Search Results (86)

The gut microbiota, acting as a critical extrinsic endocrine organ, is profoundly involved in the pathological evolution and therapeutic response of hormone-dependent malignancies. This review elucidates the core mechanisms governing the microbiota, endocrine, and immune triple-axis. Multi-omic and biochemical evidence demonstrates that microbial metabolic networks, comprising the estrobolome, androbolome, and progestobolome/corticobolome, rely on enzymatic systems such as β-glucuronidases (GUS) and steroid-17,20-desmolases to execute hormone deconjugation and structural modification, thereby modulating systemic steroid exposure. Concurrently, microbe-derived metabolites, such as secondary bile acids and purine derivatives, act as inter-kingdom messengers. These metabolites remodel the tumor immune microenvironment by antagonizing hormone receptors and activating specific signaling axes, such as the Inosine-A2AR pathway. By modulating localized immune cells like effector T cells and myeloid cells, they play a pivotal role in tumor immune evasion. Furthermore, pharmacomicrobiomics reveals a bidirectional regulation between anti-tumor agents and the gut microbiota, where endocrine and immunotherapeutic drugs can induce microbial dysbiosis, while specific gut taxa contribute to primary or acquired resistance by enzymatically inactivating drugs (e.g., reductive inactivation of Enzalutamide) or providing hormonal precursors through bypass pathways. Facing translational challenges, such as real-world microbiome complexity and the colonization resistance of indigenous flora, we propose treating the human body as a unified host–microbe holobiont system. Future research should leverage gnotobiotic models and genetic causal inference to establish functional causality. These efforts will facilitate the development of precision tools, including ubiquitin–proteasome system (UPS) modulators, microbial enzyme inhibitors, and engineered live biotherapeutics. Collectively, these systems biology strategies offer a robust framework for overcoming therapeutic resistance in hormone-dependent malignancies. Full article

Glycyrrhizic acid and its derivatives are a crucial class of glycoside terpenoids with significant pharmaceutical and food industry applications. The biotransformation of glycyrrhizin (GL) into glycyrrhetic acid 3-O-mono-β-D-glucuronide (GAMG) and glycyrrhetinic acid (GA) can enhance the production of these valuable compounds. This study aimed to develop strategies to improve the catalytic and operational stability of β-glucuronidase from wild-type Talaromyces pinophilus Li-93, previously known as Penicillium purpurogenum Li-3 (w-PGUS), for efficient GL hydrolysis. Whole cells of T. pinophilus Li-93 expressing w-PGUS were capable of directly converting GL into GAMG. To enhance enzyme stability and reusability, three polymeric supports including, polyurethane foam (PUF), loofah sponge (LS), and polyvinyl chloride (PVC), were evaluated for immobilization of w-PGUS from the fermentation medium. Among these, PUF was the most effective immobilization support, yielding higher immobilization efficiency, GAMG production, and biomass retention. Under optimized conditions (1% PUF, 1.5 g.L⁻¹ w-PGUS inoculum, pH 5.0, 36 °C, 180 rpm), the immobilized w-PGUS produced a final GAMG yield of 3.90 g.L⁻¹, achieving 67.10% immobilization efficiency within 72 h. The PUF-immobilized w-PGUS retained 37.51% of its initial activity after 10 repeated batch reactions, whereas free w-PGUS retained only 6.21%. Additionally, the storage stability of immobilized w-PGUS was significantly higher (40.22%) than that of free w-PGUS (14.74%) after 30 days. Immobilization slightly reduced the initial yield due to mass-transfer limits but enabled much higher cumulative GAMG production through improved stability and reusability. Full article

Antibiotic-resistant, biofilm-forming Escherichia coli may constitute a reservoir of antibiotic resistance and other determinants that can be transmitted to pathogenic bacteria for animals and humans. Therefore, it is crucial to reduce the incidence of these types of E. coli. The aim of this study was to determine whether essential oils from oregano, thyme, sage, and coriander, as well as the postbiotic substance PS412, can inhibit virulence factor genes possessing pig-derived E. coli. It aimed to find a new tool for the prevention and/or elimination of virulent E. coli. Altogether, 16 pig-derived E. coli from a pig farm in the region of Vojvodina (Serbia) were taxonomically identified using MALDI-TOF mass spectrometry; 14 strains (87.5%) with secure genus identification/probable species identification and 2 with highly probable genus identification. The fimA gene was detected in 62.5% of E. coli strains, and the crl gene in 87.5% of the strains. Ec3419/2 contained five analyzed genes. Five E. coli were found to form biofilm, as indicated by their growth on Congo red agar. The strains were mostly multi-resistant to antibiotics. Each E. coli strain produced the damaging enzyme, such as β-glucuronidase and/or α-chymotrypsin. However, they were susceptible to herbal essential oils (HEOs) with average inhibitory zones from 15 to 27 mm in diameter. They were also (6) susceptible to the PS412 (activity to 6400 AU/mL). The results contribute to the practical effectiveness of postbiotic substances, HEOs, and their combination as a novel approach to combating the virulence factors of E. coli. This insight also contributes to the strategy behind the One Health Concept. Full article

Gut microbial β-glucuronidase (GUS) plays a pivotal role at the microbiota—host interface by hydrolyzing glucuronide conjugates, thereby influencing xenobiotic metabolism, enterohepatic circulation, and systemic homeostasis. Dysregulated GUS activity has been increasingly linked to adverse health outcomes, including drug-induced toxicity, inflammation, and cancer. However, current literature often overlooks the enzyme’s dual role in maintaining physiological balance and promoting disease progression, as well as the multidimensional ways in which natural products interact with GUS. This work reviews recent advances in GUS research, emphasizing its structural diversity, functional complexity, and regulatory impact on host health. It also highlights the potential of natural products as precision modulators of GUS activity, capable of direct enzyme inhibition or indirect modulation through reshaping the gut microbiota. These mechanisms collectively influence drug efficacy, toxicity, and the systemic availability of endogenous metabolites. By integrating structural, pharmacological, and microbiological perspectives, this work provides a theoretical foundation for the development of microbiota-targeted therapies centered on GUS. Such approaches may support the rational design of natural product-derived inhibitors and promote their application in disease models, ultimately advancing personalized therapeutic strategies. Full article

Background: Bile acids participate in several metabolic processes, and disturbances in their circulating profiles are commonly observed in type 2 diabetes. In a cohort of older adults, individuals with diabetes exhibited markedly lower concentrations of metabolites derived from lithocholic acid. These findings prompted further evaluation of the metabolic effects of lithocholic acid. Methods: We assessed the actions of lithocholic acid in a mouse model of diabetes induced by a high-fat diet and streptozotocin. Fasting glucose, insulin levels, lipid parameters, and measures of insulin resistance were evaluated. Gut microbial composition, short-chain fatty acids, fecal enzyme activities, intestinal barrier markers, and bile acid patterns were analyzed. In vitro assays examined the direct effects of lithocholic acid on A. muciniphila and bile acid metabolism. Results: Lithocholic acid supplementation lowered fasting glucose and insulin levels and improved insulin resistance. It shifted the gut microbial community toward a healthier structure, increased the abundance of A. muciniphila, and raised short-chain fatty acid concentrations. Fecal bile salt hydrolase and β-glucuronidase activity declined, and intestinal barrier markers improved. Lithocholic acid enhanced TGR5 expression and reduced FXR signaling in the ileum. In vitro, physiologically relevant concentrations promoted A. muciniphila growth and altered microbial bile acid metabolism. Conclusions: Lithocholic acid influences the interactions among gut microbes, bile acid pathways, and host metabolic regulation. These findings suggest that this compound may have value as a dietary component that supports metabolic health in type 2 diabetes. Full article

Heparan sulfate proteoglycans (HSPGs) are essential constituents of the extracellular matrix (ECM) and cell surface, orchestrating a wide range of biological processes, such as cell adhesion, migration, proliferation, and intercellular communication. Through their highly sulfated glycosaminoglycan chains, HSPGs serve as crucial modulators of bioavailability and signaling of growth factors, cytokines, and chemokines, thereby influencing tissue homeostasis. Their dynamic remodeling is mediated by numerous enzymes, with heparanase (HPSE) playing a predominant role as the only known human endo-β-D-glucuronidase that specifically cleaves heparan sulfate chains. Beyond its well-documented enzymatic activity in ECM degradation and the release of HS-bound molecules, HPSE also exerts non-enzymatic functions that regulate intracellular signaling cascades, transcriptional programs, and immune cell behavior. Dysregulated HPSE expression or activity has been implicated in various pathological conditions, including fibrosis, chronic inflammation, cancer progression, angiogenesis, metastasis, and immune evasion, positioning this enzyme as a pivotal driver of ECM plasticity in both health and disease. This review provides an updated overview of HSPG biosynthesis, structure, localization, and functional roles, emphasizing the activity of HPSE and its impact on tissue remodeling and disease pathogenesis. We further explored its involvement in the hallmark processes of cancer, the inflammatory tumor microenvironment, and its contribution to fibrosis. Finally, we summarize current therapeutic strategies targeting HPSE, outlining their potential to restore ECM homeostasis and counteract HPSE-driven pathological mechanisms. A deeper understanding of the HSPG/HPSE axis may pave the way for innovative therapeutic interventions in cancer, inflammatory disorders, and fibrotic diseases. Full article

To discover new probiotics that can protect mammary glands from mastitis, 40 Lactobacillus (Ligilactobacillus) spp. isolates from bovine milk were subjected to a preliminary series of in vitro subtractive analyses. Antibiotic susceptibility testing was performed according to the ISO norm 10932. Many lactobacilli had elevated MIC values for kanamycin (35%), but fewer were resistant to chloramphenicol (15%), streptomycin (7.5%) and tetracycline (5%). The enzymic activities of lactobacilli were tested using an API ZYM system. Nearly 27% exhibited undesirable activities (β-glucuronidase, β-glucosidase and N-acetyl-β-glucosaminidase). The safe strains were monitored for antimicrobial activity against Staphylococcus aureus, Escherichia coli, Salmonella enteritidis, and Bacillus cereus using microtiter plates and for their ability to form biofilms using the crystal violet assay. The antimicrobial activity of lactobacilli against indicator bacteria ranged from 29 to 89% and the isolates exhibited moderate-to-high biofilm formation. Suitable strains were selected for whole-genome sequencing analysis. Antibiotic-resistance genes and putative virulence genes were not predicted in the genomic analysis. Moreover, the isolate Ligilactobacillus salivarius 48 carries genetic information responsible for bacteriocin production that is similar to that encoding salivaricin CRL1328. Our study demonstrates the safety of the above mentioned isolate, which has potential to be used as a probiotic, exerting health benefits through production of antimicrobial substances. Full article

Isoflavone synthase (IFS) is the key enzyme in isoflavonoid biosynthesis and has been functionally characterized in numerous plant species. Glycyrrhiza species, valued for their medicinal properties, accumulate flavonoids with significant physiological activities. Among these, isoflavones play crucial roles in plant growth, development and stress responses. However, the IFS gene family in Glycyrrhiza remains poorly understood. In this study, we identified 10, 9 and 9 IFS genes in G. uralensis, G. inflata and G. glabra, respectively. Phylogenetic analysis classified these genes into four distinct clades (Clade A–D). Further characterization included chromosomal localization, gene structure, conserved motifs, cis-acting elements and synteny analysis. Using yeast one-hybrid (Y1H) screening, dual-luciferase assays and an electrophoretic mobility shift assay (EMSA), these results revealed that auxin response factor 4 (GgARF4) directly binds to the isoflavone synthase 9 (GgIFS9) promoter and activates its expression. Following indole-3-acetic acid (IAA) treatment, RNA-seq revealed that in the differentially expressed genes (DEGs), the genes involved in isoflavonoid and flavonoid biosynthesis pathways were significantly enriched. The result of quantitative reverse transcription polymerase chain reaction (qRT-PCR) revealed that GgIFS9 was strongly induced by IAA. β-Glucuronidase (GUS) assays confirmed that IAA activates the expression of the GgIFS9 promoter in Nicotiana tabacum. Our findings reveal that, through GgARF4 and its downstream-activated gene GgIFS9, IAA may promote flavonoid synthesis in G. glabra. This study provides novel insights into the auxin-mediated regulation of secondary metabolism in Glycyrrhiza species. Full article

Anxiety and depression are mental disorders with significant global impact, and are especially prominent in women during times of hormonal fluctuations. The microbiota–gut–brain axis (MGB axis) has emerged as a crucial pathway in the pathogenesis of these disorders, as it directly influences the production of neurotransmitters such as serotonin (5-HT), gamma-aminobutyric acid (GABA) and dopamine (DA). In addition, they have shown estrogenic effects through enzymes such as β-glucuronidase, which modulate hormone metabolism and consequently mood. A comprehensive search of recent preclinical studies has found that probiotic intake in female rats led to significant improvements in anxiety- and depression-related behaviors. Similarly, clinical trials in certain populations, particularly women with hormonal imbalances during menopause or premenstrual syndrome, have shown promising results. However, there are still significant problems, such as the individual variability of responses and the need for controlled long-term studies. The development of specific probiotics for hormonal modulation and the implementation of personalized approaches integrating omics and neuroimaging technologies to optimize therapeutic interventions in the field of mental health are promising. Accordingly, a comprehensive search was conducted in scientific databases such as PubMed, ScienceDirect, Scopus and Web of Science. Preclinical studies investigating the estrogenic effects of different probiotic strains in animal models and in controlled clinical trials during chronic treatment were selected, excluding those studies that did not provide access to full text. Full article

Background: Polluted rivers may become reservoirs of antibiotic-resistant Escherichia coli (AREc), raising concerns about environmental health. While monitoring is crucial for recognizing their incidence and evaluating mitigation solutions, current approaches are limited due to high costs, labor-intensive methods, and a lack of standardized indicators. This study aims to identify the priority AREc as the monitoring target and evaluate the applicability of the β-glucuronidase enzyme detection method (MPR Method) as an alternative rapid method for profiling AREc in the Upper Citarum River. Methods: River water sampling was conducted along the river during two periods with varying rainfall levels. Total Escherichia coli (TEc) and twelve types of antibiotic-resistant Escherichia coli (AREc) were measured simultaneously by the Agar Method and the β-D-Glucuronidase detection (MPR Method). Results: Statistical data analyses indicate that Total Escherichia coli (TEc) concentrations in the Upper Citarum River increase during periods of higher rainfall (𝓍 = 2558 ± 360 CFU/mL). Erythromycin-resistant Escherichia coli dominates in both periods (Period I 𝓍 = 57.6 ± 25.9%, Period II 𝓍 = 49.96 ± 29.5%). However, tetracycline-resistant Escherichia coli and Extended-Spectrum β-lactamase-producing Escherichia coli (ESBL-Ec) are the most suitable indicators for AREc concentration due to their consistency and correlation with other AREc types. The MPR method achieved an accuracy of up to 87.2%, a sensitivity of 67.4%, and a specificity of 94%. Conclusion: The MPR Method was considered a better alternative for the AREc screening method, particularly in a high bacterial load aquatic environment. Full article

The fungi belonging to the genus Pleurotus can be cultivated in different substrates and represent excellent producers of several extracellular enzymes. In this study, we analyzed eleven hydrolytic enzymes of the P. eryngii and P. ostreatus secretomes, which were collected at three different growth stages after 23 days (mycelial colonization of about 50% of the substrate), 34 days (100% colonization of the substrate) and 50 days (after the first flush). Mushrooms were axenically cultivated on the same substrate. The results demonstrate that proteases, lipases, amylases, α-glucosidase, cellulases (endoglucanase, β-cellobiohydrolase and β-glucosidase) and hemicellulase (xylosidase, glucuronidase, arabinosidase and mannosidase) activities were higher in the secretomes from P. eryngii than those from P. ostreatus. Time course analysis revealed for both species a similar enzymatic activity profile, in which in the early stages of mycelium development, both species use starch as the main carbon source. Protease and lipase activities increased and remained constant during the subsequent formation of fruiting bodies, whereas cellulase and hemicellulase activities decreased after the complete mycelial colonization of the substrate. The zymographic analysis suggested the presence in the secretomes of proteolytic activities belonging to different classes. In conclusion, both mushroom species released into the secretomes a broad spectrum of hydrolytic enzymes potentially useful in various biotechnological fields. Full article

β-glucuronidase is an important hydrolase, which plays an important role in drug metabolism, clinical diagnostics, and biotransformation. This study focuses on the heterologous expression, isolation, purification, and its enzymatic properties of β-glucuronidase CpGUS from Clostridium perfringens, as well as its application in the whole-cell transformation of unconjugated bilirubin from pig bile. A recombinant E. coli BL21(DE3)/pET-28a-CpGUS was constructed for the heterologous expression of CpGUS, with the majority of the expressed enzyme being soluble. Enzymatic analysis showed that CpGUS displayed optimal activity at pH 5.0 and 45 °C, and it rapidly lost activity at pH < 4.5. Metal ions, such as Mg²⁺ and Fe²⁺, enhanced CpGUS catalysis, while Zn²⁺, K⁺, Fe³⁺, Mn²⁺, Cu²⁺, and Na⁺ inhibited it. Notably, Cu²⁺ and Fe³⁺ can significantly inhibit β-glucuronidase, resulting in the complete loss of its activity. The results of the whole-cell transformation experiment show that when E.coli BL21(DE3)/ pET-28a-CpGUS at an OD₆₀₀ of 10 was incubated at pH 5.0, a temperature of 45 °C, and a rotation speed of 200 rpm for 12 h, the hydrolysis rate of the conjugated bilirubin in pig bile reached 81.1%, the yield of unconjugated bilirubin was 76.8%, and the purity of unconjugated bilirubin was 98.2%. The three-dimensional structure of CpGUS was predicted using AlphaFold2 (AlphaFold v2.0, DeepMind Technologise Limited, London, UK), and p-Nitrophenyl-β-D-Glucuronide (pNPG) and conjugated bilirubin were then docked to the CpGUS protein model using SWISSDOCK. The best docked conformations of the CpGUS–pNPG and CpGUS–conjugated bilirubin complex systems were simulated by independent 500 ns molecular dynamics (MD) runs with the RSFF2C force field, and the binding dynamic and catalytic mechanism of each system were obtained. The results indicated that π-π stacking, hydrogen bonding, and hydrophobic interactions between the key residue Tyr472 and the benzene ring of pNPG molecules are crucial for its catalytic process. Similarly, for the binding and catalysis of conjugated bilirubin by CpGUS, the π-π stacking and hydrogen bonding and hydrophobic interactions between the sidechains of residues Phe368 and Tyr472 and the benzene ring of conjugated bilirubin play a synergistic role during its catalytic process. Their total binding free energy (∆G_bind) values were calculated to be as high as −65.05 ± 12.66 and −86.70 ± 17.18 kJ/mol, respectively. These results suggest that CpGUS possesses high binding and catalytic hydrolysis properties for both pNPG and conjugated bilirubin. Full article

Vertebrate mast cells are the first cells to initiate the inflammatory response. The origin of these highly specialised innate immunity cells in chordates is an intriguing unanswered question, and tunicates represent the best candidates to address this question for their close relationship with vertebrates. In the colonial ascidian Botrylloides leachii, a particular cell type circulates in the haemolymph, namely, ‘granular cell’, which is a distinct immunocyte from both phagocytic and cytotoxic lines. Like mast cells and unlike basophils, granular cells were labelled with anti-c-kit antibody on their plasmalemma and exhibited a high content of heparin in their granules, as revealed by various histochemical techniques. Immunohistochemistry revealed the presence of heparin and histamine inside the same granules resembling the granules of mast cells. Histoenzymatic assays revealed the presence of mast cell enzymes such as β-glucuronidase, arylsulphatase, chloroacetyl esterase, and proteases. These cells degranulated after exposure to bacteria, compound 48/80, or heterologous plasma. During exposure to bacteria, they crowd into the perivisceral sinus and then infiltrate the epithelium of the postbranchial gut, where they release the content of their granules, a behaviour remarkably similar to that of the gastric leukopedesis of mast cells. Full article

Baicalein is a unique flavonoid compound with important pharmacological activities, derived from Scutellaria baicalensis Georgi. Baicalein, as the aglycone of baicalin, is a key form for exerting pharmacological activity in vivo. β-glucuronidases (GUSs) are the enzymes involved in the conversion of baicalin to baicalein. In this study, the content of baicalein in S. baicalensis was significantly increased by 20.44% after treatment with 5% PEG6000. Seven GUSs from the glycoside hydrolase 79 family were identified through comparative transcriptome analysis. Among them, GUS1 and GUS2 were confirmed to have catalytic activity in converting baicalin to baicalein in prokaryotic and eukaryotic systems. The correlation analysis further revealed a significant positive correlation of 0.962 (p < 0.01) between the expression of GUS2 and baicalein content in six different sources of S. baicalensis. Interestingly, the presence of variable sites in the GUS1 and GUS2 genes significantly affected their catalytic efficiency in the S. baicalensis samples from the six geographic origins. These findings also provide valuable GUS biological enzyme resources for the effective synthesis of baicalein and offer new insights into the accumulation pattern of baicalein in S. baicalensis. Full article

Background and Clinical Significance: Mucopolysaccharidosis type VII (MPS VII), an ultrarare lysosomal storage disorder caused by β-glucuronidase deficiency, presents significant therapeutic challenges. Given its extreme rarity and limited treatment experience in Asian populations, documenting long-term treatment outcomes is crucial for advancing clinical knowledge and improving patient care. Case Presentation: We report a 3-year follow-up of enzyme replacement therapy (ERT) in the first Taiwanese case of MPS VII. The patient, who initially presented with hydrops fetalis and developmental delay, was diagnosed at age 4 through genetic analysis, which revealed compound heterozygous variants of c.104C > A (p.Ser35Ter) and c.1454C > T (p.Ser485Phe) on the GUSB gene. ERT with vestronidase alfa was initiated at age 6. During the follow-up period, significant clinical improvements were observed, including elimination of oxygen dependency, with BiPAP needed only during sleep; changes in mobility, with 6-min walk test distance showing an initial decline from 130 to 70 m followed by partial recovery to 95 m after multiple orthopedic surgeries; and steady progression of growth parameters showed, with height increasing from 110 to 118 cm. Urinary glycosaminoglycan (GAG) levels measured by dimethylmethylene blue spectrophotometry decreased and stabilized. The patient’s cardiac and hepatic conditions remained stable, although splenomegaly persisted. No severe adverse events were reported during ERT. Conclusions: This case demonstrates the effectiveness and safety of long-term ERT in MPS VII, particularly in improving respiratory function and physical performance. Our experience highlights the importance of early diagnosis and treatment initiation, while providing valuable insights into the management of this ultrarare disease in the Asian population. Full article