BMC Biotechnology最新文献

Patulin (PAT), a mycotoxin produced primarily by Penicillium expansum, poses significant health risks and frequently contaminates apples and apple-derived products, often exceeding permissible safety limits. This study investigated the potential of orotate phosphoribosyl transferase (URA5) to degrade PAT in apple juice under controlled conditions. PAT degradation was assessed at initial concentrations of 100 µg/L and 250 µg/L, with enzymatic treatment using 0.2 mg/mL URA5. Samples were incubated for up to 24 h, and PAT degradation was monitored at time intervals of 3, 6, 9, 12, 18, and 24 h using liquid chromatography-mass spectrometry (LC-MS). The results demonstrated a time-dependent PAT degradation, with significant reductions observed as incubation time increased. After 6 h, PAT concentrations decreased to 57.30 µg/L and 112.69 µg/L for the 100 µg/L and 250 µg/L samples, respectively. At 12 h, PAT levels in the 100 µg/L sample fell just below the permissible limit (50 µg/kg), while substantial degradation was observed in the 250 µg/L sample. By 18 h, PAT concentrations dropped further to 47.22 µg/L and 40.10 µg/L, reaching safe consumption levels. After 24 h, degradation rates reached 96.36% and 98.25%, reducing PAT levels to 30.22 µg/L and 31.48 µg/L, confirming the efficacy of URA5 in detoxifying PAT-contaminated apple juice. The findings highlight the potential application of URA5 as a biocatalyst for PAT detoxification in the fruit juice industry. Compared to existing detoxification methods, enzyme-based degradation presents a promising, environmentally friendly, cost-effective, and non-toxic alternative. Further studies should explore its feasibility in large-scale processing and its interaction with other contaminants in commercial apple juice production.

Background: The composition and roles of intestinal microbial populations have been clarified including mammals and humans however, less is understood concerning the gut microbiota of mollusks. For the first time, we investigated non-parasite transmitting freshwater snails Lanistes carinatus (L. carinatus), Cleopatra bulimoides (C. bulimoides) and Helisoma duryi (H. duryi) gut microbiota as a source of probiotic strains with anticancer potential and explore their microbial population structure.

Results: Our investigation demonstrated significant variation in microbial richness, identifying 32 bacterial phyla across the three snail species. Pseudomonadota (44-60%) and Bacteroidota (17-20%) were identified as the predominant phyla in all snails, with p value = 0.28 and 0.39, respectively in relative abundance. Distinct compositional changes were observed as L. carinatus had a greater abundance of Bacillota. H. duryi exhibited higher microbial diversity with Verrucomicrobiota and Cyanobacteria comprising 5-20% of its gut microbiota. Lysinibacillus macroides (L. macroides), Kurthia huakuii (K. huakuii) and Enterococcus faecium (E. faecium) were isolated from L. carinatus, C. bulimoides and H. duryi, respectively. L. macroides, K. huakuii and E. faecium demonstrated antimicrobial efficacy towards selected pathogenic strains. The bacterial isolates displayed significant tolerance to acidic pH and bile salts concentrations (0.3% and 0.7% w/v). The cytotoxicity of the microbial isolates secreted metabolites was examined using the MTT assay. Cytopathological changes and caspase-3 / TNF α immunohistochemistry were examined on Caco-2 cells. Results demonstrated the anticancer activity of the metabolites of the three microbial isolates on Caco2 cells where K. huakuii exhibited the highest enhancement in apoptosis and necrosis.

Conclusions: Our study identified diverse bacterial populations in freshwater snail gut microbiota with compositional differences. The isolated bacterial strains showed promising antimicrobial properties and anticancer potential, particularly K. huakuii. These results suggest that snails could be used as niche sources for beneficial bacteria with biotechnological and therapeutic applications.

Background: In China, lung cancer stands as the leading cause of cancer-related deaths, often resulting in brain metastases (BM) that severely compromise patients' quality of life and reduce survival outcomes. The delivery of drugs to the brain is further complicated by the blood-brain barrier (BBB). To address this, we developed EGFR single-chain fragment variable (scFv)-modified macrophage membrane liposomes (scFv-MML) encapsulating LPCAT1 siRNA (scFv-MML@LPCAT1si) as a targeted therapy for non-small cell lung cancer (NSCLC) BM.

Methods: EGFR scFv-pcDNA3.1(-) plasmids were transfected into RAW 264.7 cells to generate RAW 264.7-scFv cells. Macrophage membranes were isolated from these cells and used to coat liposomes (Lip) encapsulating LPCAT1 siRNA via extrusion. The cellular uptake, LPCAT1 silencing, and anti-tumor efficacy of scFv-MML@LPCAT1si were evaluated in vitro using PC9 lung cancer cells. In vivo studies were performed in a mouse model of NSCLC BM to assess tumor targeting, accumulation, and therapeutic effects.

Results: In vitro, scFv-MML@LPCAT1si exhibited superior cellular uptake and silencing of LPCAT1 expression in EGFR-positive PC9 cells compared to control liposomes, leading to increased cell apoptosis and decreased proliferation. In vivo, scFv-MML@LPCAT1si showed improved tumor targeting and accumulation in the brain, effectively slowing tumor growth and reducing body weight loss in mice with NSCLC BM. The biodistribution study revealed sustained tumor fluorescence intensity for more than 24 h after injection, with significant retention of siRNA within the tumor site. No significant systemic toxicity or organ damage was observed in mice treated with scFv-MML@LPCAT1si.

Conclusions: Our findings suggest that scFv-MML@LPCAT1si represents a promising targeted therapy for NSCLC BM, leveraging the unique properties of scFv-MML to traverse the BBB and deliver therapeutic payloads to tumor sites with high accuracy and efficiency.

Ultraviolet B (UVB) radiation severely damages human skin by causing DNA damage, oxidative stress, and collagen degradation. This study explored the photoprotective properties of Asparagus cochinchinensis extracts fermented with endophytic fungus Aspergillus aculeatus TD103. Compared to the unfermented control, TD103-fermented A. cochinchinensis demonstrated stronger radical scavenging and ferric ion reduction abilities in vivo, significantly reduced intracellular reactive oxygen species (ROS) and increased the antioxidant enzymes including heme oxygenase-1 (HO-1), superoxide dismutase (SOD), and catalase (CAT) in UVB-induced HaCaT cells. It also downregulated the expression of the AP-1 and MMP genes, reduced the content of matrix metalloproteinase (MMP-1) and increased type I procollagen amino-terminal propeptide (PINP) levels in UVB-induced HaCaT cells. Non-targeted metabolomics and HPLC quantification revealed that elevated sarsasapogenin content may critically contribute to enhanced photoprotective capacity in Asparagus cochinchinensis. The safety assessment of fungus TD103 revealed that this strain was not drug resistant and did not produce mycotoxins, thereby indicating its safety for application. Eye irritation tests demonstrated the safety profile of the fermented extract, indicating negligible irritant potential. The TD103-mediated fermentation markedly potentiated the photoprotective capacity of A. cochinchinensis, ​providing a viable biotechnological platform for sustainable cosmeceutical development targeting UV-induced skin damage.

Background: Drought is an abiotic stress that significantly reduces the yield of thyme (Thymus vulgaris). This study investigated how iron oxide nanoparticles (FeNPs), together with symbiotic bacterial (Azospirillum lipoferum) and fungal (Aspergillus oryzae) endophytes, modulate osmotic adjustment, molecular and biochemical mechanisms related to photosynthesis, and drought tolerance mechanisms in thyme.

Results: The experiment was evaluated as a factorial experiment in a completely randomized design with three replications. evaluating three treatment factors: four irrigation levels (100%, 75%, 50%, and 25% of field capacity), four FeNPs concentrations (0, 0.5, 1, and 1.5 mg L⁻¹), and three endophyte treatments (control, bacterial (EB), and fungal (EF) inoculation). At 25% FC, EB and spraying with 1 mg L^- 1 FeNPs increased Fv/Fm (maximum quantum efficiency of photosystem II), chlorophyll a, chlorophyll b, and total chlorophyll, carotenoids, relative water content (RWC), and protein levels level protein levels by 18.75%, 10.41%, 31.54%, 18.20%, 14.26%, 35.53%, and 125.22% respectively, compared to the control. At 25% FC, electrolyte leakage (EL) was increased by 47.44% with the combination of EF and 1.5 mg L^- 1 FeNPs. The highest proline accumulation at 25% FC was observed after inoculation with EF and 1 mg L^- 1 FeNPs, resulting in significant increases of 36.36% and 13.04%, respectively, compared to the control. Soluble sugar was remarkably increased by 28.57% under upon treatment with FeNPs (1.5 mg L^- 1 FeNPs). At 25% FC, EB and 1.5 mg L^- 1 FeNPs showed significant reductions of 17.33% and 37.10%, respectively, in malondialdehyde levels compared to control plants. At 50% FC, 1 mg L⁻¹ FeNPs increased Catalase by 15%, peroxidase by 31.25%, and superoxide dismutase by 43.42%, while higher concentrations reduced enzyme activities. Similarly, 1.5 mg L⁻¹ FeNPs and EB inoculation enhanced ascorbate peroxidase by 37.44% and 17.37%, respectively. FeNPs acted as abiotic stressors at low levels but became toxic at higher concentrations.

Conclusion: Our findings demonstrate that the synergistic application of FeNPs and endophytes significantly enhances drought tolerance in T. vulgaris by optimizing photosynthetic efficiency (Fv/Fm, chlorophyll content) and preserving membrane integrity (RWC, MDA reduction). These results provide a framework for leveraging nano-bio partnerships to improve crop resilience under water scarcity.

Base editing technologies allow for the precise and efficient installation of defined nucleotide substitutions into a target genome without the introduction of double strand breaks or DNA templates. Here we describe two recombinant, protein format cytosine base editors (CBEs) that efficiently catalyze the installation of cytosine-to-thymine edits, termed "Flexible" and "Precision." Flexible exhibits a wide editing window, while Precision uses a fused single-stranded DNA binding protein to narrow the editing window, lowering the risk of editing multiple cytosine residues at the target site. We show that co-transfection with uracil glycosylase inhibitor protein increases the proportion of substitutions that are C-to-T and the ratio of C-to-T editing to indel formation, thus reducing undesired editing outcomes. We use in vitro editing assays to characterize our editors and show a preference for cytosine residues preceded by thymine (TpC dinucleotides) and unmethylated cytosine residues.

Colorectal cancer is one of the deadliest forms of gastrointestinal cancer, with conventional treatments often facing significant limitations. As a result, new approaches, particularly in targeted drug delivery, have shown great promise. In this study, the COF-FA@DOX nanocarrier was developed, where covalent organic frameworks (COFs) were functionalized with folic acid (FA) and then loaded with Doxorubicin (DOX). The as-synthesized COF-FA@DOX nanocarrier was characterized using different techniques. To assess its anticancer effectiveness, MTT, flow cytometry, and scratch assays were conducted on SW480 and HUVEC cells to examine cell viability, cellular uptake, cell cycle progression, apoptosis, and cell migration, respectively. The obtained results demonstrated that the COF-FA@DOX nanocarrier was efficiently internalized by cancer cells and showed significantly higher cytotoxicity compared to other synthesized nanocarrier groups and free DOX drug. Moreover, the COF-FA@DOX nanocarrier caused cell cycle arrest, induced apoptosis, and inhibited cell migration at lower doses than the free DOX drug. Altogether, these findings suggest that the COF-FA@DOX nanocarrier is an effective and promising drug delivery system for DOX in colorectal cancer, potentially enhancing the therapeutic efficacy of DOX drug while minimizing side effects through targeted delivery. Further investigation is required to assess their efficacy in vivo and discover potential clinical applications.