3 Biotech最新文献

Butyric acids are important flavor compounds in strong-aroma Baijiu, mainly synthesized by the microbial community in cellar mud. However, the metabolic characteristics and substrate utilization profiles of many uncultured microorganisms in this fermentation system remain unclear. This study aimed to isolate and identify butyric acid-producing strains from cellar mud and investigate their metabolic characteristics. A strain designated BJN0013 (= GDMCC 1.5973 = CGMCC No. 46148) was isolated from the cellar mud and capable of producing 31.11 ± 0.94 g/L of butyric acid. Genomic analyses (16S rRNA similarity 98.24%; ANI 90.09%; dDDH 41.80%) suggested it was a genomically distinct Clostridium strain with species-level divergence. Some physiological and biochemical characterizations also supported its species-level divergence. The carbon source utilization and enzyme metabolic pathways of acid-producing bacteria isolated from cellar mud were further summarized through genomic comparison. Acetyl-CoA transferase, the key enzyme responsible for the inability of strain BJN0013 to produce caproic acid, was identified by in silico evidence. This study offered a high-yielding butyric acid-producing strain resource from cellar mud and revealing the enzymatic targets for synthesizing butyric acid to optimize Baijiu fermentation.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-026-04703-4.

Neuronal death in Alzheimer's disease (AD) is closely associated with NLRP3 inflammasome-mediated pyroptosis. This study aimed to investigate the protective effects of Aloe-emodin (AE) in an AD cellular model and to explore the underlying mechanisms involving the NLRP3 inflammasome pathway. Molecular docking simulations predicted strong binding affinities between AE and key pyroptosis-related proteins (NLRP3, ASC, Caspase-1, GSDMD), with the highest affinity observed for NLRP3. In an Aβ₂₅₋₃₅-induced AD cellular model, AE (6 µM) significantly enhanced cell viability and alleviated pyroptotic morphological changes, including cellular swelling and rupture. EdU staining and immunofluorescence analysis further revealed that AE promoted HT22 cell proliferation and reduced Aβ deposition. Moreover, assessments of plasma and mitochondrial membrane integrity, via Hoechst 33,342/PI staining and mitochondrial permeability transition pore (MPTP) assay, respectively, revealed that AE treatment reduced the population of PI-positive cells and suppressed MPTP opening. Western blot, immunofluorescence, and ELISA analyses consistently demonstrated that AE downregulated the expression of pyroptosis-related proteins (NLRP3, ASC, Caspase-1, GSDMD, GSDMD-N) and suppressed the release of inflammatory cytokines (IL-1β, IL-18, IL-6, TNF-α). The inhibitory effect of AE on the pyroptosis pathway was comparable to that of the specific NLRP3 inhibitor MCC950. These results suggest that AE exerts neuroprotective effects in the AD cellular model by inhibiting NLRP3 inflammasome activation, thereby blocking Caspase-1 and GSDMD-N activation, attenuating neuronal pyroptosis, reducing inflammatory responses, and mitigating Aβ-induced pathological damage. Collectively, these findings identify AE as a promising therapeutic candidate for AD.

Glioblastoma multiforme (GBM) is a highly aggressive type of brain cancer known for its rapid progression and treatment resistance, presenting significant challenges for effective management. This article examines the promising potential of mesoporous silica nanoparticles (MSNs) as a groundbreaking platform for both the treatment and diagnosis of this formidable disease. MSNs boast several advantageous properties, including a large surface area, customizable pore sizes, and excellent biocompatibility. These characteristics enable efficient encapsulation of therapeutic agents, controlled release, and targeted delivery directly to GBM cells. One of the key advantages of MSNs is their ability to be functionalized with specific targeting ligands, which enhances their specificity toward tumor cells, facilitates navigation through the blood-brain barrier (BBB), and helps address the issues of tumor heterogeneity and drug resistance. When integrated with multimodal therapies, such as chemotherapy, immunotherapy, and photodynamic therapy, MSNs can create synergistic effects that improve therapeutic outcomes while reducing adverse off-target effects. Additionally, MSNs are poised to enhance diagnostic capabilities, improving imaging techniques for the accurate detection and monitoring of GBM. This review consolidates recent advancements in MSN-based approaches, emphasizing their therapeutic and diagnostic potential while also discussing toxicity concerns and outlining future pathways for clinical application to ultimately enhance patient outcomes.

Chickpea (Cicer arietinum L.) productivity is heavily constrained by major biotic stresses, particularly Fusarium wilt, Ascochyta blight and Botrytis gray mold, which collectively cause significant annual yield losses worldwide. To develop a refined understanding of the genetic architecture underlying resistance to these pathogens, a comprehensive meta-analysis was conducted using 113 QTLs taken from 24 independent studies, including diverse mapping populations. This analysis led to the identification of 27 MQTLs, which represent both novel genomic regions and, crucially, refined positions of previously known QTLs with reduced confidence intervals. Four robust Breeders' MQTLs were identified on the basis of high phenotypic variance (PVE ≥ 10%), a low confidence interval (CI ≤ 2 cM) and the involvement of multiple initial QTLs. Among these breeder MQTLs, 229 candidate genes, including key players in plant defense, such as receptor-like kinases (RLKs), resistance gene analogues (RGAs) and genes for RML1A, HSPRO2 and endochitinase A, were identified. These genes were validated through qRT‒PCR expression profiling in contrasting genotypes (WR-315 and JG-62). These refined genomic regions and their associated markers provide a direct pathway for pyramiding multiple resistance QTLs through marker-assisted selection and provide a direct pathway to breed chickpea varieties with durable, broad-spectrum resistance to key fungal diseases. The integrated meta-genomic framework significantly enhances precision and utility and paves the way for the functional characterization of the underlying resistance mechanisms.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-026-04698-y.

This study establishes an in situ refolding-based high-throughput screening strategy that enables directed evolution of eukaryotic enzymes expressed as inactive inclusion bodies in Escherichia coli. Guided by bioinformatic analysis, the catalytic domain of human DNase I was selected for mutagenesis, generating a library of 1,051 variants. A streamlined workflow-comprising microplate-based cultivation, induction, cell lysis, in situ denaturation with alkaline buffer containing β-mercaptoethanol, refolding with arginine, and activity detection-allowed efficient screening directly from insoluble expression. From this library, a DNase I mutant carrying N78T and V90N substitutions exhibited a 4.1-fold increase in enzymatic activity compared with the wild type. The same strategy applied to benzonase yielded a mutant with a 40% activity improvement, demonstrating the method's generality. Collectively, these results show that in situ refolding enables rapid identification of functional mutants from eukaryotic proteins produced as inclusion bodies, thereby improving the efficiency of directed evolution for otherwise challenging enzyme targets.

Bacterial fructan-metabolizing enzymes exhibit substantial structural and mechanistic diversity to support their biotechnological uses. Recent findings on glycoside hydrolase (GH) families 32 and 68 emphasize the conserved catalytic triads, calcium-binding motifs, and domain architectures that define their reaction frameworks. Differences in carbohydrate-binding modules (CBMs), extended loops, and accessory domains contribute to variations in substrate affinity, polymer length, and the balance between polymerization and hydrolysis. Aggregated data shows that GH68 enzymes generally exhibit higher catalytic efficiencies on sucrose, while GH32 hydrolases display stronger preferences for inulin and short-chain fructooligosaccharides (ScFOS). In gut commensals, distinct fructan utilization operons provide the basis for substrate-driven cross-feeding interactions, with ScFOS typically exhibiting shorter fermentation times compared to high molecular-weight levans. Rapid progress in artificial intelligence for structural predictions, molecular dynamics simulations, and CRISPR-enabled pathway engineering now supports the rational redesign of fructan-active enzymes, enabling the generation of catalysts with customized product profiles, enhanced stability, or altered chain-length distributions. This review provides a comprehensive overview of bacterial fructan-metabolizing enzymes, integrating structural, biochemical, and ecological perspectives to establish a foundation for applying fructan-modifying enzymes to prebiotic production, food texturization, microbiome modulation, and emerging oral enzyme therapeutics.

Cervical cancer ranks as the fourth most prevalent cancer among women worldwide. The increasing incidence and mortality rates are largely attributed to limited screening infrastructure, a shortage of skilled medical personnel, and inadequate awareness-particularly in developing nations. Conventional diagnostic methods, including the Pap smear, HPV testing, histopathology, and visual inspection with acetic acid (VIA), are often prone to human error due to subjective interpretation by clinicians. To address these limitations, this study proposes a robust automated framework for cervical cancer detection and cervix type classification using advanced deep learning techniques with minimal data requirements. A total of 915 histopathology images and over 4,000 colposcopy images were collected from public repositories and local healthcare institutions. Several pre-trained convolutional models were evaluated for cervix type classification. The proposed system employs a lightweight GoogleNet architecture to identify the transformation zone, followed by an EfficientMobileNet model for cervix type classification and cancer detection. Experimental results demonstrate superior performance, achieving 96% accuracy for cervix type classification, 95% accuracy for cervical cancer detection, and 99% mean average precision (mAP) for region-of-interest (ROI) localisation. The proposed approach offers an effective and resource-efficient diagnostic solution, particularly beneficial for regions with constrained healthcare infrastructure and limited clinical expertise. The proposed model gives 97% recall, 97% specificity, 97% accuracy, 96% mAP, 97% F1-measure and 99.8% AUC, respectively, compared to existing approaches like VGG16, ResNet50, CNN, ConvNet_1, ConvNet_5 and ConvNet_10.

Lignin, the second most abundant natural polymer on Earth after cellulose, is highly resistant to degradation, particularly in cold environments. In this study, a psychrotolerant bacterial strain, Psychrobacter faecalis CLB018, was isolated from the gut microbiota of the Antarctic fish Trematomus bernacchii near Zhongshan Station, Antarctica, using lignin as the sole carbon source at 4 ℃. Strain CLB018 exhibited high-efficiency lignin degradation, achieving a 40.39% degradation rate within 120 h at 10 ℃. It also demonstrated robust enzymic activities, producing lignin peroxidase (284 U/L at 48 h), laccase (111.9 U/L at 24 h), and manganese peroxidase (41.7 U/L at 48 h), essential for converting lignin into soluble aromatic compounds. Furthermore, CLB018 displayed versatility in utilizing diverse carbon source, including proteins, carboxylates, lipids, and aromatic compounds. Genomic analysis revealed the presence of genes encoding osmoprotectant transporters, cold shock proteins, an antioxidant system, and trehalose synthesis capability, which enhance its cold adaptation. These findings demonstrate that CLB018 possesses efficient lignin-degrading capabilities at low temperatures, highlighting its potential for sustainable biotechnological applications in cold environments.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-025-04623-9.

This study aims to hydrothermally synthesize magnetic iron nanoparticles (MNPs), conjugate them with cefotaxime (CTX), evaluate their efficacy against Escherichia coli strains (AK3 and AK9), and investigate the antibacterial mechanisms involved. The synthesized MNPs exhibited a crystalline structure in X-ray diffraction (XRD), the associated functional groups were confirmed by Fourier transform infrared spectroscopy (FTIR), and the morphology was confirmed through scanning electron microscopy (SEM) and transmission electron microscopy (TEM), with particle sizes ranging from 16.4 nm to 88.2 nm. Conjugation with CTX was confirmed using UV-Vis spectroscopy, FTIR, and dynamic light scattering (DLS), which showed an increased hydrodynamic diameter. The minimum inhibitory concentration (MIC) of CTX was high against the two E. coli strains (46.87 µg/mL and 750 µg/mL), whereas MNPs-conjugated CTX showed strong inhibition at lower doses (11.71 µg/mL and 46.87 µg/mL). Sub-MIC assays demonstrated that MNPs-conjugated CTX significantly inhibited biofilm formation, reduced β-lactamase expression to 22% and 32% in the two strains, respectively, and generated high levels of Reactive Oxygen Species (ROS). Colony-forming unit (CFU) analysis showed enhanced bactericidal activity, with reductions of 1.86 log₁₀ (AK3) and 1.63 log₁₀ (AK9). MNPs-conjugated CTX exhibited minimal cytotoxicity, maintaining 100% viability in HEK293T cells at 23 and 5 µg/mL, and improved cell survival in infected co-cultures to 73.39% (AK3) and 68.97% (AK9). SEM imaging of a tooth biofilm model revealed pronounced biofilm disruption after treatment with MNPs-conjugated CTX. These findings highlight MNPs-conjugated CTX as a promising nanoformulation for treating drug-resistant E. coli infections and support its potential for future biomedical applications.

Systemic acquired resistance (SAR) and induced systemic resistance (ISR) are the defense strategies that protect plants from a broad range of pathogens. Due to the ever-evolving and dynamic microbial diversity, the SAR/ISR genes are gifted with a competent regulatory mechanism. Promoters are one of the niche regulating areas of gene expression, which harbour numerous cis regulatory elements. While there are many reports on whole genome profiling of these regulatory features, studies focussing on the defense genes alone are few. Deeper analyses of a subset of genome provides less diluted and sharper perspective. Hence, in silico profiles of putative binding sites of 31 transcription factor families (TFFs), 17 pathogenesis-related transcription factors (PRTFs), and microRNAs were compared in 162 Arabidopsis thaliana SAR/ISR gene promoters. This study revealed new insights into defense gene regulation. The AT-Hook motif, and putative binding sites for NAC and MYB TFFs and TGA1a PRTFs had the highest frequencies. Previous studies on involvement of AT-Hook motif and TGA1a in A. thaliana defense are few. Potential multi gene-targeting role of certain microRNAs, such as MIR2934, is suggested. The possibility of certain microRNAs previously known for post-transcription regulation, to be involved in transcription regulation is proposed. Though there are reports on plant transcriptomes under various biotic stresses, the reason for temporal expression pattern similarity of certain genes under multiple stresses is unclear. To understand this phenomenon, the regulatory profiles of defense genes under nine biotic stresses fetched from EFP browser were analysed. Similarly expressing gene pairs did not necessarily have the same regulatory features, indicating a stochastic correlation of temporal expression patterns and PRTFs/microRNAs putative binding sites/DNA and histone methylation sites. A vague similarity in the histone modification patterns was observed in their promoters. The findings highlighted in this study could be harnessed for molecular fine tuning of genomic pathways to mitigate broad range biotic stresses.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-026-04706-1.