This study investigated the impact of alcalase hydrolysis (10–180 min) on the structural and functional properties of millet prolamin (MP). Alcalase exhibited exceptional proteolytic efficiency, achieving a hydrolysis degree of 27.69% at 180 min, outperforming trypsin, α-chymotrypsin, and papain. Tricine-SDS-PAGE and SEM confirmed that alcalase hydrolyzed MP aggregates into small peptides (≤14 kDa) and disrupted its spherical structure. The millet prolamin hydrolysates (MPHs) showed markedly improved solubility (96.82% at 180 min), with surface hydrophobicity peaking at 30 min. MPHs also exhibited enhanced emulsifying and foaming properties, with emulsifying indices peaking at 120 min (30.14 m2/g, 98.06 min) and foaming capacity and stability peaking at 30 min (147.50%) and 120 min (86.23%), respectively. Antioxidant activity was significantly elevated, with DPPH scavenging activity nearly doubled within 10 min, ABTS scavenging activity peaking at 76.43% at 60 min, and reducing power increased. Intrinsic fluorescence, UV–Vis, and FTIR spectroscopy confirmed protein conformational transitions through increased exposure of hydrophobic and aromatic residues and changes in amide bands. These findings demonstrate alcalase hydrolysis is an efficient strategy for producing bioactive peptides from MP, supporting its potential as a versatile, high-solubility functional ingredient in plant-based formulations, particularly in the development of food emulsions and antioxidant-rich dietary supplements.

COVID-19 is an infectious disease resulting from the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which emerged as a global pandemic in 2020. Virus-like particles (VLP) are artificially generated nanoparticles composed of a subset of viral components that closely mimic the native virus in size, structure, and surface composition. These features contribute to their robust immunogenic potential, stimulating both antibody-mediated and cell-mediated immune responses. This study aimed to evaluate the expression of SARS-CoV-2 structural proteins and the VLP production using a baculovirus–insect cell system. Two monocistronic recombinant baculoviruses, each carrying either the spike (S) or nucleocapsid (N) gene of SARS-CoV-2, were employed in two different culture systems: Schott flasks and a stirred-tank bioreactor. The Sf9 cells showed substantial differences between the Schott flask and stirred tank bioreactor in terms of metabolism, virus titer, and cell death after infection. Both proteins’ expressions were confirmed in the two culture systems assessed. However, VLP formation (69.1–78.2 nm) was only confirmed in the co-infection setup with both monocistronic recombinant baculoviruses. In contrast, infection with only the S-protein baculovirus failed to produce VLP in our findings.

The unicellular green microalga Dunaliella salina is well known for being one of the best natural sources of β-carotene, a potent antioxidant and essential precursor to vitamin A. β-carotene is vital for human health because it improves immune system performance, supports vision, and lowers the risk of a number of chronic illnesses, including some forms of cancer. So far, a number of methods to increase β-carotene production in D. salina, including genetic engineering, applying abiotic stressors, plant growth regulators, and using metal nanoparticles. In this study, we examined the synergistic effects of plant growth regulators on the production of β-carotene and biomass accumulation in D. salina during various growth phases. Among the phytohormones examined, methyl jasmonate (MeJA) and indole acetic acid (IAA) were discovered to have important physiological functions in controlling the microalga’s growth and metabolism. We maximized productivity by optimizing these phytohormone concentrations using response surface methodology (RSM) with a central composite design (CCD). Our findings showed that when applied during the combination of 3 mg L−1 IAA implemented at non carotenogenic phase (the 0th day of cultivation), and 200 μM MeJA implemented on carotenogenic phase (20th day of the cultivation) significantly increased growth and carotenoid accumulation, yielding 5.2 g/L of biomass and a 2027.96 μg of β-carotene content for every 100 mg of dry cell weight.

This review provides a comprehensive overview of Litsea cubeba essential oil (EO), covering its extraction methods, compositional variation, bioactivities, and applications. It systematically examines both traditional (e.g., hydrodistillation, steam distillation) and emerging extraction techniques (e.g., ultrasound-, microwave-, enzyme-, and deep eutectic solvent-assisted methods), highlighting their respective advantages and limitations. A key focus is placed on how multiple factors—including geographical origin, harvest season, plant part, and extraction method—interact to influence the EO chemical profile and yield, which in turn determine its bioactivities. The growing market demand across food, pharmaceutical, daily chemical, and agrochemical sectors is discussed, with particular emphasis on how bioactivity-driven mechanisms (e.g., antioxidant, antimicrobial, and anti-inflammatory effects) underpin its diverse applications. While novel techniques offer improved efficiency, the review critically notes that methods using organic solvents or supercritical fluids may overestimate yield by co-extracting nonvolatiles. Moreover, a broader evaluation framework incorporating extraction efficiency, economic viability, and environmental impact is proposed. To promote resource-efficient utilization, a cascaded strategy—such as recovering kernel oils for biodiesel after EO distillation—is recommended for sustainable development. Finally, future interdisciplinary research should prioritize elucidating bioactivity mechanisms including synergistic/antagonistic interactions among EO components, enhancing formulation stability, and advancing high-purity citral separation technologies for value-added applications.

Haptoglobin is a major plasma glycoprotein and is widely utilized as a clinically relevant biomarker. However, we found that conventional single-step immunoaffinity purification using mono- or polyclonal anti-haptoglobin antibodies consistently resulted in the unintended co-elution. Through in-gel digestion and LC–MS/MS, these contaminants were identified as IgM, whose abundant high-mannose glycans might interfere with haptoglobin glycan profiling. In this study, we established a two-step purification strategy to obtain highly pure haptoglobin from human serum without immunoglobulin contamination. In the first purification step, immunoglobulins were selectively depleted via thiophilic adsorption, after which haptoglobin was isolated through an anti-haptoglobin affinity column. The resulting eluates were examined by CBB staining, western blotting, and lectin blotting, confirming that haptoglobin was the sole major component. Binding assays with hemoglobin confirmed that the purified haptoglobin preserved its intrinsic ligand-binding characteristics. Furthermore, LC–MS/MS analysis revealed no significant differences in overall glycan structures between the one-step and two-step purification methods, except for the absence of high-mannose–type glycans in the two-step–purified haptoglobin. Therefore, this two-step purification method provides a reliable approach for preparing functionally intact and immunoglobulin-free haptoglobin for biomedical and glycoproteomic studies.

The optimal hot water extraction process for a polysaccharide from Euphorbia trigona Mill. was established using single-factor and orthogonal experiments, yielding the best conditions of a 1:30 (g/mL) solid-to-liquid ratio at 90 °C for 3.5 h, with a polysaccharide yield of 5.89%. Sulfated (ETM-S) and phosphorylated (ETM-P) derivatives were successfully prepared, with degree of substitution of 0.32 and 0.48, respectively. Structural analysis confirmed the polysaccharides were acidic with complex monosaccharide compositions. Bioactivity assessments revealed that both modifications markedly improved functional properties. ETM-P exhibited significantly enhanced antioxidant activity, with IC50 values of 2.16, 1.57, and 1.79 mg/mL against DPPH, ·OH, and O2·–, respectively. Sulfation boosted inhibitory effects on α-amylase and α-glucosidase, suggesting hypoglycemic potential, and also increased pancreatic lipase inhibition, indicating a synergistic role in regulating glucose and lipid metabolism. Furthermore, ETM-S showed the strongest in vitro cytotoxicity against HepG2 cells, achieving an inhibition rate of 45.00% at 800 μg/mL, outperforming ETM-P (33.37% at 400 μg/mL) and the native polysaccharide (27.92% at 800 μg/mL). Both modified polysaccharides promoted RAW264.7 macrophage proliferation, enhanced phagocytosis, and stimulated NO release, demonstrating immunomodulatory capacity. In summary, chemical modification, especially sulfation, effectively enhances the bioactivities of polysaccharides, showing promise for regulating glycolipid metabolism and immune modulation.

The co-valorization of lactose and proteins from dairy by-products offers both environmental and technological opportunities. In this study, bifunctional biocatalysts were developed by co-immobilizing β-galactosidase and Neutrase® onto glutaraldehyde-activated chitosan for the simultaneous hydrolysis of lactose and casein. Three co-immobilization strategies were examined, and the sequential configuration in which Neutrase® was immobilized prior to β-galactosidase (D3) exhibited the highest retained activities and the most pronounced thermal stabilization for both enzymes. The D3 derivative achieved immobilization yields of 76% for both enzymes and retained approximately 60–70% of its initial activity after five reuse cycles. Thermal inactivation analysis revealed a sixfold increase in Neutrase® stability (SF = 18.2) and a corresponding rise in activation free energy from 95.9 to 103.9 kJ.mol−1, indicating increased resistance to thermal denaturation. Kinetic modeling showed that β-galactosidase exhibited a fourfold increase in apparent reaction rate constant (k2 = 2.05 × 10−2 L.g−1.min−1), indicating that enzyme co-localization enhanced the apparent catalytic rate, while proteolytic activity was moderately affected by diffusional constraints (α = 0.40). The integrated kinetic and thermodynamic analysis indicated that co-immobilization can be associated with structural stabilization effects and interaction phenomena within the chitosan matrix. These findings support a sustainable and economically feasible approach for whey valorization.

Rhizophus oryzae biomass (RB) can be incorporated into polylactic acid (PLA) to create improved biocomposites that perfectly fit the biobased circular economy model. In this study, Rhizopus oryzae EGE-F75, produced through solid-state fermentation in aluminum trays with the maximum RB production conditions (Initial moisture (%): 79.84%, Inoculum ratio (%): 28.01, Incubation time (days): 9.74) was used as reinforcement material in PLA biocomposites. Biocomposites were obtained by adding varying amounts of RB (5% and 10% by weight) to PLA using a twin screw extruder, and the physical, thermal, mechanical and morphological properties of the biocomposites were investigated. According to the results, the incorporation of RB resulted in a decrease in mechanical properties. The tensile strength of 10% RB/PLA composite decreased by 23%, while the flexural modulus value of the composite increased by 6% compared to PLA. Furthermore, the addition of RB to PLA delayed the thermal decomposition of PLA and reduced its thermal conductivity. Introducing 5 wt% of RB to the PLA matrix reduced the thermal conductivity of the PLA matrix by 79%. Therefore, RB may not serve as an ideal filler for enhancing the mechanical properties and thermal stability of biocomposites, but it does offer potential as an alternative material for thermal insulation.

In this study, iron nanoparticles (YBFeNPs) were green-synthesized using the aqueous extract (YB-E) of wild cherry laurel (Prunus laurocerasus L.) fruit to evaluate their potential applications in food safety. The synthesized YBFeNPs were investigated through Fourier-transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, transmission electron microscopy, and energy-dispersive X-ray spectroscopy analyses. YBFeNPs showed higher total phenolic content, total flavonoid content, and total antioxidant capacity compared to the original extract, suggesting that plant metabolites enhance NP bioactivity. Antioxidant capacity was further validated by DPPH, ABTS, and FRAP assays, which demonstrated strong radical scavenging and reducing power. The antibacterial activity of YBFeNPs was assessed against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Enterococcus faecalis, revealing concentration-dependent effects with the highest inhibition zone of 14.5 ± 0.4 mm for E. coli. The chemical composition of the YB-E extract was analyzed using liquid chromatography-tandem mass spectrometry, identifying quinic acid, a polyol, and phenolic compounds such as chlorogenic acid (1.45 mg/g) and naringenin (0.02 mg/g). Additionally, the adsorption capacities of YBFeNPs were investigated against patulin and hydroxymethylfurfural (HMF), toxic compounds posing significant threats to food safety. The NPs demonstrated effective adsorption of both patulin and HMF, achieving maximum adsorption rates of 61.05% and 46.12%, respectively, at a concentration of 5.0 g/L after 240 min.

Wheat starch, the primary carbohydrate reserve in wheat grain, plays a crucial role in food formulation and industrial applications due to its functional versatility. This review provides a comprehensive overview of recent advances in wheat starch research, focusing on its isolation, modification, characterization, and patent trends. Various isolation methods, including wet milling, dry milling, and centrifugation, are discussed in terms of yield, purity, and suitability for downstream processing. Modification strategies—physical, chemical, and enzymatic—are examined for their ability to enhance thermal stability, solubility, pasting behavior, and functional properties. Characterization techniques, such as scanning electron microscopy, X-ray diffraction, differential scanning calorimetry, and rheological measurements, are highlighted as essential tools for understanding structural and functional changes. In addition, an analysis of recent patents reveals growing innovation in wheat starch isolation, modification, and application, reflecting industrial demand for tailored starch-based materials. This review serves as a valuable resource for food scientists and technologists, offering guidance for selecting suitable processing and analytical methods. Future research should emphasize sustainable processing, energy-efficient technologies, and novel applications that leverage wheat starch’s renewable and biodegradable nature.