Peptidomics profiling and biological activities of grape pomace protein hydrolysates.

Grape pomace protein isolate was hydrolysed by Alcalase, Flavourzyme and Protease either individually or in combination to produce hydrolysates with antihypertensive and antimicrobial properties. The degree of hydrolysis (DH) ranged between 22 and 52 % for Protease and Flavourzyme, respectively. Among all treatments, hydrolysates prepared using Flavourzyme exhibited the highest angiotensin-converting enzyme inhibitory (ACEi) activity, with an IC50 value of 91 µg/mL. The peptidomics analysis revealed that the peptides identified in Flavourzyme hydrolysate presented molecular features compatible with its bioactivity, like a high density of ACEi sequences per peptide. The hydrolysates were also able to inhibit the growth of Escherichia coli in a range between 9 and 54 % for Alcalase and Alcalase + Flavourzyme, respectively. Peptides in the most active hydrolysate evidenced a high occurrence of proline residues, which is a structural feature of some antimicrobial peptides.

Structure, antioxidant activity, and neuroprotective effect of black soybean (Glycine max (L.) merr.) protein hydrolysates.

The potential biological properties of protein hydrolysates have generated considerable research interest. This study was to hydrolyze black soybean protein (BSP) using five different commercial enzymes, and elucidate the influence of these enzymes on the structure and biological activities of the resulting hydrolysates. Enzymatic treatment changed secondary and tertiary structures of BSP, decreased particle size, α-helix and ß-sheet. Alcalase hydrolysate had the highest hydrolytic degree (29.84 %), absolute zeta potential (38.43 mV), the smallest particle (149.87 nm) and molecular weight (<3 kDa). In silico revealed alcalase hydrolysate had the strongest antioxidant potential. This finding was further validated through the lowest IC50 (mg/mL) in DPPH (2.67), ABTS (0.82), Fe2+ chelating (1.33) and·OH (1.12). Moreover, cellular antioxidant assays showed alcalase hydrolysate had the strongest cytoprotective effects on H2O2-induced PC12 cells. These results suggest BSPEHs, especially those prepared by alcalase, have potential as bioactive ingredients for nutrition, healthcare and food industry.

Enzymatic hydrolysis pretreatment combined with glycosylation for soybean protein isolate applying in dual-protein yogurt.

This research investigated the viability of replacing milk protein with glycosylated soybean protein isolate (SPI) treated with different enzymatic hydrolysis times (0, 10, 20, 30, 40, and 50 min) in yogurt. The results showed that enzymatic hydrolysis pretreatment combined with glycosylation for SPI exhibited elevated grafting and solubility. Additionally, the high solubility of SPI (94.77 %) at 40 min facilitates the preparation of dual-protein yogurt (DPY). Compared to ESPI0-G, DPY that incorporates ESPI40-G through partial substitution of milk protein is capable of forming a denser and more stable gel matrix. Especially, the syneresis of DPY40 was reduced by 7.61 % compared to DPY0, which more closely approximates the texture properties of traditional yogurt. Meanwhile, glycosylated SPI treated with enzymatic hydrolysis can effectively degrade the beany flavor and slightly bitter taste in DPY. This study could provide a solid theoretical basis for the broader application and industrialization of plant-based yogurt.

Study on the effect of phenoxyethanol-citric acid pretreatment for the enzymatic hydrolysis of bamboo residues.

This study investigated the biphasic phenoxyethanol-citric acid (PECA) pretreatment for bamboo residues (BRs) and its corresponding effects on the enzymatic hydrolysis performance. It is found that increasing the concentration of citric acid in the pretreatment system from 2.5% to 15% greatly enhanced the delignification and xylan removal for BRs. Consequently, the enzymatic hydrolysis yield of pretreated BRs significantly enhanced, increasing from 12.4% to 58.2% and 28.0%72.4% when the concentration of citric acid was increased from 2.5% to 15.0% at 160°C and 170°C, respectively. The characterization results from cellulose crystallinity, accessibility, and hydrophobicity of pretreated bamboo residues indicated that their changes possessed a beneficial performance on the enzymatic hydrolysis yield, which could result from the synergistic removal of lignin and xylan. The Chrastil model analysis showed that pretreatment at higher conditions resulted in the pretreated BRs possessing weaker diffusion resistance for cellulase, which is attributed to its higher enzymatic hydrolysis yield.

On-site cellulase production by Trichoderma reesei RutC-30 to enhance the enzymatic saccharification of ball-milled corn stover.

Cellulases are essential for the enzymatic saccharification of lignocellulose. They play a crucial role in breaking down the structure of lignocellulose to obtain fermentable sugars. In this study, we conducted on-site cellulase production by Trichoderma reesei RutC-30 through submerged fermentation. The effects of carbon source, nitrogen source, KH2PO4, and mineral elements on cellulase production were evaluated using the hydrolyzed total sugar concentration of ball-milled corn stover as an indicator. The optimal fermentation medium conditions for cellulase production were determined through orthogonal experimental design analysis. Additionally, by optimizing culture conditions, including inoculation, pH, and bottling volume, we achieved a total sugar concentration of 92.25 g/L. After the optimization, the FPA, CMCA, protein, and total sugar concentration increased by 75.49 %, 18.43 %, 89.71 %, and 17.83 %, respectively. Furthermore, corn stover pretreated by different methods was applied to induce cellulase production. Ball-milled and steam-exploded corn stover was identified as suitable incubation carbon sources with total sugar concentration up to 94.31 g/L. Our work exploits the cellulase induced by lignocellulose and then applies it to lignocellulose, enabling the customization and providing a reference for the production of cellulase with corn stover as an inducer.

Integrative approach to assessing bioactivity from hempseed protein isolate extracted and dehydrated by different methods: Synergising in silico prediction and in vitro validation.

This study demonstrated a comprehensive workflow combining in silico screening and prediction with in vitro validation to investigate the bioactivity of hempseed protein isolate (HPI) extracted and dehydrated using different methods. By adopting an in silico approach, 13 major proteins of HPI were hydrolysed by 20 selected enzymes, leading to the prediction of 20 potential bioactivities. With papain hydrolysis, dipeptidyl peptidase-IV (DPP4) and angiotensin-converting enzyme (ACE) inhibitory activities emerged as having the highest potential. In vitro experiments confirmed these predictions, with DPP4 and ACE inhibitory activities displaying IC50 values of 0.32-0.42 mg/mL and 6.8-9.17 µg/mL, respectively. A strong correlation (r2 = 0.96) was observed between in vitro protein inhibitory results and in silico predicted data. This study demonstrated an effective integrative approach for predicting bioactive peptides in food protein, providing valuable guidance on its processing to create value-added products.

Bioprocess to produce biostimulants/biofertilizers based on microalgae grown using piggery wastewater as nutrient source.

In the present work, two downstream processes - high-pressure homogenization at 100 (HPH-100) and 1200 bar (HPH-1200), and enzymatic hydrolysis (EH) - were tested to produce biostimulant extracts from Tetradesmus obliquus grown in piggery wastewater at two concentrations (12.8 and 88.3 g/L). Extracts before and after centrifugation (C) were evaluated in four bioassays using garden cress (germination), mung bean (auxin-like activity), and cucumber (auxin- and cytokinin-like activity) relative to distilled water. The initial microalgal culture, without any treatment, had the best germination results (162 % at 0.2 g/L) and the only one that showed cytokinin-like activity (141 % at 0.5 g/L). In both auxin-like bioassays, the HPH-1200 + C and EH + C originated high values (186 and 155 % for cucumber, 290 and 285 % for mung bean, respectively). For mung bean, the HPH-1200 achieved the highest auxin-like effect (378 %). Finally, the extracted biomass contained essential nutrients for biofertilization, complementing the biostimulant extracts for sustainable agriculture application.

High-solids saccharification of non-pretreated citrus peels through tailored cellulase.

Citrus peels, characterized by their low lignin and high sugar content, have been drawing increasing attention as a valuable lignocellulosic biomass with significant potential in biorefinery. Notably, in this study, the citrus waste was found to be enzymatically accessible without any pretreatment. Moreover, to promote the high-solids saccharification of the citrus peels, a tailored cellulase cocktail was formulated by response surface methodology (RSM), along with a fed-batch strategy aiming to obtain a high substrate loading. The study resulted in an optimized cellulase cocktail (7.08 U/g DM of ß-glucosidase, 164.17 U/g DM of hemicellulase, 47.38 mg/g DM of sophorolipid, and 64.68 mg/g DM of Tween 80) and achieved solids loading of 22 % with a total sugar concentration of 123.84 g/L, corresponding to a yield of 93.12 % (65.28 % in batch operation). These findings provided essential validation for the efficient utilization of citrus waste, ensuring them promising potential as feedstock for sugar platforms.

[Development of selective method for identifying mebeverine in hair]. / Razrabotka selektivnoi metodiki opredeleniya mebeverina v volosakh.

OBJECTIVE: To develop the method for identification of mebeverine and its metabolites in biological materials (urine and hair) for solution of the problem of possible cross-reaction with psychoactive substances during preliminary studies by immunochromatographic method. MATERIAL AND METHODS: The study was carried out using «Mebeverine¼ tablets (NAO «Severnaya Zvezda¼, Russia), enzymes, namely trypsin, chymotrypsin and hyaluronidase, papain. The obtained extracts were analyzed using HELC-MS/MS on a modular liquid Nexera XR chromatograph with a tandem LCMS-8050 (Shimadzu) mass spectrometer. Guinea pigs weighing about 500 gr were used in modelling the long-term use of mebeverine. The drug was administered per os, once to 5 guinea pigs at a dosage of 200 mg calculated as an animal weighing 500 gr and daily urine was collected. The wool was collected 30 days after regular administration. RESULTS AND CONCLUSION: The main mebeverine's metabolites have been determined and native mebeverine and its marker metabolites (mebeverine and veratric acids) have been found in urine while taking this medicine in therapeutic doses for medical purposes. The application of enzymatic hydrolysis of hair method allowed to establish that hair tissue accumulates only a native molecule of mebeverine, therefore, only by results of hair analysis it is possible to conclude, if there was pharmacological use of this medicine or psychoactive substances from the group of phenylalkilamines. The effectiveness of enzymatic hydrolysis by hyaluronidase to isolate mebeverine from hair has been shown.

Eco-friendly technologies for obtaining antioxidant compounds and protein hydrolysates from edible insect Tenebrio molitor beetles.

The functional properties of edible insects can be explored by a joint use of novel technologies. This work applied varied pre-treatments (ultra-sound-assisted extraction, UAE; microwave-assisted extraction, MAE; temperature-assisted extraction, TAE; CO2-assisted extraction) and solvents (water, ethanol, water:ethanol) in Tenebrio molitor beetles to enhance the extraction of phenolic compounds with antioxidant activity. An enzymatic hydrolysis (EH) was performed in wet and treated biomasses to determine the protein hydrolysis. Higher phenolic compounds and antioxidant activity was released after MAE using water as solvent compared to the other treatments and solvents. Treatments decreased 32 %, 19 % and 30 % the protein, chitin and lipids content. EH improved protein and amino acids hydrolysis in the MAE-treated insects, followed by UAE and TAE treatments. In conclusion, MAE was the most effective to release phenolics and antioxidant activity from T. molitor beetles, while using MAE followed by EH improved protein and amino acids hydrolysis, envisioning valuable applications for this insect biomass.

The Potential of Glucosinolates and Their Hydrolysis Products as Inhibitors of Cytokine Storms.

A cytokine storm is an intense inflammatory response characterized by the overproduction of proinflammatory cytokines, resulting in tissue damage, and organ dysfunction. Cytokines play a crucial role in various conditions, such as coronavirus disease, in which the immune system becomes overactive and releases excessive levels of cytokines, including interleukins, tumor necrosis factor-alpha (TNF-α), and interferon-gamma (IFN-γ). This anomalous response often leads to acute respiratory distress syndrome (ARDS), disseminated intravascular coagulation (DIC), and multiple organ injury (MOI). Glucosinolates are plant secondary metabolites predominantly found in Brassica vegetables, but are also present in other species, such as Moringa Adens and Carica papaya L. When catalyzed by the enzyme myrosinase, glucosinolates produce valuable products, including sulforaphane, phenethyl isothiocyanate, 6-(methylsulfinyl) hexyl isothiocyanate, erucin, goitrin, and moringin. These hydrolyzed products regulate proinflammatory cytokine production by inhibiting the nuclear factor kappa-light-chain-enhancer of activated B-cell (NF-κB) signaling pathway and stimulating the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. This action can alleviate hyperinflammation in infected cells and modulate cytokine storms. In this review, we aimed to examine the potential role of glucosinolates in modulating cytokine storms and reducing inflammation in various conditions, such as coronavirus disease. Overall, we found that glucosinolates and their hydrolysis products can potentially attenuate cytokine production and the onset of cytokine storms in diseased cells. In summary, glucosinolates could be beneficial in regulating cytokine production and preventing complications related to cytokine storms.

Enhancement of hydrogen production in dark fermentation of corn stover hydrolysates through composite electric field pretreatment: Improving enzymatic efficiency and regulating microbial metabolic balance.

This study investigates the effects of composite electric field pretreatment (CEP) on hydrogen production from corn stover enzymatic hydrolysates in dark fermentation. The findings reveal that under optimal conditions, the CEP group achieved a cumulative hydrogen yield of 77.3  ±â€¯â€¯2.6 mL/g TS, marking a 55.3 % increase compared to the control group without the electric field. CEP significantly enhances the fermentation capacity of enzymatic hydrolysates during the acid-stage of dark fermentation by disrupting the lignin structure and optimizing cellulase hydrolysis efficiency. Additionally, pH self-regulation is facilitated through the interaction between volatile fatty acids (VFAs) and ammonia nitrogen. Microbial community analysis revealed that CEP shifts the metabolic balance between Clostridium_sensu_stricto_1 and Lactococcus, leading to increased hydrogen yield and concentration during acid fermentation. Notably, Terrisporobacter exhibited a superior acid-producing capability compared to Bacteroides during the dark fermentation of enzymatic hydrolysates. This study provides a new perspective for the practical application of corn stover.

Optimizing ß-Lactoglobulin antigenicity through single enzyme hydrolysis: Exploring structural changes and effects on linear epitopes.

ß-lactoglobulin (ß-LG) is the major allergen in dairy products, but research on the optimal conditions for antigen reduction in ß-LG using different enzymes remains limited. Therefore, this study aims to investigate the antigenicity, structural characteristics, and peptide distribution of advantageous protease hydrolysates capable of eliminating the allergenic epitopes of ß-LG selected via bioinformatics tools. The results showed that under optimal enzymatic hydrolysis conditions, the antigen reduction rates for the four advantageous proteases acting on ß-LG were 47.37 % (pepsin), 33.54 % (chymotrypsin A), 38.71 % (papain), and 45.91 % (stem bromelain), respectively. The four proteases effectively degraded ß-LG, causing shorter peptide chain formation, reduced content of highly ordered α-helix, decreased fluorescence intensity, and lower surface hydrophobicity. Furthermore, they cleaved the linear epitopes of ß-LG into peptides of varying sizes, leading to different antigen reduction rates among the hydrolysates. These findings provide a theoretical basis for developing targeted enzymatic hydrolysis technologies and low-allergenicity dairy-based materials.

Maximizing microbial activity and synergistic interaction to boost biofuel production from lignocellulosic biomass.

Addressing global environmental challenges and meeting the escalating energy demands stand as two pivotal issues in the current landscape. Lignocellulosic biomass emerges as a promising renewable bio-energy source capable of fulfilling the world's energy requirements on a large scale. One of the most important steps in lowering reliance on fossil fuel and lessening environmental effect is turning lignocellulosic biomass into biofuel. As carbon-neutral substitutes for traditional fuel, biofuel offer a solution to environmental concerns compared to conventional fuel. Effective utilization of lignocellulosic biomass is imperative for sustainable development. Ongoing research focuses on exploring the potential of various microorganisms and their co-interactions to synthesize diverse biofuels from different starting materials, including lignocellulosic biomass. Co-culture techniques demonstrate resilience to nutrient scarcity and environmental fluctuations. By utilising a variety of carbon sources, microbes can enhance their adaptability to environmental stressors and potentially increase productivity through their symbiotic interactions. Furthermore, compared to single organism involvement, co-interactions allow faster execution of multistep processes. Lignocellulosic biomass serves as a primary substrate for pre-treatment, fermentation, and enzymatic hydrolysis processes. This review primarily delves into the pretreatment, enzymatic hydrolysis process and the biochemical pathways involved in converting lignocellulosic biomass into bioenergy.

Fractionation of lignin and fermentable sugars from wheat straw using an alkaline hydrogen peroxide/pentanol biphasic pretreatment.

To breakthrough the delignification saturation point (DSP) of alkaline hydrogen peroxide (AHP) pretreatment, a biphasic AHP/pentanol (AHPP) pretreatment was proposed in this work. The temperature and H2O2 concentration were evaluated. Under the optimal conditions (110 °C, 2h, 4% H2O2), 70.73% of lignin was removed, which was increased by 11.65% than the traditional AHP pretreatment, indicating successful overcoming of the DSP by adding pentanol. 85.74% and 88.62% of glucan and xylan digestibility were achieved, respectively, which increased by 7.41% and 5.87% as compared to AHP pretreatment. Furthermore, the lignin extracted from the organic phase accounted for 38.51% of the delignification, and it had a low molecular weight, effectively preserving the ß-O-4 bonds. Finally, satisfied pentanol recovery (77.91%) and delignification (57.19%) along with excellent glucan (76.11%) and xylan (77.52%) digestibility were reached after fourth recycling of AHPP pretreatment. Therefore, AHPP pretreatment was a promising method for biomass valorization within biorefinery concept.

Marine bioactive peptides with anticancer potential, a narrative review.

In this paper, we explore marine bioactive peptides with anticancer potential sourced from various marine organisms, including tunicates, sea sponges, and mollusks. Peptides like Stylisin and Papuamides have been isolated, identified, and modified to enhance their activity, with many advancing to clinical trials due to their diverse biological activities, promising prospects in medicine. Enzymatic hydrolysis is a favored method for extracting peptides from marine proteins, particularly from sponges known for their rich bioactive compounds. Compounds such as Jaspamide and Homophymins exhibit potent cytotoxic activity against cancer cells, underscoring their therapeutic potential. Additionally, peptides from ascidians and mollusks, such as Aplidine and Kahalalide F, demonstrate significant anticancer properties. This study also explores peptides influencing apoptosis, microtubule dynamics, and angiogenesis, providing insights into potential mechanisms for cancer treatment. While peptides like Neovastat and mycothiazole target known pathways, others such as patellamides act through unknown mechanisms, highlighting the intricate interactions of marine peptides with cancer cells. Overall, marine-derived peptides show promise as valuable candidates for developing novel anticancer therapies.

Exploring the seas for cancer cures: the promise of marine-derived bioactive peptide.

Marine environments harbor a wealth of bioactive peptides with potential anticancer properties, sourced from diverse organisms like tunicates, sea sponges, and mollusks. Through isolation, identification, and modification, peptides such as Stylisin and Papuamides have shown enhanced activity and progressed to clinical trials, underscoring their therapeutic promise. Enzymatic hydrolysis emerges as a favored method for peptide extraction from marine proteins, with sponges identified as particularly rich sources. Compounds like Jaspamide and Homophymins exhibit potent cytotoxic activity against cancer cells, highlighting their therapeutic potential. Additionally, peptides from ascidians and mollusks, including Aplidine and Kahalalide F, demonstrate significant anticancer properties. The study delves into peptides affecting apoptosis, microtubule dynamics, and angiogenesis inhibition, offering insights into potential cancer treatment mechanisms. Marine-derived peptides hold great promise as valuable candidates for novel anticancer therapies, with ongoing research aimed at unlocking their full therapeutic benefits.

Antioxidant Peptides and Protein Hydrolysates from Tilapia: Cellular and In Vivo Evidences for Human Health Benefits.

Antioxidant peptides derived from aquatic organisms have attracted tremendous research interest due to their potential applications in human health. Tilapia is one of the most widely farmed aquaculture species globally. The current understanding of tilapia-derived antioxidant peptides is gradually expanding. This review discusses the current knowledge of peptides and protein hydrolysates derived from tilapia muscle, skin, and scales, whose antioxidant capacity has been validated in various cellular and in vivo models. To date, at least 16 peptides and several hydrolysates have been identified from tilapia that protect human and non-human cell models against oxidative injury. Tilapia hydrolysates and peptide mixtures have also shown protective effects in animal models of oxidative stress-associated diseases and exercise-induced oxidative injury and fatigue. The key mechanisms of tilapia hydrolysates and peptide mixtures involve enhancing antioxidant enzyme activities and suppressing radical production. Notably, such hydrolysates also exerted additional in vivo functions, such as anti-inflammatory, anti-diabetic, wound healing, and antiaging properties. Taken together, tilapia-derived antioxidant peptides and hydrolysates represent a valuable source of functional ingredients for applications in functional food, dietary supplements, and therapeutic applications. Continued research into their health benefits is warranted in the future.

Antioxidant Activity of Protein Hydrolysates from Redlip Mullet (Chelon haematocheilus) Muscle and Byproducts.

Fish muscle and byproducts represent a valuable source of bioactive compounds, with their protein hydrolysates exhibiting noteworthy antioxidant properties. This study assessed the antioxidant activity of protein hydrolysates derived from the muscle and byproducts of redlip mullet (Chelon haematocheilus), utilizing different proteases (Neutrase, Alcalase, and Protamex). Hydrolysates were prepared from various parts of the fish, including muscle (white and red meat) and byproducts (frames, head, viscera, fins, skin, and scales). The enzymatic hydrolysis resulted in the highest degree of hydrolysis, achieving 83.24 ± 1.45% for skin at 60 min and 82.14 ± 4.35% for head at 30 min, when treated with Neutrase. Frames treated with Neutrase exhibited the highest protein concentration, measured at 1873.01 ± 71.11 µg/mL at 15 min. Significantly, skin hydrolysates treated with Protamex showed the highest DPPH• scavenging activity (70.07 ± 3.99% at 120 min), while those treated with Alcalase demonstrated the highest ABTS• scavenging activity (93.47 ± 0.02% at 15 min). The highest superoxide dismutase (SOD) activity (92.01 ± 1.47%) was observed in head hydrolysates treated with Protamex after 90 min. These results suggest that C. haematocheilus protein hydrolysates possess significant antioxidant activity within a short time frame of less than 120 min.

Decarbonizing paper mill sludge waste into micro and nanofibrillated cellulose via enzyme hydrolysis and dual asymmetric centrifugation.

The production of micro- and nano-fibrillated cellulose (MNFC) from paper mill sludge (PS) using simple enzymatic and mechanical treatments has been evaluated for their performance as value-added materials in packaging and other applications. Sludge from a US paper mill was analyzed for the viability of this conversion process. The enzymatic treatment was conducted at variable concentrations using an enzyme cocktail of exoglucanase and endoglucanase, followed by mechanical dual asymmetric centrifugation (DAC) treatment. The presence of inorganics and lignin facilitated the mechanical defibrillation of the fibers, making the MNFC production more energy-efficient. The prepared MNFC was characterized for fibrillation, charge, crystallinity, and surface morphology. Scanning electron microscopic (SEM) images show the highly fibrillated MNFC and the distribution of inorganic nanoparticles on the fiber surface. The X-ray diffractometric (XRD) analysis shows 44-65 % crystallinity. Furthermore, MNFC-based films derived from PS demonstrated excellent strength and flexibility, making them suitable for packaging and other applications. Overall, this conversion approach can save the paper industry millions of dollars in disposal costs while upcycling waste and reducing greenhouse gas (GHG) emissions associated with waste streams.