Search Results (669)

This study identified the optimal enzymatic treatment for improving the foaming characteristics of oat globulin, and alkaline protease was found to be the most effective enzyme. The impact of alkaline protease on the foaming properties and structural changes in oat globulin was explored. The results show that the foaming capacity of oat globulin hydrolysates is negatively correlated with surface hydrophobicity and positively correlated with the degree of hydrolysis. The results of circular dichroism (CD) and size-exclusion chromatography (SEC) indicate that hydrolysis generated smaller, disordered peptides. Under equilibrium conditions at a 2% concentration, a reduction of 1.62 mN/m in surface tension and an increase of 3.82 μm in foam film thickness were observed. These peptides reduce surface tension between air and water, forming larger, thicker, and more stable foams. Compared to untreated oat globulin, the foaming capacity of hydrolyzed ones increased by 87.17%. Under comparable conditions, these findings demonstrate that limited hydrolyzed oat globulin exhibits potential as an effective plant-based foaming agent up to a degree of hydrolysis of 15.06%. Full article

Consumer interest in vegetarian, ethical, and clean-label foods is reviving the use of plant-derived milk coagulants. Cardosins from Cynara cardunculus (“thistle”) are aspartic proteases with strong clotting activity, yet their technological impact in cheese remains under-explored. This study compared a commercial thistle extract (PC) with traditional bovine rennet rich in chymosin (AC) during manufacture and 60-day ripening of Caciotta cheese. Classical compositional assays (ripening index, texture profile, color, solubility) were integrated with scanning electron microscopy, three-dimensional surface reconstruction, and descriptive sensory analysis. AC cheeses displayed slower but sustained proteolysis, yielding a higher and more linear ripening index, softer body, greater solubility, and brighter, more yellow appearance. Imaging revealed a continuous protein matrix with uniformly distributed, larger pores, consistent with a dairy-like sensory profile dominated by milky and umami notes. Conversely, PC cheeses underwent rapid early proteolysis that plateaued, producing firmer, chewier curds with lower solubility and darker color. Micrographs showed a fragmented matrix with smaller, heterogeneous pores; sensory evaluation highlighted vegetal, bitter, and astringent attributes. The data demonstrate that thistle coagulant can successfully replace animal rennet but generates cheeses with distinct structural and sensory fingerprints. The optimization of process parameters is therefore required when targeting specific product styles. Full article

Microbe–microbe interactions have been explored at the molecular level to a lesser degree than plant–pathogen interactions, primarily due to the economic impact of crop losses caused by pathogenic microorganisms. Effector proteins are well known for their role in disease development in many plant–pathogen pleinteractions, but there is increasing evidence showing their involvement in other types of interaction, including microbe–microbe interactions. Through the use of LC-MS/MS sequencing, effector candidates were identified in the in vitro interaction between a banana pathogen, Pseudocercospora fijiensis and a biological control agent, Trichoderma harzianum. The diverse interaction secretome revealed various glycoside hydrolase families, proteases and oxidoreductases. T. harzianum secreted more proteins in the microbial interaction compared to P. fijiensis, but its presence induced the secretion of more P. fijiensis proteins that were exclusive to the interaction secretome. The interaction secretome, containing 256 proteins, was screened for effector candidates using the algorithms EffHunter and WideEffHunter. Candidates with common fungal effector motifs and domains such as LysM, Cerato-platanin, NPP1 and CFEM, among others, were identified. Homologs of true effectors and virulence factors were found in the interaction secretome of T. harzianum and P. fijiensis. Further characterization revealed a potential novel effector of T. harzianum. Full article

Plant latex is a rich source of proteolytic enzymes with potential biomedical applications, particularly in hemostasis. Among them, thrombin-like enzymes (TLEs) have garnered interest in their ability to mimic thrombin by catalyzing the conversion of fibrinogen to fibrin, facilitating clot formation. While TLEs from snake venoms have been well-characterized and applied clinically, their plant-derived counterparts remain underexplored. This review critically examines the structural and functional characteristics of TLEs from plant latex, comparing them to animal-derived TLEs and evaluating their role in both procoagulant and fibrinolytic processes. Emphasis is placed on dual fibrinogenolytic and fibrinolytic activities exhibited by latex proteases, which often vary with concentration, incubation time, and protease type. In vitro coagulation assays and electrophoretic analyses are discussed as critical tools for characterizing their multifunctionality. By addressing the knowledge gaps and proposing future directions, this paper positions plant latex proteases as promising candidates for development in localized hemostatic and thrombolytic therapies. Full article

Gingival inflammation compromises the integrity of the gingival epithelium and the underlying tissues, highlighting the need for adjuvant therapies with immunomodulatory and healing properties. Casearia sylvestris, a medicinal plant known as guaçatonga, is traditionally used to treat inflammatory lesions. This study aimed to investigate the effects of C. sylvestris on the synthesis of pro- and anti-inflammatory, proteolytic, and antioxidant molecules and on wound healing in epithelial cells. A human telomerase-immortalized gingival keratinocyte cell line (TIGKs) was used, and cells were exposed to Escherichia coli lipopolysaccharide (LPS) in the presence and absence of C. sylvestris extract, its diterpene-concentrated fraction, and its clerodane diterpene casearin J for 24 h and 48 h. Gene expression and protein synthesis were analyzed by RT-qPCR and ELISA, respectively. Nitric oxide (NO) and NF-κB activation were analyzed by Griess reaction and immunofluorescence, respectively. Additionally, cell viability was evaluated by alamarBlue^® assay, and an automated scratch assay was used for wound healing. LPS significantly increased the expression of cytokines (TNF-α, IL-1β, IL-6, IL-8, IL-10, IL-17), proteases (MMP-1 and MMP-13), iNOS as well as NO synthesis, and triggered NF-κB nuclear translocation. It also reduced IL-4 expression, cell viability, and cellular wound repopulation. Treatment with C. sylvestris derivatives significantly abrogated all aforementioned LPS-induced effects by 80–100%. Furthermore, even at higher concentrations, C. sylvestris did not affect cell viability, thus proving the safety of its derivatives. C. sylvestris exerts anti-inflammatory, antiproteolytic, and antioxidant effects on gingival keratinocytes, highlighting its potential as a valuable adjunct in the prevention and treatment of periodontal diseases. Full article

Soybean meal (SBM) is extensively used as a predominant protein source in animal feed. However, raw soybean cannot be directly utilized in animal feed, due to the presence of the Kunitz trypsin inhibitor (KTi) and the Bowman–Birk protease inhibitor (BBi). These antinutritional factors inhibit the digestive enzymes in animals, trypsin and chymotrypsin, resulting in poor animal performance. To inactivate the activity of protease inhibitors, SBM is subjected to heat processing, a procedure that can negatively impact the soybean protein quality. Thus, it would be beneficial to develop soybean varieties with little or no trypsin inhibitors. In this study, we report on the creation of experimental soybean lines with significantly reduced levels of Bowman–Birk protease inhibitors. RNA interference (RNAi) technology was employed to generate several transgenic soybean lines. Some of these BBi knockdown soybean lines showed significantly lower amounts of both trypsin and chymotrypsin inhibitor activities. Western blot analysis revealed the complete absence of BBi in selected RNAi-derived lines. RNA sequencing (RNAseq) analysis demonstrated a drastic reduction in the seed-specific expression of BBi genes in the transgenic soybean lines during seed development. Confocal fluorescence immunolabeling studies showed that the accumulation of BBi was drastically diminished in BBi knockdown lines compared to wild-type soybeans. The absence of BBi in the transgenic soybean did not alter the overall protein, oil, and sulfur amino acid content of the seeds compared to wild-type soybeans. The seed protein from the BBi knockdown lines were more rapidly hydrolyzed by trypsin and chymotrypsin compared to the wild type, indicating that the absence of BBi enhances protein digestibility. Our study suggests that these BBi knockdown lines could be a valuable resource in order for plant breeders to incorporate this trait into commercial soybean cultivars, potentially enabling the use of raw soybeans in animal feed. Full article

Drought stress poses a major constraint on global crop productivity. Although aspartic proteases (APs) are primarily characterized in plant disease resistance, their roles in abiotic stress adaptation remain largely unexplored. Here, we demonstrate that rice (Oryza sativa) OsAP4 critically regulates drought stress tolerance at the seedling stage. Genetic manipulation through overexpression (OsAP4-OE) or CRISPR knockout (OsAP4-KO) resulted in significantly reduced or enhanced stress tolerance compared to wild-type plants, respectively. Through integrated approaches including yeast two-hybrid, bimolecular fluorescence complementation, pull-down, co-immunoprecipitation, and protein degradation assays, we established that OsAP4 physically interacts with and destabilizes OsCATA/OsCATC, two catalase enzymes responsible for reactive oxygen species (ROS) scavenging. Importantly, OsAP4 modulates ROS production under drought stress treatment conditions. Together, these findings reveal a novel OsAP4-OsCATA/OsCATC regulatory module governing rice drought stress responses. Full article

Rapeseed meal, a byproduct of oil extraction, is increasingly recognised as a valuable source of plant protein and health-promoting peptides. This study aimed to identify key proteins in cold-pressed rapeseed meal and assess their potential to release bioactive peptides through in silico hydrolysis using plant-derived proteases, namely papain, bromelain, and ficin. Proteomic profiling via two-dimensional electrophoresis and MALDI-TOF/TOF mass spectrometry revealed cruciferin as the dominant protein, along with other metabolic and defence-related proteins. In silico digestion of these sequences using the BIOPEP database generated thousands of peptide fragments, of which over 50% were predicted to exhibit bioactivities, including ACE and DPP-IV inhibition, as well as antioxidant, neuroprotective, and anticancer effects. Among the evaluated enzymes, bromelain exhibited the highest efficacy, yielding the greatest quantity and diversity of bioactive peptides. Notably, peptides with antihypertensive and antidiabetic properties were consistently identified across all of the protein and enzyme variants. Although certain rare functions, such as anticancer and antibacterial activities, were observed only in specific hydrolysates, their presence underscores the broader functional potential of peptides derived from rapeseed. These findings highlight the potential of rapeseed meal as a sustainable source of functional ingredients while emphasising the necessity for experimental validation to confirm the predicted bioactivities. Full article

Deubiquitination plays a pivotal role in regulating plant responses to abiotic stress, growth, and development. Among the deubiquitinase (DUB) families, ubiquitin-specific proteases (UBPs) constitute the largest group. Despite this, limited research has been conducted on the functional characteristics of the UBP gene family in soybean (Glycine max). In this study, we identified 52 UBP gene family members in soybean, all of which harbored UCH (ubiquitin C-terminal hydrolase) domains with short yet evolutionarily conserved Cys-box and His-box. These genes were phylogenetically classified into 14 distinct groups; GmUBP genes within the same group shared analogous patterns of conserved domains and motifs. Moreover, a synteny analysis reveals that the GmUBP family has undergone extensive gene duplication events and shares a close evolutionary relationship with Arabidopsis thaliana. We conducted a focused analysis on GmUBP7, which is a gene exhibiting high expression levels in soybean seeds. Intriguingly, this gene exhibited several haplotypes in natural soybean varieties, with significant differences being observed in relation to seed traits, such as 100-seed weight, total fatty acid content, and protein content among different haplotypes. Collectively, the findings from this study provide a foundation for the functional characterization of GmUBP genes, offering new insights into the regulatory network underlying seed development in soybean. Full article

Plant-parasitic nematodes represent a significant threat to agriculture, causing substantial economic losses worldwide. Among the biological alternatives for their control, the genus Trichoderma has emerged as a promising solution for suppressing various nematode species. This article reviews key studies on the interaction between Trichoderma spp. and plant-parasitic nematodes, highlighting the most studied species such as Trichoderma harzianum, Trichoderma longibrachiatum, Trichoderma virens, and Trichoderma viride, mainly against the genera Meloidogyne, Pratylenchus, Globodera, and Heterodera. Trichoderma spp. act through mechanisms such as mycoparasitism, antibiosis, competition for space in the rhizosphere, production of lytic enzymes, and modulation of plant defense responses. They also produce metabolites that affect nematode mobility, reproduction, and survival, such as gliotoxin, viridin and cyclosporine A. In addition, they secrete enzymes such as chitinases, proteases, lipases, and glucanases, which degrade the cuticle of nematodes and their eggs. Furthermore, Trichoderma spp. induce systemic resistance in plants through modulation of phytohormones such as jasmonic acid, ethylene, salicylic acid and auxins. The use of Trichoderma in integrated nematode management enables its application in combination with crop rotation, organic amendments, plant extracts, and resistant varieties, thereby reducing the reliance on synthetic nematicides and promoting more sustainable and climate-resilient agriculture. Full article

Plant growth-promoting rhizobacteria (PGPR) have significant potential for enhancing soil quality and plant growth; however, their agricultural application is limited by challenges such as immobilization and desiccation vulnerability. Background: This study addressed PGPR solid formulation by applying JetCutter-assisted immobilization technology to PGPR strains isolated from the rhizosphere of hazelnut (Corylus avellana). Methods: Four immobilized PGPR strains were evaluated under controlled greenhouse conditions: Serratia proteamaculans, Pseudomonas mohnii, Pseudomonas baetica, and Bacillus safensis. Their effects on root development, gas exchange parameters, dissolved organic carbon (DOC), and soil enzymatic activities (phosphatase, urease, protease, and β-glucosidase) were assessed. Principal component analysis (PCA) was used to identify the top-performing strain. Results: Treatment with encapsulated bacteria resulted in a 27% increase in DOC compared to controls (p < 0.05), while phosphatase and urease activities increased by 35% and 28%, respectively. Root length and volume improved by 18% and 22%, respectively, with PCA identifying P. baetica as the most effective strain. Conclusions: Immobilized Gram-negative PGPR strains enhanced root development and soil biochemical activity in hazelnuts, whereas B. safensis enhanced photosynthesis but had minimal impact on soil properties. These results highlight functional differences and support the use of PGPR immobilization to promote early plant establishment. Full article

Proteins play pivotal roles in safeguarding plants against numerous biotic and abiotic stresses. Understanding their biological functions and mechanisms of action is essential for advancing plant biology, agriculture, and biotechnology. This review considers the diversity and potential applications of plant defense proteins including pathogenesis-related (PR) proteins, chitinases, glucanases, protease inhibitors, lectins, and antimicrobial peptides. Recent advances, such as the omics technologies, have enabled the discovery of new plant defense proteins and regulatory networks that govern plant defense responses and unveiled numerous roles of plant defense proteins in stress perception, signal transduction, and immune priming. The molecular affinities and enzymatic activities of plant defense proteins are essential for their defense functions. Applications of plant defense proteins span agriculture, biotechnology, and medicine, including the development of resistant crop varieties, bio-based products, biopharmaceuticals, and functional foods. Future research directions include elucidating the structural bases of defense protein functions, exploring protein interactions with ligands and other proteins, and engineering defense proteins for enhanced efficacy. Overall, this review illuminates the significance of plant defense proteins against biotic stresses in plant biology and biotechnology, emphasizing their potential for sustainable agriculture and environmental management. Full article

This review presents information obtained over the past 10 years on the methods to control the widespread worldwide phytopathogen Pectobacterium carotovorum subsp. carotovorum (Pcc). This bacterium is among the ten most dangerous phytopathogens; it affects a wide range of cultivated plants: vegetables, ornamental and medicinal crops, both during vegetation and during the storage of fruits. Symptoms of Pcc damage include the wilting of plants, blackening of vessels on leaves, stems and petioles. At the flowering stage, the stem core gradually wilts and, starting from the root, the stem breaks and the plant dies. Pcc is a rod-shaped, non-capsule and endospore-forming facultative anaerobic Gram-negative bacterium with peritrichous flagellation. Pcc synthesizes bacteriocins—carocins. The main virulence factors of Pcc are the synthesis of N-acyl-homoserine lactone (AHL) and plant cell wall-degrading enzymes (PCWDEs) (pectinases, polygalacturonases, cellulases, and proteases). Diagnostic methods for this phytopathogen include polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), multilocus genotyping of strain-specific genes and detection of unique volatile organic compounds (VOCs). The main methods to control this microorganism include the use of various chemicals (acids, phenols, esters, salts, gases), plant extracts (from grasses, shrubs, trees, and algae), antagonistic bacteria (Bacillus, Pseudomonas, Streptomyces, and lactic acid bacteria), viruses (including a mixture of bacteriophages), and nanomaterials based on metals and chitosan. Full article

This study aims to identify the optimal conditions—host, plant material, seasonality, and agricultural practices—for isolating and developing a collection of culturable endophytic microorganisms to support sustainable Olea europaea L. cultivation. Samples were collected from three Sicilian olive cultivars (‘Nocellara del Belice’, ‘Nocellara Etnea’, and ‘Nocellara Messinese’) and six wild olive accessions across different phenological phases and under organic and conventional agronomic management. Endophytes were isolated from leaves and twigs using a culture-dependent approach, and their taxonomic diversity and plant-growth-promoting (PGP) traits were analyzed. A total of 133 endophytic isolates were identified, spanning bacterial (Proteobacteria, Firmicutes, and Actinobacteria) and fungal (Ascomycota and Basidiomycota) phyla. Wild olive trees contributed more than cultivated varieties to enriching the diversity and composition of culturable endophyte collection as well as twigs instead of leaves. Winter sampling allowed to implement the taxonomic genera of olive endophyte collection. Both farming systems favored an increase in the composition of microbial collection, though organic farming systems supported greater microbial richness. Functional analysis highlighted key PGP traits in a selection of bacterial isolates, including indole-3-acetic acid and siderophore production, nitrogen fixation, and antifungal activity. Bacillus spp. dominated enzymatic activities, such as amylase, protease, and lipase production, as well as antifungal activity against the olive fungal pathogen Neofusicoccum vitifusiforme. This research highlights the significant diversity and functional potential of Mediterranean olive endophytes. Our findings emphasize the role of native microbial communities as bio-inoculants, promoting plant growth, nutrient uptake, and disease resistance. These insights lay the groundwork for developing targeted olive-microbial consortia for biocontrol and stress tolerance applications. Full article

Bradysia difformis Frey (Diptera: Sciaridae) is a fungus gnat that poses a significant threat to greenhouse cultures, and is attracted to soils devoid of peat. Fungal strains from the German Collection of Microorganisms and Cell Culture (DSMZ), such as Beauveria bassiana, Metarhizium flavoviride, Mucor hiemalis, and Niesslia tinuis, as well as Serendipita indica, were screened for entomopathogenic activity against B. difformis and their capacity to colonize Petunia hybrida cv. “Mitchell” and Ocimum basilicum plants. The survival rates of Bradysia difformis (three instar larvae) treated with Metarhizium flavoviride were 45.33% at 14 days following inoculation with 1 × 10⁶ spores/mL of each fungal strain, when compared to others. We concluded that the fungal strain M. flavoviride could serve as an entomopathogenic fungus with the highest virulence against B. difformis larvae. Although M. flavoviride did not show a beneficial effect as an endophyte, interestingly, the strain Niesslia tinuis exhibited plant growth benefits in Petunia hybrida cv. “Mitchell” by enhancing its shoot length up to 13.18 ± 0.72 cm, whereas the control treatment had a shoot length up to 10.68 ± 0.39. Enzymatic assays confirmed the ability of M. flavoviride to produce cuticle-degrading enzymes such as chitinase and protease. Together, these findings highlight the potential of EMPF—particularly M. flavoviride—as a sustainable biocontrol tool well-suited for peat-free horticultural systems, offering an eco-friendly alternative to chemical insecticides where fungus gnat pressure is typically high. Full article