Biotechnology and applied biochemistry最新文献

This study investigates the potential of three synthesized ferrocenylmethylaniline derivatives (FMBA, FMAA, and FMA) as inhibitors of α-amylase, a key enzyme involved in the pathophysiology of diabetes. In vitro inhibition assays demonstrated that FMBA and FMAA exhibited significantly lower IC50 values of 9.23 and 11.23 µM, respectively, compared to 259 µM for the standard drug acarbose (ARE). Molecular docking studies supported these findings, with FMBA showing the highest binding affinity (∆G of -7.33 kcal/mol), followed by FMAA (-6.44 kcal/mol) and FMA (-5.85 kcal/mol), outperforming ARE (-4.88 kcal/mol). ADMET analysis suggested favorable pharmacokinetic and safety profiles for FMBA and FMAA, reinforcing their potential as viable drug candidates. To further assess the stability and dynamics of the enzyme-ligand interactions, molecular dynamics simulations were conducted, showing that FMBA and FMAA formed significantly more stable complexes with α-amylase compared to ARE, as indicated by low root mean square deviation (RMSD) values of 0.156 and 0.164 nm, respectively, compared to 0.359 nm for ARE. Root mean square fluctuation (RMSF) analysis revealed consistent stability at key active site residues. Additional analyses of radius of gyration (R_g) and solvent-accessible surface area (SASA) supported the compact and stable nature of the complexes. Frontier molecular orbital (FMO) analysis showed smaller HOMO-LUMO energy gaps for FMBA and FMAA, suggesting greater reactivity and potential biological activity. Molecular electrostatic potential (MEP) surface analysis highlighted key reactive sites, with high negative potential localized on the carbonyl groups of FMBA and FMAA, and high positive potential in regions favoring hydrogen bonding. These findings underscore the potential of FMBA and FMAA as promising antidiabetic agents and support their further development as therapeutic candidates.

Astaxanthin, a high-value carotenoid with potent antioxidant and anti-inflammatory activities, is increasingly in demand in various industries. This study reports the successful enhancement of astaxanthin and zeaxanthin production in Paracoccus sp. LL1 through random mutagenesis using ethyl methanesulfonate (EMS). To induce genetic diversity, EMS mutagenesis was employed, followed by the selection of mutants that exhibited increased carotenoid production. The top-performing mutants showed a significant 2.76-fold increase in astaxanthin and a 10.14-fold increase in zeaxanthin compared to the wild-type strain when treated with the optimal EMS concentration of 0.5%. The effects of initial glucose concentration and inoculum size on astaxanthin and zeaxanthin production were evaluated, and production was higher when glucose was 2% and inoculum size was 10%. Our findings demonstrate the potential of Paracoccus sp. LL1 as a promising alternative to traditional astaxanthin-producing organisms, such as Haematococcus pluvialis, offering advantages including faster growth, simpler cultivation requirements, and genetic tractability. This approach not only enhances carotenoid production but also highlights the novelty of using Paracoccus sp. LL1, a less-explored strain, for high-yield production of both astaxanthin and zeaxanthin. These results suggest that Paracoccus sp. LL1 could serve as an efficient platform for industrial-scale carotenoid production through metabolic engineering and random mutagenesis, providing a viable alternative to current production systems.

Acute pancreatitis (AP) is a prevalent inflammatory disorder of the pancreas, with emerging evidence implicating aberrant acinar cell glycolysis in disease progression. Although TIGAR (TP53-induced glycolysis and apoptosis regulator), a key glycolytic regulator, has been implicated in various pathological processes, its role in AP remains unexplored. In this study, we established an AP mouse model through intraperitoneal injection of high-dose caerulein. AP mice exhibited downregulated pancreatic TIGAR expression accompanied by enhanced glycolysis. In vitro, primary pancreatic acinar cells were stimulated with LPS to mimic the inflammatory microenvironment. TIGAR overexpression effectively mitigated LPS-induced reductions in cell viability, inflammatory cytokine expression, reactive oxygen species (ROS) production, and glycolytic activation. Notably, LAMP2 (lysosome-associated membrane protein 2) knockdown abolished the protective effects of TIGAR against LPS-induced ROS, inflammation, and glycolytic flux. Mechanistically, TIGAR suppressed LPS-induced glycolytic activation by upregulating LAMP2 expression, thereby inhibiting PI3K/Akt pathway activation. Consistently, the glycolytic inhibitor 2-DG reversed the detrimental effects of TIGAR knockdown on cell viability and inflammatory responses. Furthermore, both preventive and therapeutic AAV8-TIGAR administration attenuated AP progression in mice. In conclusion, our findings demonstrate that TIGAR protects against AP progression by modulating the LAMP2/PI3K/Akt axis to maintain glycolytic homeostasis, highlighting its potential as a therapeutic target for AP.

The rapid and accurate detection of COVID-19 (coronavirus disease 2019) from normal and pneumonia chest x-ray images is essential for timely diagnosis and treatment. The overlapping features in radiology images make it challenging for radiologists to distinguish COVID-19 cases. This research study investigates the effectiveness of combining local binary pattern (LBP) and histogram of oriented gradients (HOG) features with machine learning algorithms to differentiate COVID-19 from normal and pneumonia cases using chest x-rays. The proposed hybrid fusion model "RadientFusion-XR" utilizes LBP and HOG features with shallow learning algorithms. The proposed hybrid HOG-LBP fusion model, RadientFusion-XR, detects COVID-19 cases from normal and pneumonia classes. This fusion model provides a comprehensive representation, enabling more precise differentiation among the three classes. This methodology presents a promising and efficient tool for early COVID-19 and pneumonia diagnosis in clinical settings, with potential integration into automated diagnostic systems. The findings highlight the potential of this hybrid feature extraction and a shallow learning approach to improve diagnostic accuracy in chest x-ray analysis significantly. The hybrid model using LBP and HOG features with an ensemble model achieved an exceptional accuracy of 99% for binary class (COVID-19, normal) and 97% for multi-class (COVID-19, normal, pneumonia), respectively. These results demonstrate the efficacy of our hybrid approach in enhancing feature representation and achieving superior classification accuracy. The proposed RadientFusion-XR model with hybrid feature extraction and shallow learning approach significantly increases the accuracy of COVID-19 and pneumonia diagnoses from chest x-rays. The interpretable nature of RadientFusion-XR, alongside its effectiveness and explainability, makes it a valuable tool for clinical applications, fostering trust and enabling informed decision-making by healthcare professionals.

Biocontrol plays a pivotal role in mitigating biotic stress and promoting plant growth by utilizing beneficial microorganisms that exhibit antagonistic activity against phytopathogens. Xanthomonas campestris is a bacterial pathogen known to cause diseases such as bacterial black leaf spots in various economically important crops. Therefore, in the current study to identify effective biocontrol agents, over 30 endophytic isolates from various tissues of Azadirachta indica were examined for their antagonistic activity against X. campestris pv. vesicatoria. Among these, two potential isolates, Bacillus safensis (strain LE8) and Pseudomonas lactis (strain LE11), based on their inhibitory effects, were subsequently selected for further analysis to contribute to sustainable agricultural practices. The in vitro as well in vivo treatments to tomato leaves with these potential isolates showed both preventive as well as curative effects. The current investigation confirmed a notable reduction in disease symptoms, showcasing their effectiveness as a biocontrol agent. Our findings highlight the beneficial impact of endophytic bacteria as a biocontrol, providing a sustainable alternative to pesticides.

Studies that focus on turning biodiesel byproducts into valuable products have recently garnered increasing attention. This investigation highlights the utilization of waste substrates for a circular economy approach using glycerine pitch as the main carbon source to produce biodegradable poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [(P(3HB-co-3HV)] by Cupriavidus malaysiensis USMAA1020. The primary use of glycerine pitch, along with several parameters that may affect the growth and biosynthesis of copolymers, are investigated. The utilization of glycerine pitch along with 1-pentanol and oleic acid has the most effective effect on bacterial growth and copolymer accumulation. Response surface methodology (RSM) was used to optimize these variables. The polyhydroxyalkanoate (PHA) content increased up to 77.7 wt% from 68.9 wt%, with residual dry cell weight (RDCW) of 6.5 g/L from 4.5 g/L. The optimal conditions for producing various compositions of the 3HVs were also determined. The three selected copolymer compositions were P(3HB-co-4%3HV), P(3HB-co-11%3HV), and P(3HB-co-18%3HV). Moreover, varying the copolymer compositions produced distinct polymer characteristics. According to this study, P(3HB-co-3HV) with variable properties can be produced for a range of applications using glycerine pitch as a potential primary carbon source. In addition to reducing the cost of production, this would enhance efficient waste management.

Amylases function as hydrolytic enzymes, facilitating the decomposition of starch molecules and other associated polymers. These enzymes are found ubiquitously across all domains of life. Amylase dominates the enzyme market in terms of sales because of its extensive utilization in the starch processing field and its wide-ranging applications across the food, textile, and pharmaceutical sectors. Microorganisms are primarily used to produce amylase; they are readily available, flexible, and easy to employ. Fruit and vegetable wastes (FAVWs) containing proteins and lipids add to the detrimental effects on the environment. However, this waste offers cost-effective alternatives for manufacturing value-added products through the synthesis of industrially essential enzymes by microorganisms. The most recent advancements in biocatalytic systems aim to improve the catalytic efficiency of commercially available enzymes or generate new enzymes with unique features. This study emphasizes the valorization of FAVW to derive amylase, recent advancements in the use of enzyme immobilization approaches for sustainable development, and their application in the present scenario.

Diabetes mellitus (DM), which can result in a number of problems such as cataracts, neuropathy, retinopathy, nephropathy, and several cardiovascular illnesses, continues to be a growing issue despite major advancements in treatment approaches. Numerous scientists have targeted the polyol pathway as a target for intervention since it includes aldose reductase (ALR2, AR (E.C.1.1.1.21)), a crucial enzyme. Oxidative damage, NADPH depletion, and intracellular sorbitol buildup result from the overactivation of ALR2 brought on by hyperglycemia. Interest in creating novel ALR2 inhibitors (ALR2Is) with enhanced therapeutic characteristics has increased as a result of this circumstance. The amazing biological capabilities of isoxazole molecules led us to look into the biological properties of isoxazole and related compounds. We examined these isoxazoles' binding affinities and interactions in the ALR2 active site using thorough in vitro and in silico techniques. In comparison to the reference pharmaceutical epalrestat (EPR, K_I 232.70 ± 15.51 nM), our results demonstrate that these isoxazoles efficiently inhibit ALR2 at nanomolar doses, with inhibition constants (K_I) ranging from 12.13 ± 1.24 nM to 89.51 ± 4.68 nM. Important interactions between these isoxazoles and ALR2 are highlighted by the combined in vitro and in silico studies, indicating their potential as therapeutic agents against a range of pathological diseases. Furthermore, these substances that have ALR2 inhibitory properties could be useful as stand-in treatments or preventative measures for diabetes problems.

Antibiotic resistance, which renders existing antibiotics ineffective against bacterial infections, is among the top-most pressing global public health challenges. A promising strategy to combat bacterial infection without inducing the occurrence of drug resistance is by disrupting quorum sensing (QS)-a complex communication circuit that bacterial pathogens employ to regulate their virulence. Therefore, QS inhibitors have emerged in recent times as potential therapeutic agents against bacterial infections. S-Ribosylhomocysteinase (LuxS) is one particularly attractive target in the QS pathway, which synthesizes the signaling molecule that mediates interspecies bacterial communication called autoinducer-2 (AI-2). In this study, we used computational chemistry and drug discovery techniques, molecular docking, drug-likeness, toxicity prediction studies, and interaction profiling to identify bioactive phytochemicals from Annona muricata plant extract as potential anti-QS agents against LuxS. Screening a library of 123 natural acetogenins from A. muricata, we identified gigantetronenin and isoannonacin as promising LuxS inhibitors. The potential inhibitory activity of these compounds against LuxS suggests that they could be explored as QS inhibitors with broad-spectrum activity against bacterial pathogens. These findings highlight the potential of gigantetronenin and isoannonacin as novel therapeutic candidates for combating bacterial infections through QS inhibition.