RESUMEN

Hydrogen polysulfides (H2Sn) have emerged as critical physiological mediators that are closely associated with hydrogen sulfide (H2S) signaling. H2Sn exhibit greater nucleophilicity than H2S while also having electrophilic characteristics, enabling unique activities such as protein S-persulfidation. Despite their physiological importance, mechanisms and reactivities of H2Sn remain inadequately explored due to their inherent instability in aqueous environments. Consequently, there is a need to develop biocompatible methods for controlled H2Sn generation to elucidate their behaviors in biological contexts. Herein, we present a dual enzyme system (containing glucose oxidase (GOx) and chloroperoxidase (CPO)) with thioglucose as the substrate to facilitate the controlled release of H2Sn. Fluorescence measurements with SSP4 and the trapping studies allowed us to confirm the production of H2Sn. Such a method may be useful in elucidating the reactivity of hydrogen polysulfides in biological systems as well as provide a potential delivery of H2Sn to target sites for biological applications.

Asunto(s)

Cloruro Peroxidasa , Glucosa Oxidasa , Sulfuros , Glucosa Oxidasa/metabolismo , Glucosa Oxidasa/química , Cloruro Peroxidasa/metabolismo , Cloruro Peroxidasa/química , Sulfuros/química , Sulfuros/metabolismo , Sulfuro de Hidrógeno/metabolismo , Sulfuro de Hidrógeno/química , Aspergillus niger/enzimología

RESUMEN

Halogenated phenazine meroterpenoids are a structurally unusual family of marine actinobacterial natural products that exhibit antibiotic, antibiofilm, and cytotoxic bioactivities. Despite a lack of established phenazine halogenation biochemistry, genomic analysis of Streptomyces sp. CNZ-289, a prolific lavanducyanin and C2-halogenated derivative producer, suggested the involvement of vanadium-dependent haloperoxidases. We subsequently discovered lavanducyanin halogenase (LvcH), characterized it in vitro as a regioselective vanadium-dependent chloroperoxidase, and applied it in late-stage chemoenzymatic synthesis.

Asunto(s)

Cloruro Peroxidasa , Halogenación , Vanadio , Cloruro Peroxidasa/metabolismo , Cloruro Peroxidasa/química , Vanadio/química , Estructura Molecular , Streptomyces/química , Estereoisomerismo , Fenazinas/química , Fenazinas/farmacología , Fenazinas/síntesis química , Antibacterianos/farmacología , Antibacterianos/química , Antibacterianos/síntesis química

RESUMEN

The initiation of this study relies on a targeted genome-mining approach to highlight the presence of a putative vanadium-dependent haloperoxidase-encoding gene in the deep-sea hydrothermal vent fungus Hortaea werneckii UBOCC-A-208029. To date, only three fungal vanadium-dependent haloperoxidases have been described, one from the terrestrial species Curvularia inaequalis, one from the fungal plant pathogen Botrytis cinerea, and one from a marine derived isolate identified as Alternaria didymospora. In this study, we describe a new vanadium chloroperoxidase from the black yeast H. werneckii, successfully cloned and overexpressed in a bacterial host, which possesses higher affinity for bromide (Km = 26 µM) than chloride (Km = 237 mM). The enzyme was biochemically characterized, and we have evaluated its potential for biocatalysis by determining its stability and tolerance in organic solvents. We also describe its potential three-dimensional structure by building a model using the AlphaFold 2 artificial intelligence tool. This model shows some conservation of the 3D structure of the active site compared to the vanadium chloroperoxidase from C. inaequalis but it also highlights some differences in the active site entrance and the volume of the active site pocket, underlining its originality.

Asunto(s)

Ascomicetos , Cloruro Peroxidasa , Exophiala , Respiraderos Hidrotermales , Cloruro Peroxidasa/genética , Cloruro Peroxidasa/química , Cloruro Peroxidasa/metabolismo , Exophiala/metabolismo , Saccharomyces cerevisiae/metabolismo , Vanadio/metabolismo , Inteligencia Artificial , Ascomicetos/genética

RESUMEN

Chloroperoxidase (CPO) is a haeme-thiolate enzyme able to catalyse the halogenation and oxidation of a wide range of organic substrates. In this work, the CPO-catalysed chlorination and bromination reaction of natural estrogens was characterised. Estradiol, estrone and equiline were efficiently converted to halogenated compounds in the presence of chloride or bromide and hydrogen peroxide. The catalytic efficiency of CPO in this reaction is similar to that measured for other aromatic substrates; as expected the bromination reaction proceeds more efficiently than the chlorination reaction. Three major products were detected for chlorination of estradiol; two of them were monohalogenated compounds while a third product was a dihalogenated compound at positions 2 and 4 of the aromatic ring A. Chlorinated compounds are not substrates for tyrosinase, suggesting that the halogenated form of estrogens is less susceptible to form o-quinones.

Asunto(s)

Cloruro Peroxidasa , Bromuros , Catálisis , Cloruro Peroxidasa/química , Cloruro Peroxidasa/metabolismo , Cloruros , Estradiol , Estrógenos , Estrona , Halogenación , Peróxido de Hidrógeno , Monofenol Monooxigenasa , Quinonas

RESUMEN

Recently, enzyme dynamic therapy (EDT) has drawn much attention as a new type of dynamic therapy. However, the selection of suitable nanocarriers to deliver chloroperoxidase (CPO) and enhancement of the level of hydrogen peroxide (H2 O2 ) in the tumor microenvironment (TME) are critical factors for improving the efficiency of EDT. In this study, a rapidly decomposing nanocomposite is designed using tetra-sulfide-bond-incorporating dendritic mesoporous organosilica (DMOS) as a nanocarrier, followed by loading CPO and sodium-hyaluronate-modified calcium peroxide nanoparticles (CaO2 -HA NPs). The nanocomposite can effectively generate singlet oxygen (1 O2 ) for tumor therapy without any exogenous stimulus via trimodal-enhanced EDT, including DMOS-induced depletion of glutathione (GSH), H2 O2 compensation from CaO2 -HA NPs in mildly acidic TME, and oxidative stress caused by overloading of Ca2+ . As tetra-sulfide bonds are sensitive to GSH, DMOS can generate hydrogen sulfide (H2 S) gas as a new kind of H2 S gas nanoreactor. Additionally, the overloading of Ca2+ can cause tumor calcification to accelerate in vivo tumor necrosis and promote computed tomography imaging efficacy. Therefore, a novel H2 S gas, EDT, and Ca2+ -interference combined therapy strategy is developed.

Asunto(s)

Cloruro Peroxidasa/química , Portadores de Fármacos/química , Peróxido de Hidrógeno/química , Sulfuro de Hidrógeno/química , Nanocompuestos/química , Neoplasias/terapia , Animales , Cloruro Peroxidasa/metabolismo , Liberación de Fármacos , Activación Enzimática , Femenino , Glutatión/química , Humanos , Ácido Hialurónico/química , Peróxido de Hidrógeno/farmacología , Ratones Endogámicos BALB C , Estrés Oxidativo , Peróxidos/química , Porosidad , Dióxido de Silicio/química , Oxígeno Singlete/química , Sulfuros/química , Propiedades de Superficie , Microambiente Tumoral

RESUMEN

Halloysite nanotube (HNT) is a natural bio-compatible and stable nanomaterial available in abundance at low-cost. In this work, HNT was modified by two strategies to make it suitable for supporting immobilization of chloroperoxidase (CPO). Firstly, Fe3O4 nanoparticles were deposited on HNT, so magnetic separation can be used instead of centrifugation. Then, the magnetic HNT was modified by 3-aminopropyltriethoxysilane (APTES), which can provide amine group on surface of HNT and meanwhile inhibit the agglomeration of magnetic HNT. Then, HNT-Fe3O4 -APTES was linked with branched polyethyleneimine (PEI) to provide more amino for binding with enzyme. The so-prepared CPO@HNT-Fe3O4-APTES-PEI showed enhanced enzyme loading, reusability, improved thermal stability and tolerance to organic solvents than free CPO. For example, after 10 repeated uses, CPO@HNT- Fe3O4-APTES-PEI can maintain 92.20% of its original activity compared with 65.12% of activity of CPO@HNT-APTES-PEI and 45.69% of activity of CPO@HNT. The kinetic parameters indicated the affinity and specificity of immobilized enzyme to substrate was increased. CPO@HNT-Fe3O4-APTES-PEI was very efficient when it was applied in the degradation of pesticides mesotrione in wastewater. The degradation efficiency can reach 90% within 20 min at range of 5-40 µmol·L-1. These results ensure the potential practical application of this bio-materials in wastewater treatment.

Asunto(s)

Ascomicetos/enzimología , Cloruro Peroxidasa/química , Arcilla/química , Enzimas Inmovilizadas/química , Óxido Ferrosoférrico/química , Proteínas Fúngicas/química , Nanotubos/química , Plaguicidas/química , Aguas Residuales/química

RESUMEN

Over 5000 halogenated natural products have been reported so far, many of these arising from the marine environment. The introduction of a halogen into a molecule can significantly impact its bioavailability and bioactivity. More recently enzymatic halogenation has been used to enable late stage functionalisation through site-selective halogenation and cross-coupling. Halogenases are becoming increasingly valued tools. This review outlines the various classes of halogenases that have been discovered, and examines these from both a structural and a mechanistic perspective, reflecting upon the many recent advances in halogenase discovery.

Asunto(s)

Cloruro Peroxidasa , Halogenación , Cloruro Peroxidasa/química , Cloruro Peroxidasa/metabolismo , Especificidad por Sustrato

RESUMEN

The inefficiency of cyanide/HCN (CN) binding with heme proteins (under physiological regimes) is demonstrated with an assessment of thermodynamics, kinetics, and inhibition constants. The acute onset of toxicity and CN's mg/Kg LD50 (µM lethal concentration) suggests that the classical hemeFe binding-based inhibition rationale is untenable to account for the toxicity of CN. In vitro mechanistic probing of CN-mediated inhibition of hemeFe reductionist systems was explored as a murburn model for mitochondrial oxidative phosphorylation (mOxPhos). The effect of CN in haloperoxidase catalyzed chlorine moiety transfer to small organics was considered as an analogous probe for phosphate group transfer in mOxPhos. Similarly, inclusion of CN in peroxidase-catalase mediated one-electron oxidation of small organics was used to explore electron transfer outcomes in mOxPhos, leading to water formation. The free energy correlations from a Hammett study and IC50/Hill slopes analyses and comparison with ligands ( CO/ H 2 S/ N 3 - ) $\left( {\text{CO}}/{{{{\text{H}}_{2}}\text{S}}/{\text{N}_{3}^{\text{-}}}\;}\; \right)$ provide insights into the involvement of diffusible radicals and proton-equilibriums, explaining analogous outcomes in mOxPhos chemistry. Further, we demonstrate that superoxide (diffusible reactive oxygen species, DROS) enables in vitro ATP synthesis from ADP+phosphate, and show that this reaction is inhibited by CN. Therefore, practically instantaneous CN ion-radical interactions with DROS in matrix catalytically disrupt mOxPhos, explaining the acute lethal effect of CN.

Asunto(s)

Cianuros/toxicidad , Hemo/química , Hemoproteínas/antagonistas & inhibidores , Hemoglobinas/antagonistas & inhibidores , Mitocondrias/efectos de los fármacos , Adenosina Trifosfato/biosíntesis , Adenosina Trifosfato/química , Adenosina Trifosfato/metabolismo , Sitios de Unión , Catalasa/metabolismo , Catálisis , Respiración de la Célula/efectos de los fármacos , Respiración de la Célula/fisiología , Cloruro Peroxidasa/química , Cianuros/química , Complejo IV de Transporte de Electrones/química , Complejo IV de Transporte de Electrones/metabolismo , Hemo/antagonistas & inhibidores , Hemo/metabolismo , Hemoproteínas/química , Hemoproteínas/metabolismo , Hemoglobinas/química , Peroxidasa de Rábano Silvestre/metabolismo , Hidróxidos/química , Cinética , Ligandos , Mitocondrias/química , Mitocondrias/enzimología , Mitocondrias/metabolismo , Oxidación-Reducción , Especies Reactivas de Oxígeno/metabolismo , Estirenos/química , Estirenos/farmacología , Superóxidos/química , Termodinámica

RESUMEN

Neutrophils can responsively release reactive oxygen species (ROS) to actively combat infections by exogenous stimulus and cascade enzyme catalyzed bio-oxidation. A supramolecular nanogel is now used as an artificial neutrophil by enzymatic interfacial self-assembly of peptides (Fmoc-Tyr(H2 PO3 )-OH) with magnetic nanoparticles (MNPs) and electrostatic loading of chloroperoxidase (CPO). The MNPs within the nanogel can elevate H2 O2 levels in cancer cells under programmed alternating magnetic field (AMF) similar to the neutrophil activator, and the loaded CPO within protective peptides nanolayer converts the H2 O2 into singlet oxygen (1 O2 ) in a sustained manner for neutrophil-inspired tumor therapy. As a proof of concept study, both the H2 O2 and 1 O2 in cancer cells increase stepwise under a programmed alternating magnetic field. An active enzyme dynamic therapy by magnetically stimulated oxygen stress and sustained enzyme bio-oxidation is thus shown with studies on both cells and animals.

Asunto(s)

Cloruro Peroxidasa/metabolismo , Nanopartículas de Magnetita/química , Nanogeles/química , Animales , Antineoplásicos/química , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cloruro Peroxidasa/química , Humanos , Peróxido de Hidrógeno/química , Peróxido de Hidrógeno/metabolismo , Campos Magnéticos , Ratones , Nanogeles/uso terapéutico , Nanogeles/toxicidad , Neoplasias/tratamiento farmacológico , Neoplasias/mortalidad , Neoplasias/patología , Neutrófilos/química , Neutrófilos/inmunología , Tamaño de la Partícula , Péptidos/química , Oxígeno Singlete/química , Oxígeno Singlete/metabolismo , Electricidad Estática , Tasa de Supervivencia , Trasplante Heterólogo

RESUMEN

BACKGROUND: Tetrabromobisphenol (TBBPA), a flame retardant compound, is considered a ubiquitous pollutant, with potential impact on the environment and human health. Several technologies have been applied to accelerate its degradation and minimize environmental impacts. Due to its aromaticity character, peroxidase enzymes may be employed to carry out its transformation in mild conditions. Therefore, the purpose of this work was to determine the capacity of the enzyme chloroperoxidase (CPO) to oxidize TBBPA in several water samples. METHODS: The oxidation capacity of CPO was evaluated in catalytic conditions using water samples from surface and groundwater, as well as effluents from wastewater treatment plants. The biocatalytic performance of CPO was improved due to its immobilization on nanofibers composed of polyvinyl alcohol and chitosan (PVA/chitosan). RESULTS: Free and immobilized CPO were able to transform more than 80% in short reaction times (60 min); producing more biodegradable and less toxic products. Particularly, the immobilized enzyme was catalytically active in a wider range of pH than the free enzyme with the possibility of reusing it up to five times. CONCLUSIONS: The biocatalytic oxidation of TBBPA under environmental conditions is highly efficient, even in complex media such as treated effluents of wastewater treatment plants.

Asunto(s)

Cloruro Peroxidasa/química , Enzimas Inmovilizadas/química , Retardadores de Llama , Nanofibras/química , Bifenilos Polibrominados/química , Contaminantes Ambientales/química , Oxidación-Reducción , Aguas Residuales

RESUMEN

Chloroperoxidase (CPO) is a hybrid of two different families of enzymes, peroxidases and P450s. However, it is poorly understood on CPO's multiple catalytic functions. Herein, phenol was selected as a model substrate to investigate the multiple catalytic roles of CPO. Results showed that phenol was readily transformed into a variety of brominated organic compounds (BOCs) via the CPO-mediated oxidative process. A total of 16 BOCs were identified using gas and liquid chromatography coupled with mass spectrometry. Possible reaction pathways could be attributable to four CPO-mediated processes, including bromination, radical coupling, intramolecular cyclization and debromination. Higher bromide concentrations and lower pH conditions both facilitated the formation of brominated products. While a higher bromination capacity was observed in pH 3.0 solutions, CPO-mediated radical couplings were more favorable at pH 5.0 and 6.0. Although CPO might catalyze chlorination when chloride and bromide coexisted in the solution, BOCs were the dominant products of CPO-mediated phenol oxidation. Results of this study suggest that various catalytic roles of CPO may contribute to the biotic formation of BOCs in the natural environment.

Asunto(s)

Bromuros/química , Cloruro Peroxidasa/química , Hidrocarburos Bromados/química , Fenol/química , Catálisis , Cromatografía Liquida , Halogenación , Estructura Molecular , Oxidación-Reducción , Espectrometría de Masas en Tándem

RESUMEN

A multitude of industrial processes are catalyzed by two or more enzymes working together in a cascade way. However, designing efficient enzymatic cascade reactions is still a challenge. In this work, a TiO2 thin film with mesoporous pores was prepared and used as carrier for co-immobilization of chloroperoxidase (CPO) and glucose peroxidase (GOx). By adjusting the dosage of hexadecyltrimethylammonium bromide (CTAB) and the ratio of the two enzymes, CPO and GOx were well distributed and positional orientated to their own appropriate pores to form an ordered "occupation" based on a "feet in right shoes" effect. Moreover, when the pore size was controlled around 12 nm, the enzymes aggregation was inhibited so as to avoid the decrease of activity of enzyme; The catalytic performance of TiO2-GOx and CPO composites was evaluated by the application of decolorization of Orange G dye in a cascaded manner. The oxidant H2O2 needed by CPO is generated in situ through glucose oxidation by GOx. Upon co-immobilization of CPO and GOx on the same carrier, a large increase in the initial catalytic efficiency was detected when compared to an equimolar mixture of the free enzymes, which was four times greater. Moreover, the affinity of the enzyme toward substrate binding was improved according to the kinetic assay. The thermal stability of TiO2-GOx and CPO composites were greatly improved than free enzymes. The TiO2-GOx and CPO composites can be easily separated from the reaction media which facilitate its recycle use.

Asunto(s)

Compuestos Azo/química , Cloruro Peroxidasa/química , Enzimas Inmovilizadas/química , Membranas Artificiales , Peroxidasas/química , Titanio/química , Oxidación-Reducción , Porosidad

RESUMEN

As the first line of innate immune cells to migrate towards tumour tissue, neutrophils, can immediately kill abnormal cells and activate long-term specific adaptive immune responses. Therefore, the enzymes mediated elevation of reactive oxygen species (ROS) bioinspired by neutrophils can be a promising strategy in cancer immunotherapy. Here, we design a core-shell supramolecular hybrid nanogel via the surface phosphatase triggered self-assembly of oligopeptides around iron oxide nanoparticles to simulate productive neutrophil lysosomes. The cascade reaction of superoxide dismutase (SOD) and chloroperoxidase (CPO) within the bioinspired nanogel can convert ROS in tumour tissue to hypochlorous acid (HOCl) and the subsequent singlet oxygen (1O2) species. Studies on both cells and animals demonstrate successful 1O2-mediated cell/tumour proliferation inhibition, making this enzyme therapy capable for treating tumours without external energy activation.

Asunto(s)

Carcinoma Hepatocelular/terapia , Hidrogeles/uso terapéutico , Neoplasias Hepáticas/terapia , Nanomedicina/métodos , Oxígeno Singlete/metabolismo , Animales , Carcinoma Hepatocelular/inmunología , Carcinoma Hepatocelular/mortalidad , Carcinoma Hepatocelular/patología , Cloruro Peroxidasa/química , Cloruro Peroxidasa/metabolismo , Femenino , Óxido Ferrosoférrico/química , Células Hep G2 , Humanos , Hidrogeles/química , Ácido Hipocloroso/metabolismo , Hígado/patología , Neoplasias Hepáticas/mortalidad , Neoplasias Hepáticas/patología , Lisosomas/inmunología , Lisosomas/metabolismo , Masculino , Nanopartículas del Metal/química , Nanopartículas del Metal/uso terapéutico , Nanopartículas del Metal/ultraestructura , Ratones , Ratones Endogámicos NOD , Ratones Desnudos , Ratones SCID , Microscopía Electrónica de Rastreo , Neutrófilos/inmunología , Neutrófilos/metabolismo , Superóxido Dismutasa/química , Superóxido Dismutasa/metabolismo , Resultado del Tratamiento , Ensayos Antitumor por Modelo de Xenoinjerto

RESUMEN

Vanadium-dependent haloperoxidases are a class of enzymes that catalyze oxidation reactions with halides to form halogenated organic products and water. These enzymes include chloroperoxidase and bromoperoxidase, which have very different protein sequences and sizes, but regardless the coordination environment of the active sites is surprisingly constant. In this manuscript, the comparison of the coordination chemistry of V-containing-haloperoxidases of the trigonal bipyramidal geometry was done by data mining. The catalytic cycle imposes changes in the coordination geometry of the vanadium to accommodate the peroxidovanadium(V) intermediate in an environment we describe as a distorted square pyramidal geometry. During the catalytic cycle, this intermediate converts to a trigonal bipyramidal intermediate before losing the halogen and forming a tetrahedral vanadium-protein intermediate. Importantly, the catalysis is facilitated by a proton-relay system supplied by the second sphere coordination environment and the changes in the coordination environment of the vanadium(V) making this process unique among protein catalyzed processes. The analysis of the coordination chemistry shows that the active site is very tightly regulated with only minor changes in the coordination geometry. The coordination geometry in the protein structures deviates from that found for both small molecules crystalized in the absence of protein and the reported functional small molecule model compounds. At this time there are no examples reported of a structurally similar small molecule with the geometry observed for the peroxidovanadium(V) in the active site of the vanadium-containing haloperoxidases.

Asunto(s)

Cloruro Peroxidasa , Halógenos , Peroxidasas , Vanadio , Animales , Catálisis , Dominio Catalítico , Cloruro Peroxidasa/química , Cloruro Peroxidasa/metabolismo , Halógenos/química , Halógenos/metabolismo , Humanos , Oxidación-Reducción , Peroxidasas/química , Peroxidasas/metabolismo , Vanadio/química , Vanadio/metabolismo

RESUMEN

This paper examines the influence of the proximal pockets of cytochrome P450CAM and chloroperoxidase (CPO) on the relative favorability of catalytic epoxidation and allylic hydroxylation of olefins, a type of alkene oxidation selectivity. The study employs quantum mechanical models of the active site to isolate the proximal pocket's influence on the barrier for the selectivity-determining step for each reaction, using cyclohexene and cis-ß-methylstyrene as substrates. The proximal pocket is found to preference epoxidation by 2-5 kcal/mol, the largest value being for CPO, converting the active heme-thiolate moiety from being intrinsically hydroxylation-selective to being intrinsically epoxidation-selective. This theoretical study, the first to correctly predict these enzymes' preference for epoxidation of allylic substrates, strongly suggests that the proximal pocket is the key determinant of alkene oxidation selectivity. The selectivity for epoxidation can be rationalized in terms of the proximal pocket's modulation of the thiolate's electron "push" and consequent influence on the heme redox potential and the basicity of the trans ligand.

Asunto(s)

Alquenos/química , Cloruro Peroxidasa/química , Sistema Enzimático del Citocromo P-450/química , Dominio Catalítico , Cloruro Peroxidasa/metabolismo , Sistema Enzimático del Citocromo P-450/metabolismo , Enlace de Hidrógeno , Simulación de Dinámica Molecular , Oxidación-Reducción , Teoría Cuántica , Especificidad por Sustrato

RESUMEN

Vanadium-dependent haloperoxidases in seaweeds, cyanobacteria, fungi, and possibly phytoplankton play an important role in the release of halogenated volatile compounds in the environment. These halocarbons have effects on atmospheric chemistry since they cause ozone depletion. In this chapter, a survey is given of the different sources of these enzymes, some of their properties, the various methods to isolate them, and the bottlenecks in purification. The assays to detect and quantify haloperoxidase activity are described as well as their kinetic properties. Several practical tips and pitfalls are given which have not yet been published explicitly. Recent developments in research on structure and function of these enzymes are reviewed. Finally, the application of vanadium-dependent haloperoxidases in the biosynthesis of brominated and other compounds is discussed.

Asunto(s)

Organismos Acuáticos/metabolismo , Cloruro Peroxidasa/aislamiento & purificación , Pruebas de Enzimas/métodos , Yoduro Peroxidasa/aislamiento & purificación , Peroxidasas/aislamiento & purificación , Organismos Acuáticos/química , Cloruro Peroxidasa/química , Cloruro Peroxidasa/metabolismo , Tecnología Química Verde/métodos , Yoduro Peroxidasa/química , Yoduro Peroxidasa/metabolismo , Peroxidasas/química , Peroxidasas/metabolismo

RESUMEN

Polyhalogenated carbazoles (PHCs) are a class of emerging organic contaminants that have received increasing concern due to their widespread distribution and dioxin-like toxicity. Although previous studies have suggested possible natural sources of PHCs in the environment, the formation pathways are poorly understood. Here we explored the production of PHCs from halogenation of carbazole in the presence of Br- and/or Cl- under the catalysis of chloroperoxidase (CPO) isolated from the marine fungus Caldariomyces fumago. Overall, a total of 25 congeners including mono-to tetra-substituted chlorinated, brominated, and mixed halogenated carbazoles (with substitution patterns of -BrCl, -BrCl2, -BrCl3, -Br2Cl, -Br2Cl2, and -Br3Cl) were produced from the reactions under various conditions. The PHC product profiles were apparently dependent on the halide concentrations. In the CPO-mediated chlorination of carbazole, 3-mono- and 3,6-dichlorocarbazoles predominated in the formation products. In addition to the less abundant mixed halogenated carbazoles (-Br2Cl), 1,3,6-tri- and 1,3,6,8-tetrabromocarbazoles were the dominant products in reactions containing both Br- and Cl-. The CPO-catalyzed halogenation of carbazole could take place in pH 3-7, but the formation products were pH dependent. Results of this study suggest that CPO-catalyzed halogenation of carbazole may play an important role in the natural formation of PHCs.

Asunto(s)

Carbazoles/química , Cloruro Peroxidasa/química , Modelos Químicos , Carbazoles/toxicidad , Catálisis , Dioxinas , Ambiente , Halogenación , Bifenilos Policlorados , Dibenzodioxinas Policloradas

RESUMEN

A novel ZnO nanowire/macroporous SiO2 composite was used as a support to immobilize chloroperoxidase (CPO) by in situ cross-linking method. An anionic bi-epoxy compound was synthesized and used as a long-chained anionic cross-linker, and it was adsorbed on the surface of ZnO nanowires through static interaction before reaction with CPO, creating a new approach to change the structure, property, and catalytic performance of the produced cross-linking enzyme aggregates (CLEAs) of CPO. The immobilized CPO showed high activity in the decolorization of three azo dyes. The effect of various conditions such as the loading amount of CPO, solution pH, temperature, and dye concentration was optimized on the decolorization. Under optimized conditions, the decolorization percentage of Acid Blue 113, Direct Black 38, and Acid Black 10 BX reached as high as 95.4, 92.3, and 89.1%, respectively. The immobilized CPO exhibited much better thermostability and resistance to pH inactivation than free CPO. The storage stability and reusability were greatly improved through the immobilization. It was found from the decolorization of Acid Blue 113 that 83.6% of initial activity retained after incubation at 4 °C for 60 days and that 80.9% of decolorization efficiency retained after 12 cycles of reuses.

Asunto(s)

Compuestos Azo/química , Cloruro Peroxidasa/química , Enzimas Inmovilizadas/química , Nanocompuestos/química , Dióxido de Silicio/química , Óxido de Zinc/química

RESUMEN

It is well established that the majority of chlorinated organic substances found in the terrestrial environment are produced naturally. The presence of these compounds in soils is not limited to a single ecosystem. Natural chlorination is also a widespread phenomenon in grasslands and agricultural soils typical for unforested areas. These chlorinated compounds are formed from chlorination of natural organic matter consisting of very complex chemical structures, such as lignin. Chlorination of several lignin model compounds results in the intermediate formation of trichloroacetyl-containing compounds, which are also found in soils. These decay, in general, through a haloform-type reaction mechanism to CHCl3 . Upon release into the atmosphere, CHCl3 will produce chlorine radicals through photolysis, which will, in turn, lead to natural depletion of ozone. There is evidence that fungal chloroperoxidases able to produce HOCl are involved in the chlorination of natural organic matter. The objective of this review is to clarify the role and source of the various chloroperoxidases involved in the natural formation of CHCl3 .

Asunto(s)

Cloruro Peroxidasa/metabolismo , Compuestos de Cloro/síntesis química , Cloroformo/síntesis química , Ambiente , Cloruro Peroxidasa/química , Compuestos de Cloro/química , Compuestos de Cloro/metabolismo , Cloroformo/química , Cloroformo/metabolismo , Hongos/química , Hongos/enzimología , Fotólisis , Suelo/química

RESUMEN

Chloroperoxidase (CPO), a heme-thiolate protein, from Caldariomyces fumago catalyzes a plethora of reactions including halogenation, dismutation, epoxidation, and oxidation. Although all CPO-catalyzed reactions go through a common intermediate, compound I, different mechanisms are followed in subsequent transformations. To understand the mechanism of CPO-catalyzed halide-dependent degradation of orange G, the role of halide and pH was systematically investigated. It is revealed that formation and protonation of compound X, a long-sought after hypochlorite heme adduct intermediate existed during CPO-catalyzed halide-dependent reactions, significantly lowers the reaction barrier and increases the efficiency of CPO-catalyzed orange G degradation. The extremely acidic optimal reaction pH suggests the protonation of a residue, presumably, Glu 183 in CPO catalysis. Halide dependent studies showed that Kcat is higher in the presence of Br(-) than in the presence of Cl(-). The degradation products of orange G indicate the cleavage at a single position of orange G, demonstrating a high regioselectivity of CPO-catalyzed degradation. Based on our kinetic, NMR and QM/MM studies, the mechanism of CPO-catalyzed orange G degradation was proposed.

Asunto(s)

Ascomicetos/enzimología , Compuestos Azo/química , Cloruro Peroxidasa/química , Proteínas Fúngicas/química , Modelos Químicos , Catálisis , Cinética , Resonancia Magnética Nuclear Biomolecular/métodos