Search Results (65)

To improve the bioavailability of yak by-products, novel antioxidant peptides were prepared and identified from yak skin hydrolysate. The results showed that the ultrafiltration fraction of a molecular weight of less than 1 kDa had the strongest free radical scavenging activity. A total of 219 novel peptides were identified by mass spectrometry and five antioxidant peptides were screened based on molecular docking with Keap1 (LMGPR, GFDGD, FGFDGDF, GHNGLDGL, and GPAGPQGPR). These peptides may bind with Keap1 competitively and exert antioxidant effects by activating the Nrf2/ARE pathway. After synthesis, FGFDGDF showed a better free radical scavenging ability and protective effect on H₂O₂-induced oxidative damage of HepG2 cells among these peptides. The pretreatment of peptides could enhance the activity of intracellular antioxidant enzymes and reduce the level of malondialdehyde and IL-8. This study provides a scientific basis for the application of yak skin peptide as a novel antioxidant in functional food. Full article

Background/Objectives: This study investigated whether enzymatic hydrolysis enhances the cognitive benefits of HongJam (steamed mature silkworms) and explored the underlying mechanisms. A marker compound of enzyme-treated HongJam was also identified to support quality control. Methods and Results: Mice were supplemented with Golden Silk HongJam (GS) or its enzyme hydrolysates (GS-EHS). Behavioral tests showed both improved fear-aggravated memory, with GS-EHS producing similar or greater effects at lower doses. GS-EHS activated the cyclic AMP response element binding protein/brain-derived neurotrophic factor signaling pathway and mitigated scopolamine-induced mitochondrial dysfunction by enhancing mitochondrial complex activity and ATP production. It also increased esterase activity, reduced reactive oxygen species, and modulated programmed cell death by suppressing apoptosis while promoting autophagy and unfolded protein response pathways. These changes led to reduced endoplasmic reticulum stress and neuroinflammation. Mass spectrometry identified glycine-tyrosine dipeptide as a potential bioactive marker. Conclusions: GS-EHS enhances cognitive function by improving mitochondrial activity, reducing oxidative stress, and regulating programmed cell death. Enzymatic hydrolysis appears to increase the bioavailability of active compounds, making GS-EHS effective at lower doses. The glycine–tyrosine dipeptide may serve as a marker compound for standardizing GS-EHS based on its cognitive-enhancing properties. Full article

Increasing the cell mass used as an inoculum is an effective strategy for enhancing productivity in alcoholic fermentation processes. In batch processes without cell recycling, such as those used in maize ethanol production, this objective can be achieved through two main approaches: (i) increasing the amount of commercially acquired dry cell mass or (ii) extending the propagation time. In this study, an economic assessment of both approaches was carried out, considering the Brazilian industrial context of maize ethanol production. Fermentation assays demonstrated that specific substrate consumption decreases with increasing initial cell concentration, following a hyperbolic model. This experimental behavior was used to simulate different operational scenarios and estimate productivity gains and economic impacts. The results showed that both strategies increase ethanol production and revenue, although the associated costs vary significantly. Based on this model, productivity and revenue gains were estimated for both approaches. The findings suggest that extending the propagation time is the most economically viable strategy to increase the initial cell concentration, even in scenarios where the plant lacks existing infrastructure and additional equipment investments are required. The analysis also accounted for operational costs associated with increased energy consumption during extended aeration time. Full article

The objective of this study was to determine the effects of growth-related myopathies, i.e., normal, wooden breast (WB), white striping (WS), and the combined lesions of WS and WB (WS + WB), on the molecular response of Caco-2 cells. A total of 24 cooked chicken breasts (n = 6 per myopathy) was subjected to an in vitro digestion using an enzymatic process mimicking human gastrointestinal digestion. Based on peptidomics, in vitro protein digestion of the abnormal samples, particularly WB meat, resulted in more peptides with lower molecular mass relative to those of normal samples. The cooked meat hydrolysates obtained at the end of the digestion were applied to a Caco-2 cell model for 4 h. The cell viability of treated normal and abnormal samples was not different (p ≥ 0.05). Absolute transcript abundances of genes associated with primary oxidative stress response, including nuclear factor erythroid 2 like 2, superoxide dismutase, and hypoxia-inducible factor 1 were determined using a droplet digital polymerase chain reaction. No significant differences in transcript abundance of those genes in Caco-2 cells were demonstrated between normal and the abnormal samples (p ≥ 0.05). Overall, the findings supported that, compared to normal meat, the cooked chicken meat with growth-related myopathies might be digested and absorbed to a greater extent. The cooked abnormal meat did not exert significant transcriptional impacts regarding oxidative stress on the human epithelial Caco-2 cells. Full article

Walnut milk residues (WMR) were investigated for the first time through their phenolic characterization including soluble (free, esterified, and etherified) phenolics and those released from their insoluble-bound form (insoluble-bound phenolic hydrolysates, IBPHs) and their antioxidant properties. Free phenolics were recovered and alkaline or acid hydrolysis were used to recover the remaining phenolic fractions. Total phenolic compounds (TPCs) and their antioxidant activity were analyzed by Folin–Ciocalteu, FRAP, and ORAC methods, respectively. Soluble phenolics (free + esterified + etherified fractions) showed a higher TPC (275.3 mg GAE 100 g⁻¹ dw) and antioxidant activity (FRAP: 138.13 µmol TE g⁻¹ dw; ORAC: 45.41 µmol TE g⁻¹ dw) with respect to the IBPH. There was a significant correlation between TPC and FRAP and ORAC values regardless of the fraction and tested sample. Phenolic acids and flavonoids were identified and quantified by ultra-performance liquid chromatography–electrospray tandem mass spectrometry (UPLC-ESI-MS/MS). Gallic acid, mainly in the free form (3061.0 µg 100 g⁻¹), was the most representative, followed by biochanin A, identified for the first time in a walnut product and mostly present in the fraction released from the esterified form (593.75 µg 100 g⁻¹). No detrimental cytotoxic impact on Caco-2 cells was observed. Hence, WMR could be considered a potential source for the development of nutraceutical and/or antioxidant food additives. Full article

Baker’s yeast is a key starting material for producing extracts with diverse compositions and applications. This study investigates the effect of pulsed electric field (PEF) pretreatment, which induces irreversible electropermeabilization, on the enzymatic hydrolysis of yeast. Cell suspensions were exposed to monopolar rectangular pulses in a continuous flow system followed by 4 h of incubation with Alcalase at concentrations of 0.2% and 0.5%. PEF pretreatment significantly improved enzymatic hydrolysis, with maximum intracellular content recovery under electrical conditions resulting in outlet temperatures of 56–58 °C. The released protein reached 163.7 ± 13 mg per gram of dry cell weight (DCW). SDS-PAGE analysis showed that the extracts predominantly contained peptides with molecular masses below 4.7 kDa. The phenolic content was comparable to that of cell lysates obtained after mechanical disruption. The free α-amino nitrogen content and total antioxidant activity reached 218.2 ± 26 mg/gDCW and 53.4 ± 4.6 mg TE/gDCW, respectively, representing 3.2-fold and 2.65-fold increases compared to cell lysates. The hydrolysates from PEF-pretreated cells demonstrated a positive effect on the proliferation of the human keratinocyte cell line HaCat. The obtained data lead to the conclusion that PEF pretreatment is a promising approach to enhance the production of yeast hydrolysates with various applications. Full article

Background/Objectives: When prooxidants outweigh antioxidants, oxidative stress can occur, causing an accumulation of reactive oxygen species (ROS). This process can lead to cellular damage and plays a role in the development of numerous health conditions. This study aimed to investigate the cytoprotective effects on differentiated Caco-2 cells of hydrolysates derived from the red Californian worm (WH) and their fractions, identify the peptides responsible for this effect, and elucidate the mechanisms involved. Methods: The WH was obtained through hydrolysis with Alcalase 2.4 L and subsequently fractionated to two fractions (F > 3 kDa and F < 3 kDa) using a ceramic membrane with a molecular weight cutoff of 3 kDa. The peptides found in the F < 3 kDa fraction, demonstrating the highest cytoprotective activity, were then sequenced via liquid chromatography-mass spectrometry analysis (LC-MS/MS), and molecular docking was conducted to elucidate the underlying antioxidant mechanisms. Results: The hydrolysate of Eisenia foetida and its F < 3 kDa fraction exhibited no cytotoxicity, protected the cells from H₂O₂-induced oxidative stress (50% increase viability), preserved cell viability by restoring their redox status (ROS: 20% decrease, and glutathione (GSH): recovered to basal control levels) and cell cycle distribution, and decreased apoptosis (16%). Twenty-eight peptides were identified, with five showing antioxidant activity through stable interactions with myeloperoxidase (MPO) and Kelch-like ECH-associated protein 1 (Keap-1), KPEDWDDR being the peptide that presented the highest affinity with both molecules (−7.9 and −8.8 kCal/mol, respectively). Conclusions: These results highlight the WH as a potential source of bioactive peptides for the management of oxidative stress. Full article

The production of inflammatory cytokines such as tumor necrosis factor (TNF)-α by activated macrophage cells plays an important role in the development of intestinal inflammation. The present study investigated the anti-inflammatory effect of the protein hydrolysates prepared from the jack bean (JBPHs), Canavalia ensiformis (L.) DC, using the enzyme Alcalase, in a murine macrophage model, RAW 264.7 cells, which were stimulated by lipopolysaccharides. JBPHs reduced the TNF-α expression at the protein and mRNA levels through the downregulation of cellular signaling pathways involved in nuclear factor kappa B (NF-κB), extracellular signal-regulated kinase (ERK), and p38. A combination of mass spectrometry and in silico approaches identified 10 potential anti-inflammatory peptides in the JBPHs, including LFLLP and DFFL. Interestingly, while LFLLP targeted the NF-κB pathway, DFFL targeted p38 and ERK to suppress the TNF-α production in the RAW 264.7 cells. In addition, LFLLP and DFFL were localized in the cytosol of the cells. These results demonstrated that LFLLP and DFFL were incorporated by RAW 264.7 cells and, at least in part, contributed to the reduction in TNF-α by JBPHs. These peptides isolated from JBPHs may well be utilized as new alternatives to alleviate intestinal inflammation. Full article

This paper introduces an enzymatic approach to estimate internal mass-transfer resistances during food digestion studies. Cellulase has been used to degrade starch cell walls (where cellulose is a significant component) and reduce the internal mass-transfer resistance, so that the starch granules are released and hydrolysed by amylase, increasing the starch hydrolysis rates, as a technique for measuring the internal mass-transfer resistance of cell walls. The estimated internal mass-transfer resistances for granular starch hydrolysis in a beaker and stirrer system for simulating the food digestion range from 2.2 × 10⁷ m⁻¹ s at a stirrer speed of 100 rpm to 6.6 × 10⁷ m⁻¹ s at 200 rpm. The reaction rate constants for cellulase-treated starch are about three to eight times as great as those for starch powder. The beaker and stirrer system provides an in vitro model to quantitatively understand external mass-transfer resistance and compare mass-transfer and reaction rate kinetics in starch hydrolysis during food digestion. Particle size analysis indicates that starch cell wall degradation reduces starch granule adhesion (compared with soaked starch samples), though the primary particle sizes are similar, and increases the interfacial surface area, reducing internal mass-transfer resistance and overall mass-transfer resistance. Dimensional analysis (such as the Damköhler numbers, Da, 0.3–0.5) from this in vitro system shows that mass-transfer rates are greater than reaction rates. At the same time, SEM (scanning electron microscopy) images of starch particles indicate significant morphology changes due to the cell wall degradation. Full article

Sacha inchi (Plukenetia volubilis L.) is an under-exploited crop with great potential due to its nutritional and medicinal characteristics. A Sacha inchi protein isolate (SII), obtained from defatted Sacha inchi flour (SIF), was hydrolyzed by Bioprotease LA 660 under specific conditions. The hydrolysates were characterized chemically, and their digestibility and antioxidant capacity were evaluated by in vitro cell-free experiments to select the hydrolysate with major antioxidant activity. Sacha inchi protein hydrolysate at 20 min (SIH20B) was selected, and the anti-inflammatory capacity was evaluated by RT-qPCR and ELISA techniques, using two different doses in monocytes THP-1 stimulated with lipopolysaccharide (LPS). The results obtained showed that the in vitro administration of SIH20B down-regulated the TNF-α gene and reduced the release of this cytokine, whereas the anti-inflammatory cytokines IL-10 and IL-4 were up-regulated in LPS-stimulated monocytes and co-administrated with SIH20B. The peptides contained in SIH20B were identified, and the 20 more relatively abundant peptides with a mass by 1 kDa were subjected to in silico analysis to hypothesize those that could be responsible for the bioactivity reported in the hydrolysate. From the identified peptides, the peptides AAGALKKFL and LGVKFKGGL, among others, are proposed as the most biologically actives. In conclusion, SIH20B is a novel, natural source of high-value-added biopeptides that could be used as an ingredient in formulations of food or nutraceutical compounds. Full article

Copper-chelated chitosan microgels were investigated as an immobilized metal affinity chromatography (IMAC) phase for peptide separation. The copper-crosslinked chitosan beads were shown to strongly interact with a range of amino acids, in a wide range of pH and saline conditions. The beads exhibited an affinity that seemed to depend on the isoelectric point of the amino acid, with the extent of uptake increasing with decreasing isoelectric point. This selective interaction with anionic amino acids resulted in a significant relative enrichment of the supernatant solution in cationic amino acids. The beads were then studied as a novel fractionation system for complex milk hydrolysates. The copper chitosan beads selectively removed larger peptides from the hydrolysate aqueous solution, yielding a solution relatively enriched in medium and smaller peptides, which was characterized both quantitatively and qualitatively by size exclusion chromatography (SEC). Liquid chromatography–mass spectrometry (LCMS) work provided comprehensive data on a peptide sequence level and showed that a depletion of the anionic peptides by the beads resulted in a relative enrichment of the cationic peptides in the supernatant solution. It could be concluded that after fractionation a dramatic relative enrichment in respect to small- and medium-sized cationic peptides in the solution, characteristics that have been linked to bioactivities, such as anti-microbial and cell-penetrating properties. The results demonstrate the use of the chitosan copper gel bead system in lab scale fractionation of complex hydrolysate mixtures, with the potential to enhance milk hydrolysate bioactivity. Full article

The production of olive oil has important economic repercussions in Mediterranean countries but also a considerable impact on the environment. This production generates enormous quantities of waste and by-products, which can be exploited as new raw materials to obtain innovative ingredients and therefore make the olive production more sustainable. In a previous study, we decided to foster olive seeds by generating two protein hydrolysates using food-grade enzymes, alcalase (AH) and papain (PH). These hydrolysates have shown, both in vitro and at the cellular level, antioxidant and antidiabetic activities, being able to inhibit the activity of the DPP-IV enzyme and modulate the secretion of GLP-1. Given the multifunctional behavior of peptides, both hydrolysates displayed dual hypocholesterolemic activity, inhibiting the activity of HMGCoAR and impairing the PPI of PCSK9/LDLR, with an IC₅₀ equal to 0.61 mg/mL and 0.31 mg/mL for AH and PH, respectively. Furthermore, both samples restored LDLR protein levels on the membrane of human hepatic HepG2 cells, increasing the uptake of LDL from the extracellular environment. Since intestinal bioavailability is a key component of bioactive peptides, the second objective of this work is to evaluate the capacity of AH and PH peptides to be transported by differentiated human intestinal Caco-2 cells. The peptides transported by intestinal cells have been analyzed using mass spectrometry analysis, identifying a mixture of stable peptides that may represent new ingredients with multifunctional qualities for the development of nutraceuticals and functional foods to delay the onset of metabolic syndrome, promoting the principles of environmental sustainability. Full article

This study explores the mechanical properties, as well as the water-reducing and setting delay mechanism, of a novel xylonic acid-based water reducer applied to cementitious materials. Four xylonic acid water reducers were synthesized in this study: XACa (PX) from pure xylose, XACa (HS) from hemicellulose hydrolysate, XANa (PX) from pure xylose, and XANa (HS) from hemicellulose hydrolysate. These were generated through the whole-cell catalysis of Gluconobacter oxydans bacteria, using pure xylose and hemicellulose hydrolysate as substrates. The findings indicate that the xylonic acid-based water reducer can attain a water-reducing capability between 14% and 16% when the dosage (expressed as a mass fraction of cement) is roughly 0.2%. In initial and final setting tests, XACa (PX) demonstrated a pronounced retarding influence at admixture levels below 0.15%, reaching its apex at 0.10%. This delayed the initial setting time by 76% and the final setting time by 136% relative to the control group. However, a slight pro-setting effect was noted beyond a 0.2% dosage. In the compressive and flexural tests of concrete, under the same slump, the XA group improved its mechanical properties by 5% to 10% compared to the SodiuM lignosulfonate (SL) group. In the air content and chloride ion migration resistance tests, the XA group reduced the air content by 38% compared to the SL group, but also increased the data of rapid chloride migration (D_RCM) by 16%. Characterization studies revealed that the carboxyl and hydroxyl groups in xylonic acid undergo chemisorption with the Si-O bonds on the surface of cement particles. These groups interact with the Si-O bonds on cement particles, contributing to water-reducing effects and delaying the setting process by impeding Ca²⁺ ion aggregation in the calcium-silicate-hydrate gel. Its significant water-reducing effect, adjustable setting time, and excellent mechanical and durability properties suggest its viability as an alternative to lignosulfonate series water-reducing agents. Full article

The production of carotenoids by microbial organisms has gained significant interest due to the growing demand for natural products. Among the non-model oleaginous red yeasts, Rhodotorula toruloides stands out as an appealing host for natural carotenoid production. R. toruloides possesses the natural ability to metabolize a wide range of substrates, including lignocellulosic hydrolysates, and convert them into lipids and carotenoids. In this study, we focused on utilizing xylose, the main component of hemicellulose, as the major substrate for R. toruloides. We conducted a comprehensive kinetic evaluation to examine the impact of aeration and agitation on carotenoid production. Results in stirred-tank reactor demonstrated that under milder conditions (300 rpm and 0.5 vvm), R. toruloides accumulated over 70% of its cell mass as lipids. Furthermore, the highest carotenoid yields were achieved at high agitation rates (700 rpm), with carotenoid levels reaching nearly 120 µg/mL. Several carotenoids were identified, including β-carotene, γ-carotene, torularhodin, and torulene, with β-carotene being the major carotenoid, accounting for up to 70% of the total carotenoid content. The carotenoid-rich extract produced by R. toruloides under evaluated conditions was successfully incorporated into soap formulations, demonstrating the addition of antioxidant properties. This work provides a comprehensive understanding of xylose conversion into natural carotenoids by R. toruloides, presenting a promising avenue for their application in cosmetics. Furthermore, this study highlights the potential of a renewable and cost-effective approach for carotenoid production in the soap industry. Full article

Meat and bone meal (MBM) is a product of the rendering industry, which is looking for high-value applications of rendered animal proteins (RAP). The objective of this research was to utilize MBM as a nitrogen source to produce astaxanthin (AX) by Xanthophyllomyces dendrorhous and quantify the bioavailability of MBM as a potential substitution of commercial nitrogen sources (i.e., yeast extract and peptone). To conduct yeast fermentation under the optimal glucose loading, the C/N ratio was optimized to achieve maximum AX content. MBM was hydrolyzed by using proteinase and alkaline (Ca(OH)₂) for 4, 8, and 16 h with different enzyme and alkaline loadings to produce MBM hydrolysates (MBMHs). The MBMHs were directly fermented by X. dendrorhous under the optimum glucose concentration. Experimentally, the optimum medium contained 40 g/L glucose, 5 g/L peptone, and 3 g/L yeast extract, where AX content of 3.69 mg/g dry cell mass was achieved. MBMHs were used by X. dendrorhous as a nitrogen source, while fermentation with lyophilized MBMHs was generated using proteinase K. This resulted in a maximum AX content of 1.58 mg/g dry cell mass. This research exhibits the feasibility of using MBM as a nitrogen source to produce AX with X. dendrorhous. Full article