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20 pages, 1861 KB  
Article
An Exploratory Study of the Nutritional Composition and Caco-2 Safety Assessment of Elche Date Flour and Its Green Hydroethanolic Extracts
by Katarzyna Dawidowicz, Sergio Martinez-Terol, Estrella Sayas-Barberá, José Ángel Pérez-Álvarez, Francisco J. Marti-Quijal, Patricia Roig and Juan Manuel Castagnini
Foods 2025, 14(22), 3908; https://doi.org/10.3390/foods14223908 (registering DOI) - 15 Nov 2025
Abstract
The Elche palm grove (Spain) produces large surpluses of fresh date fruits due to low industrial processing and strict market standards. This exploratory study assessed the potential of these fruits as sustainable ingredients through the production of freeze-dried date flour and its green [...] Read more.
The Elche palm grove (Spain) produces large surpluses of fresh date fruits due to low industrial processing and strict market standards. This exploratory study assessed the potential of these fruits as sustainable ingredients through the production of freeze-dried date flour and its green hydroethanolic extracts. Computer vision analysis of nine local cultivars (D1–D9) revealed broad chromatic and phenotypic diversity. Mineral and heavy metal analyses in the flour indicated high nutritional value and overall safety: D8 was richest in Mg (1.23 mg/g), P (0.78 mg/g), Fe (15.32 mg/kg), Zn (9.20 mg/kg), Cu (5.22 mg/kg), and Se (68 µg/kg), while D4 showed the highest K (22.1 mg/g) and D1 the highest Ca (1.94 mg/g). Lead and cadmium were highest in D8 and arsenic in D1, although all values remained within the regulatory limits. Hydroethanolic extracts exhibited remarkable compositional variability: D4 and D5 had the greatest carbohydrates (737.70 ± 55.79 mg/g DM), D8 and D9 the highest proteins (up to 40.31 ± 1.33 mg/g DM), and D2 and D8 the highest carotenoids (up to 36.44 ± 1.55 μg/g DM). D8 also showed the highest phenolics (13.98 ± 2.93 mg GAE/g DM) and antioxidant capacity. Cytotoxicity assays in Caco-2 cells showed no significant effects up to 1000 µg/mL. These preliminary findings suggest that green-extracted date fractions may combine nutritional richness, antioxidant potential, and biological safety, providing a basis for future studies on their application as natural and sustainable food ingredients. Full article
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34 pages, 8474 KB  
Article
Integrated Metabolomic and Transcriptomic Profiles Provide Insights into the Molecular Mechanisms in Modulating Female Flower of Coconut (Cocos nucifera L.)
by Lilan Lu, Yuan Zhang, Zhiguo Dong, Weibo Yang and Ruoyun Yu
Agriculture 2025, 15(22), 2336; https://doi.org/10.3390/agriculture15222336 - 10 Nov 2025
Viewed by 172
Abstract
Coconut yield and quality are significantly affected by multiple female inflorescences (MFF), which disrupt flower differentiation balance. To elucidate the molecular mechanisms, we compared MFF with normal female inflorescences (NFF) using phenotypic, morphological, physiological, and multi-omics approaches. The results revealed that MFF exhibited [...] Read more.
Coconut yield and quality are significantly affected by multiple female inflorescences (MFF), which disrupt flower differentiation balance. To elucidate the molecular mechanisms, we compared MFF with normal female inflorescences (NFF) using phenotypic, morphological, physiological, and multi-omics approaches. The results revealed that MFF exhibited altered flower structures. MFF showed elevated iron (Fe), nitrogen (N), sulfur (S), potassium (K), calcium (Ca), zinc (Zn), proline (Pro), catalase (CAT), malondialdehyde (MDA), abscisic acid (ABA), and jasmonic acid (JA), but reduced molybdenum (Mo), soluble sugar (SS), soluble protein (SP), superoxide dismutase (SOD), peroxidase (POD), indole acetic acid (IAA), zeatin riboside (ZR), and gibberellic acid (GA). We detected 445 differentially expressed genes (DEGs) mainly enriched in ABA, ETH, BR, and JA pathways in MFF compared to NFF. We identified 144 differentially accumulated metabolites (DAMs) primarily in lipids and lipid-like molecules, phenylpropanoids and polyketides, as well as organic acids and derivatives in the comparison of MFF and NFF. Integrated analysis linked these to key pathways, e.g., “carbon metabolism”, “carbon fixation in photosynthetic organisms”, “phenylalanine, tyrosine, and tryptophan biosynthesis”, “glyoxylate and dicarboxylate metabolism”, “glycolysis/gluconeogenesis”, “pentose and glucuronate interconversions”, “flavonoid biosynthesis”, “flavone and flavonol biosynthesis”, “pyruvate metabolism”, and “citrate cycle (TCA cycle)”. Based on our results. the bHLH137, BHLH062, MYB (CSA), ERF118, and MADS2 genes may drive MFF formation. This study provides a framework for understanding coconut flower differentiation and improving yield. Full article
(This article belongs to the Section Crop Genetics, Genomics and Breeding)
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18 pages, 800 KB  
Article
Durum Wheat Kernel: Influence of the Genotype and Environment on the Mineral Profile of Grains and Ashes
by José Moreira, Sara Rodrigo, Nuno Pinheiro, Rita Costa, Armindo Costa, José Dôres, Manuel Patanita, Benvindo Maçãs, Roberta Leitão, Mauro Guerra and Ana Sofia Bagulho
Plants 2025, 14(22), 3414; https://doi.org/10.3390/plants14223414 - 7 Nov 2025
Viewed by 290
Abstract
Thirteen genotypes of durum wheat were grown in two different environments in Portugal. Grain and ash mineral profile, as well as protein content, test weight, and grain ash content were evaluated. Genotype, environment, and their interaction explains the variation in the quality traits, [...] Read more.
Thirteen genotypes of durum wheat were grown in two different environments in Portugal. Grain and ash mineral profile, as well as protein content, test weight, and grain ash content were evaluated. Genotype, environment, and their interaction explains the variation in the quality traits, with the environment having the highest influence. Mineral profile analysis was performed by the μ-EDXRF system: macroelements (K, P, Ca, Cl, and S) represented 99% of the total concentration detected in the grain samples, while microelements represented up to 2% of the total concentration when analyzing the ash samples (Fe, Mn, Zn, Cu, Si, Rb, Sr, and Ti). Almost every element found in the grain and ash analysis was affected by the environment. Only K and Ca in the grain had higher concentrations in the environment with water scarcity, while the concentrations of all the detected elements except for Si and Sr were higher in the ashes in this environment. Regarding the genotype, P, S, and Cu grain concentrations were not affected by the environment. The highest grain mineral concentration was found for Gingão, suggesting a better mineral uptake and/or translocation-to-grain capacity. However, regarding the technological quality, most of the genotypes presented ash content values above the maximum specified threshold. Full article
(This article belongs to the Section Plant Genetics, Genomics and Biotechnology)
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18 pages, 8678 KB  
Article
Mulberrin Alleviates Renal Ischemia–Reperfusion by Inhibiting Ferroptosis and Oxidative Stress Through Sirt3 Activation
by Qiangmin Qiu, Zhan Chen, Wenbin Yang, Yujie Zhou, Nan Jiang, Jiahao Jiang, Dalin He, Yifan Lu, Bo Yu, Tao Qiu and Jiangqiao Zhou
Biomedicines 2025, 13(11), 2687; https://doi.org/10.3390/biomedicines13112687 - 31 Oct 2025
Viewed by 372
Abstract
Background: Renal ischemia–reperfusion (I/R) injury represents a principal etiologic factor in acute kidney injury (AKI), in which ferroptosis plays a critical role. Mulberrin (Mul), a prenylated flavonoid with antioxidative properties, has an as-yet undefined role in renal I/R injury. Methods: We established a [...] Read more.
Background: Renal ischemia–reperfusion (I/R) injury represents a principal etiologic factor in acute kidney injury (AKI), in which ferroptosis plays a critical role. Mulberrin (Mul), a prenylated flavonoid with antioxidative properties, has an as-yet undefined role in renal I/R injury. Methods: We established a mouse renal IRI model and an HK-2 H/R system. Renal function, histological injury, oxidative stress, ferroptosis markers, and mitochondrial function were assessed. The role of Sirtuin 3 (Sirt3) in Mul-mediated effects was further examined using siRNA knockdown in HK-2 cells. Results: The administration of Mul led to a marked improvement in renal function, lessened tubular injury, and reduced apoptosis in IRI mice. Mul also restored GSH levels, decreased MDA and Fe2+ accumulation, and normalized expression of ferroptosis-related proteins, thereby suppressing ferroptosis. In H/R-injured HK-2 cells, Mul restored mitochondrial membrane potential, increased ATP production, and reduced ROS accumulation. Mechanistically, Mul markedly upregulated Sirt3 expression, and silencing Sirt3 abolished its antioxidant and anti-ferroptosis effects, confirming the essential role of Sirt3 in Mul-mediated protection. Conclusions: Our findings underscore Mul’s therapeutic promise in acute kidney injury and provide a mechanistic foundation for interventions directed at the Sirt3–ferroptosis pathway to safeguard renal function. Full article
(This article belongs to the Special Issue From Pathogenesis to Therapies: Innovations in Kidney Disease)
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19 pages, 9464 KB  
Article
Characterisation of SapYZUs891@Fe/Mn-MOF Provides Insight into the Selection of Temperate Phage and Nanozyme for the Rapid and Sensitive Colourimetric Detection of Viable Staphylococcus aureus in Food Products
by Wenyuan Zhou, Wenjuan Li, Yeling Han, Aiping Deng, Yajie Li, Qin Hu, Lei Yuan, Guoqiang Zhu and Zhenquan Yang
Foods 2025, 14(21), 3726; https://doi.org/10.3390/foods14213726 - 30 Oct 2025
Viewed by 377
Abstract
Although phage@nanozymes have proven to be a rapid, precise, and cost-effective method for detecting pathogens in food, the basis of phage and nanozyme selection remains poorly understood. In this study, a novel colourimetric biosensor utilising the temperate phage SapYZUs891 and an Fe/Mn-MOF nanozyme [...] Read more.
Although phage@nanozymes have proven to be a rapid, precise, and cost-effective method for detecting pathogens in food, the basis of phage and nanozyme selection remains poorly understood. In this study, a novel colourimetric biosensor utilising the temperate phage SapYZUs891 and an Fe/Mn-MOF nanozyme was constructed and assessed for its efficacy in detecting Staphylococcus aureus in food products. Notably, SapYZUs891 exhibited a high titre, broad host range, and strong pH and thermal stability. Moreover, the bimetallic Fe/Mn-MOF nanozyme exhibited an enhanced oxidase-mimicking ability, greater affinity, and a higher reaction rate. The biosensor had a detection time of 19 min, a detection limit of 69 CFU/mL, and a recovery rate between 92.52% and 121.48%, signifying its high reliability and accuracy in identifying S. aureus. This sensor distinguishes between viable and non-viable bacteria and demonstrates resistance to interferent bacterial and food compounds, likely attributable to the particular receptor-binding proteins of SapYZUs891 that bind to the teichoic acid wall on the S. aureus. These results indicated that the SapYZUs891@Fe/Mn-MOF is suitable for the rapid visual assessment of S. aureus. Moreover, the highly sensitive and specific detection system holds significant potential for extended application in on-site screening of S. aureus contamination within food processing environments. Full article
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17 pages, 1080 KB  
Review
Metal–Organic Frameworks for Enzyme Modulation in Protein Kinase and Phosphatase Regulation—Mechanisms and Biomedical Applications
by Azizah Alamro and Thanih Balbaied
Kinases Phosphatases 2025, 3(4), 21; https://doi.org/10.3390/kinasesphosphatases3040021 - 30 Oct 2025
Viewed by 343
Abstract
Metal–organic frameworks (MOFs) have been increasingly recognized as promising platforms for enzyme modulation, owing to their tunable porosity, high surface area, and versatile chemical functionality. In this review, the potential of MOFs for the inhibition and modulation of protein kinases and phosphatases—key regulators [...] Read more.
Metal–organic frameworks (MOFs) have been increasingly recognized as promising platforms for enzyme modulation, owing to their tunable porosity, high surface area, and versatile chemical functionality. In this review, the potential of MOFs for the inhibition and modulation of protein kinases and phosphatases—key regulators of cellular signaling and disease progression—is examined. The structural fundamentals of MOFs are outlined, followed by a discussion of common synthesis strategies, including solvothermal, microwave-assisted, sonochemical, and mechanochemical methods. Emphasis is placed on how synthesis conditions influence critical features such as particle size, crystallinity, surface chemistry, and functional group accessibility, all of which impact biological performance. Four primary mechanisms of MOF–enzyme interaction are discussed: surface adsorption, active site coordination, catalytic mimicry, and allosteric modulation. Each mechanism is linked to distinct physicochemical parameters, including pore size, surface charge, and metal node identity. Special focus is given to biologically relevant metal centers such as Zr4+, Ce4+, Cu2+, Fe3+, and Ti4+, which have been shown to contribute to both MOF stability and enzymatic inhibition through Lewis acid or redox-mediated mechanisms. Recent in vitro studies are reviewed, in which MOFs demonstrated selective inhibition of disease-relevant enzymes with minimal cytotoxicity. Despite these advancements, several limitations have been identified, including scalability challenges, limited physiological stability, and potential off-target effects. Strategies such as post-synthetic modification, green synthesis, and biomimetic surface functionalization are being explored to overcome these barriers. Through an integration of materials science, coordination chemistry, and molecular biology, this review aims to provide a comprehensive perspective on the rational design of MOFs for targeted enzyme inhibition in therapeutic contexts. Full article
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14 pages, 8136 KB  
Article
Snake Venom C-Type Lectin-like Protein Vaa-Snaclec-3/2 Efficiently Prevents Carotid Artery Thrombosis in a Mouse Model Without Compromising Blood Coagulation
by Monika C. Žužek, Igor Križaj, Miran Brvar, Tomaž Trobec, Simona Kranjc Brezar, Mojca Dobaja Borak, Adrijana Leonardi, Kity Požek, Milka Vrecl and Robert Frangež
Toxins 2025, 17(11), 523; https://doi.org/10.3390/toxins17110523 - 23 Oct 2025
Viewed by 515
Abstract
Platelets play pivotal roles in thromboembolic diseases, such as myocardial infarction and ischemic stroke. In patients envenomed by the snake Vipera a. ammodytes (Vaa), pronounced and transient thrombocytopenia without bleeding is observed. We previously showed that Vaa-snaclec-3/2, the snake venom [...] Read more.
Platelets play pivotal roles in thromboembolic diseases, such as myocardial infarction and ischemic stroke. In patients envenomed by the snake Vipera a. ammodytes (Vaa), pronounced and transient thrombocytopenia without bleeding is observed. We previously showed that Vaa-snaclec-3/2, the snake venom C-type lectin-like protein, mediates this effect ex vivo. Here, we extended our study of the antithrombotic potential of this protein in vivo using a mouse model of ferric chloride (FeCl3)-induced carotid artery thrombosis. Prior to inducing thrombus formation, the mice received 1, 5, 10, 20, or 50 μg/kg Vaa-snaclec-3/2 intravenously. Afterward, the arterial blood flow was monitored with a perivascular Doppler probe. Additionally, the platelet count in the peripheral venous blood; tail bleeding time; and liver, lung, kidney, spleen, and heart histology were evaluated. The lowest dose of Vaa-snaclec-3/2 that we showed to cause severe thrombocytopenia and completely inhibit FeCl3-induced thrombus formation was 20 µg/kg. This dose prolonged the median tail bleeding time from 86.5 to 153.5 s but did not induce acute spontaneous hemorrhage, as demonstrated by histological analysis. Histology revealed no signs of apoptosis, necrosis or other degenerative changes in the inspected organs of mice exposed to 20 μg/kg Vaa-snaclec-3/2. Platelet clusters were observed only in the lungs, which appear to be the primary site of platelet sequestration and the cause of thrombocytopenia. Taken together, our findings highlight the high potential of Vaa-snaclec-3/2 as a safe and effective antithrombotic agent for the transient prevention of thrombosis in acute clinical settings. Full article
(This article belongs to the Special Issue Animals Venom in Drug Discovery: A Valuable Therapeutic Tool)
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16 pages, 820 KB  
Article
Sustainable Recovery of Valuable Constituents from Octopus (Octopus vulgaris) Cooking Liquor
by Ricardo Prego, Antonio Cobelo-García, Marcos Trigo, Susana Calvo and Santiago P. Aubourg
Sustainability 2025, 17(21), 9391; https://doi.org/10.3390/su17219391 - 22 Oct 2025
Viewed by 283
Abstract
Cooking liquor (CL) from marine species processing has been reported to include a wide range of valuable constituents. In this study, the chemical composition of CL from octopus (Octopus vulgaris) processing, with and without a filtration process, was analysed. Regarding non-filtered [...] Read more.
Cooking liquor (CL) from marine species processing has been reported to include a wide range of valuable constituents. In this study, the chemical composition of CL from octopus (Octopus vulgaris) processing, with and without a filtration process, was analysed. Regarding non-filtered CL, values of 15.30, 0.29, 8.85 and 174.53 g·L−1 CL for protein, lipids, ash, and total volatile base-nitrogen (TVB-N), respectively, were detected. The most abundant fatty acids (FAs) (g·100 g−1 total FAs) were C16:0 (37.8), C18:0 (20.8), and C22:6ω3 (13.4). Values of 0.40 and 2.10 were obtained for polyunsaturated FA/saturated FA and ω3 FA/ω6 FA ratios. Macroelement content varied from 0.036 (Ca) to 1.81 (Na) g·L−1 CL. For microelements, values ranged between 0.0015 (Co) and 1.95 (As) mg·L−1 CL. Industrial filtration of CL led to decreased values of protein, lipid, ash, TVB-N, and C22:5ω3; in contrast, an increased presence of C14:0, C18:1ω9, C20:1ω9, and C22:1ω9 was detected. Filtration led to a ca. 50% decrease in macroelement presence. For microelements, this process led to losses of 20–40% (Ba, Pb), 40–60% (As, Fe, Mn), 60–70% (Co, Zn), and 84% (Cd). This study provides a first comprehensive characterisation of octopus cooking liquor as a potential source of bioactive compounds. Full article
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28 pages, 2175 KB  
Article
The Effect of Chelated Trace Mineral Supplementation in the Form of Proteinates on Broiler Performance Parameters and Mineral Excretion: A Meta-Analysis
by Laurann Byrne, Stephen Ross, Jules Taylor-Pickard and Richard Murphy
Animals 2025, 15(21), 3062; https://doi.org/10.3390/ani15213062 - 22 Oct 2025
Viewed by 600
Abstract
Comprehensive Meta-Analysis (CMA) software, using data from 64 global studies (288 dietary assessments, 194,356 broilers) evaluated the effects of substituting inorganic trace minerals (ITM) with proteinate trace minerals (PTM) in broiler diets at various inclusion levels. Replacing ITM with PTM at equivalent (100%) [...] Read more.
Comprehensive Meta-Analysis (CMA) software, using data from 64 global studies (288 dietary assessments, 194,356 broilers) evaluated the effects of substituting inorganic trace minerals (ITM) with proteinate trace minerals (PTM) in broiler diets at various inclusion levels. Replacing ITM with PTM at equivalent (100%) or reduced (11–80%) levels improved performance metrics, showing reduced total feed intake (FI) (−6 g/bird), lower average daily feed intake (ADFI) (−0.43 g/bird), higher average daily gain (ADG) (+0.36g), greater body weight gain (BWG) (+4.29 g/bird), higher final body weight (BW) (+7.50 g/bird), improved feed conversion ratio (FCR) (−1.26%), and lower mortality (−10.95%), all significant (p < 0.05). Median mineral inclusion reductions of 40% Cu, 59.82% Fe, 41.41% Mn, and 34.67% Zn had no adverse effects, instead enhancing outcomes. Across 17 studies (25,144 broilers, 85 dietary assessments), mineral excretion decreased significantly with PTM versus ITM by 16% Cu, 14% Fe, 21% Mn, and 15% Zn (p < 0.001). When PTM replaced ITM at 50–80% inclusion, further benefits were observed, including lower total FI (−7 g/bird), lower ADFI (−1.07 g/bird), higher ADG (+1.67), higher BWG (+2.65 g/bird), lower FCR (−4.50%) and lower mortality (−11.09%) with mineral inclusion reductions of 17% Cu, 42.16% Fe, 42.89% Mn, and 50% Zn. Meta-regression identified significant influences (p < 0.05) from study variables such as strain, study duration, and region. Life cycle assessment modelling demonstrated PTM inclusion lowered gross carbon emissions by 3.5% and lower emission intensities per unit live weight of both feed use and overall lifecycle by 4.5% and 4.1%, respectively on diets of high and low soybean meal inclusion. Overall, replacing ITM with PTM in broiler diets can promote production performance of broilers and lower mineral excretion levels while contributing to a lower CFP. Full article
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20 pages, 3670 KB  
Article
Discovery of a Novel Anticoagulant Cystine Knot Peptide from Spider Venom Gland Transcriptome
by Jinai Gao, Di Yang, Wanting Wang, Xiaoshan Huang, Ruiyin Guo, Kaixun Cao, Qiumin Lu, Ziyi Wang, Ren Lai and Juan Li
Int. J. Mol. Sci. 2025, 26(20), 10154; https://doi.org/10.3390/ijms262010154 - 19 Oct 2025
Cited by 1 | Viewed by 528
Abstract
The development of effective anticoagulants remains a critical need in modern medicine, particularly for preventing and treating thromboembolic disorders, such as arterial thrombosis and deep vein thrombosis (DVT), as well as complications like ischemic stroke. This study identifies a cysteine-knotted peptide GC38 (sequence: [...] Read more.
The development of effective anticoagulants remains a critical need in modern medicine, particularly for preventing and treating thromboembolic disorders, such as arterial thrombosis and deep vein thrombosis (DVT), as well as complications like ischemic stroke. This study identifies a cysteine-knotted peptide GC38 (sequence: GCSGKGARCAPSKCCSGLSCGRHGGNMYKSCEWNWKTG) derived from the venom gland transcriptome of the Macrothele sp. spider, which exerts thrombus-inhibitory effects by potentiating activated protein C (APC) activity. In vitro assays reveal that GC38 enhances APC activity, prolongs plasma clotting time, and shows no significant cytotoxicity or hemolytic activity. Mechanistically, GC38 interacts allosterically with APC; biolayer interferometry (BLI) confirms this direct interaction, with a dissociation constant KD of 6.16 μM. Additionally, three in vivo thrombosis models (FeCl3-induced arterial occlusion, stasis-induced DVT, and cortical photothrombotic stroke) consistently demonstrated that GC38 was effective in alleviating thrombus formation, with tail-bleeding assays confirming its low hemorrhagic risk. Collectively, our findings position GC38 as a pioneering spider venom-derived lead molecule that addresses dual arterial and venous antithrombotic actions. This opens new avenues for developing spider venom-derived peptides as therapeutic agents targeting intravascular coagulation in arteries and veins. Full article
(This article belongs to the Special Issue Molecular Mechanisms of Animal Toxins, Venoms and Antivenoms 2.0)
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25 pages, 3623 KB  
Article
Fusarium proliferatum PSA-3 Produces Xylanase-Aggregate to Degrade Complex Arabinoxylan
by Kanlaya Thattha, Lakha Salaipeth, Saengchai Akeprathumchai, Ken-Lin Chang, Takashi Watanabe and Paripok Phitsuwan
Catalysts 2025, 15(10), 988; https://doi.org/10.3390/catal15100988 - 16 Oct 2025
Viewed by 618
Abstract
Xylanolytic enzymes of the Fusarium species are closely associated with pathogenesis, where they soften plant cell walls to facilitate infection and nutrient uptake. This study investigated the xylanolytic system of Fusarium proliferatum PSA-3, a strain isolated from mango leaves showing dark spot symptoms. [...] Read more.
Xylanolytic enzymes of the Fusarium species are closely associated with pathogenesis, where they soften plant cell walls to facilitate infection and nutrient uptake. This study investigated the xylanolytic system of Fusarium proliferatum PSA-3, a strain isolated from mango leaves showing dark spot symptoms. When cultivated on rice straw under solid-state fermentation, PSA-3 produced high xylanase activity against rye arabinoxylan (50.2 U) and beechwood xylan (56.8 U). Partial purification by ion-exchange and gel-filtration chromatography yielded a large xylanase aggregate (158 kDa), which appeared as a smear at the top of the gel under native conditions. Mild denaturation resolved the aggregate into at least four active proteins of ~25, 35, 48, and 63 kDa, indicating that multiple xylanases assemble into a functional aggregate. The aggregate retained activity across pH 4.0–8.0, with an optimum at pH 5.0 and 50 °C, and was resistant to Ni2+, Fe2+, Co2+, and β-mercaptoethanol, but inhibited by SDS. Hydrolysis of xylo-oligosaccharides (DP 2–6), purified xylans, and plant-derived xylans confirmed predominantly endo-type action with debranching activity toward A2XX and A2,3XX. These findings reveal a natural xylanase aggregate in F. proliferatum, providing a potential mechanism for efficient degradation of arabinoxylan-rich cell walls and offering targets for antifungal strategies and biotechnological applications. Full article
(This article belongs to the Special Issue 15th Anniversary of Catalysts: The Future of Enzyme Biocatalysis)
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22 pages, 2455 KB  
Article
Bioactive Potential of Protein Extracts Derived from Dried Wolffia globosa on In Vitro Antioxidant Activities and Pro-Inflammatory Cytokine Production
by Ruttiros Khonkarn, Krai Daowtak, Oranit Kraseasintra, Thitiya Luetragoon, Kanchana Usuwanthim, Kraisorn Taynawa and Kanokphon Chanphong
Molecules 2025, 30(20), 4092; https://doi.org/10.3390/molecules30204092 - 15 Oct 2025
Viewed by 1018
Abstract
This study investigated the contamination, composition, and functional properties of Wolffia globosa from northern Thailand. The results showed that the heavy metal content of dried W. globosa complied with Thai regulations, ensuring its safety. Its proximate analysis revealed high protein levels with lysine, [...] Read more.
This study investigated the contamination, composition, and functional properties of Wolffia globosa from northern Thailand. The results showed that the heavy metal content of dried W. globosa complied with Thai regulations, ensuring its safety. Its proximate analysis revealed high protein levels with lysine, leucine, and phenylalanine as the principal essential amino acids. The protein was effectively extracted using the alkaline extraction method, followed by precipitation induced by acid or heat. The precipitates and supernatants resulting from various acid- or heat-induced protein precipitation were obtained. The highest protein content was found in the pH 3 precipitate (51.15 ± 6.71%). In contrast, the pH 5 supernatant exhibited the most potent antioxidant activities (2.22 ± 0.05 mmol Trolox/mg and 4.55 ± 0.18 mmol Fe2+/mg), as determined by ABTS and FRAP assays, respectively. Additionally, a strong correlation was observed between phenolic content and antioxidant activity. Both supernatant and precipitate protein extracts from W. globosa exhibited no cytotoxicity in THP-1 cells and displayed anti-inflammatory effects by decreasing the production of IL-1β and IL-6. They also downregulated phospho-NF-κB, phospho-IκB-α, and COX-2, consistent with reduced NF-κB pathway activation. These findings position W. globosa as a promising, sustainable plant-based protein with bioactive and functional properties, making it a viable candidate for functional food formulations that enhance dietary health and add value to local agricultural resources. Full article
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15 pages, 558 KB  
Article
Benefits and Trade-Offs of Long-Term Organic Fertilization Substitution: Wheat Grain Nutrition and Heavy Metal Risks in an 11-Year Field Trial
by Yumin Liu, Xiaolin Zhou, Zishuang Li, Lei Ma, Yan Li, Huanyu Zhao, Yu Xu and Deshui Tan
Agronomy 2025, 15(10), 2369; https://doi.org/10.3390/agronomy15102369 - 10 Oct 2025
Viewed by 396
Abstract
Optimizing organic fertilizer substitution is essential for enhancing the sustainability of agriculture and achieving a balance between crop productivity, nutritional quality, and environmental safety. Here, we conducted an 11-year field experiment to evaluate the effects of substituting 50% of mineral fertilizers with pig [...] Read more.
Optimizing organic fertilizer substitution is essential for enhancing the sustainability of agriculture and achieving a balance between crop productivity, nutritional quality, and environmental safety. Here, we conducted an 11-year field experiment to evaluate the effects of substituting 50% of mineral fertilizers with pig manure (PM) or cattle manure (CM) on the nutritional quality of wheat grain, heavy metal (HM) accumulation, and associated human health risks. The yield and protein content were highest in the mineral fertilizer (MF) treatment, and grain micronutrients (Fe, Mn, Cu, Zn) were 6.7–13.8% higher under organic substitution (PM/CM) than in the MF treatment. The Ni, Pb, and As contents were 35.4–43.0% higher in the PM treatment than in the MF treatment, which stems from the higher HM content in pig manure. Health risk assessments indicated that the Hazard Index (HI) for children exceeded 1 in the PM treatment, primarily due to As, which accounted for 69.6% of the HI. All treatments remained within safe thresholds, although As and Pb posed detectable carcinogenic health risks. The higher levels of Ni and As in pig manure likewise led to a significant increase in the health risk associated with the PM treatment compared to the MF treatment. We developed a novel Grain Quality Index (GQI) that combined nutrient and HM data, which indicated that the nutritional quality of wheat grain was similar in the CM and MF treatments. The GQI was 9.1% lower in the PM treatment than in the MF treatment. These findings suggest that the substitution of mineral fertilizer with cow manure can help achieve a balance between yield, nutrition, and safety, and more stringent regulation of HMs is required for the use of pig manure. Our findings provide actionable insights with implications for sustainable wheat production policies. Full article
(This article belongs to the Special Issue Nutrient Enrichment and Crop Quality in Sustainable Agriculture)
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26 pages, 1369 KB  
Article
Effects of Free and Conjugated Methionine on Growth, Meat Quality, Mineral Profile, and Shell Strength in Garden Snails (Cornu aspersum)
by Anna Rygało-Galewska, Klara Piotrowska, Magdalena Matusiewicz, Damian Bień, Monika Łukasiewicz-Mierzejewska, Zbigniew Skibko, Andrzej Borusiewicz and Tomasz Niemiec
Animals 2025, 15(19), 2922; https://doi.org/10.3390/ani15192922 - 8 Oct 2025
Viewed by 627
Abstract
The present study examined the impact of adding methionine (Met) and its conjugated form (Met-Met) on Cornu aspersum snails. The primary focus was on the animals’ growth performance, the chemical composition of their carcass (whole body without the shell), the mineral profile, and [...] Read more.
The present study examined the impact of adding methionine (Met) and its conjugated form (Met-Met) on Cornu aspersum snails. The primary focus was on the animals’ growth performance, the chemical composition of their carcass (whole body without the shell), the mineral profile, and the mechanical properties of their shells. In two experiments conducted under controlled laboratory conditions, diets supplemented with varying levels of Met addition (0.3, 0.6, 1.4 g/kg feed) were used, and the effects of free methionine, Met-Met and their mixture (1.4 g/kg feed) were compared. The study incorporated measurements of body weight, shell width, and mortality of snails. Analyses encompassing protein, fat, sulphur amino acids, glutathione levels, oxidative stress indices (DPPH, TAC, TBARS), and macro- and micronutrient content of carcass and shells were conducted. The findings demonstrated that adding 1.4 g Met/kg feed significantly enhanced the shells’ weight gain (+56% vs. Control), shell weight (+56%) and crushing force (+135%). Furthermore, an increase in the Met content of the carcass was observed (+18%), along with elevated carcass Ca (+28%) and P (+30%) and higher shell Ca (+12%) and Zn (+87%), alongside reduced carcass Fe (−38%) and Cu (−19%). In Experiment II, the Met-Met group exhibited the highest carcass weight (+16% vs. Control), the greatest carcass-to-body weight ratio, and the highest proportion of mature individuals (+27%). Moreover, Met-Met supplementation improved Cu absorption and retention in the carcass (+19%). Also, the results suggest that the conjugated form of methionine may improve Cu absorption and storage in the carcass (+19%). The study’s findings indicate that methionine addition, especially in Met-Met form, can substantially impact the efficiency of C. aspersum farming, enhancing both the productivity outcomes and the quality of the product. That is particularly important in increasing the shell’s mechanical resistance and the carcass’s nutritional value. Full article
(This article belongs to the Section Animal Nutrition)
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Article
Predicting the Structure of Hydrogenase in Microalgae: The Case of Nannochloropsis salina
by Simone Botticelli, Cecilia Faraloni and Giovanni La Penna
Hydrogen 2025, 6(4), 77; https://doi.org/10.3390/hydrogen6040077 - 2 Oct 2025
Viewed by 486
Abstract
The production of green hydrogen by microalgae is a promising strategy to convert energy of sun light into a carbon-free fuel. Many problems must be solved before large-scale industrial applications. One solution is to find a microalgal species that is easy to grow, [...] Read more.
The production of green hydrogen by microalgae is a promising strategy to convert energy of sun light into a carbon-free fuel. Many problems must be solved before large-scale industrial applications. One solution is to find a microalgal species that is easy to grow, easy to manipulate, and that can produce hydrogen open-air, thus in the presence of oxygen, for periods of time as long as possible. In this work we investigate by means of predictive computational models, the [FeFe] hydrogenase enzyme of Nannochloropsis salina, a promising microcalga already used to produce high-value products in salt water. Catalysis of water reduction to hydrogen by [FeFe] hydrogenase occurs in a peculiar iron-sulfur cluster (H-cluster) contained into a conserved H-domain, well represented by the known structure of the single-domain enzyme in Chlamydomonas reinhardtii (457 residues). By combining advanced deep-learning and molecular simulation methods we propose for N. salina a two-domain enzyme architecture hosting five iron-sulfur clusters. The enzyme organization is allowed by the protein size of 708 residues and by its sequence rich in cysteine and histidine residues mostly binding Fe atoms. The structure of an extended F-domain, containing four auxiliary iron-sulfur clusters and interacting with both the reductant ferredoxin and the H-domain, is thus predicted for the first time for microalgal [FeFe] hydrogenase. The structural study is the first step towards further studies of the microalga as a microorganism producing pure hydrogen gas. Full article
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