks and when reused 10 times, without evident activity loss. Furthermore, the films displayed a good linear response range (0.1–0.8 mM) and a good limit of detection (LOD, 33 μM), thus being appropriate for the estimation of glucose concentration in real samples through a smartphone application. [ABSTRACT FROM AUTHOR]

Large quantities of wine lees are produced annually by the wine industry. The high phenolic content makes them unsuitable for disposal in the environment or animal feed without a suitable treatment. In this study, wine lees were treated by Pleurotus ostreatus in submerged cultivation, producing a high-value biomass and elevated levels of laccase, an important industrial enzyme. Biomass and laccase production reached 21 g/L and 74,000 Units/L, respectively, at the optimal conditions of initial pH 6.0, 20% v/v wine lees, 30 g/L glucose, and 20 g/L yeast extract, while decolorization and dephenolization rates of the waste were over 90%. The mycelial biomass was rich in proteins and essential amino acids reaching up to 43% and 16% per dry weight, respectively. Carbohydrates and lipids were the second richest bioactive compound in biomass, with values of 29.4 ± 2.7% and 29.5 ± 2.7%, respectively. The crude laccase in the culture supernatant was purified via a simple two-step purification procedure by 4.4-fold with a recovery of 44%. The molecular weight of the enzyme was determined to be 62 kDa via SDS electrophoresis. Enzyme activity was optimal at pH 5.0 and 70 °C. The activation energy of the enzyme was calculated at a value of 20.0 ± 0.2 kJ/mol. The pH stability and thermostability of the purified laccase were studied. The enzyme was remarkably stable at pH 8.0 and at temperatures up to 40 °C. The thermal inactivation energy of the enzyme was determined to be 76.0 ± 1.2 kJ/mol. The thermodynamic parameters (ΔH*, ΔG*, and ΔS*) for the thermal deactivation of the purified laccase at a temperature range of 20–60 °C were: 73.8 ≤ ΔH* ≤ 74.3 kJ·mol−1, 98.7 ≤ ΔG* ≤ 101.9 kJ·mol−1, and −90.5 ≤ ΔS* ≤ −84.3 J·mol−1·K−1. Wine lees could be ideal substrates of fungal cultivation for laccase production and biomass with a high protein content in an eco-friendlier way. [ABSTRACT FROM AUTHOR]

In this work, we investigated the effect of multi-walled carbon nanotubes (MWCNTs) and bio-graphene (bG) on the structure and activity of glucose oxidase (GOx), as well as on the performance of the respective electrochemical glucose biosensors. Various spectroscopic techniques were applied to evaluate conformational changes in GOx molecules induced by the presence of MWCNTs and bG. The results showed that MWCNTs induced changes in the flavin adenine dinucleotide (FAD) prosthetic group of GOx, and the tryptophan residues were exposed to a more hydrophobic environment. Moreover, MWCNTs caused protein unfolding and conversion of α-helix to β-sheet structure, whereas bG did not affect the secondary and tertiary structure of GOx. The effect of the structural changes was mirrored by a decrease in the activity of GOx (7%) in the presence of MWCNTs, whereas the enzyme preserved its activity in the presence of bG. The beneficial properties of bG over MWCNTs on GOx activity were further supported by electrochemical data at two glucose biosensors based on GOx entrapped in chitosan gel in the presence of bG or MWCNTs. bG-based biosensors exhibited a 1.33-fold increased sensitivity and improved reproducibility for determining glucose over the sweat-relevant concentration range of glucose. [ABSTRACT FROM AUTHOR]

The current study proposes an efficient coating methodology for the development of low-density polyethylene (LDPE) films with prolonged bioactivity for food packaging applications. Three natural phenolic-based substances were incorporated at optimized concentrations in methyl-cellulose-based solutions and used as coatings on LDPE films. The amount of surfactant/emulsifier was optimized to control the entrapment of the bioactive substances, minimizing the loss of the substances during processing, and offering prolonged bioactivity. As a result, the growth of Escherichia coli was substantially inhibited after interaction with the coated films, while coated films presented excellent antioxidant activities and maintained their mechanical performance after coating. Considerable bioactivity was observed after up to 7 days of storage in sealed bags in the case of carvacrol- and thymol-coated films. Interestingly, films coated with olive-leaf extract maintained a high level of antimicrobial and antioxidant properties, at least for 40 days of storage. [ABSTRACT FROM AUTHOR]