Biotechnology and applied biochemistry最新文献

L-arginine is a vital amino acid metabolized by L-arginase, an enzyme with growing biotechnological and therapeutic significance, particularly in cancer treatment. This study presents a comprehensive in silico characterization of the L-arginase gene isolated from Alcaligenes aquatilis BC2, a bacterium originating from Ethiopian soda lakes. The analysis identified a 336-amino-acid enzyme predicted to be stable, soluble, and extracellular. Homology modeling generated a reliable hexameric 3D structure with high stereochemical quality, validated through multiple structural assessment tools. Phylogenetic and conserved domain analyses confirmed the enzyme's evolutionary placement within the Alcaligenes genus and highlighted preserved functional motifs. Molecular docking predicted a strong binding affinity (-7.1 kcal mol^-1) for L-arginine, with ligand enzyme complex stabilized by a dense network of hydrogen bonds and electrostatic interactions within the active site. These findings elucidate the structural basis of the enzyme's function and underscore its potential for future experimental validation and therapeutic applications.

Acute myeloid leukemia (AML) is a rapidly progressing blood cancer with poor survival rates, necessitating aggressive treatment strategies like chemotherapy. Doxorubicin (DOXO) is commonly used but is limited by severe side effects, including myeloablation, which involves the depletion of bone marrow cells leading to immunosuppression and heightened infection risk. This study explores the potential of omega-3 polyunsaturated fatty acids (n-3 PUFAs), specifically docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), to enhance the efficacy of DOXO against AML cells while mitigating some of its toxicities. The results show that DHA and EPA increase the DOXO-induced apoptosis in KG1a cells and greater accumulation in the sub-G1 phase, suggesting enhanced cell death. TUNEL assays confirmed increased DNA fragmentation, whereas mRNA analysis revealed upregulation of apoptosis and cell cycle regulation genes. Importantly, DHA and EPA also reduced the hemolytic activity of DOXO, suggesting a protective effect against chemotherapy-associated side effects. These findings suggest that DHA and EPA could enhance the anti-leukemic impact of DOXO, potentially reducing the need for high-dose chemotherapy and alleviating risks like myeloablation, offering a promising adjunct strategy for AML treatment.

α-Galactosidase enzymes have a variety of applications in numerous areas, including medicine, energy, food, feed, prebiotics, and paper pulp production. A single α-galactosidase gene was identified in the Geobacillus kaustophilus genome through the analysis of the carbohydrate-active enzymes (CAZY) database. The objective of this study was to clone, express heterologously, purify, and biochemically characterize the α-galactosidase enzyme encoded in the G. kaustophilus genome. In order to achieve this, a codon-optimized synthetic gene encoding the α-galactosidase enzyme was cloned into the pQE30 plasmid and expressed in Escherichia coli BL21 DE3 pLysS. The biochemical characterization of G. kaustophilus α-galactosidase revealed that the purified enzyme exhibited optimal activity at 40°C and pH 6.0 in citrate buffer, with demonstrable activity over a broad range of pH and temperature. Furthermore, the GKαGal enzyme exhibited a V_max value of 41.09 U/mg and a 0.248 mM K_m value towards 4-nitrophenyl-α-d-galactopyranoside (pNPGal). In essence, the produced α-galactosidase enzyme has potential applications in the degradation of lignocellulosic biomass, disaccharides production, and in medical contexts.

Leukemia continues to provide a significant therapeutic challenge due to relapse, medication resistance, and treatment-associated toxicity, which frequently hinder sustained disease management. Rhizomes of ginger (Zingiber officinale) possess bioactive phenolics, notably 6-gingerol and 6-shogaol derivatives, which have demonstrated antileukemia efficacy in preclinical models. This study rigorously assesses the evidence regarding ginger-derived preparations and isolated compounds in both acute and chronic leukemia models, focusing on recurring mechanisms and translational viability. In leukemia cell line investigations and sparse resistant-model data, ginger-related interventions are consistently linked to diminished viability and the induction of mitochondrial apoptosis, typically indicated by alterations in Bax/Bcl-2 ratios, PARP breakage, and caspase-related measurements. Numerous studies indicate redox modulation, often characterized by elevated intracellular reactive oxygen species in leukemic cells, coupled with diminished pro-survival signaling, such as PI3K/Akt, as indicated by decreased pAkt and survivin levels. The suggested immunomodulatory and anti-inflammatory effects, encompassing alterations in NK-cell activity and cytokines like TNF-α and IL-6, are inadequately substantiated within leukemia-specific immunological contexts. Interpretation is limited by the variability in extract composition and chemical characterisation, inconsistent dose and exposure circumstances, dependence on endpoint markers without causative manipulation, and a lack of leukemia-specific clinical data. Ginger-derived compounds exhibit multi-target biological activity that necessitates further exploration through standardized and chemically defined preparations, pharmacokinetic and pharmacodynamic characterization, clinically relevant exposure benchmarks, and meticulously designed leukemia-focused translational and early-phase clinical studies to elucidate safety, efficacy, and compatibility with current therapies.

Ovarian Cancer is a leading cause of mortality among women globally, primarily due to lack of specific and sensitive early-stage diagnostic tools. This study aims to identify hub genes associated with recurrent, late-stage, and metastatic tumors as potential prognostic biomarkers and drug targets. Gene expression data from eight National Center for Biotechnology Information (NCBI)-Gene Expression Omnibus (GEO) datasets were categorized by recurrence, tumor-stage, and metastasis. Differential gene expression and enrichment analyses were performed. Hub genes were identified by protein-protein interaction networks and validated by the University of Alabama at Birmingham Cancer Data Analysis Portal (UALCAN), GEPIA2, pROC, and Kaplan-Meier plotter databases. Genetic alterations, immune cell infiltration, miRNA prediction, and drug-gene interactions were assessed using cBioPortal, CIBERSORTx, Encyclopedia of RNA Interactomes (ENCORI), and Drug-Gene Interaction Database (DGIdb), respectively. Eight hub genes (FN1, COL1A1, COL1A2, COL3A1, POSTN, LUM, IGF1, and CXCL8) were identified, with COL1A2 common across all tumor categories. Note that 19.6% of cases showed mutations in these genes, primarily COL3A1. Overexpression of most hub genes and reduced expression of CXCL8 correlated with worse survival outcomes. COL1A1 and FN1 showed strong diagnostic ability. Late-stage tumors showed elevated M2 macrophages and neutrophils. hsa-miR-29a-3p, hsa-miR-29b-3p, and hsa-miR-29c-3p were identified as the most interactive miRNAs. Ocriplasmin and pamidronate were identified as potential therapeutics. Our findings highlight the therapeutic relevance of these hub genes and identify them as potential drug targets and prognostic biomarkers in ovarian cancer.

We present a novel, cost-effective sensor for carcinoembryonic antigen (CEA) detection utilizing a pencil graphite electrode (PGE) in combination with electrochemical impedance spectroscopy (EIS), which offers high sensitivity and selectivity. Anti-CEA/AuNPs/PGE was successfully illustrated as a label-free impedimetric immunosensor for the detection of CEA. Through EIS, we observed distinct impedance changes upon CEA binding, enabling real-time detection with high reproducibility and low interference from non-target molecules. Due to its satisfying impedimetric response, this new immunosensor demonstrated that it can be used for high-performance detection of CEA with a wide linear range extending from 13.2 to 1 × 10⁵ pg mL^-1, with correlation coefficients (R²) of 0.9923. The PGE's excellent conductive properties and surface stability allowed for the successful detection of CEA at low concentrations, demonstrating a detection limit of 4.4 pg mL^-1, which is competitive with existing, more costly alternatives. The sensor's robust performance in spiked artificial urine samples indicates its potential for practical application in point-of-care cancer diagnostics, especially in resource-limited environments. The developed electrochemical biosensor holds promise for accurately detecting CEA in urine samples, offering a precise technique that could find valuable application in clinical tumor detection.

Multidrug-resistant tumor cells pose significant challenges in cancer treatment. Alternative strategies such as targeted gene silencing and the use of compounds with minimal cytotoxicity toward normal cells are therefore of great interest. Antimicrobial peptides (AMPs) have demonstrated anticancer potential due to their physicochemical properties. In lung cancer, overexpression of AKT serine/threonine kinase 1 (AKT1) promotes abnormal tumor growth and progression. In this study, we synthesized chitosan-based nanoparticles (CSNPs) co-loaded with Pseudomonas aeruginosa peptide from strain P3 (PAP3) (an AMP) and siRNA targeting the AKT1 gene, and evaluated their anticancer activity at the cellular and molecular levels. Characterization of the CSNPs revealed a nanoscale structure, low polydispersity index, and moderate encapsulation efficiency for both peptide and siRNA. Evaluation using L929 cells confirmed PAP3's nontoxic profile, while a dose-dependent anticancer effect against A549 cells was observed. Delivery of encapsulated peptide, siRNA, and their combination increased cell death and induced morphological changes in A549 cells. Gene expression analysis showed upregulation of pro-apoptotic markers (Bax and Caspase-3) and downregulation of the anti-apoptotic marker Bcl2, indicating promising anticancer properties of the engineered compound. In conclusion, co-delivery of PAP3 and AKT1-targeting siRNA via CSNPs demonstrates potential for future anticancer therapies.

In this study, we report biogenic synthesis of silver nanoparticles (AgNPs) using polyextremophile bacteria Deinococcus radiodurans. Optical and structural properties of the green synthesized silver nanoparticles were investigated by various techniques, including Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy-dispersive x-ray (EDX) spectroscopy, and UV-visible (UV-Vis) absorption spectroscopy. The AgNPs were entrapped in calcium alginate beads and were used for photo decolorization of various charged pollutant dyes, under solar irradiation. In this study, cationic (methylene blue [MB], methyl green [MG]) and anionic (methyl orange [MO]) dyes were used as model dyes. Both AgNPs in suspension and those entrapped in beads could degrade all the three dyes with 100% degradation efficiency in suspension and slightly lower efficiency with beads. The photocatalytic activity of immobilized AgNPs in fabricated column model demonstrates potential application for the removal of dyes from effluents, contributing ultimately to ecological cleanup process and facilitated in recovery and reprocessing. The nanocomposites retained a significant amount of photocatalytic efficiency even after four reuse cycles, and the degradation efficiency followed the order: MB > MO > MG. Additionally, phytotoxicity and cytotoxicity assay was performed to show significant reduction in toxicity of nanoparticle (NP)-assisted degraded cationic and anionic dyes, thereby substantiating the nontoxic nature of the degraded dye. The efficiency of beads entrapped silver NPs as a viable option, for the degradation of harmful organic dyes from the environment, is established in the present study.

The escalating global demand for biopharmaceuticals is placing increasing strain on conventional production systems, highlighting the need for innovative and sustainable alternatives. Industrial byproducts, produced extensively across pharmaceutical and allied sectors, remain an underexploited resource with significant potential to reduce production costs and strengthen circular economy integration. This review systematically explores the sources and classification of industrial wastes relevant to biopharmaceutical manufacturing, while addressing critical regulatory, safety, and quality considerations for their adoption. Emerging biotechnological strategies-such as microbial fermentation, enzymatic biotransformation, and synthetic biology-driven metabolic engineering-are evaluated for their ability to convert industrial residues into high-value therapeutic products. Representative case studies demonstrate the feasibility of these approaches, including the utilization of agro-industrial waste for therapeutic enzymes, marine-derived residues for bioactive compounds, and fermentation byproducts for vaccine components. Environmental and economic implications are assessed through life cycle analysis (LCA) and cost-benefit evaluations, underscoring the alignment of waste valorization with sustainable manufacturing principles. Despite these opportunities, technological limitations, stringent quality and standardization requirements, and complex policy and ethical challenges remain substantial barriers. Future perspectives highlight the role of green bioprocessing, artificial intelligence (AI), and automation in optimizing waste-to-medicine pathways, alongside the long-term vision of achieving zero-waste biopharmaceutical production. By positioning industrial byproducts as valuable feedstocks, this review underscores their transformative potential in driving sustainable, resilient, and responsible healthcare manufacturing.

Cow dung is a low-cost lignocellulosic biomass generated in large quantities across India, yet remains underutilized and contributes to environmental pollution when improperly managed. In this study, cellulose was isolated from cow dung using two different approaches a green, microbially-assisted natural extraction process under dark conditions with mild chemical uses, and a chemically driven Soxhlet-assisted method employing alkali oxidative pretreatment. The physicochemical characteristics of the isolated cellulose were examined using the Van Soest compositional protocol, FTIR spectroscopy, UV-Vis analysis, CHNS elemental profiling, and SEM imaging. The Soxhlet route produced a cellulose yield of 3.65% ± 0.2%, with purity of 28.68% cellulose and 3.68% lignin, whereas the natural method resulted in a yield of 3.55% ± 0.3%, with purity of 26.31% cellulose and 8.9% lignin. Soxhlet extraction enabled more effective delignification and improved fiber defibrillation, while the natural method, despite of lower lignin removal rate, preserved the structural integrity of the cellulose and offered substantial sustainability advantages by reducing chemical consumption and energy requirements. These findings highlight cow dung as a viable renewable feedstock for cellulose-based biomaterials and demonstrate that low-resource, environmentally benign extraction strategies can support decentralized and rural circular bio-economy initiatives.