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1.
Mikrochim Acta ; 191(7): 368, 2024 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-38833176

RESUMO

A colorimetric analysis platform has been successfully developed based on FeCo-NC dual-atom nanozyme (FeCo-NC DAzyme) for the detection of organophosphorus pesticides (OPPs). The FeCo-NC DAzyme exhibited exceptional oxidase-like activity (OXD), enabling the catalysis of colorless TMB to form blue oxidized TMB (oxTMB) without the need for H2O2 involvement. By combining acid phosphatase (ACP) hydrolase with FeCo-NC DAzyme, a "FeCo-NC DAzyme + TMB + ACP + SAP" colorimetric system was constructed, which facilitated the rapid detection of malathion. The chromogenic system was applied to detect malathion using a smartphone-based app and an auxiliary imaging interferogram device for colorimetric measurements, which have a linear range of 0.05-4.0 µM and a limit of detection (LOD) as low as 15 nM in real samples, comparable to UV-Vis and HPLC-DAD detection methods. Overall, these findings present a novel approach for convenient, rapid, and on-site monitoring of OPPs.


Assuntos
Colorimetria , Limite de Detecção , Praguicidas , Smartphone , Colorimetria/métodos , Praguicidas/análise , Compostos Organofosforados/análise , Compostos Organofosforados/química , Malation/análise , Malation/química , Oxirredutases/química , Ferro/química , Fosfatase Ácida/análise , Fosfatase Ácida/química , Benzidinas
2.
PLoS One ; 19(6): e0297215, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38875297

RESUMO

Dianthus barbatus linn. is widely used in gardens, mainly as flower beds and flower borders. The effects of different gradients of P on the growth and root morphology of Dianthus barbatus were studied to explore its morphological and physiological responses and adaptive strategies. Hence, this study provides a theoretical basis and practical guidance for D. barbatus production. Two soil substrates, namely loess and vegetable soil, and five phosphorus concentration gradients were set; no phosphorus application was used as the control. The morphology and physiology of D. barbatus were also investigated. Low-to-medium- and low-phosphorus treatments promoted the growth of D. barbatus in the above and underground parts of the plants grown on both substrates. Chlorophyll content, flower quantity, and acid phosphatase activity in the rhizosphere soil were significantly increased in the H1 and H2 treatments of loess and in the C4 treatment of vegetable soil. Thus, D. barbatus seems to reduce the damage caused by phosphorus stress by increasing chlorophyll content and root acid phosphatase activity. The latter was significantly higher in vegetable soil than in loess. Vegetable soil was more conducive to D. barbatus growth than loess.


Assuntos
Clorofila , Dianthus , Fósforo , Raízes de Plantas , Solo , Fósforo/metabolismo , Solo/química , Clorofila/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Raízes de Plantas/efeitos dos fármacos , Dianthus/crescimento & desenvolvimento , Dianthus/metabolismo , Dianthus/fisiologia , Fosfatase Ácida/metabolismo , Flores/metabolismo , Flores/crescimento & desenvolvimento , Rizosfera
3.
J Hazard Mater ; 474: 134867, 2024 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-38861900

RESUMO

Organic phosphorus (Po) is a large component of soil P, but it is often unavailable for plant uptake. Purple acid phosphatases (PAP) can hydrolyze a wide range of Po, playing an important role in Po utilization by plants. In this study, we investigated a novel secretary PvPAP1 from the As-hyperaccumulator Pteris vittata, which can effectively utilize exogenous Po, including adenosine triphosphate (ATP) and phytate. Unlike other PAP, PvPAP1 was abundantly-expressed in P. vittata roots, which was upregulated 3.5-folds under P-deprivation than P-sufficient conditions. When expressed in tobacco, its activity in the roots of PvPAP1-Ex lines was ∼8 folds greater than that in wild-type (WT) plants. Besides, PvPAP1 exhibited its secretory ability as evidenced by the sapphire-blue color on the root surface after treating with 5-bromo-4-chloro-3-indolyl phosphate. In a long-term experiment using sand media, PvPAP1-expressing tobacco plants showed 25-30 % greater root biomass than WT plants when using ATP as the sole P source. This is because PvPAP1-expression enhanced its phosphatase activity by 6.5-9.2 folds in transgenic tobacco, thereby increasing the P contents by 39-41 % in its roots under ATP treatment and 9.4-30 % under phytate treatment. The results highlight PvPAP1 as a novel secreted phosphatase crucial for external Po utilization in P. vittata, suggesting that PvPAP1 has the potential to serve as a valuable gene resource for enhancing Po utilization by crop plants.


Assuntos
Nicotiana , Fósforo , Ácido Fítico , Raízes de Plantas , Pteris , Ácido Fítico/metabolismo , Nicotiana/metabolismo , Nicotiana/genética , Nicotiana/crescimento & desenvolvimento , Fósforo/metabolismo , Pteris/metabolismo , Pteris/genética , Pteris/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Hidrólise , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Fosfatase Ácida/metabolismo , Fosfatase Ácida/genética , Arsênio/metabolismo , Regulação da Expressão Gênica de Plantas
4.
Huan Jing Ke Xue ; 45(6): 3523-3532, 2024 Jun 08.
Artigo em Chinês | MEDLINE | ID: mdl-38897772

RESUMO

In this study, the effects of four types of amendments on effective Cd and Cd content in different parts of prickly ash soil and soil enzyme activity were studied, which provided scientific basis for acidification improvement of purple soil and heavy metal pollution control. A field experiment was conducted. Six treatments were set up:no fertilizer (CK), only chemical fertilizer (F), lime + chemical fertilizer (SF), organic fertilizer + chemical fertilizer (OM), biochar + chemical fertilizer (BF), and vinasse biomass ash + chemical fertilizer (JZ). Soil pH; available Cd (DTPA-Cd); Cd content in branches, leaves, shells, and seeds of Zanthoxylum; as well as the activities of catalase (S-CAT), acid phosphatase (S-ACP), and urease (S-UE) in different treatments were studied, and their relationships were clarified. The results showed following:① The two treatments of vinasse biomass ash + chemical fertilizer and lime + chemical fertilizer significantly increased soil pH (P < 0.05) to 3.39 and 2.25 units higher than that in the control, respectively. Compared with that in the control treatment, the content of available Cd in soil under vinasse biomass ash + chemical fertilizer and lime + chemical fertilizer treatment decreased by 28.91 % and 20.90 %, respectively. ② The contents of Cd in leaves, shells, and seeds of Zanthoxylum were decreased by 31.33 %, 30.24 %, and 34.01 %, respectively. The Cd enrichment ability of different parts of Zanthoxylum was different, with the specific performances being leaves > branches > seeds > shells. Compared with that of the control, the enrichment coefficient of each part of Zanthoxylum treated with vinasse biomass ash + chemical fertilizer decreased significantly(P < 0.05)by 27.54 %-40.0 %. ③ The changes in catalase and urease activities in soil treated with amendments were similar. Compared with those in the control group, the above two enzyme activities were significantly increased by 191.26 % and 199.50 %, respectively, whereas the acid phosphatase activities were decreased by 16.45 %. Correlation analysis showed that soil available Cd content was significantly negatively correlated with soil pH value(P < 0.01), S-CAT and S-UE enzyme activities were significantly positively correlated with soil pH(P < 0.01), and the soil available Cd content was significantly negatively correlated (P < 0.01); the S-ACP enzyme showed the complete opposite trends. The application of lime and vinasse biomass ash to acidic purple soil had the most significant effect on neutralizing soil acidity. It was an effective measure to improve acidic purple soil and prevent heavy metal pollution by reducing the effective Cd content in soil and improving the soil environment while inhibiting the absorption and transfer of Cd in various parts of Zanthoxylum.


Assuntos
Cádmio , Fertilizantes , Poluentes do Solo , Solo , Poluentes do Solo/metabolismo , Cádmio/metabolismo , Solo/química , Urease/metabolismo , Zanthoxylum/química , Zanthoxylum/metabolismo , Fosfatase Ácida/metabolismo , Catalase/metabolismo , Disponibilidade Biológica , Óxidos/química , Compostos de Cálcio/química , Carvão Vegetal/química
5.
Anat Histol Embryol ; 53(3): e13054, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38735037

RESUMO

Identifying and analysing distinct blood cells is crucial for the diagnosis and treatment of diseases in the field of biomedicine. The present study was undertaken to study the cytomorphological and cytochemical characteristics of the blood cells of Zoar, a non-descript indigenous breed of chicken extensively reared under backyard poultry farming in Mizoram, India. For this study, 2 mL of blood samples were aseptically collected from the wings veins of 12 chickens and were processed for light microscopic study under standard protocols. The matured erythrocytes were elliptical, while the immature erythrocytes appeared oval. The heterophils were positive for SBB (SBB), Periodic Acid Schiff (PAS), acid phosphatase, alkaline phosphatase and Arylsulphatase while the eosinophils were positive for SBB, PAS, alkaline phosphatase, cytochrome oxidase and peroxidase. The basophils of were positive for toluidine blue while the thrombocytes were positive for PAS. These cytochemical and cytoenzymatic staining properties plays a very important role in diagnosis, differentiation, and classification of leukaemias.


Assuntos
Galinhas , Eosinófilos , Eritrócitos , Animais , Galinhas/anatomia & histologia , Índia , Eritrócitos/citologia , Eosinófilos/citologia , Células Sanguíneas/citologia , Plaquetas/citologia , Fosfatase Alcalina/sangue , Basófilos/citologia , Fosfatase Ácida/sangue , Complexo IV da Cadeia de Transporte de Elétrons/análise
6.
Int J Biol Macromol ; 270(Pt 2): 132222, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38729468

RESUMO

Fungal polysaccharides have been explored by many for both structural studies and biological activities, but few studies have been done on the extracellular polysaccharides of Dictyophora rubrovalvata, so a new exopolysaccharide was isolated from Dictyophora rubrovalvata and its structure and its immunological activity were investigated. The crude exopolysaccharide (EPS) was purified by DEAE52 cellulose and Sephadex G-200 to obtain a new acidic polysaccharide (DR-EPS). DR-EPS (2.66 × 103 kDa) was consisted mainly of mannose, glucose, galactose and glucuronic acid with a molar ratio of 1: 0.86: 0.20: 0.01. In addition, DR-EPS increased the phagocytic activity of RAW264.7 cells up to 2.67 times of the blank control group. DR-EPS improved intracellular nucleic acid and glycogen metabolism as observed by AO and PAS staining. DR-EPS(40 µg/mL) promoted NO production up to 30.66 µmol, enhanced acid phosphatase (ACP) and superoxide dismutase (SOD) activities, with activity maxima of 660 U/gprot and 96.27 U/mgprot, respectively, and DR-EPS (160 µg / mL) significantly increased the lysozyme content as 2.73 times of the control group. The good immunological activity of extracellular polysaccharides of Dictyophora rubrovalvata provides directions for the use of fermentation broths.


Assuntos
Polissacarídeos Fúngicos , Camundongos , Animais , Células RAW 264.7 , Polissacarídeos Fúngicos/farmacologia , Polissacarídeos Fúngicos/química , Polissacarídeos Fúngicos/isolamento & purificação , Óxido Nítrico/metabolismo , Fatores Imunológicos/farmacologia , Fatores Imunológicos/química , Fatores Imunológicos/isolamento & purificação , Fagocitose/efeitos dos fármacos , Agentes de Imunomodulação/farmacologia , Agentes de Imunomodulação/química , Agentes de Imunomodulação/isolamento & purificação , Superóxido Dismutase/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/imunologia , Fosfatase Ácida/metabolismo
7.
Chemosphere ; 359: 142288, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38750729

RESUMO

Helicoverpa armigera, a ubiquitous polyphagous pest, poses a significant threat to global agriculture, causing substantial economic losses and demonstrating resistance to synthetic pesticides. This study investigates the potential of emamectin benzoate (EMB), an avermectin derivative, as an effective control agent against H. armigera. The larvae of the NBII-MP-NOC-01 strain of H. armigera were reared on an artificial diet. The impact of dietary EMB was examined on four midgut enzymes; alanine aminotransferase (ALT), aspartate aminotransferase (AST), acid phosphatase (ACP), and alkaline phosphatase (ALP). Results showed a dose-dependent and time-dependent reduction in ALT and AST activity, while an initial increase and subsequent decline in ACP and ALP activity at higher EMB concentrations. Computational modelling of enzyme structures and molecular docking studies revealed differential binding of EMB with the midgut enzymes. The strongest interaction was observed between EMB and ALT residues, contrasting with weakest interactions observed with AST. The study also showed that decreased activity of transaminases in H. armigera caused by EMB may be because of stability-activity trade-off, while in phosphatases reverse may be the case. This research provides crucial insights into the biochemical responses and the intricate insecticide-enzyme interactions in H. armigera caused by EMB exposure. This study lays the foundation for further research aimed at developing environmentally friendly approaches for managing H. armigera, addressing the challenges associated with conventional pesticides.


Assuntos
Fosfatase Ácida , Alanina Transaminase , Fosfatase Alcalina , Aspartato Aminotransferases , Inseticidas , Ivermectina , Larva , Simulação de Acoplamento Molecular , Mariposas , Animais , Ivermectina/análogos & derivados , Ivermectina/toxicidade , Larva/efeitos dos fármacos , Mariposas/efeitos dos fármacos , Inseticidas/toxicidade , Inseticidas/química , Inseticidas/metabolismo , Fosfatase Alcalina/metabolismo , Fosfatase Ácida/metabolismo , Alanina Transaminase/metabolismo , Aspartato Aminotransferases/metabolismo , Helicoverpa armigera
8.
Sci Adv ; 10(19): eadm7515, 2024 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-38728394

RESUMO

The nonpolymorphic major histocompatibility complex E (MHC-E) molecule is up-regulated on many cancer cells, thus contributing to immune evasion by engaging inhibitory NKG2A/CD94 receptors on NK cells and tumor-infiltrating T cells. To investigate whether MHC-E expression by cancer cells can be targeted for MHC-E-restricted T cell control, we immunized rhesus macaques (RM) with rhesus cytomegalovirus (RhCMV) vectors genetically programmed to elicit MHC-E-restricted CD8+ T cells and to express established tumor-associated antigens (TAAs) including prostatic acidic phosphatase (PAP), Wilms tumor-1 protein, or Mesothelin. T cell responses to all three tumor antigens were comparable to viral antigen-specific responses with respect to frequency, duration, phenotype, epitope density, and MHC restriction. Thus, CMV-vectored cancer vaccines can bypass central tolerance by eliciting T cells to noncanonical epitopes. We further demonstrate that PAP-specific, MHC-E-restricted CD8+ T cells from RhCMV/PAP-immunized RM respond to PAP-expressing HLA-E+ prostate cancer cells, suggesting that the HLA-E/NKG2A immune checkpoint can be exploited for CD8+ T cell-based immunotherapies.


Assuntos
Antígenos de Neoplasias , Linfócitos T CD8-Positivos , Antígenos HLA-E , Animais , Humanos , Masculino , Fosfatase Ácida , Apresentação de Antígeno/imunologia , Antígenos de Neoplasias/imunologia , Vacinas Anticâncer/imunologia , Linfócitos T CD8-Positivos/imunologia , Linhagem Celular Tumoral , Citomegalovirus/imunologia , Antígenos de Histocompatibilidade Classe I/imunologia , Antígenos de Histocompatibilidade Classe I/metabolismo , Macaca mulatta , Mesotelina
9.
Ying Yong Sheng Tai Xue Bao ; 35(3): 631-638, 2024 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-38646750

RESUMO

Litter input triggers the secretion of soil extracellular enzymes and facilitates the release of carbon (C), nitrogen (N), and phosphorus (P) from decomposing litter. However, how soil extracellular enzyme activities were controlled by litter input with various substrates is not fully understood. We examined the activities and stoichiometry of five enzymes including ß-1,4-glucosidase, ß-D-cellobiosidase, ß-1,4-N-acetyl-glucosaminidase, leucine aminopeptidase and acidic phosphatase (AP) with and without litter input in 10-year-old Castanopsis carlesii and Cunninghamia lanceolata plantations monthly during April to August, in October, and in December 2021 by using an in situ microcosm experiment. The results showed that: 1) There was no significant effect of short-term litter input on soil enzyme activity, stoichiometry, and vector properties in C. carlesii plantation. In contrast, short-term litter input significantly increased the AP activity by 1.7% in May and decreased the enzymatic C/N ratio by 3.8% in August, and decreased enzymatic C/P and N/P ratios by 11.7% and 10.3%, respectively, in October in C. lanceolata plantation. Meanwhile, litter input increased the soil enzymatic vector angle to 53.8° in October in C. lanceolata plantations, suggesting a significant P limitation for soil microorganisms. 2) Results from partial least squares regression analyses showed that soil dissolved organic matter and microbial biomass C and N were the primary factors in explaining the responses of soil enzymatic activity to short-term litter input in both plantations. Overall, input of low-quality (high C/N) litter stimulates the secretion of soil extracellular enzymes and accelerates litter decomposition. There is a P limitation for soil microorganisms in the study area.


Assuntos
Carbono , Cunninghamia , Fagaceae , Nitrogênio , Fósforo , Microbiologia do Solo , Solo , Solo/química , Cunninghamia/crescimento & desenvolvimento , Cunninghamia/metabolismo , Carbono/metabolismo , Carbono/análise , Nitrogênio/metabolismo , Nitrogênio/análise , Fósforo/metabolismo , Fósforo/análise , Fagaceae/crescimento & desenvolvimento , Fagaceae/metabolismo , Leucil Aminopeptidase/metabolismo , Celulose 1,4-beta-Celobiosidase/metabolismo , Ecossistema , Folhas de Planta/metabolismo , Folhas de Planta/química , Acetilglucosaminidase/metabolismo , Fosfatase Ácida/metabolismo , beta-Glucosidase/metabolismo , China
10.
Spectrochim Acta A Mol Biomol Spectrosc ; 315: 124263, 2024 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-38593539

RESUMO

Fluorescence analysis has attracted much attention due to its rapidity and sensitivity. The present work describes a novel fluorescence detection method for acid phosphatase (ACP) on the basis of inner-filter effect (IFE), where MnO2 nanosheets (MnO2 NSs) and vitamin B2 (VB2) are served as absorbers and fluorophores, respectively. In the absence of ACP, the absorption band of MnO2 NSs overlaps well with the excitation band of VB2, resulting in effective IFE and inhibition of VB2 fluorescence. In the presence of ACP, 2-phospho-L-ascorbic acid trisodium salt (AAP) is hydrolyzed to generate ascorbic acid (AA), which efficiently trigger the reduction of MnO2 NSs into Mn2+ ions, causing the weakening of the MnO2 NSs absorption band and the recovery of VB2 fluorescence. Further investigation indicates that the fluorescence recovery degree of VB2 increases with the increase of ACP concentration. Under selected experimental conditions, the proposed method can achieve sensitive detection of ACP in the ranges of 0.5-4.0 mU/mL and 4.0-15 mU/mL along with a limit of detection (LOD) as low as 0.14 mU/mL. Finally, this method was successfully applied for the detection of ACP in human serum samples with satisfactory recoveries in the range of 95.0 %-108 %.


Assuntos
Fosfatase Ácida , Limite de Detecção , Compostos de Manganês , Nanoestruturas , Óxidos , Espectrometria de Fluorescência , Compostos de Manganês/química , Óxidos/química , Espectrometria de Fluorescência/métodos , Humanos , Fosfatase Ácida/sangue , Fosfatase Ácida/metabolismo , Fosfatase Ácida/análise , Nanoestruturas/química , Ácido Ascórbico/análise , Ácido Ascórbico/farmacologia
11.
Int J Biol Macromol ; 266(Pt 2): 131339, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38574925

RESUMO

The AcPase exhibits a specific activity of 31.32 U/mg of protein with a 728-fold purification, and the yield of the enzyme is raised to 3.15 %. The Zn2+-dependent AcPase showed a purification factor of 1.34 specific activity of 14 U/mg of proteins and a total recovery of 5.14. The SDS-PAGE showed a single band corresponding to a molecular weight of 18 kDa of AcPase and 29 kDa of Zn2+-dependent AcPase. The AcPase enzyme has shown a wide range of substrate specificity for p-NPP, phenyl phosphate and FMN, while in the case of ZnAcPase α and ß-Naphthyl phosphate and p-NPP were proved to be superior substrates. The divalent metal ions like Mg2+, Mn2+, and Ca2+ increased the activity, while other substrates decreased the enzyme activity. The Km (0.14 mM) and Vmax (21 µmol/min/mg) values of AcPase were higher than those of Zn2+-AcPase (Km = 0.5 mM; Vmax = 9.7 µmol/min/mg). The Zn2+ ions activate the Zn2+-AcPase while Fe3+, Al3+, Pb2+, and Hg2+ showed inhibition on enzyme activity. Molybdate, vanadate and phosphate were found to be competitive inhibitors of AcPase with Ki values 316 µM, 185 µM, and 1.6 mM, while in Zn2+-AcPase tartrate and phosphate also showed competitive inhibition with Ki values 3 mM and 0.5 mM respectively.


Assuntos
Fosfatase Ácida , Encéfalo , Galinhas , Zinco , Animais , Zinco/química , Especificidade por Substrato , Fosfatase Ácida/metabolismo , Fosfatase Ácida/química , Fosfatase Ácida/isolamento & purificação , Encéfalo/enzimologia , Cinética , Concentração de Íons de Hidrogênio , Peso Molecular
12.
World J Microbiol Biotechnol ; 40(6): 171, 2024 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-38630327

RESUMO

A histidine acid phosphatase (HAP) (PhySc) with 99.50% protein sequence similarity with PHO5 from Saccharomyces cerevisiae was expressed functionally with the molecular mass of ∼110 kDa through co-expression along with the set of molecular chaperones dnaK, dnaJ, GroESL. The purified HAP illustrated the optimum activity of 28.75 ± 0.39 U/mg at pH 5.5 and 40 ˚C. The Km and Kcat values towards calcium phytate were 0.608 ± 0.09 mM and 650.89 ± 3.6 s- 1. The half-lives (T1/2) at 55 and 60 ˚C were 2.75 min and 55 s, respectively. The circular dichroism (CD) demonstrated that PhySc includes 30.5, 28.1, 21.3, and 20.1% of random coils, α-Helix, ß-Turns, and ß-Sheet, respectively. The Tm recorded by CD for PhySc was 56.5 ± 0.34˚C. The molecular docking illustrated that His59 and Asp322 act as catalytic residues in the PhySc. MD simulation showed that PhySc at 40 ˚C has higher structural stability over those of the temperatures 60 and 80 ˚C that support the thermodynamic in vitro investigations. Secondary structure content results obtained from MD simulation indicated that PhySc consists of 34.03, 33.09, 17.5, 12.31, and 3.05% of coil, helix, turn, sheet, and helix310, respectively, which is almost consistent with the experimental results.


Assuntos
Magnésio , Simulação de Dinâmica Molecular , Radioisótopos , Proteínas de Saccharomyces cerevisiae , Fosfatase Ácida/genética , Saccharomyces cerevisiae/genética , Histidina , Simulação de Acoplamento Molecular , Proteínas de Saccharomyces cerevisiae/genética
13.
Microb Biotechnol ; 17(4): e14404, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38588312

RESUMO

Acid phosphatases are enzymes that play a crucial role in the hydrolysis of various organophosphorous molecules. A putative acid phosphatase called FS6 was identified using genetic profiles and sequences from different environments. FS6 showed high sequence similarity to type C acid phosphatases and retained more than 30% of consensus residues in its protein sequence. A histidine-tagged recombinant FS6 produced in Escherichia coli exhibited extremophile properties, functioning effectively in a broad pH range between 3.5 and 8.5. The enzyme demonstrated optimal activity at temperatures between 25 and 50°C, with a melting temperature of 51.6°C. Kinetic parameters were determined using various substrates, and the reaction catalysed by FS6 with physiological substrates was at least 100-fold more efficient than with p-nitrophenyl phosphate. Furthermore, FS6 was found to be a decamer in solution, unlike the dimeric forms of crystallized proteins in its family.


Assuntos
Fosfatase Ácida , Extremófilos , Fosfatase Ácida/metabolismo , Extremófilos/genética , Extremófilos/metabolismo , Hidrólise , Sequência de Aminoácidos , Especificidade por Substrato , Concentração de Íons de Hidrogênio
14.
Tuberculosis (Edinb) ; 146: 102498, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38461765

RESUMO

Drug resistance to tuberculosis (TB) has become an obstacle in eliminating tuberculosis. The transmission of drug-resistant TB from patients increases the incidence of primary drug-resistant (DR) TB in individuals who are in close contact. Therefore, it is necessary to incorporate an immunological approach into preventive therapy. This study focuses on the activity of lysosomal enzymes, oxygen bursts, and the attachment ability of macrophages among individuals diagnosed with active drug-resistant TB compared with close contacts with latent TB or healthy cases. We measured macrophage oxygen burst ability (Water-soluble tetrazolium salt (WST) test, Nitric Oxide production, and myeloperoxidase activity) and the degradative ability of lysosomes (activity of the ß-glucuronidase and acid phosphatase enzymes). Six active DR-TB patients and 18 close-contact cases (8 Latent Tuberculosis Infection (LTBI); 10 healthy) were recruited at Universitas Indonesia Hospital. The macrophage attachment of the LTBI group was higher than in the other groups. NO production, myeloperoxidase activity, ß-glucuronidase, and acid phosphatase were higher in the active DR-TB group. A negative correlation was uncovered between phagocytosis and NO production, myeloperoxidase activity, and lysosomal enzymes. The difference in macrophage function is expected to be a further reference in active DR-TB treatment or preventive therapy.


Assuntos
Tuberculose Latente , Mycobacterium tuberculosis , Tuberculose Resistente a Múltiplos Medicamentos , Tuberculose , Humanos , Tuberculose/diagnóstico , Tuberculose Resistente a Múltiplos Medicamentos/tratamento farmacológico , Macrófagos , Glucuronidase , Óxido Nítrico , Fosfatase Ácida , Peroxidase
15.
Appl Spectrosc ; 78(6): 633-643, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38529537

RESUMO

The precise regulation of nanoenzyme activity is of great significance for application to biosensing analysis. Herein, the peroxidase-like activity of carbon dots was effectively modulated by doping phosphorus, which was successfully employed for sensitive, selective detection of acid phosphatase (ACP). Phosphorus-doped carbon dots (P-CDs) with excellent peroxidase-like activity were synthesized by a one-pot hydrothermal method, and the catalytic activity could be easily modulated by controlling the additional amount of precursor phytic acid. P-CDs could effectively catalyze the oxidation of colorless 3,3',5,5'-tetramethylbenzidine (TMB) to blue TMB oxidation products in the presence of hydrogen peroxide. While ACP was able to catalyze the hydrolysis of L-ascorbyl-2-phosphate trisodium salt (AAP) to produce ascorbic acid (AA), which inhibited the peroxidase-like activity of P-CDs, by combining P-CDs nanoenzymes and ACP-catalyzed hydrolysis the colorimetric method was established for ACP detection. The absorbance variation showed a good linear relationship with ACP concentration in the range of 0.4-4.0 mU/mL with a limit of detection at 0.12 mU/mL. In addition, the method was successfully applied to detect ACP in human serum samples with recoveries in the range of 98.7-101.6%. The work provides an effective strategy for regulating nanoenzymes activity and a low-cost detection technique for ACP.


Assuntos
Fosfatase Ácida , Carbono , Colorimetria , Limite de Detecção , Fósforo , Pontos Quânticos , Colorimetria/métodos , Carbono/química , Pontos Quânticos/química , Humanos , Fosfatase Ácida/análise , Fosfatase Ácida/sangue , Fosfatase Ácida/química , Fósforo/química , Benzidinas/química , Peroxidase/química , Peroxidase/metabolismo , Peróxido de Hidrogênio/química , Peróxido de Hidrogênio/análise , Oxirredução , Ácido Ascórbico/análise , Ácido Ascórbico/química , Ácido Ascórbico/sangue , Ácido Ascórbico/análogos & derivados
16.
Physiol Plant ; 176(2): e14247, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38499953

RESUMO

Oilseed rape (Brassica napus) is one of the most important oil crops in the world and shows sensitivity to low phosphorus (P) availability. In many soils, organic P (Po) is the main component of the soil P pool. Po must be mineralised to Pi through phosphatases, and then taken up by plants. However, the relationship between root-secreted acid phosphatases (APase) and root morphology traits, two important P-acquisition strategies in response to P deficiency, is unclear among B. napus genotypes. This study aimed to understand their relationship and how they affect P acquisition, which is crucial for the sustainable utilisation of agricultural P resources. This study showed significant genotypic variations in root-secreted APase activity per unit root fresh weight (SAP) and total root-secreted APase activity per plant (total SAP) among 350 B. napus genotypes. Seed yield was positively correlated with total SAP but not significantly correlated with SAP. Six root traits of 18 B. napus genotypes with contrasting root biomass were compared under normal Pi, low Pi and Po. Genotypes with longer total root length (TRL) reduced SAP, but those with shorter TRL increased SAP under P deficiency. Additionally, TRL was important in P-acquisition under three P treatments, and total SAP was also important in P-acquisition under Po treatment. In conclusion, trade-offs existed between the two P-acquisition strategies among B. napus genotypes under P-deficient conditions. Total SAP was an important root trait under Po conditions. These results might help to breed B. napus with greater P-acquisition ability under low P availability conditions.


Assuntos
Brassica napus , Fósforo , Brassica napus/genética , Fosfatase Ácida/genética , Fenótipo , Genótipo , Solo
17.
Sci Total Environ ; 926: 171916, 2024 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-38522536

RESUMO

Dredging wastewater discharge is a significant environmental concern for mariculture near mangrove ecosystems. However, little attention has been paid to its effects on the soil physical-chemical properties and enzyme activities in mangrove habitats. This study compared the soil physical-chemical properties and enzyme activities in the polluted area that received dredging wastewater from a shrimp pond with those in the control area without wastewater to explore the effects of wastewater discharge on the soil physical-chemical properties and enzyme activities. Variations in soil physical-chemical properties and enzyme activities across different tidal flat areas and depths were also examined. The polluted area exhibited lower soil salinity (10.47 ± 0.58 vs. 15.64 ± 0.54) and moisture content (41.85 ± 1.03 % vs. 45.81 ± 1.06 %) than the control area. Wastewater discharge increased soil enzyme activities, (acid phosphatase, protease, and catalase), resulting in higher inorganic nitrogen (13.20 ± 0.00 µg g-1 vs. 11.60 ± 0.03 µg g-1) but lower total nitrogen (0.93 ± 0.01 mg g-1 vs. 1.62 ± 0.11 mg g-1) in the contaminated zone. From the control to polluted area, there was an approximate increase of 0.43 and 0.83 mg g-1 in soil total phosphorus and soluble phosphate, driven by increased acid phosphatase. However, soil humus and organic matter decreased by 0.04 and 1.22 %, respectively, because of wastewater discharge. The impact of wastewater discharge on the soil physical-chemical properties and enzyme activities was most pronounced in the landward and surface soil layers (0-5 cm). The results showed that wastewater discharge altered soil physical-chemical properties and enzyme activities, accumulating soil bioavailable nutrients (inorganic nitrogen and soluble phosphate), but at the cost of reduced soil quality, especially organic matter, further adversely affecting the overall health of mangrove ecosystems. Prioritizing the management of wastewater discharged from mariculture adjacent to mangrove forests is crucial for mangrove conservation.


Assuntos
Ecossistema , Solo , Solo/química , Águas Residuárias , Lagoas , Áreas Alagadas , Fosfatos , Fosfatase Ácida , Nitrogênio/análise
18.
Plant Physiol Biochem ; 208: 108389, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38377886

RESUMO

Purple acid phosphatases (PAPs) are involved in activating the rhizosphere's organic phosphorus (P) and promoting P recycling during plant development, especially under the long-term P deficiency conditions in acid soil. However, the function of BnaPAPs in response to P deficiency stress in Brassica napus has rarely been explored. In this study, we found that the acid phosphatase activities (APA) of rapeseed shoot and root increased under P deficienct conditions. Genome-wide identification found that 82 PAP genes were unevenly distributed on 19 chromosomes in B. napus, which could be divided into eight subfamilies. The segmental duplication events were the main driving force for expansion during evolution, and the gene structures and conserved motifs of most members within the same subfamily were highly conservative. Moreover, the expression levels of 37 and 23 different expressed genes were induced by low P in leaf and root, respectively. BnaA09.PAP10a and BnaC09.PAP10a were identified as candidate genes via interaction networks. Significantly, both BnaPAP10a overexpression lines significantly increased root-related APA and total phosphate concentration under P deficiency and ATP supply conditions, thereby improving plant growth and root length. In summary, our results provided a valuable foundation for further study of BnaPAP functions.


Assuntos
Brassica napus , Brassica napus/metabolismo , Família Multigênica , Homeostase , Fosfatase Ácida/genética , Fosfatase Ácida/metabolismo , Fosfatos/metabolismo , Regulação da Expressão Gênica de Plantas , Filogenia , Proteínas de Plantas/metabolismo
19.
Physiol Plant ; 176(1): e14197, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38344855

RESUMO

Increased acid phosphatase (APase) activity is a prominent feature of tomato (Solanum lycopersicum) responses to inorganic phosphate (Pi) restriction. SlPHL1, a phosphate starvation response (PHR) transcription factor, has been identified as a positive regulator of low Pi (LP)-induced APase activity in tomato. However, the molecular mechanism underlying this regulation remains to be elucidated. Here, SlPHL1 was found to positively regulate the LP-induced expression of five potential purple acid phosphatase (PAP) genes, namely SlPAP7, SlPAP10b, SlPAP12, SlPAP15, and SlPAP17b. Furthermore, we provide evidence that SlPHL1 can stimulate transcription of these five genes by binding directly to the PHR1 binding sequence (P1BS) located on their promoters. The P1BS mutation notably weakened SlPHL1 binding to the promoters of SlPAP7, SlPAP12, and SlPAP17b but almost completely abolished SlPHL1 binding to the promoters of SlPAP10b and SlPAP15. As a result, the transcriptional activation of SlPHL1 on SlPAP10b and SlPAP15 was substantially diminished. In addition, not only did transient overexpression of either SlPAP10b or SlPAP15 in tobacco leaves increase APase activity, but overexpression of SlPAP15 in Arabidopsis and tomato also increased APase activity and promoted plant growth. Subsequently, two SPX proteins, SlSPX1 and SlSPX4, were shown to physically interact with SlPHL1. Moreover, SlSPX1 inhibited the transcriptional activation of SlPHL1 on SlPAP10b and SlPAP15 and negatively regulated the activity of APase. Taken together, these results demonstrate that SlPHL1-mediated LP signaling promotes APase activity by activating the transcription of SlPAP10b and SlPAP15, which may provide valuable insights into the mechanisms of tomato response to Pi-limited stress.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Solanum lycopersicum , Fosfatos , Solanum lycopersicum/genética , Fatores de Transcrição/metabolismo , Arabidopsis/metabolismo , Fosfatase Ácida/genética , Fosfatase Ácida/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Arabidopsis/metabolismo
20.
J Colloid Interface Sci ; 661: 1060-1069, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38335790

RESUMO

Herein, a novel dual-mode probe for organophosphorus pesticides (OPs) colorimetric and photothermal detection was developed based on manganese modified porphyrin metal-organic framework (PCN-224-Mn). PCN-224-Mn had excellent oxidase-like activity and oxidized colorless 3,3,5,5-tetramethylbenzidine (TMB) to blue-green oxidation state TMB (oxTMB), which exhibited high temperature under near-infrared irradiation. l-ascorbate-2-phosphate was hydrolyzed by acid phosphatase to produce ascorbic acid, which weakened colorimetric and photothermal signals by impacting oxTMB generation. The presence of OPs blocked the production of ascorbic acid by irreversibly inhibiting the activity of acid phosphatase, causing the restoration of chromogenic reaction and the increase of temperature. Under the optimal conditions, the probe showed a good linear response to OPs in the concentration range of 5 âˆ¼ 10000 ng/mL, using glyphosate as the analog. The detection limits of glyphosate in colorimetric mode and photothermal mode were 1.47 ng/mL and 2.00 ng/mL, respectively. The probe was successfully used for sensitive identification of OPs residues in tea, brown rice, and wheat flour. This work proposes a simple and reliable colorimetric/photothermal platform for OPs identification, which overcomes the problem that single-mode detection probes are susceptible to external factors, and has broad application potential in the field of food safety.


Assuntos
Benzidinas , Estruturas Metalorgânicas , Praguicidas , Compostos Organofosforados , Colorimetria , Farinha , Triticum , Ácido Ascórbico/química , Fosfatase Ácida
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