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1.
Chemosphere ; 286(Pt 2): 131687, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34343919

RESUMO

Jute sticks obtained after the extraction of jute fiber are an excellent biomass feedstock with a significant amount of carbohydrates that makes it an attractive resource for sustainable energy generation. However, the high lignin content in the jute stick hinders the cellulosic component of the cell wall from enzymatic hydrolysis.This work demonstrates the lignin degradation of jute stick biomass by Trametes maxima laccase in the presence of mediator Hydroxybenzotriazole and improvement in its subsequent saccharification. Lignin component in jute stick is reduced by 21.8% in a single reaction treatment with laccase-mediator compared to the untreated jute stick sample used as control. The yield of fermentable sugar is increased by 19.5% that verifies enhanced saccharification after lignin removal. Delignification of jute stick was corroborated through different analytical techniques. The Pyrolysis gas chromatography/mass spectrometry results further confirms abundance of S lignin unit in the jute stick compared to the H and G unit and modification in lignin polymer as a change in the syringyl-to-guaiacyl ratio. Hence, this work demonstrates that jute stick can be effectively delignified using biocatalyst-mediator system and utilized as biomass source, thus contributing in circular bio-economy through waste valorization.


Assuntos
Lacase , Açúcares , Biomassa , Carboidratos , Hidrólise , Lignina , Polyporaceae , Trametes
2.
Bioresour Technol ; 343: 126114, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34648963

RESUMO

The success of establishing bioeconomies replacing current economies based on fossil resources largely depends on our ability to degrade recalcitrant lignocellulosic biomass. This study explores the potential of employing various enzymes acting synergistically on previously pretreated agricultural side streams (corn bran, oat hull, soluble and insoluble oat bran). Degrees of synergy (oligosaccharide yield obtained with the enzyme combination divided by the sum of yields obtained with individual enzymes) of up to 88 were obtained. Combinations of a ferulic acid esterase and xylanases resulted in synergy on all substrates, while a laccase and xylanases only acted synergistically on the more recalcitrant substrates. Synergy between different xylanases (glycoside hydrolase (GH) families 5 and 11) was observed particularly on oat hulls, producing a yield of 57%. The synergistic ability of the enzymes was found to be partly due to the increased enzyme stability when in combination with the substrates.


Assuntos
Lacase , Lignina , Hidrolases de Éster Carboxílico , Endo-1,4-beta-Xilanases , Humanos , Oligossacarídeos
3.
Food Chem ; 372: 131332, 2022 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-34818742

RESUMO

Poor solubility of proteins negatively affects their functional properties and greatly limits their application. Enzymatic cross-linking with polysaccharides can improve solubility and functional properties of proteins. The enzymes used include transglutaminase, laccase and peroxidase. Therefore, this work introduces the cross-linking mechanisms of these enzymes and the characterization techniques, the improved properties and the potential applications of the enzymatically-synthesized protein-polysaccharide conjugate. Transglutaminase catalyzes the formation of a new peptide bond and thus works on amino-containing polysaccharides to conjugate with proteins. However, laccase and peroxidase catalyze oxidation of various compounds with phenol and aniline structures. Therefore, these two enzymes can catalyze the conjugate reaction between proteins and feruloylated polysaccharides which are widely distributed in cereal bran. Compared with the unmodified protein, the enzymatically-synthesized protein-polysaccharide conjugate usually has higher solubility and better functional properties. Thus, it is inferred that enzymatic conjugation with polysaccharide molecules can extend the application of proteins.


Assuntos
Polissacarídeos , Proteínas , Lacase , Oxirredução , Solubilidade
4.
Sci Total Environ ; 802: 150005, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34525729

RESUMO

Humification processes of phenolic pollutants may play a profound role in environment purification and plant growth. However, little literature is performed to explore exoenzyme-driven humification to polymerize 17ß-estradiol (E2) and humic constituents (HCs), and the effects of their polymeric precipitates on plant growth are usually overlooked. Herein, E2 conversion and radish (Raphanus sativus L.) growth were systematically investigated under humification mediated by extracellular laccase (EL) of Trametes versicolor. Results disclosed that EL-assisted humification achieved a wonderful E2 conversion efficiency (>99%) within 2-h, but the presence of HCs such as humic acid (HA), vanillic acid (VA), and ferulic acid (FA) impeded E2 elimination significantly. Compared with HC-free, the kinetics constants declined by 2.84-, 5.72-, and 5.22-fold with HA, VA, and FA present, respectively. Intriguingly, three close-knit self/cross-linked precipitates (i.e., E2-HA, E2-VA, and E2-FA hybrid precipitates) in dark gray, dark brown, and deep yellow were created after a continuous humification by phenolic radical-initiated polymerization mechanisms. The formation of these humified precipitates was extremely effective on circumventing phytotoxicity caused by monomeric E2, VA, or FA. Furthermore, they acted as humic-like organic fertilizers, accelerating seed germination, root elongation, and enhancing NaCl-tolerance of radish through the combination of oxygen-contained functional components and auxin structural analogues with unstable and stubborn carbon skeletons. This is the first study reporting the pollution purification and plant growth promotion in EL-activated humification. Our findings frame valuable perspectives regarding the natural detoxification and carbon sequestration of phenolic pollutants and the application of their polymeric precipitates in global crop production.


Assuntos
Poluentes Ambientais , Lacase , Substâncias Húmicas/análise , Polyporaceae , Trametes
5.
PLoS One ; 16(10): e0259205, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34705877

RESUMO

Phenolic compounds are chemical precursor building blocks of soil organic matter. Their occurrence can be inhibitory to certain enzymes present in soil, thereby influencing the rate of decomposition of soil organic matter. Microbe-derived phenoloxidases (laccases) are extracellular enzymes capable of degrading recalcitrant polyphenolic compounds. In this study, our aim was to investigate the relationships between phenoloxidase enzyme activity, organic carbon content and microbial abundance in the context of long-term anthropogenically amended soils. To achieve this, we used a series of complementary biochemical analytical methods including gas chromatography, enzyme assays and solid-state Carbon-13 Cross Polarisation Magic-Angle Spinning Nuclear Magnetic Resonance Spectroscopy (13C CPMAS NMR). Using several anthrosols found in St Andrews (Scotland, UK) that had been subjected to intense anthropogenic modification since the medieval period (11th century AD) to present-day, we were able to scope the impact of past waste disposal on soils. The long-term anthropogenic impact led to organic matter-rich soils. Overall, phenoloxidase activity increased by up to 2-fold with soil depth (up to 100 cm) and was inversely correlated with microbial biomass. Solid-state 13C NMR characterisation of carbon species revealed that the observed decline in soil organic matter with depth corresponded to decreases in the labile organic carbon fractions as evidenced by changes in the O/N-alkyl C region of the spectra. The increase in phenoloxidase activity with depth would appear to be a compensatory mechanism for the reduced quantities of organic carbon and lower overall nutrient environment in subsoils. By enzymatically targeting phenolic compounds, microbes can better utilise recalcitrant carbon when other labile soil carbon sources become limited, thereby maintaining metabolic processes.


Assuntos
Proteínas de Bactérias/metabolismo , Monofenol Mono-Oxigenase/metabolismo , Polifenóis/análise , Microbiologia do Solo , Solo/química , Carbono/análise , Carbono/metabolismo , Lacase/metabolismo , Microbiota , Polifenóis/metabolismo
6.
Molecules ; 26(20)2021 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-34684746

RESUMO

(+)-Catechin-laccase oxidation dimeric standards were hemi-synthesized using laccase from Trametes versicolor in a water-ethanol solution at pH 3.6. Eight fractions corresponding to eight potential oxidation dimeric products were detected. The fractions profiles were compared with profiles obtained with two other oxidoreductases: polyphenoloxidase extracted from grapes and laccase from Botrytis cinerea. The profiles were very similar, although some minor differences suggested possible dissimilarities in the reactivity of these enzymes. Five fractions were then isolated and analyzed by 1D and 2D NMR spectroscopy. The addition of traces of cadmium nitrate in the samples solubilized in acetone-d6 led to fully resolved NMR signals of phenolic protons, allowing the unambiguous structural determination of six reaction products, one of the fractions containing two enantiomers. These products can further be used as oxidation markers to investigate their presence and evolution in wine during winemaking and wine ageing.


Assuntos
Catequina/química , Lacase/química , Vitis/química , Biomarcadores , Botrytis/enzimologia , Botrytis/metabolismo , Ressonância Magnética Nuclear Biomolecular/métodos , Oxirredução , Fenóis , Polyporaceae/enzimologia , Relação Estrutura-Atividade , Trametes/enzimologia , Vitis/metabolismo , Vinho/análise
7.
PLoS One ; 16(10): e0258043, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34653213

RESUMO

Fungal diseases in agronomically important plants such as grapevines result in significantly reduced production, pecuniary losses, and increased use of environmentally damaging chemicals. Beside the well-known diseases, there is an increased interest in wood-colonizing fungal pathogens that infect the woody tissues of grapevines. In 2015, a traditional isolation method was performed on grapevine trunks showing symptoms of trunk diseases in Hungary. One isolate (T15142) was identified as Kalmusia longispora (formerly Dendrothyrium longisporum) according to morphological and phylogenetic analyses. To evaluate the pathogenicity of this fungus on grapevines, artificial infections were carried out under greenhouse and field conditions, including the CBS 824.84 and ex-type CBS 582.83 strains. All isolates could be re-isolated from inoculated plants; however, varying virulence was observed among them in terms of the vascular necrosis caused. The incidence and severity of this symptom seemed to be congruent with the laccase-producing capabilities of the isolates. This is the first report on the ability of Kalmusia longispora to cause symptoms on grapevines, and on its possible dependence on laccase secretion.


Assuntos
Ascomicetos , Lacase/metabolismo , Doenças das Plantas/microbiologia , Vitis/microbiologia , Ascomicetos/isolamento & purificação , Ascomicetos/metabolismo , Ascomicetos/patogenicidade , Virulência
8.
J Agric Food Chem ; 69(44): 13113-13124, 2021 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-34696587

RESUMO

The small subunit, ssPOXA3a/b, and the large subunit, POXA3, are indispensable components of typical heterodimeric laccase (Lacc2) in white rot fungi. However, the enzymatic and biological functions of ssPOXA3a/b remain unclear. The present study revealed that neither ssPOXA3a nor ssPOXA3b per se has a catalytic ability, whereas their combination with POXA3 (and especially ssPOXA3b) enhances the activity, thermostability, and pH stability of POXA3. In Pleurotus eryngii var. ferulae, there was no regulatory relationship between ssPOXA3a/b and POXA3 at the transcriptional level. However, sspoxa3a/b overexpression had a negative feedback effect on lacc6 transcription. By contrast, poxa3 transcripts had no effect on any other laccase isoenzyme. Overexpression of sspoxa3a/b resulted in small fungal pellets, thin mycelial walls, and facilitated laccase secretion. However, poxa3 overexpression had no influence on pellet morphology. Collectively, this work elucidated the functions of ssPOXA3a/b and laid an empirical foundation for the development of high-yield laccase.


Assuntos
Ferula , Pleurotus , Lacase/genética , Pleurotus/genética
9.
Biomacromolecules ; 22(11): 4501-4509, 2021 11 08.
Artigo em Inglês | MEDLINE | ID: mdl-34601873

RESUMO

Biografting is a promising and ecofriendly approach to meet various application requirements of products. Herein, a popular green enzyme, laccase, was adopted to graft a hydrophobic phenolic compound (lauryl gallate, LG) onto chitosan (CTS). The resultant chitosan derivate (Lac/LG-CTS) was systematically analyzed by Fourier transform infrared (FTIR), grafting efficiency, scanning probe microscopy (SPM), and X-ray diffraction (XRD). This grafting technique produced a multifunctional chitosan copolymer with remarkably enhanced antioxidant property, hydrophobicity, and moisture barrier property. Furthermore, the swelling capacity and acid solubility of the copolymer film decreased significantly, although the tensile strength and elongation were slightly weakened as compared to those of native chitosan. These results suggest that the Lac/LG-CTS holds great potential as a food-packaging material, preservative agent, or edible coating material.


Assuntos
Quitosana , Antioxidantes , Catálise , Ácido Gálico/análogos & derivados , Interações Hidrofóbicas e Hidrofílicas , Lacase
10.
Ecotoxicol Environ Saf ; 226: 112823, 2021 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-34597843

RESUMO

To alleviate the risk of textile effluent, the development of highly effective bioremediation strategies for synthetic dye removal is needed. Herein, we aimed to assess whether intensified bioactivity of Bacillus pumilus ZB1 by oxidative stress could improve the removal of textile dyes. Methyl methanesulfonate (MMS) induced oxidative stress significantly promoted laccase expression of B. pumilus ZB1. Both the level of hydrogen dioxide and superoxide anion showed a significant positive correlation with laccase activity (RSQ = 0.963 and 0.916, respectively) along with the change of MMS concentration. The regulation of laccase expression was closely related to oxidative stress. The overexpressed laccase in the supernatant improved the decolorization of synthetic dyes (16.43% for Congo Red, 54.05% for Crystal Violet, and 41.61% for Reactive Blue 4). Laccase was subsequently expressed in E. coli. Investigation of the potential of bacterial laccase in dye remediation using Congo Red showed that an effective degradation of azo dye could be achieved with laccase treatment. Laccase remediation alleviated the cytotoxicity of Congo Red to human hepatocytes. In silico study identified eight amino acid residues of laccase involved in binding with Congo Red. Overall, regulation of oxidative stress towards bacterium can be used as a promising approach for the improvement of bacterial bioactivity in synthetic dye remediation.


Assuntos
Corantes , Lacase , Biodegradação Ambiental , Vermelho Congo , Escherichia coli/metabolismo , Humanos , Lacase/genética , Lacase/metabolismo , Estresse Oxidativo
11.
Int J Mol Sci ; 22(19)2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34639017

RESUMO

Bisphenol (BPA) is a key ingredient in the production of epoxy resins and some types of plastics, which can be released into the environment and alter the endocrine systems of wildlife and humans. In this study, the ability of the fungus M. roridumIM 6482 to BPA elimination was investigated. LC-MS/MS analysis showed almost complete removal of BPA from the growth medium within 72 h of culturing. Products of BPA biotransformation were identified, and their estrogenic activity was found to be lower than that of the parent compound. Extracellular laccase activity was identified as the main mechanism of BPA elimination. It was observed that BPA induced oxidative stress in fungal cells manifested as the enhancement in ROS production, membranes permeability and lipids peroxidation. These oxidative stress markers were reduced after BPA biodegradation (72 h of culturing). Intracellular proteome analyses performed using 2-D electrophoresis and MALDI-TOF/TOF technique allowed identifying 69 proteins in a sample obtained from the BPA containing culture. There were mainly structural and regulator proteins but also oxidoreductive and antioxidative agents, such as superoxide dismutase and catalase. The obtained results broaden the knowledge on BPA elimination by microscopic fungi and may contribute to the development of BPA biodegradation methods.


Assuntos
Compostos Benzidrílicos/metabolismo , Biodegradação Ambiental , Fungos/metabolismo , Fenóis/metabolismo , Compostos Benzidrílicos/química , Biomassa , Biotransformação , Cinética , Lacase/metabolismo , Oxirredução , Fenóis/química
12.
Colloids Surf B Biointerfaces ; 208: 112147, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34634655

RESUMO

Laccase from Aspergillus sp. (LC) was immobilized within Fe-BTC and ZIF-zni metal organic frameworks through a one-pot synthesis carried out under mild conditions (room temperature and aqueous solution). The Fe-BTC, ZIF-zni MOFs, and the LC@Fe-BTC, LC@ZIF-zni immobilized LC samples were characterized by X-ray diffraction, scanning electron microscopy, Fourier transform infrared spectroscopy, and thermogravimetric analysis. The kinetic parameters (KM and Vmax) and the specific activity of the free and immobilized laccase were determined. Immobilized LCs resulted in a lower specific activity compared with that of the free LC (7.7 µmol min-1 mg-1). However, LC@ZIF-zni was almost 10 times more active than LC@Fe-BTC (1.32 µmol min-1 mg-1 vs 0.17 µmol min-1 mg-1) and only 5.8 times less active than free LC. The effect of enzyme loading showed that LC@Fe-BTC had an optimal loading of 45.2 mg g-1, at higher enzyme loadings the specific activity decreased. In contrast, the specific activity of LC@ZIF-zni increased linearly over the loading range investigated. The storage stability of LC@Fe-BTC was low with a significant decrease in activity after 5 days, while LC@ZIF retained up to 50% of its original activity after 30 days storage. The difference in activity and stability between LC@Fe-BTC and LC@ZIF-zni is likely due to release of Fe3+ and the low stability of Fe-BTC MOF. Together, these results indicate that ZIF-zni is a superior support for the immobilization of laccase.


Assuntos
Estruturas Metalorgânicas , Aspergillus/metabolismo , Estabilidade Enzimática , Enzimas Imobilizadas/metabolismo , Cinética , Lacase/metabolismo
13.
Molecules ; 26(19)2021 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-34641596

RESUMO

Laccase-based biocatalytic reactions have been tested with and without mediators and optimized in the oxidation of allylbenzene derivatives, such as methyl eugenol taken as a model substrate. The reaction primarily consisted in the hydroxylation of the propenyl side chain, either upon isomerization of the double bond or not. Two pathways were then observed; oxidation of both allylic alcohol intermediates could either lead to the corresponding α,ß-unsaturated carbonyl compound, or the corresponding benzaldehyde derivative by oxidative cleavage. Such a process constitutes a green equivalent of ozonolysis or other dangerous or waste-generating oxidation reactions. The conversion rate was sensitive to the substitution patterns of the benzenic ring and subsequent electronic effects.


Assuntos
Derivados de Alilbenzenos/química , Derivados de Alilbenzenos/metabolismo , Lacase/química , Lacase/metabolismo , Ozônio/metabolismo , Biocatálise , Eugenol/análogos & derivados , Eugenol/metabolismo , Concentração de Íons de Hidrogênio , Oxirredução , Ozônio/química , Especificidade por Substrato , Temperatura , Fatores de Tempo
14.
Enzyme Microb Technol ; 152: 109934, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34688090

RESUMO

The phenolic compound catechol has found various applications in the industry but is often discharged untreated in industrial effluents. Catechol is highly toxic and adversely affects the environment. This has increased extensive investigation into elucidating the effects of various synthetic elements or different biocatalysts on catechol, thereby leading the way to its bioremediation. Hence, an electrochemical-based study on catechol in the presence of the enzyme laccase could provide a basic understanding of the unique characteristics exhibited by catechol, thus facilitating a distinct perspective to its subsequent treatment and removal. The present study focuses on the electrochemical characterization of catechol based on the oxidation of laccase and the redox mediator 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). Catechol exhibited distinct electrochemical behavior across various concentrations. The unique electroactive nature of ABTS assisted in the polymerization of catechol which was found to be concentration-dependent. Laccase produced a higher oxidation-reduction rate, thereby producing a much more stable condition for the polymerization of catechol. However, with the laccase-mediator system (LMS), the catechol polymerization rate was distinctly higher and more gradual with the enzyme utilizing the electroactive species produced by ABTS to increase the electron transfer and producing a combinatorial impact on the phenolic compound. This study could rightly serve as the building block in developing future technologies like wastewater treatment and biosensors for catechol bioremediation.


Assuntos
Lacase , Trametes , Benzotiazóis , Catecóis , Lacase/metabolismo , Oxirredução , Polimerização , Polyporaceae , Ácidos Sulfônicos , Trametes/metabolismo
15.
Molecules ; 26(20)2021 Oct 19.
Artigo em Inglês | MEDLINE | ID: mdl-34684902

RESUMO

Lignin derivatives have potential as antioxidants in advanced packaging materials through their ability to scavenge oxygen in reactions catalyzed by phenol-oxidizing enzymes, such as laccase. The effects of size fractionation of lignosulfonates on laccase-catalyzed reactions were investigated in experiments with aqueous solutions, films, and coated paperboard. Four industrial lignosulfonate preparations were compared: Feed (unfractionated), Prod (5-60 kDa enriched), Conc (≥60 kDa enriched), and Perm (≤60 kDa enriched). Extraction of lignosulfonates from films showed that the enzymic reaction increased the average molecular weight from <10,000 to up to 66,000. The enzymatic reaction resulted in an increase in the water contact angle of the films from the range 25-49° to 56-81°. The four preparations showed relatively small differences with regard to their ability to scavenge oxygen in aqueous solution and in experiments with coated paperboards in sealed chambers. Coatings with lignosulfonates where the contents of low-molecular weight material had been reduced (i.e., Prod and Conc) showed improved water resistance after the enzymic reaction. Thus, in both aqueous and solid media, fractionation of lignosulfonates had little effect on oxygen scavenging, but fractionation was beneficial for other reasons, such as improved cross-linking resulting in higher molecular weight and superior water resistance.


Assuntos
Lacase/química , Lignina/análogos & derivados , Oxigênio/química , Antioxidantes/química , Catálise , Excipientes/química , Lignina/química , Peso Molecular , Oxirredução , Fenóis/química , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Água/química
16.
Enzyme Microb Technol ; 150: 109865, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34489024

RESUMO

In this study, we cross-linked aminated Thermothelomyces thermophilus laccase onto Immobead 150P epoxy carrier, and achieved an immobilization yield of 99.84 %. The optimum temperature and pH values for the oxidation of ABTS by laccase were determined to be 70 °C and pH 3.0. After 6 h at 50 °C, laccase activity was diminished by about 13 % in the free form and 28 %, in the immobilized form. Km values for both free and cross-linked laccase were 0.051 and 0.567 mM, whereas Vmax values were 2.027 and 0.854 µmol. min-1, respectively. The immobilized laccase was able to preserve its full activity for 6 weeks, retaining approximately 95 % and 78 % of its initial activity after 8 and 20 weeks, respectively. The contact angles were two-fold higher when the laccase enzyme was occupied in the biografting reaction, revealing that the hydrophobic compound bonded stably onto beechwood samples.


Assuntos
Enzimas Imobilizadas , Lacase , Estabilidade Enzimática , Enzimas Imobilizadas/metabolismo , Concentração de Íons de Hidrogênio , Cinética , Lacase/metabolismo , Sordariales , Temperatura
17.
Enzyme Microb Technol ; 150: 109890, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34489043

RESUMO

In this work, we elucidated the interactions between Myceliophthora thermophila laccase and deep eutectic solvent (DES) by crystallographic and kinetics analyses. Four types of DESs with different hydrogen bond acceptor (HBA) and hydrogen bond donor (HBD), including lactic acid: betaine, glycerol: choline chloride, lactic acid: choline chloride and glycerol: betaine was used. The results revealed that different DES have different effects on laccase activity. Lactic acid-betaine (2:1) DES has shown to enhance laccase activity up to 300 % at a concentration ranged from 2% to 8% v/v, while glycerol: choline chloride and lactic acid: choline chloride DES choline chloride-based DES have found to possess inhibitory effects on laccase under the same concentration range. Detailed kinetic study showed that glycerol: choline chloride DES is a S-parabolic-I-parabolic mixed non-competitive inhibitor, where conformational changes can occur. The crystal structures of laccase with lactic acid: choline chloride DES (LCDES) were obtained at 1.6 Å. Crystallographic analysis suggested that the addition of LCDES causes changes in the laccase active site, but the increase in water molecules observed in the resulting crystal prevented laccase from experiencing drastic structural change. Fluorescence and circular dichroism spectroscopies were also applied to determine the effects of DES on the structural conformation of laccase. The results have confirmed that the presence of DES can trigger changes in the local environments of the amino acids in the active site of laccase which contributes to the changes in its activity and stability.


Assuntos
Colina , Lacase , Ligação de Hidrogênio , Solventes , Sordariales
18.
J Environ Manage ; 299: 113619, 2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34467865

RESUMO

By virtue of screening, purification, and properties characterization, this study captures a new pH- and temperature-stable laccase, designated Galacc-F, from Ganoderma australe for dye bioremediating applications. The enzyme was purified to homogeneity by salt precipitation, ionic exchange, and size exclusion chromatography with a final specific activity of 22.214 U mg-1, yielding a purification fold of 23.989 and recovery of 38.44%. Its molecular weight was estimated to be 48.0 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, zymography, Sephadex G-100 column, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, which confirmed its monomeric nature. Galacc-F exhibited high levels of activity and stability over wide ranges of pH (5.0-8.0) and temperature (10-60 °C), which are highly valuable properties in industrial processes. Broad substrate specificity was observed, wherein a better affinity was found for 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) with a low value of Km (164.137 µM) and higher kcat/Km ratio (1.663 s-1 µM-1). Activity was stimulated by Cu2+ and ß-mercaptoethanol but inhibited by ethylenediaminetetraacetic acid, diethylpyrocarbonate, iodoacetic acid, phenylmethylsulfonyl fluoride, and Hg2+, indicating that Galacc-F is a metalloprotease containing a typical histidine-cysteine-serine catalytic triad. It had high tolerance to surfactants, oxidants, and salts. Additionally, a fabricated protocol for native Galacc-F immobilization onto Fe3O4@Chitosan composite nanoparticles using glutaraldehyde as a crosslinker was developed. Most importantly, the enzyme was determined to be ideal for use in efficient treatment of dye effluents as compared with the laccases requiring redox mediators.


Assuntos
Ganoderma , Lacase , Biodegradação Ambiental , Corantes , Estabilidade Enzimática , Ganoderma/metabolismo , Concentração de Íons de Hidrogênio , Cinética , Lacase/metabolismo , Temperatura , Têxteis
19.
Bioresour Technol ; 341: 125906, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34523564

RESUMO

This study aims to explore the impacts of the Fenton-like reaction on hydrogen peroxide, hydroxyl radicals, humic substance (HS) formation, laccase activity and microbial communities during composting to optimize composting performances. The results indicated that the activity of laccase in the presence of the Fenton-like reaction (HC) (35.92 U/g) was significantly higher than that in the control (CP) (29.56 U/g). The content of HS in HC (151.91 g/kg) was higher than that in CP (131.73 g/kg), and amides, quinones, aliphatic compounds and aromatic compounds were promoted to form HS in HC by 2D-FTIR-COS analysis. Proteobacteria contributed most greatly to AA1 at phylum level, Pseudomonas and Sphingomonas abundances increased in HC. Redundancy analysis indicated that there was a strong positive correlation among the Fenton-like reaction, laccase and HS. Conclusively, the Fenton-like reaction improved the activity of laccase, promoted the formation of HS and enhanced the quality of compost.


Assuntos
Compostagem , Substâncias Húmicas/análise , Lacase , Esterco , Solo
20.
Talanta ; 235: 122736, 2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34517604

RESUMO

Methods to improve the sensitivity of electrochemical sensors based on catalytic reactions generally require adscititious or pre-modified catalysts, which make the sensitive detection of sensors extremely challenging. This is because the activity of the catalyst is susceptible to the storage and modification process, such as aggregation during storage or loss of active sites during multi-step modification, which impairs the performance of the sensor. To solve this thorny issue, a novel electrochemical sensor based on a process-formed laccase-like catalyst was constructed for sensitive detection of tumor markers. Cu2+-polydopamine (CuPDA) combined with antibody (Ab2) were employed as copper-containing immunoprobe, which released Cu(Ⅱ) ions under acidic stimulation. Cu(Ⅱ) ions coordinate with the self-assembly cationic diphenylalanine-glutaraldehyde nanospheres (CDPGA) to form a laccase-like catalyst, which had stronger catalytic activity than laccase. The freshly formed catalyst was immediately used to degrade the polyhydroquinone-reduced graphene oxide (PHQ-rGO) composite, resulting in a significant reduction in the current signal. The PHQ-rGO composite plays dual roles of signal substance and substrate on the sensing interface. The proposed electrochemical sensor demonstrated wide linearity for the determination of a model analyte, human epididymis protein 4 (HE4), from 1 pg mL-1 to 100 ng mL-1, and the detection limit was as low as 0.302 pg mL-1 (S/N = 3), which had good consistency with that of electrochemiluminescence method. This process-formed catalyst approach will have potential reference significance for the construction of other sensors.


Assuntos
Técnicas Eletroquímicas , Grafite , Biomarcadores Tumorais , Catálise , Cobre , Humanos , Lacase
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