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1.
Microb Cell Fact ; 23(1): 88, 2024 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-38519954

RESUMO

BACKGROUND: The halophilic bacterium Halomonas elongata is an industrially important strain for ectoine production, with high value and intense research focus. While existing studies primarily delve into the adaptive mechanisms of this bacterium under fixed salt concentrations, there is a notable dearth of attention regarding its response to fluctuating saline environments. Consequently, the stress response of H. elongata to salt shock remains inadequately understood. RESULTS: This study investigated the stress response mechanism of H. elongata when exposed to NaCl shock at short- and long-time scales. Results showed that NaCl shock induced two major stresses, namely osmotic stress and oxidative stress. In response to the former, within the cell's tolerable range (1-8% NaCl shock), H. elongata urgently balanced the surging osmotic pressure by uptaking sodium and potassium ions and augmenting intracellular amino acid pools, particularly glutamate and glutamine. However, ectoine content started to increase until 20 min post-shock, rapidly becoming the dominant osmoprotectant, and reaching the maximum productivity (1450 ± 99 mg/L/h). Transcriptomic data also confirmed the delayed response in ectoine biosynthesis, and we speculate that this might be attributed to an intracellular energy crisis caused by NaCl shock. In response to oxidative stress, transcription factor cysB was significantly upregulated, positively regulating the sulfur metabolism and cysteine biosynthesis. Furthermore, the upregulation of the crucial peroxidase gene (HELO_RS18165) and the simultaneous enhancement of peroxidase (POD) and catalase (CAT) activities collectively constitute the antioxidant defense in H. elongata following shock. When exceeding the tolerance threshold of H. elongata (1-13% NaCl shock), the sustained compromised energy status, resulting from the pronounced inhibition of the respiratory chain and ATP synthase, may be a crucial factor leading to the stagnation of both cell growth and ectoine biosynthesis. CONCLUSIONS: This study conducted a comprehensive analysis of H. elongata's stress response to NaCl shock at multiple scales. It extends the understanding of stress response of halophilic bacteria to NaCl shock and provides promising theoretical insights to guide future improvements in optimizing industrial ectoine production.


Assuntos
Diamino Aminoácidos , Halomonas , Cloreto de Sódio/farmacologia , Cloreto de Sódio/metabolismo , Halomonas/genética , Halomonas/metabolismo , Pressão Osmótica , Perfilação da Expressão Gênica , Peroxidases/metabolismo
2.
Anal Chim Acta ; 1297: 342386, 2024 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-38438231

RESUMO

In this work, we developed a new strategy to fabricate a series of transition metallic nanoparticles (NPs) embedded on B, N co-doped carbon nanotubes (CNTs) arrays modified flexible carbon fiber electrodes (M@BNCNTs/CF, M = Co, Fe, Ni) via facile inkjet printing assisted with chemical vapor deposition using Ionic liquid as solvent of printing ink and heteroatom dopants. Furthermore, Pt NPs via impregnation-thermal reduction process was anchored on the surface of Co@BNCNTs/CF (Pt-Co@BNCNTs/CF), which holds enhanced peroxidase-like activity and could be directly used as freestanding electrode to detect H2O2, exhibiting a low detection limit of 0.19 µM with wide linear range (0.5 µM-9.4 mM), and high sensitivity (1679 µA cm-2 mM-1). The excellent sensing performance of Pt-Co@BNCNTs/CF is attributed to the Pt, Co NPs anchored on CNTs with great catalytic activity, and the doping B, N would cause graphitic carbon with more defects to improve its inherent reactivity toward H2O2. Besides, CNTs arrays with high surface area also enlarge the exposure of active sites. Moreover, the Pt-Co@NBCNTs/CF microelectrode has been successfully applied in monitoring H2O2 secreted from human colonic cancer cells and normal colonic epithelial cells, which could offer crucial data for distinguishing various cell types and identifying cancer cells from normal cells. This work opens a new horizon to fabricate flexible miniaturized sensing device for extracellular analysis and offers an extended strategy to fabricate other metallic NPs embedded in heteroatoms doped CNTs functionalized flexible fiber electrode, by choosing diverse metal ions and ILs as inkjet printing precursors.


Assuntos
Nanotubos de Carbono , Humanos , Microeletrodos , Peróxido de Hidrogênio , Transporte Biológico , Peroxidases
3.
ACS Nano ; 18(11): 8083-8098, 2024 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-38456744

RESUMO

Active polymetallic atomic clusters can initiate heterogeneous catalytic reactions in the tumor microenvironment, and the products tend to cause manifold damage to cell metabolic functions. Herein, bimetallic PtPd atomic clusters (BAC) are constructed by the stripping of Pt and Pd nanoparticles on nitrogen-doped carbon and follow-up surface PEGylation, aiming at efficacious antineoplastic therapy through heterogeneous catalytic processes. After endocytosed by tumor cells, BAC with catalase-mimic activity can facilitate the decomposition of endogenous H2O2 into O2. The local oxygenation not only alleviates hypoxia to reduce the invasion ability of cancer cells but also enhances the yield of •O2- from O2 catalyzed by BAC. Meanwhile, BAC also exhibit peroxidase-mimic activity for •OH production from H2O2. The enrichment of reactive oxygen species (ROS), including the radicals of •OH and •O2-, causes significant oxidative cellular damage and triggers severe apoptosis. In another aspect, intrinsic glutathione (GSH) peroxidase-like activity of BAC can indirectly upregulate the level of lipid peroxides and promote ferroptosis. Such deleterious redox dyshomeostasis caused by ROS accumulation and GSH consumption also results in immunogenic cell death to stimulate antitumor immunity for metastasis suppression. Collectively, this paradigm is expected to inspire more facile designs of polymetallic atomic clusters in disease therapy.


Assuntos
Antineoplásicos , Ferroptose , Neoplasias , Humanos , Peróxido de Hidrogênio , Espécies Reativas de Oxigênio , Apoptose , Peroxidases , Antineoplásicos/farmacologia , Catálise , Glutationa , Linhagem Celular Tumoral , Neoplasias/tratamento farmacológico , Microambiente Tumoral
4.
Environ Geochem Health ; 46(3): 102, 2024 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-38433158

RESUMO

Explosives are perilous and noxious to aquatic biota disrupting their endocrinal systems. Supplementarily, they exhibit carcinogenic, teratogenic and mutagenic effects on humans and animals. Henceforth, the current study has been targeted to biotransform the explosive, 2, 4, 6 trinitrophenol (TNP) by wetland peroxidase from Streptomyces coelicolor. A total peroxidase yield of 20,779 mg/l with 51.6 folds of purification was observed. In silico molecular docking cum in vitro appraisals were accomplished to assess binding energy and interacting binding site residues of peroxidase and TNP complex. TNP required a minimal binding energy of-6.91 kJ/mol and was subjected to biodeterioration (89.73%) by peroxidase in purified form, with 45 kDa and a similarity score of 34 by MASCOT protein analysis. Moreover, the peroxidase activity was confirmed with Zymogram analysis. Characterization of peroxidase revealed that optimum values of pH and temperature as 6 and 40 °C, respectively, with their corresponding stability varying from 3.5 to 7. Interestingly, the kinetic parameters such as Km and Vmax on 2,2'-azino-bis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and H2O2 were 19.27 µm and 0.41 µm/min; 21.4 µm and 0.1 µm/min, respectively. Among the diverse substrates, chemicals and trace elements, ABTS (40 mM), citric acid (5 mM) and Fe2+ (5 mM) displayed the highest peroxidase activity. Computational docking and in vitro results were corroborative and UV-Vis spectroscopy, HPLC, FTIR and GC-MS indicated the presence of simple metabolites of TNP such as nitrophenols and benzoquinone, showcasing the efficacy of S. coelicolor peroxidase to biotransform TNP. Henceforth, the current study offers a promising channel for biological treatment of explosive munitions, establishing a sustainable green earth.


Assuntos
Benzotiazóis , Peróxido de Hidrogênio , Peroxidase , Picratos , Ácidos Sulfônicos , Animais , Humanos , Simulação de Acoplamento Molecular , Peroxidases , Corantes
5.
ACS Appl Bio Mater ; 7(3): 1778-1789, 2024 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-38437514

RESUMO

Inspired by the two kinds of naturally occurring peroxidases (POD) with vanadium or heme (iron)-based active catalytic centers, we have developed a dual metal-based nanozyme with dual V and Fe-based active catalytic centers. Co-doping of graphene with heteroatoms has a synergistic effect on the catalytic properties of the nanomaterial as the distances of migration of the substrates drastically reduce. However, a few studies have reported the codoping of heterometallic elements in the graphene structure due to the complexity of the synthesis procedures. Herein, we report the synthesis of in situ doped bimetallic VNFe@C mesoporous graphitic spheroids nanozyme via pyrolysis without the assistance of any template assisted method. The Prussian-blue analog-based precursor material was synthesized by a facile one-step low-temperature synthesis procedure. The bimetallic spheroids showed an excellent affinity toward H2O2, with a Km value of 0.26 mM when compared to 0.436 for the natural POD, which is much better than the natural POD, which was utilized to detect tumor cells in vitro through the intracellular H2O2 produced by these cells under high oxidative stress. The VNFe@C mesoporous spheroids generate dual reactive oxygen species, including the •OH and •O2H- radicals, in the presence of H2O2, which are responsible for the POD-like activity of these nanozymes, while the bimetallic V/Fe doping plays a synergistic role in the enhancement of the activity of codoped graphitic spheroids.


Assuntos
Grafite , Peroxidase , Peróxido de Hidrogênio , Peroxidases , Catálise
6.
Biosens Bioelectron ; 253: 116161, 2024 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-38457864

RESUMO

We herein describe a novel electrochemical strategy to detect hydrogen peroxide (H2O2) by utilizing the peroxidase-mimicking activity of cerium oxide nanoparticles (CeO2 NP) and reduced graphene oxide (rGO). Particularly, CeO2 NP/rGO nanocomposites were deposited on the commercial electrode by a very convenient and direct electrochemical reduction of graphene oxide. Due to the peroxidase-mimicking activity of CeO2 NP and the outstanding electrochemical properties of reduced graphene oxide, the reduction current of H2O2 was greatly enhanced. Based on this strategy, we reliably determined H2O2 down to 1.67 µM with excellent specificity and further validated its practical capabilities by robustly detecting H2O2 present in heterogeneous human serum samples. We believe that this work could serve as a new facile platform for H2O2 detection.


Assuntos
Técnicas Biossensoriais , Cério , Grafite , Nanocompostos , Humanos , Peróxido de Hidrogênio , Grafite/química , Cério/química , Nanocompostos/química , Peroxidases , Técnicas Eletroquímicas
7.
BMC Med Genomics ; 17(1): 77, 2024 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-38515109

RESUMO

BACKGROUND: Cancer-associated fibroblasts (CAFs) play a crucial role in the tumor microenvironment of lung adenocarcinoma (LUAD) and are often associated with poorer clinical outcomes. This study aimed to screen for CAF-specific genes that could serve as promising therapeutic targets for LUAD. METHODS: We established a single-cell transcriptional profile of LUAD, focusing on genetic changes in fibroblasts. Next, we identified key genes associated with fibroblasts through weighted gene co-expression network analysis (WGCNA) and univariate Cox analysis. Then, we evaluated the relationship between glutathione peroxidase 8 (GPX8) and clinical features in multiple independent LUAD cohorts. Furthermore, we analyzed immune infiltration to shed light on the relationship between GPX8 immune microenvironment remodeling. For clinical treatment, we used the tumor immune dysfunction and exclusion (TIDE) algorithm to assess the immunotherapy prediction efficiency of GPX8. After that, we screened potential therapeutic drugs for LUAD by the connectivity map (cMAP). Finally, we conducted a cell trajectory analysis of GPX8+ CAFs to show their unique function. RESULTS: Fibroblasts were found to be enriched in tumor tissues. Then we identified GPX8 as a key gene associated with CAFs through comprehensive bioinformatics analysis. Further analysis across multiple LUAD cohorts demonstrated the relationship between GPX8 and poor prognosis. Additionally, we found that GPX8 played a role in inducing the formation of an immunosuppressive microenvironment. The TIDE method indicated that patients with low GPX8 expression were more likely to be responsive to immunotherapy. Using the cMAP, we identified beta-CCP as a potential drug-related to GPX8. Finally, cell trajectory analysis provided insights into the dynamic process of GPX8+ CAFs formation. CONCLUSIONS: This study elucidates the association between GPX8+ CAFs and poor prognosis, as well as the induction of immunosuppressive formation in LUAD. These findings suggest that targeting GPX8+ CAFs could potentially serve as a therapeutic strategy for the treatment of LUAD.


Assuntos
Adenocarcinoma de Pulmão , Fibroblastos Associados a Câncer , Neoplasias Pulmonares , Humanos , Adenocarcinoma de Pulmão/genética , Fibroblastos , Imunoterapia , Neoplasias Pulmonares/genética , Microambiente Tumoral , Prognóstico , Peroxidases
8.
Dalton Trans ; 53(12): 5624-5631, 2024 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-38440932

RESUMO

Noble metal nanomaterials have been widely demonstrated to possess intrinsic enzyme-like properties and have been increasingly applied in the fields of analysis and biomedicine. However, current exploration of high-activity noble metal nanozymes is still far from adequate. The construction of hollow structures and adjustment of their elemental composition are effective ways to improve the specific activity (SA) of nanozymes. In this study, trimetallic PtPdAu hollow nanorods (HNRs) were developed using a galvanic replacement reaction and Kirkendall effect. The catalytic experiment showed that the PtPdAu HNRs possessed outstanding peroxidase-like performance and their SA value was up to 563.71 U mg-1, which is remarkable among various previously reported nanozymes and higher than that of monometallic or bimetallic counterparts with similar structure and size prepared in this study. Electron paramagnetic resonance (EPR)measurements showed that the PtPdAu HNRs could contribute to the formation of hydroxyl radicals (˙OH) in catalyzing hydrogen peroxide. When using PtPdAu HNRs as a nanozyme in the colorimetric detection of H2O2 and ascorbic acid (AA), the limits of detection were as low as 1.8 µM and 0.068 µM, respectively. This study demonstrates that PtPdAu HNRs are high-activity nanozymes and have the potential to be applied in the field of analysis.


Assuntos
Nanotubos , Peroxidase , Peroxidase/química , Colorimetria , Peróxido de Hidrogênio/química , Peroxidases/química , Corantes/química
9.
Anal Chem ; 96(11): 4673-4681, 2024 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-38451931

RESUMO

Perfluorooctanesulfonic acid potassium salt (PFOS) residues in ecosystems over long periods are of increasing concern and require a selective and stable optical probe for monitoring. Herein, two functional groups (-F and -NH2) with opposite electronic modulation ability were introduced into Fe/Zn-BDC (denoted as Fe/Zn-BDC-F4 and Fe/Zn-BDC-NH2, respectively) to tailor the coordination environment of the Fe metal center, further regulating the nanozyme activity efficiently. Notably, the peroxidase-like activity is related to the coordination environment of the nanozymes and obeys the following order Fe/Zn-BDC-F4 > Fe/Zn-BDC > Fe/Zn-BDC-NH2. Based on the excellent peroxidase-like activity of Fe/Zn-BDC-F4 and the characteristics of being rich in F atoms, a rapid, selective, and visible colorimetric method was developed for detecting PFOS with a detection limit of 100 nM. The detection mechanism was attributed to various interaction forces between Fe/Zn-BDC-F4 and PFOS, including electrostatic interactions, Fe-S interactions, Fe-F bonds, and halogen bonds. This work not only offers new insights into the atomic-scale rational design of highly active nanozymes but also presents a novel approach to detecting PFOS in environmental samples.


Assuntos
Ecossistema , Potássio , Colorimetria , Peroxidases , Zinco
10.
Nat Commun ; 15(1): 2558, 2024 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-38519509

RESUMO

Encapsulins are self-assembling protein nanocompartments capable of selectively encapsulating dedicated cargo proteins, including enzymes involved in iron storage, sulfur metabolism, and stress resistance. They represent a unique compartmentalization strategy used by many pathogens to facilitate specialized metabolic capabilities. Encapsulation is mediated by specific cargo protein motifs known as targeting peptides (TPs), though the structural basis for encapsulation of the largest encapsulin cargo class, dye-decolorizing peroxidases (DyPs), is currently unknown. Here, we characterize a DyP-containing encapsulin from the enterobacterial pathogen Klebsiella pneumoniae. By combining cryo-electron microscopy with TP and TP-binding site mutagenesis, we elucidate the molecular basis for cargo encapsulation. TP binding is mediated by cooperative hydrophobic and ionic interactions as well as shape complementarity. Our results expand the molecular understanding of enzyme encapsulation inside protein nanocompartments and lay the foundation for rationally modulating encapsulin cargo loading for biomedical and biotechnological applications.


Assuntos
Proteínas de Bactérias , Peroxidase , Proteínas de Bactérias/metabolismo , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/metabolismo , Microscopia Crioeletrônica , Peroxidases/metabolismo
11.
Mikrochim Acta ; 191(4): 195, 2024 03 13.
Artigo em Inglês | MEDLINE | ID: mdl-38478128

RESUMO

Transition metal carbides modified by Au nanoparticles (Au/Ti3C2) were synthesized and developed as a colorimetric sensor for the determination of H2O2 and ampicillin. The surface electrical properties of Ti3C2 were changed, and Au nanoparticles (AuNPs) and gold growth solution were synthesized simultaneously. Au/Ti3C2 was obtained by seed growth method with AuNPs modified on the surface of transition metal carbides, nitrides or carbon-nitrides (Ti3C2 MXenes). The synthesized AuNPs and Ti3C2 had no peroxidase-like activity, but Au/Ti3C2 had. The peroxidase catalytic mechanism was due to electron transfer. The peroxidase activity of Au/Ti3C2 can be utilized for the determination of H2O2. The linear range of Au/Ti3C2 for H2O2 was 1-60 µM, and the detection limit was 0.12 µM (S/N = 3). A colometric sensor for ampicillin detection based on Au/Ti3C2 was further constructed since S in ampicillin formed an Au-S bond with Au/Ti3C2, leading to the weakening of its peroxidase-like property. The change of peroxidase-like property attenuated oxidation of TMB, and the ampicillin content was inversely proportional to the concentration of oxidized TMB, and the blue color of solution faded, which enabled the determination of ampicillin. The linear range for ampicillin was 0.005-0.5 µg mL- 1, and the detection limit was 1.1 ng mL- 1 (S/N = 3). The sensor was applied to the detection of ampicillin in milk and human serum.


Assuntos
Ouro , Nanopartículas Metálicas , Humanos , Ouro/química , Colorimetria/métodos , Peróxido de Hidrogênio/química , Nanopartículas Metálicas/química , Titânio , Peroxidase/química , Peroxidases
12.
Microb Cell Fact ; 23(1): 76, 2024 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-38461254

RESUMO

BACKGROUND: Aspergillus niger ATCC 20611 is an industrially important fructooligosaccharides (FOS) producer since it produces the ß-fructofuranosidase with superior transglycosylation activity, which is responsible for the conversion of sucrose to FOS accompanied by the by-product (glucose) generation. This study aims to consume glucose to enhance the content of FOS by heterologously expressing glucose oxidase and peroxidase in engineered A. niger. RESULTS: Glucose oxidase was successfully expressed and co-localized with ß-fructofuranosidase in mycelia. These mycelia were applied to synthesis of FOS, which possessed an increased purity of 60.63% from 52.07%. Furthermore, peroxidase was expressed in A. niger and reached 7.70 U/g, which could remove the potential inhibitor of glucose oxidase to facilitate the FOS synthesis. Finally, the glucose oxidase-expressing strain and the peroxidase-expressing strain were jointly used to synthesize FOS, which content achieved 71.00%. CONCLUSIONS: This strategy allows for obtaining high-content FOS by the multiple enzymes expressed in the industrial fungus, avoiding additional purification processes used in the production of oligosaccharides. This study not only facilitated the high-purity FOS synthesis, but also demonstrated the potential of A. niger ATCC 20611 as an enzyme-producing cell factory.


Assuntos
Aspergillus niger , Aspergillus , beta-Frutofuranosidase , Aspergillus niger/genética , Glucose Oxidase/genética , Oligossacarídeos , Peroxidases , Glucose
13.
Anal Chim Acta ; 1298: 342408, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38462333

RESUMO

BACKGROUND: In vitro screening strategies based on the inhibition of α-glucosidase (GAA) activity have been widely used for the discovery of potential antidiabetic drugs, but they still face some challenges, such as poor enzyme stability, non-reusability and narrow range of applicability. To overcome these limitations, an in vitro screening method based on GAA@GOx@Cu-MOF reactor was developed in our previous study. However, the method was still not satisfactory enough in terms of construction cost, pH stability, organic solvent resistance and reusability. Thence, there is still a great need for the development of in vitro screening methods with lower cost and wider applicability. RESULTS: A colorimetric sensing strategy based on GAA/(Au-Au/IrO2)@Cu(PABA) cascade catalytic reactor, which constructed through simultaneous encapsulating Au-Au/IrO2 nanozyme with glucose oxidase-mimicking and peroxidase-mimicking activities and GAA in Cu(PABA) carrier with peroxidase-mimicking activity, was innovatively developed for in vitro screening of GAA inhibitors in this work. It was found that the reactor not only exhibited excellent thermal stability, pH stability, organic solvent resistance, room temperature storage stability, and reusability, but also possessed cascade catalytic performance, with approximately 12.36-fold increased catalytic activity compared to the free system (GAA + Au-Au/IrO2). Moreover, the in vitro GAA inhibitors screening method based on this reactor demonstrated considerable anti-interference performance and detection sensitivity, with a detection limit of 4.79 nM for acarbose. Meanwhile, the method owned good reliability and accuracy, and has been successfully applied to the in vitro screening of oleanolic acid derivatives as potential GAA inhibitors. SIGNIFICANCE: This method not only more effectively solved the shortcomings of poor stability, narrow scope of application, and non-reusability of natural enzymes in the classical method compared with our previous work, but also broaden the application scope of Au-Au/IrO2 nanozyme with glucose oxidase and peroxidase mimicking activities, and Cu(PABA) carrier with peroxidase mimicking activity, which was expected to be a new generation candidate method for GAA inhibitor screening.


Assuntos
Ácido 4-Aminobenzoico , Inibidores de Glicosídeo Hidrolases , Inibidores de Glicosídeo Hidrolases/farmacologia , Glucose Oxidase , Reprodutibilidade dos Testes , Colorimetria/métodos , Peroxidases , Solventes , Peróxido de Hidrogênio
14.
Mikrochim Acta ; 191(4): 192, 2024 03 12.
Artigo em Inglês | MEDLINE | ID: mdl-38467931

RESUMO

FeOx-TiO2@Carbon hybrid structure materials (FeOx-TiO2@CHs) with high peroxidase (POD)-like activity have been prepared by one-pot hydrothermal method. Based on the excellent POD activity of FeOx-TiO2@CHs, one pot colorimetric detection for glucose was constructed by using TMB as substrate with the synergistic reaction of glucose oxidase; the linear range and the limit of detection (LOD) are 25 ~ 1000 and 1.77 µM, respectively. Using this method, the glucose in serum real samples was detected with satisfactory results, and the results are consistent with that of the glucometer method in the hospital. The recovery in diabetic and artificial urine samples was 95.71 ~ 104.67% and 99.01 ~ 103.16%, respectively. The mechanism of the catalytic colorimetric reaction was also investigated by multiple measurements, and the results indicated that superoxide anions (O2•-) between FeOx-TiO2@CHs and substrate play a main role, but a small quantity of hydroxyl radical •OH and singlet oxygen 1O2 is also generated simultaneously. The one-pot reaction method is simple and fast; the detection process only requires a simple mixing, which is suitable for application in special environment.


Assuntos
Glucose , Peroxidase , Peroxidase/química , Carbono/química , Colorimetria/métodos , Peróxido de Hidrogênio/química , Peroxidases/química , Corantes
15.
Biosens Bioelectron ; 252: 116134, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38417287

RESUMO

Herein, novel nanozyme mimics MoO3/MIL-125-NH2 were reported and conjugated with bacteriophages as a new electrochemical probe for high sensitivity and specific electrochemical detection of staphylococcus aureus. The excellent peroxidase-like activity of MoO3/MIL-125-NH2 composites was attributed to the integration of MIL-125-NH2 with MoO3, which can boost the generation of superoxide radicals (O• 2-) and thus promote the oxidation of TMB in the presence of H2O2. In this work, two bacteriophages named SapYZU04 and SapYZU10 were isolated from sewage samples by using staphylococcus aureus YZUsa12 as the host. In comparison, MoO3/MIL-125-NH2@SapYZU04 was selected as a recognition agent. The DPV current declined linearly with staphylococcus aureus YZUsa12 concentration in the range of 101-108 CFU mL-1, with a low detection limit of 16 CFU mL-1 (S/N = 3). 20 strains including 13 host strains and 7 non-host strains were used to evaluate the selectivity of the proposed sensor. Regardless of the differences in the degrees of lytic performance for phage SapYZU04, all selected host strains can be screened with merely the same DPV current. Host spectrum-oriented bacteriophage sensing is of great importance for the practical application of bacteriophage-based biosensors in the future.


Assuntos
Bacteriófagos , Técnicas Biossensoriais , Infecções Estafilocócicas , Humanos , Staphylococcus aureus , Peróxido de Hidrogênio , Peroxidases
16.
Nano Lett ; 24(9): 2912-2920, 2024 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-38391386

RESUMO

Nanozymes with peroxidase-like activity have been extensively studied for colorimetric biosensing. However, their catalytic activity and specificity still lag far behind those of natural enzymes, which significantly affects the accuracy and sensitivity of colorimetric biosensing. To address this issue, we design PdSn nanozymes with selectively enhanced peroxidase-like activity, which improves the sensitivity and accuracy of a colorimetric immunoassay. The peroxidase-like activity of PdSn nanozymes is significantly higher than that of Pd nanozymes. Theoretical calculations reveal that the p-d orbital hybridization of Pd and Sn not only results in an upward shift of the d-band center to enhance hydrogen peroxide (H2O2) adsorption but also regulates the O-O bonding strength of H2O2 to achieve selective H2O2 activation. Ultimately, the nanozyme-linked immunosorbent assay has been successfully developed to sensitively and accurately detect the prostate-specific antigen (PSA), achieving a low detection limit of 1.696 pg mL-1. This work demonstrates a promising approach for detecting PSA in a clinical diagnosis.


Assuntos
Técnicas Biossensoriais , Peróxido de Hidrogênio , Masculino , Humanos , Antígeno Prostático Específico , Imunoensaio/métodos , Antioxidantes , Peroxidases , Colorimetria/métodos , Técnicas Biossensoriais/métodos
17.
Chemosphere ; 346: 140557, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38303399

RESUMO

Single-atom nanozymes (SANs) are nanomaterials-based nanozymes with atomically dispersed enzyme-like active sites. SANs offer improved as well as tunable catalytic activity. The creation of extremely effective SANs and their potential uses have piqued researchers' curiosity due to their advantages of cheap cost, variable catalytic activity, high stability, and large-scale production. Furthermore, SANs with uniformly distributed active centers and definite coordination structures offer a distinctive opportunity to investigate the structure-activity correlation and control the geometric and electrical features of metal centers. SANs have been extensively explored in photo-, thermal-, and electro-catalysis. However, SANs suffer from the following disadvantages, such as efficiency, non-mimicking of the 3-D complexity of natural enzymes, limited and narrow range of artificial SANs, and biosafety aspects. Among a quite limited range of artificial SANs, the peroxidase action of SANs has attracted significant research attention in the last five years with the aim of producing reactive oxygen species for use in cancer therapy, and water treatment among many other applications. In this review, we explore the recent progress of different SANs as peroxidase mimics, the role of the metal center in enzymatic activity, possible prospects, and underlying limitations in real-time applications.


Assuntos
Materiais Biomiméticos , Nanoestruturas , Materiais Biomiméticos/química , Nanoestruturas/química , Peroxidase , Catálise , Peroxidases
18.
Anal Chim Acta ; 1293: 342271, 2024 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-38331555

RESUMO

BACKGROUND: Recently, the enzymatic cascade reactions during the cellular process are widely used for fabricating robust biosensors and they have attracted extensive attention in analyzing various clinical biomarkers. The enzymatic cascades analysis is commonly based on the peroxidase (POD)/oxidase coupled system. However, the requirement of harsh acidic environment, poor stability and interference from the oxidase further limit their analytical practicability. Herein, novel chromogenic nanomaterials with H2O2 sensitive features are urgently required to replace the POD nanozyme in enzymatic cascades based bioanalysis. RESULTS: Herein, oxygen deficient MoO3-x with H2O2 sensitive features and near-infrared (NIR) absorption band have been ultra-fast synthesized and utilized for the enzymatic cascades analysis of α-Glucosidase's activity, and inhibitors screening. With the addition of 4-nitrophenyl-α-d-glucopyranoside, the simultaneous presence of α-Glucosidase and glucose oxidase (GOx) would fade their dark blue color and decrease the NIR absorption. The α-Glucosidase's activity can be analyzed by the absorption at 770 nm, and their limit of detection is 8 × 10-5 U/mL, indicating the superior performance of the proposed colorimetric assay. Moreover, this proposed α-Glucosidase assay is further utilized for inhibitors screening. Moreover, the activity of α-Glucosidase can also be analyzed by the smartphone and microplate reader through the agarose-based colorimetric portable kit. SIGNIFICANCE: This MoO3-x involved enzymatic cascades assay would facilitate for the development of bio-analysis related to H2O2 generation or consumption. Moreover, this bio-analysis strategy will contribute to the development of other H2O2 sensitive chromogenic nanomaterials for the analysis of certain biomolecules and biological enzymes.


Assuntos
Nanoestruturas , alfa-Glucosidases , Peróxido de Hidrogênio/análise , Peroxidases , Peroxidase , Corantes , Oxigênio , Colorimetria
19.
Nat Commun ; 15(1): 1239, 2024 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-38336829

RESUMO

Currently available genetically encoded H2O2 probes report on the thiol redox state of the probe, which means that they reflect the balance between probe thiol oxidation and reduction. Here we introduce the use of the engineered heme peroxidase APEX2 as a thiol-independent chemogenetic H2O2 probe that directly and irreversibly converts H2O2 molecules into either fluorescent or luminescent signals. We demonstrate sensitivity, specificity, and the ability to quantitate endogenous H2O2 turnover. We show how the probe can be used to detect changes in endogenous H2O2 generation and to assess the roles and relative contributions of endogenous H2O2 scavengers. Furthermore, APEX2 can be used to study H2O2 diffusion inside the cytosol. Finally, APEX2 reveals the impact of commonly used alkylating agents and cell lysis protocols on cellular H2O2 generation.


Assuntos
Peróxido de Hidrogênio , Peroxidases , Heme , Oxirredução , Peroxidases/química , Peroxidases/metabolismo , Compostos de Sulfidrila
20.
Int J Mol Sci ; 25(4)2024 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-38396991

RESUMO

Low-temperature chilling is a major abiotic stress leading to reduced rice yield and is a significant environmental threat to food security. Low-temperature chilling studies have focused on physiological changes or coding genes. However, the competitive endogenous RNA mechanism in rice at low temperatures has not been reported. Therefore, in this study, antioxidant physiological indices were combined with whole-transcriptome data through weighted correlation network analysis, which found that the gene modules had the highest correlation with the key antioxidant enzymes superoxide dismutase and peroxidase. The hub genes of the superoxide dismutase-related module included the UDP-glucosyltransferase family protein, sesquiterpene synthase and indole-3-glycerophosphatase gene. The hub genes of the peroxidase-related module included the WRKY transcription factor, abscisic acid signal transduction pathway-related gene plasma membrane hydrogen-ATPase and receptor-like kinase. Therefore, we selected the modular hub genes and significantly enriched the metabolic pathway genes to construct the key competitive endogenous RNA networks, resulting in three competitive endogenous RNA networks of seven long non-coding RNAs regulating three co-expressed messenger RNAs via four microRNAs. Finally, the negative regulatory function of the WRKY transcription factor OsWRKY61 was determined via subcellular localization and validation of the physiological indices in the mutant.


Assuntos
MicroRNAs , Oryza , RNA Longo não Codificante , Oryza/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Antioxidantes , Perfilação da Expressão Gênica , MicroRNAs/genética , MicroRNAs/metabolismo , Redes Reguladoras de Genes , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Peroxidases/genética , Superóxido Dismutase/genética
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