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Trametes hirsuta GMA-01 was cultivated in a culture medium supplemented with orange waste, starch, wheat bran, yeast extract, and salts. The fungus produced several holoenzymes, but the laccase levels were surprisingly high. Given the highlighted applicability of laccases in various biotechnological areas with minimal environmental impact, we provided a strategy to increase its production using response surface methodology. The immobilization of laccase into ionic supports (CM-cellulose, DEAE-agarose, DEAE-cellulose, DEAE-Sephacel, MANAE-agarose, MANAE-cellulose, and PEI-agarose) was found to be efficient and recuperative, showcasing the technical prowess of research. The crude extract laccase (CE) and CM-cellulose-immobilized crude extract (ICE) showed optimum activity in acidic conditions (pH 3.0) and at 70 °C for the CE and 60 °C for the ICE. The ICE significantly increased thermostability at 60 °C for the crude extract, which retained 21.6% residual activity after 240 min. The CE and ICE were successfully applied to sugarcane bagasse hydrolysis, showing 13.83 ± 0.02 µmol mL−1 reducing sugars after 48 h. Furthermore, the CE was tested for dye decolorization, achieving 96.6%, 71.9%, and 70.8% decolorization for bromocresol green, bromophenol blue, and orcein, respectively (0.05% (w/v) concentration). The properties and versatility of T. hirsuta GMA-01 laccase in different biotechnological purposes are interesting and notable, opening several potential applications and providing valuable insights into the future of biotechnological development. [ABSTRACT FROM AUTHOR]

BACKGROUND: To discover novel fungicide candidates, five series of novel norbornene hydrazide, bishydrazide, oxadiazole, carboxamide and acylthiourea derivatives (2a‐2t, 3a‐3f, 4a‐4f, 5a‐5f and 7a‐7f) were designed, synthesized and assayed for their antifungal activity toward seven representative plant fungal pathogens. RESULTS: In the in vitro antifungal assay, some title norbornene derivatives presented good antifungal activity against Botryosphaeria dothidea, Sclerotinia sclerotiorum and Fusarium graminearum. Especially, compound 2b exhibited the best inhibitory activity toward B. dothidea with the median effective concentration (EC50) of 0.17 mg L−1, substantially stronger than those of the reference fungicides boscalid and carbendazim. The in vivo antifungal assay on apples revealed that 2b had significant curative and protective effects, both of which were superior to boscalid. In the preliminary antifungal mechanism study, 2b was able to injure the surface morphology of hyphae, destroy the cell membrane integrity and increase the intracellular reactive oxygen species (ROS) level of B. dothidea. In addition, 2b could considerably inhibit the laccase activity with the median inhibitory concentration (IC50) of 1.02 μM, much stronger than that of positive control cysteine (IC50 = 35.50 μM). The binding affinity and interaction mode of 2b with laccase were also confirmed by molecular docking. CONCLUSION: This study presented a promising lead compound for the study of novel laccase inhibitors as fungicidal agrochemicals, which demonstrate significant anti‐B. dothidea activity and laccase inhibitory activity. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Stems are prone to bending or lodging due to inadequate stem strength, which seriously reduces the cut-flower ornamental quality of herbaceous peony (Paeonia lactiflora Pall.). Plant LACCASE (LAC), a copper-containing polyphenol oxidase, has been shown to participate in the polymerization process of monolignols; however, the role of LAC in regulating the stem strength of P. lactiflora remains unclear. Here, the full-length cDNA of PlLAC15, which demonstrated a positive association with stem strength, was isolated. It consisted of 1790 nucleotides, encoding 565 amino acids that had four typical laccase copper ion-binding domains. Moreover, PlLAC15 was highly expressed in the stem, and its expression level gradually significantly increased during stem development. Furthermore, PlLAC15 was found to be localized specifically to the cell wall, and its recombinant protein exhibited laccase activity. Additionally, the role of PlLAC15 in regulating the stem strength of P. lactiflora was confirmed by transgenic studies. When PlLAC15 was overexpressed in tobacco, stem strength increased by more than 50%, S-lignin was significantly deposited, and the lignification degree of stem xylem fiber cells increased. These results suggested that PlLAC15 facilitated S-lignin deposition to enhance stem strength in P. lactiflora, which would provide precious information that benefits future exploration of stem bending or lodging resistance in plants. [ABSTRACT FROM AUTHOR]

Background and purpose: A biosensor is composed of two main components: a biological sensing element and a transducer, which together determine the concentration of the analyte by facilitating a biological reaction near the transducer to improve detection capabilities. Compared to traditional analyte measurement and biochemical control methods, biosensors offer advantages such as reusability, fast response times, and high specificity. Laccase is a critical enzyme within the oxidoreductase family, with catalytic properties particularly effective toward ortho- and para-diphenols, depending on the producing organism. Unlike many enzymes that act exclusively on a specific substrate, laccase catalyzes the oxidation of a broad range of substrates, including diphenols, polyphenols, diamines, methoxyphenols, aromatic amines, and ascorbate. Protein tyrosine phosphatases are a class of enzymes that remove phosphate groups from tyrosine residues in proteins. This project aimed to design and develop an enzyme-based biosensor using modified screen-printed carbon electrodes with high sensitivity and selectivity for accurate and rapid dephostatin measurement. Materials and methods: The biosensor was fabricated by immobilizing laccase on the electrode surface. First, the surface of the screen-printed carbon electrode was modified with a graphene oxide/polyaniline nanocomposite, after which the laccase enzyme was immobilized on the modified surface using EDC/NHS linkers. Electrochemical characteristics of the electrodes were assessed using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Optimal parameters, including oxidation potential, pH, temperature, and concentration, were calibrated. At an oxidation potential of 0.45 V relative to the Ag/AgCl electrode, the current response for each sample was measured with the biosensor. Calibration curves were generated, and measurements were obtained by interpolating the amperometric signals from dephostatin solutions. Dephostatin standards were then introduced to the diluted Nescafe sample (phosphate buffer, 0.05 M, pH 6.5) without any pretreatment, and recovery rates were calculated. Results: By immobilizing graphene oxide and laccase on the electrode surface, the modified electrode exhibited increased cathodic and anodic currents compared to the unmodified electrode in K₄Fe(CN)₆/K₃Fe(CN)₆ solution. Due to the presence of graphene nanoparticles on the electrode surface, the electron transfer rate improved, as indicated by a 20 µA increase. The electrode response was linear across a concentration range of 10 to 900 nM, with a detection limit of 6 nM. Conclusion: Since protein tyrosine phosphatases play essential roles in cell signaling pathways and disease development, the use of dephostatin as an inhibitor has gained interest. Therefore, an accurate and rapid method for dephostatin measurement is crucial in the pharmaceutical industry. The prepared biosensor, which incorporates laccase enzyme and graphene nanoparticles stabilized with a chitosan nanocomposite, allows for precise dephostatin measurement. The laccase enzyme immobilized on the electrode surface can specifically oxidize the dephostatin molecule, producing a unique electrode response for this molecule.

Abstract Laccase, the selectively lignin degrader, vital to the initiation of lignocellulosic deconstruction was immobilized onto activated agarose beads to increase its reuse potential. Laccase cross-linked beads (~ 3.42 mm) recorded a specific activity of 23 Umg− 1, retaining about 80.43% enzyme activity after 45 days of storage. The immobilization yield and efficiency were 89% and 97% respectively. The equilibrium data fitted the Freundlich equation (R2 = 0.9987) demonstrating multilayer adsorption and the presence of Cu, Fe, and S in the elemental analysis of immobilized beads established effective binding between activated agarose beads and the laccase protein. Characterization studies of the immobilized laccase-treated crop residues revealed significant differences in the lignin polymer after each treatment cycle. An increase in digestibility of 26.21% and 7.62% was observed in paddy and finger millet-treated straws respectively, over the controls corroborating efficient lignin depolymerization. The propitious performance of laccase beads authenticated in the batch enzymatic reactor to treat crop residues paves headway as a sustainable green technology in the deconstruction of crop residues for use as ruminant feed, augmenting productivity.

Owing to the production of lignin-modifying enzymes (LMEs), white-rot fungi (WRF) such as polypores are potent organisms in the biodegradation of xenobiotic pollutants. The nonspecific function of LMEs including laccase and manganese peroxidase (MnP), has enabled the use of WRF in biotechnological applications, particularly in bioremediation. In this study, 12 strains from nine white-rot basidiomycete genera viz., Ganoderma, Inocutis, Irpex, Lentinus, Lenzites, Oxyporus, Peniophora, Sanghuangporus, and Trametes were isolated from Iran and identified using morphological and molecular tools. The enzyme activity of laccase and manganese peroxidase that directly correlated with the biodegradation were determined, and the strains with the highest enzyme activities were evaluated for their ability to degrade 400 mg/L of anthracene over 28 days. Gas chromatography with flame ionization detector (GC-FID) revealed that four polypores viz., Trametes versicolor v21te, T. versicolor v22da, T. hirsuta, and Oxyporus sp. degraded 64%, 52%, 34%, and 20% of the anthracene, respectively. According to our analysis, the examined indigenous WRF are potentially useful candidates for the development of new mycoremediation techniques to degrade polycyclic aromatic hydrocarbons (PAHs).

Abstract With the development of industry and modern manufacturing, nondegradable low-density polyethylene (LDPE) has been widely used, posing a rising environmental hazard to natural ecosystems and public health. In this study, we isolated a series of LDPE-degrading fungi from landfill sites and carried out LDPE degradation experiments by combining highly efficient degrading fungi in pairs. The results showed that the mixed microorganisms composed of Alternaria sp. CPEF-1 and Trametes sp. PE2F-4 (H-3 group) had a greater degradation effect on heat-treated LDPE (T-LDPE). After 30 days of inoculation with combination strain H-3, the weight loss rate of the T-LDPE film was approximately 154% higher than that of the untreated LDPE (U-LDPE) film, and the weight loss rate reached 0.66 ± 0.06%. Environmental scanning electron microscopy (ESEM) and Fourier transform infrared spectroscopy (FTIR) were used to further investigate the biodegradation impacts of T-LDPE, including the changes on the surface and depolymerization of the LDPE films during the fungal degradation process. Our findings revealed that the combined fungal treatment is more effective at degrading T-LDPE than the single strain treatment, and it is expected that properly altering the composition of the microbial community can help lessen the detrimental impact of plastics on the environment.

Laccase is a multicopper oxidase enzyme that oxidizes a variety of substrates, including polyphenols and polycyclic aromatic hydrocarbons (PAHs). It catalyzes the four-electron reduction of molecular oxygen that results in the production of water as a by-product. Thus, laccase can play an important role in environmental care. Previously, we have successfully expressed Trametes trogii laccase (TtLcc1) in the yeast Saccharomyces cerevisiae. In this work, we have expressed in yeast another laccase, LacA from Trametes sp. AH28-2, and tested its function on PAHs. Yeast cells engineered to produce the two laccases performed efficient PAH degradation. Both TtLcc1 and LacA led to the construction of spatiotemporal fluorescence-pulse generators when combined with a benzoate/salicylate yeast biosensor in a two-population system. Moreover, laccases returned a visual output signal in yeast synthetic circuits—upon reacting with ABTS (2,2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid)). Thus, in S. cerevisiae, laccases are a powerful alternative to fluorescent reporter proteins.

Endocrine disruptors such as bisphenol A (BPA) adversely affect the environment and human health. Laccases are used for the efficient biodegradation of various persistent organic pollutants in an environmentally safe manner. However, the direct application of free laccases is generally hindered by short enzyme lifetimes, non-reusability, and the high cost of a single use. In this study, laccases were immobilized on a novel magnetic three-dimensional poly(ethylene glycol) diacrylate (PEGDA)–chitosan (CS) inverse opal hydrogel (LAC@MPEGDA@CS@IOH). The immobilized laccase showed significant improvement in the BPA degradation performance and superior storage stability compared with the free laccase. 91.1% of 100 mg/L BPA was removed by the LAC@MPEGDA@CS@IOH in 3 hr, whereas only 50.6% of BPA was removed by the same amount of the free laccase. Compared with the laccase, the outstanding BPA degradation efficiency of the LAC@MPEGDA@CS@IOH was maintained over a wider range of pH values and temperatures. Moreover, its relative activity of was maintained at 70.4% after 10 cycles, and the system performed well in actual water matrices. This efficient method for preparing immobilized laccases is simple and green, and it can be used to further develop ecofriendly biocatalysts to remove organic pollutants from wastewater. [Display omitted] [ABSTRACT FROM AUTHOR]

The ability of laccases to work effectively in properly selected aqueous-organic solutions significantly expands the scope of their application, making them commercially attractive and more competitive. However, it must be considered that molecules of organic solvent are very aggressive towards enzymes and destroy weak non-covalent interactions that maintain the spatial structure of the protein. In this work, we quantitatively described the influence of the nature of water-miscible organic solvents and their concentration on the activity and stability of Trametes versicolor laccase during the oxidation of ABTS (2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) through the maximum rates and the Michaelis constants KM. The obtained kinetic parameters make it possible to compare the effect of various organic solvents on laccase and select the most optimal one, which minimizes the decrease in enzyme activity, maintains its stability and allows repeated use. Among the solvents studied, polyhydroxy compounds have the least negative effect on laccase activity; however, in all cases, a high concentration of organic solvent (≥30% (v/v)) leads to suppression of enzymatic activity. It has been shown that the longer the alkyl chain (hydrophobic fragment of the structure), the stronger the negative effect of alcohols on enzymes. The most toxic solvent for Trametes versicolor laccase is isopropanol. In addition, numerous examples of various uses of Trametes versicolor in aquatic-organic environments are discussed here. [ABSTRACT FROM AUTHOR]

Objectives: This study aimed to produce an engineered recombinant laccase from extremophilic Halalkalibacterium halodurans C-125 (Lac-HhC-125) with higher protein yield, into a more active conformation and with properties that meet the fundamental needs of biotechnological application. Results: The rLac-HhC125 was partially purified by size exclusion chromatography and concentrated by ultrafiltration (10 kDa) with a yield of 57.6%. Oxidation reactions showed that adding 2 mM CuSO4 to the assay solution led to activating the laccase. To increase its initial activity, the rLac-HhC125 was treated at 50 °C for 20 min before the assays, improving its performance by fourfold using the syringaldazine as a substrate. When treated with EDTA, methanol, ethanol, and DMSO, the rLac-HhC125 maintained more than 80% of its original activity. Interestingly, the acetonitrile induced a twofold activity of the rLac-HhC125. The putative rLac-HhC125 demonstrated a capability of efficient transformation of different organic compounds at pH 6, known as dye precursors, into coloured molecules. Conclusion: The rLac-HhC125 was active at high temperatures and alkaline pH, exhibited tolerance to organic solvents, and efficiently transformed different hydroxy derivatives into coloured compounds, which indicates that it can be used in various biotechnological processes. [ABSTRACT FROM AUTHOR]

Abstract A novel approach for the synthesis of 1,3,4-oxa(thia)diazole aryl thioethers through a biocatalytic strategy has been introduced. By leveraging Myceliophthora thermophila laccase (Novozym 51003) as a catalyst, catechol undergoes oxidation to ortho-quinone, facilitating subsequent 1,4-thia-Michael addition reactions. The method offers efficiency and mild reaction conditions, demonstrating promise for sustainable synthesis pathways in organic chemistry. Using this approach, 13 new derivatives of 2,5-disubstituted-1,3,4-oxa(thia)diazole aryl thioethers, with a yield of 46–94%, were synthesized.

Alginate is a naturally occurring polymer derived from brown algae biomass, which has numerous applications in various fields. Chemical modification of alginate is widely used to improve alginate’s physicochemical properties and provide new potential for multiple applications. In this article, we modified alginate with L-DOPA, using periodate oxidation and reductive amination, to obtain more suitable biopolymer for biocatalyst immobilization and hydrogel formation. Obtained modified alginate was used for the immobilization of laccase on cell walls. For this purpose, laccase from Streptomyces cyaneus was expressed on the surface of Saccharomyces cerevisiae EBY100 cells. The obtained cell wall laccase was immobilized within L-DOPA-alginate beads by crosslinking the L-DOPA-alginate with calcium ions and laccase. The effect of additional crosslinking of beads by green light-induced photopolymerization with eosin Y was investigated. The immobilized laccase systems were used for dye decolorization and investigated in multiple treatment processes. Beads with L-DOPA-alginate with a higher degree of modification (5.0 mol%) showed higher enzymatic activity and better decolorization efficiency than those with a lower degree of modification (2.5 mol%). Obtained immobilized biocatalysts are suitable for decolorizing dye Evans Blue due to their high efficiency and reusability.

Abstract Dopamine is one of the significant neurotransmitters and its monitoring in biological fluids is a critical issue in healthcare and modern biomedical technology. Here, we have developed a dopamine biosensor based on surface plasmon resonance (SPR). For this purpose, the carboxymethyl dextran SPR chip was used as a surface to immobilize laccase as a bioaffinity recognition element. Data analysis exhibited that the acidic pH value is the optimal condition for dopamine interaction. Calculated kinetic affinity (KD) (48,545 nM), obtained from a molecular docking study, showed strong association of dopamine with the active site of laccase. The biosensor exhibited a linearity from 0.01 to 189 μg/ml and a lower detection limit of 0.1 ng/ml (signal-to-noise ratio (S/N) = 3) that is significantly higher than the most direct dopamine detecting sensors reported so far. Experiments for specificity in the presence of compounds that can co-exist with dopamine detection such as ascorbic acid, urea and l-dopa showed no significant interference. The current dopamine biosensor with high sensitivity and specificity, represent a novel detection tool that offers a label-free, simple procedure and cost effective monitoring system.

Heavy metal contamination is one of the key environmental complications. Due to some disadvantages of conventional methods, the use of active organisms is becoming more popular technique to remove it. In the present study, primarily 35 bacterial strains were discovered in metal containing media. After being identified resistance power to different copper concentrations (100–1000 mg/l), JRHM33 had the highest level of resistance up to 1000 mg/l of copper. Using the 16S rRNA sequencing, bacterial strain JRHM33 was discovered and revealed 99% similarity to pseudomonas aeruginosa. Sequencing and bioinformatics study using conserved domain analysis supported the laccase gene is present in JRHM33 and has classification as a member of the multicopper oxidase superfamily, which has reduction capacity of metal ions. Analysis of phenotype microarray (PM) technology provides an insight into the metabolic profiling of microbial cell into Pseudomonas aeruginosa JRHM33. Furthermore, Using the central composite design of response surface methodology (CCD-RSM), the successive optimization of the process parameters was attempted for the maximum reduction of the copper. Maximum 68.71% Cu reduction was achieved at 6.71 pH, 90.61 min of incubation time, 5 ml of inoculum size, and 113 rpm of agitation. The generated model has R2 value of 0.9834, indicating that the ANOVA gave it a very significant result. The findings of the validation experiment showed a remarkable similarity between the projected and experimental results. It is determined that bacterial strains isolated from metal-contaminated effluent employ their natural capacity to change toxic heavy metals into less harmful or nontoxic forms.

AbstractLaccase has a wide range of substrates and the oxidation product is water, providing an ideal choice for the biological degradation and decolorization of dyeing wastewater. This study extracted laccase from the fruiting body of Agaricus bisporus, then used o-phthalaldehyde (OPA) as a cross-linking agent and added surface-aminated Fe3O4 nanoparticles (NH2-nanoFe3O4) to prepare magnetic cross-link enzyme aggregates (MCLEAs) of laccase, which was characterized by scanning electron microscope and vibrating sample magnetometer. The optimal temperature for the catalytic reaction of the MCLEAs was 35 °C, and the optimal pH was 3.0. The stability to temperature pH and storage stability were all enhanced. The immobilized laccase was utilized to treat amido black 10B solution, and the decolorization rate reached 92.48%. After five cycles of use, the MCLEAs of laccase could maintain 42.91% of the relative activity and 32.31% of the relative degradation ability to amido black 10B. [ABSTRACT FROM AUTHOR]

The development of strategies allowing either the production of high value phenolics, or the isolation of properties‐enhanced materials from technical lignins represents a fundamental step in the industrial upcycling of technical lignins. Both aims are met by the strategy presented in the present work, relying on the coupling of solvent‐based fractionation with the oxidative action of a new type of alkaline‐stable genetically modified bacterial laccase. The described approach succeeded in the tandem, high‐yield and selective isolation of valuable lignin‐monomeric compounds (MCs) and high molecular weight and hydrophobicity‐tailored polymerised materials (PMs) from two technical lignins, namely softwood kraft lignin (SKL), and wheat straw organosolv lignin (WSL). With respect to MCs, higher yields as compared to similar studies (up to 17.2 mg/g) were achieved. PMs from SKL samples where characterised by an almost quadrupled Mw, while in the case of WSL the Mw was approximately doubled. Noteworthy, the reaction conditions were optimized in terms of reaction temperature, time, enzymatic loading, and alkalinity for the selective production of single MCs. Most interestingly, technical lignins as well as their fractions and the PMs deriving from their laccase‐catalysed oxidation showed increased hydrophobicity. [ABSTRACT FROM AUTHOR]

Known for its tunable conductivity and stability, Polyaniline (PANI) is a valuable polymer for electronics and sensing devices. Challenges in solubility have been addressed by creating sulfonated PANI (SPANI), enhancing its practical use. Synthesizing SPANI from sulfonated aniline is intricate, but laccase biocatalysis offers an eco-conscious solution, effective even against high redox potential obstacles. This research monitored the Trametes versicolor laccase-induced oxidation of 3-ABSa via UV-vis spectroscopy, with a notable peak at 565 nm signifying SPANI synthesis, effective even at suboptimal pH. Mediators further boost this process. Moreover, NMR and spectroelectrochemistry confirm the green synthesis of SPANI by laccase, hinting that pH fine-tuning could improve yields, alongside the concurrent creation of azobenzene derivatives. [ABSTRACT FROM AUTHOR]

Laccases (EC 1.10.3.2) are multicopper oxidases with the capability to oxidize diverse phenolic and non-phenolic substrates. While the molecular mechanism of their activity towards phenolic substrates is well-established, their reactivity towards non-phenolic substrates, such as polycyclic aromatic hydrocarbons (PAHs), remains unclear. To elucidate the oxidation mechanism of PAHs, particularly the activation mechanism of the sp2 aromatic C–H bond, we conducted a density functional theory investigation on the oxidation of two PAHs (anthracene and benzo[a]pyrene) using an extensive model of the T1 copper catalytic site of the fungal laccase from Trametes versicolor. [ABSTRACT FROM AUTHOR]

The diversity of Ganoderma remains largely unexplored, with little information available due to fungiphobia and the morphological plasticity of the genus. To address this gap, an ongoing study aims to collect and identify species with this genus using nuclear ribosomal DNA regions called the “Internal Transcribed Spacer” (ITS1-5.8S-ITS2 = ITS). In this study, a new species, Ganoderma segmentatum sp. nov., was discovered on the dead tree trunk of the medicinal plant, Vachellia nilotica. The species was identified through a combination of morpho-anatomical characteristics and phylogenetic analyses. This new species was closely related to Ganoderma multipileum, G. mizoramense, and G. steyaertanum, with a 99% bootstrap value, forming a distinct branch in the phylogenetic tree. Morphologically, G. segmentatum can be distinguished by its frill-like appearance on the margin of basidiome. Wilt or basal stem rot, a serious disease of trees caused by Ganoderma species and V. nilotica, is brutally affected by this disease, resulting in substantial losses in health and productivity. This Ganoderma species severely damages V. nilotica through deep mycelial penetration in the upper and basal stems of the host species. Pathogenic observational descriptions of G. segmentatum on dead tree trunks showed the exudation of viscous reddish-brown fluid from the basal stem portion, which gradually extended upward. Symptoms of this disease include decay, stem discoloration, leaf drooping, and eventual death, which severely damaged the medicinal tree of V. nilotica. [ABSTRACT FROM AUTHOR]

The experiment aimed to study the screening, identification, and degradation efficiency of efficient lignindegrading strain in rape straw. A highly efficient lignin degrading strain MS25 was screened from the returned soil of rapeseed straw and identified as a Penicillium fungus. Based on single factor experiments and response surface methodology, the enzyme production characteristics was invstigated and the conditions of MS25 strain was optimized, and the lignin degradation ability was tested. The results showed that the optimal time for the strain to produce laccase (Lac) was 5 days, and the time to produce peroxidase (Lip) and manganese peroxidase (MnP) was 7 days. The best temperature was 50 ℃, the shaking table speed was 140 r/min, the carbon to nitrogen ratio was 15, and the most suitable pH value for producing MnP was 5, while the most suitable pH values for producing Lip and Lac were 6. The optimal enzyme production conditions after optimization were temperature of 50.22 ℃, carbon to nitrogen ratio of 12.29, and pH value of 6.21. Under these conditions, the activities of Lip, MnP, and Lac enzymes were 1 984.89, 774.87, and 291.74 U/L, respectively, which were 33.97%, 33.28%, and 40.35% higher than before optimization. Under optimal conditions, the lignin degradation rates of straw and fungal bran of the strains were 44.67% and 43.29%, respectively. The study indicates that strain MS25 can effectively degrade lignin and can be used as a high-quality microorganism for rape straw fermentation, and can be specifically applied in the field of feed production. [ABSTRACT FROM AUTHOR]

Laccases are polyphenol oxidase enzymes and form the enzyme complex known for their role in wood decomposition and lignin degradation. The present study aimed to systematically review the state-of-the-art trends in scientific publications on laccase enzymes of the last 10 years. The main aspects checked included the laccase-producing fungal genera, the conditions of fungal growth and laccase production, the methods of immobilization, and potential applications of laccase. After applying the systematic search method 177 articles were selected to compound the final database. Although various fungi produce laccase, most studies were Trametes and Pleurotus genera. The submerged fermentation (SmF) has been the most used, however, the use of solid-state fermentation (SSF) appeared as a promising technique to produce laccase when using agro-industrial residues as substrates. Studies on laccase immobilization showed the covalent bonding and entrapment methods were the most used, showing greater efficiency of immobilization and a high number of enzyme reuses. The main use of the laccase was in bioremediation, especially in the discoloration of dyes from the textile industry and the degradation of pharmaceutical waste. Implications and consequences of all these findings in biotechnology and environment, as well as the trends and gaps of laccase research were discussed. [ABSTRACT FROM AUTHOR]

Cryptococcus neoformans is an opportunistic pathogenic fungus that produces melanin during infection, an important virulence factor in Cryptococcal infections that enhances the ability of the fungus to resist immune defense. This fungus can synthesize melanin from a variety of substrates, including L-DOPA (L-3,4-dihydroxyphenylalanine). Since melanin protects the fungus from various stress factors such as oxidative, nitrosative, extreme heat and cold stress; we investigated the effects of environmental conditions on melanin production and survival. In this study, we investigated the effects of different pH values (5.6, 7.0 and 8.5) and temperatures (30 °C and 37 °C) on melanization and cell survival using a microtiter plate-based melanin production assay and an oxidative stress assay, respectively. In addition, the efficacy of compounds known to inhibit laccase involved in melanin synthesis, i.e., tunicamycin, β-mercaptoethanol, dithiothreitol, sodium azide and caspofungin on melanization was evaluated and their sensitivity to temperature and pH changes was measured. The results showed that melanin content correlated with pH and temperature changes and that pH 8.5 and 30 °C, were best for melanin production. Besides that, melanin production protects the fungal cells from oxidative stress induced by hydrogen peroxide. Thus, changes in pH and temperature drastically alter melanin production in C. neoformans and it correlates with the fungal survival. Due to the limited antifungal repertoire and the development of resistance in cryptococcal infections, the investigation of environmental conditions in the regulation of melanization and survival of C. neoformans could be useful for future research and clinical phasing. [ABSTRACT FROM AUTHOR]

Laccases are important and valuable enzymes with a great potential for biotechnological applications. In this study, two novel laccases, LacHU1 and LacHU2, from Alternaria sp. HU have been purified and characterized. The molecular mass of each isoenzyme was ~66 kDa. LacHU1 laccases was yellow and had no typical blue oxidase spectra and LacHU2 had a blue color and characteristic absorption spectra. The catalytic efficiency of LacHU1 for most substrates was higher than that of LacHU2 laccase. Both isoenzymes effectively oxidize flavonoids. Alternaria sp. laccases were successfully immobilized on magnetic nanoparticles. The thermostability of immobilized laccases increased and optimal pH shifted to more alkaline compared to the free laccases. Potential applications of laccases from Alternaria sp. HU are in the oxidation of flavonoids in cotton or in water treatment processes. [ABSTRACT FROM AUTHOR]

The climate-change-coupled fungal burden in crop management and the need to reduce chemical pesticide usage highlight the importance of finding sustainable ways to control Aspergillus flavus. This study examines the effectiveness of 50 Pseudomonas isolates obtained from corn rhizospheres against A. flavus in both solid and liquid co-cultures. The presence and quantity of aflatoxin B1 (AFB1) and AFB1-related compounds were determined using high-performance liquid chromatography–high resolution mass spectrometry analysis. Various enzymatic- or non-enzymatic mechanisms are proposed to interpret the decrease in AFB1 production, accompanied by the accumulation of biosynthetic intermediates (11-hydroxy-O-methylsterigmatocystin, aspertoxin, 11-hydroxyaspertoxin) or degradation products (the compounds C16H10O6, C16H14O5, C18H16O7, and C19H16O8). Our finding implies the upregulation or enhanced activity of fungal oxidoreductases and laccases in response to bacterial bioactive compound(s). Furthermore, non-enzymatic reactions resulted in the formation of additional degradation products due to acid accumulation in the fermented broth. Three isolates completely inhibited AFB1 or any AFB1-related compounds without significantly affecting fungal growth. These bacterial isolates supposedly block the entire pathway for AFB1 production in the fungus during interaction. Apart from identifying effective Pseudomonas isolates as potential biocontrol agents, this work lays the foundation for exploring new bacterial bioactive compounds. [ABSTRACT FROM AUTHOR]

Maize (Zea mays L.) may be infected by Fusarium verticillioides and F. proliferatum, and consequently contaminated with fumonisins (FBs), as well as the co-products of bioethanol intended for animal feed. Laccase enzymes have a wide industrial application such as mycotoxin degradation. The aims were to isolate and identify fungal laccase-producing strains, to evaluate laccase production, to determine the enzymatic stability under fermentation conditions, and to analyse the effectiveness in vitro of enzymatic extracts (EEs) containing laccases in degrading FB1. Strains belonging to Funalia trogii, Phellinus tuberculosus, Pleurotus ostreatus, Pycnoporus sanguineus and Trametes gallica species showed laccase activity. Different isoforms of laccases were detected depending on the evaluated species. For the FB1 decontamination assays, four enzymatic activities (5, 10, 15 and 20 U/mL) were tested, in the absence and presence of vanillic acid (VA) and 2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPO) as redox mediators (1 and 10 mM). Trametes gallica B4-IMICO-RC EE was the most effective strain in buffer, achieving a 60% of FB1 reduction. Laccases included in EEs remained stable at different alcoholic degrees in maize steep liquor (MSL), but no significant FB1 reduction was observed under the conditions evaluated using MSL. This study demonstrate that although laccases could be good candidates for the development of a strategy to reduce FB1, further studies are necessary to optimise this process in MSL. [ABSTRACT FROM AUTHOR]

Recently, prokaryotic laccases from lactic acid bacteria (LAB), which can degrade biogenic amines, were discovered. A laccase enzyme has been cloned from Oenococcus oeni, a very important LAB in winemaking, and it has been expressed in Escherichia coli. This enzyme has similar characteristics to those previously isolated from LAB as the ability to oxidize canonical substrates such as 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,6-dimethoxyphenol (2,6-DMP), and potassium ferrocyanide K4[Fe(CN6)], and non-conventional substrates as biogenic amines. However, it presents some distinctiveness, the most characteristic being its psychrophilic behaviour, not seen before among these enzymes. Psychrophilic enzymes capable of efficient catalysis at low temperatures are of great interest due to their potential applications in various biotechnological processes. In this study, we report the discovery and characterization of a new psychrophilic laccase, a multicopper oxidase (MCO), from the bacterium Oenococcus oeni. The psychrophilic laccase gene, designated as LcOe 229, was identified through the genomic analysis of O. oeni, a Gram-positive bacterium commonly found in wine fermentation. The gene was successfully cloned and heterologously expressed in Escherichia coli, and the recombinant enzyme was purified to homogeneity. Biochemical characterization of the psychrophilic laccase revealed its optimal activity at low temperatures, with a peak at 10 °C. To our knowledge, this is the lowest optimum temperature described so far for laccases. Furthermore, the psychrophilic laccase demonstrated remarkable stability and activity at low pH (optimum pH 2.5 for ABTS), suggesting its potential for diverse biotechnological applications. The kinetic properties of LcOe 229 were determined, revealing a high catalytic efficiency (kcat/Km) for several substrates at low temperatures. This exceptional cold adaptation of LcOe 229 indicates its potential as a biocatalyst in cold environments or applications requiring low-temperature processes. The crystal structure of the psychrophilic laccase was determined using X-ray crystallography demonstrating structural features similar to other LAB laccases, such as an extended N-terminal and an extended C-terminal end, with the latter containing a disulphide bond. Also, the structure shows two Met residues at the entrance of the T1Cu site, common in LAB laccases, which we suggest could be involved in substrate binding, thus expanding the substrate-binding pocket for laccases. A structural comparison of LcOe 229 with Antarctic laccases has not revealed specific features assigned to cold-active laccases versus mesophilic. Thus, further investigation of this psychrophilic laccase and its engineering could lead to enhanced cold-active enzymes with improved properties for future biotechnological applications. Overall, the discovery of this novel psychrophilic laccase from O. oeni expands our understanding of cold-adapted enzymes and presents new opportunities for their industrial applications in cold environments. [ABSTRACT FROM AUTHOR]

Increasing grain yield is a major goal of breeders due to the rising global demand for food. We previously reported that the miR397–LACCASE (OsLAC) module regulates brassinosteroid (BR) signaling and grain yield in rice (Oryza sativa). However, the precise roles of laccase enzymes in the BR pathway remain unclear. Here, we report that OsLAC controls grain yield by preventing the turnover of TRANSTHYRETIN-LIKE (OsTTL), a negative regulator of BR signaling. Overexpressing OsTTL decreased BR sensitivity in rice, while loss-of-function of OsTTL led to enhanced BR signaling and increased grain yield. OsLAC directly binds to OsTTL and regulates its phosphorylation-mediated turnover. The phosphorylation site Ser226 of OsTTL is essential for its ubiquitination and degradation. Overexpressing the dephosphorylation-mimic form of OsTTL (OsTTLS226A) resulted in more severe defects than did overexpressing OsTTL. These findings provide insight into the role of an ancient laccase in BR signaling and suggest that the OsLAC–OsTTL module could serve as a target for improving grain yield. Laccase controls grain size and yield in rice by preventing the turnover of a negative regulator of brassinosteroid signaling. [ABSTRACT FROM AUTHOR]

A stable and efficient biocatalyst was prepared by encapsulating Trametes versicolor laccase using an acrylic acid-grafted β-cyclodextrin hydrogel (Lac-CD-PAA). Scanning electron microscopy and nitrogen adsorption-desorption experiments showed that there were regularly distributed channels in the spongy Lac-CD-PAA. In addition, a large number of mesopores and macropores existed in the wall of the hydrogel lamellae. This network structure reduced the diffusion resistance of the hydrogel to the target substrate. The relative activity of the resulting Lac-CD-PAA could be maintained at 35.8% after six cycles of use. Lac-CD-PAA exhibited higher thermal and chemical stability compared to free laccase. The negative charge on the surface of Lac-CD-PAA gives it the ability to pretreat cationic dyes. In six consecutive methylene blue decolorization tests, Lac-CD-PAA decolorized better than free laccase. The results showed that the prepared β-cyclodextrin-based composite hydrogel was a good carrier for laccase. [ABSTRACT FROM AUTHOR]

In angiosperms, seed size is a critical trait that is influenced by the complex interplay between the endosperm and seed coat. The HAIKU (IKU) pathway, involving the transcription factor WRKY10, plays a crucial role in regulating seed size in Arabidopsis thaliana. However, the downstream targets of WRKY10 and their roles in seed size determination remain largely unexplored. Here, we identified LACCASE2 (LAC2), a laccase gene involved in lignin biosynthesis, as a new downstream target of WRKY10. We observed that the expression of LAC2 was upregulated in the mini3 mutant, which is defective in WRKY10. We demonstrated that WRKY10 directly binds to the promoter of miR397a, activating its expression. miR397a, in turn, represses the expression of LAC2. Genetic analyses revealed that a mutation in LAC2 or overexpression of miR397a partially rescued the small seed phenotype of the MINISEED3 (MINI3) mutant mini3. Conversely, the overexpression of LAC2 in the wild type led to a decrease in seed size. These findings suggest that LAC2 functions as a negative regulator of seed size, and its expression is modulated by WRKY10 through miR397a. Our study uncovers a novel WRKY10-miR397a-LAC2 pathway that regulates seed size in Arabidopsis, providing new insights into the complex regulatory network governing seed development in plants. [ABSTRACT FROM AUTHOR]

Laccase (LAC), a copper-containing polyphenol oxidase, is an important pathogenic factor of pathogenic fungi, and has been identified as an important virulence factor in numerous pathogenic fungi. LAC is encoded by a gene family and belongs to the class of multicopper oxidases. The study aimed to identify the LAC genes in Athelia bombacina (Link) Pers, and their interactions with the host. The expression levels of the LAC genes were quantified using RT-qPCR. The LAC activity, level of malondialdehyde (MDA) and activities of protective enzymes in 'Huangguan' pears during the interaction were measured. The AbLac4 gene deletion mutant strain was constructed. Six LAC genes were identified in A. bombacina, distributed across three chromosomes. Interspecies collinearity analysis suggested that LAC genes could serve as crucial pathogenic factors in A. bombacina. The LAC gene family can be classified into three distinct subgroups. Among the subgroups, variations were observed in their characteristic sequences and conserved motifs. However, the LAC genes within the same subgroup exhibited a high degree of conservation. The genes showed diverse expression profiles, with their promoters harboring multiple stress-responsive elements. Signal peptide prediction showed that all LAC proteins, with the exception of the AbLac3 protein, possessed signal peptides, indicating that they are secretory proteins. The subcellular localization analysis showed that all LAC proteins may be localized extracellularly. RT-qPCR revealed differential expression patterns among LAC genes; specifically, AbLac1 and AbLac4 exhibited distinct expression dynamics during the infection process. The LAC activity first increased and then decreased, with the highest increase rate occurring in the early stage of culture. The MDA content and catalase (CAT) activity at the inoculated site were found to be significantly higher than the uninoculated control. In addition, the deletion of AbLac4 gene reduced the growth rate and pathogenic ability of A. bombacina. This investigation found that AbLac1 and AbLac4 may play pivotal roles in mediating host interactions, and the fruit may combat pathogen infection through increasing the activities of CAT, phenylalanine ammonia lyase and peroxidase. This study provides valuable new insights into the pathogenic mechanisms of A. bombacina, significantly contributing to the field. [ABSTRACT FROM AUTHOR]

Copyright of Mycosystema is the property of Mycosystema Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Mycosystema is the property of Mycosystema Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Mycosystema is the property of Mycosystema Editorial Board and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The persistence and ubiquity of polycyclic aromatic hydrocarbons (PAHs) in the environment necessitate effective remediation strategies. Hence, this study investigated the potential of purified Laccases, TlFLU1L and TpFLU12L, from two indigenous fungi Trichoderma lixii FLU1 (TlFLU1) and Talaromyces pinophilus FLU12 (TpFLU12), respectively for the oxidation and detoxification of anthracene. Anthracene was degraded with vmax values of 3.51 ± 0.06 mg/L/h and 3.44 ± 0.06 mg/L/h, and Km values of 173.2 ± 0.06 mg/L and 73.3 ± 0.07 mg/L by TlFLU1L and TpFLU12L, respectively. The addition of a mediator compound 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) to the reaction system significantly increased the degradation of anthracene, with up to a 2.9-fold increase in vmax value and up to threefold decrease in Km values of TlFLU1L and TpFLU12L. The GC–MS analysis of the metabolites suggests that anthracene degradation follows one new pathway unique to the ABTS system—hydroxylation and carboxylation of C-1 and C-2 position of anthracene to form 3-hydroxy-2-naphthoic acid, before undergoing dioxygenation and side chain removal to form chromone which was later converted into benzoic acid and CO2. This pathway contrasts with the common dioxygenation route observed in the free Laccase system, which is observed in the second degradation pathways. Furthermore, toxicity tests using V. parahaemolyticus and HT-22 cells, respectively, demonstrated the non-toxic nature of Laccase-ABTS-mediated metabolites. Intriguingly, analysis of the expression level of Alzheimer's related genes in HT-22 cells exposed to degradation products revealed no induction of neurotoxicity unlike untreated cells. These findings propose a paradigm shift for bioremediation by highlighting the Laccase-ABTS system as a promising green technology due to its efficiency with the discovery of a potentially less harmful degradation pathway, and the production of non-toxic metabolites. [ABSTRACT FROM AUTHOR]

As a typical sulfonylurea herbicide, nicosulfuron is mainly used to control grass weeds and some broadleaf weeds in corn fields. However, as the amount of use continues to increase, it accumulates in the environment and eventually becomes harmful to the ecosystem. In the present study, a new metallic nanomaterial, δ-MnO2, was prepared, which not only has a similar catalytic mechanism as laccase but also has a significant effect on pesticide degradation. Therefore, the bicatalytic property of MnO2 can be utilized to improve the remediation of nicosulfuron contamination. Firstly, MnO2 nanomaterials were prepared by controlling the hydrothermal reaction conditions, and immobilized laccase was prepared by the adsorption method. Next, we investigate the effects of different influencing factors on the effect of immobilized laccase, MnO2, and free laccase on the degradation of nicosulfuron in water and soil. In addition, we also analyze the metabolic pathway of nicosulfuron degradation in immobilized laccase and the bicatalytic mechanism of MnO2. The results demonstrated that the degradation rate of nicosulfuron in water by immobilized laccase was 88.7%, and the optimal conditions were 50 mg/L, 25 h, 50 °C, and pH 5. For nicosulfuron in soil, the optimal conditions for the degradation by immobilized laccase were found to be 151.1 mg/kg, 46 °C, and pH 5.9; under these conditions, a degradation rate of 90.1% was attained. The findings of this study provide a theoretical reference for the immobilized laccase treatment of sulfonylurea herbicide contamination in water and soil. [ABSTRACT FROM AUTHOR]

The effective recovery of the immobilized enzymes using magnetic carriers has led to growing interest in this technology. The objective of this research was to evaluate the efficiency of immobilized laccase on magnetized multiwall carbon nanotubes (m-MWCNTs) in terms of stability and reusability. Laccases were efficiently adsorbed onto magnetized multiwall carbon nanotubes (m-MWCNTs) synthesized using water. The concentration of 7 mg laccase/mL was found to be ideal for immobilization. The optimal activity of both free and immobilized laccases was observed at pH 5, while for the latter, the optimal temperature was shifted from 40 to 50 °C. Compared to the free laccase, the immobilized laccase exhibited a greater range of stability at more extreme temperatures. At the fourth cycle of reactions, the immobilized laccase exhibited more than 60% relative activity in terms of reusability. Based on the fourier-transform infrared spectroscopy (FTIR) peak at 2921 cm−1, saccharification of paddy straw using immobilized laccase verified lignin degradation. The easy recovery of the immobilized laccase on m-MWCNTs lends credence to its potential use in biomass hydrolysis. [ABSTRACT FROM AUTHOR]

Copyright of Packaging & Food Machinery is the property of Packaging & Food Machinery Magazine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The complex mixture of pollutants found in industrial effluents creates a significant environmental challenge. Enzymatic treatment offers a promising alternative to traditional physiochemical techniques. With an emphasis on laccase enzymes, this review examines the potential of both free and immobilized biocatalysts for the treatment of industrial wastewater, especially from pulp and paper mills. Laccase has become one of the most popular options for bioremediation because of its broad substrate specificity and capacity to break down different kinds of organic pollutants. The different methods of laccase immobilization, including adsorption, encapsulation and cross-linking are examined, exploring their advantages and limitations. The benefits of immobilization for enzyme stability, reusability and operational efficiency in wastewater treatment processes are further discussed. By comparing the advantages and limitations of free and immobilized biocatalysts, this review aims to provide valuable insights for the development of efficient and sustainable enzymatic strategies for industrial wastewater treatment. Furthermore, this paper offers future possibilities for using biocatalyst in pulp and paper waste treatment. [ABSTRACT FROM AUTHOR]

This study investigated the oxidation of 5-hydroxymethylfurfural (HMF) to produce valuable products such as 2,5-furandicarboxaldehyde (DFF), 5-formyl-2-furancarboxylic acid (FFCA) and 2,5-furandicarboxylic acid (FDCA) through (photo) chemical and enzymatic catalysis. Laccases from different sources, including Aspergillus spp., Pseudomonas cepacea, and Trametes versicolor, were evaluated for their catalytic activity in the HMF oxidation. Laccase from Trametes versicolor (LacTV) emerged as the most effective catalyst, achieving complete conversion of HMF under specific pH conditions. The reaction mechanisms were explored, revealing a preference for the primary alcohol oxidation pathway over the aldehyde, leading to the formation of DFF and subsequent conversion to FFCA and FDCA. The impact of substrate concentration on HMF conversion was examined, revealing optimal conversion at lower HMF concentrations (<100 mM) and reduced performance at higher concentrations (>100 mM). The study also examined the influence of blue light (430 nm) on this reaction, revealing the dependence on light exposure for gC3N4 based catalysts and a negative effect for LacTV. Furthermore, the study introduces a modular flow chemical platform that utilizes Continuous Stirred Tank Reactors (CSTR) in a cascade configuration, the optimization of HMF oxidation. The implementation of this innovative approach has led to the effective synthesis of FDCA, demonstrating an impressive 40-fold increase in productivity compared to the traditional batch system. These results present significant potential for advancing green chemistry and sustainable chemical synthesis, introducing novel possibilities for environmentally friendly HMF oxidation processes. [ABSTRACT FROM AUTHOR]

Enzymatic polymerization of lignosulfonate (LS) has a high potential for various applications ranging from coatings to adhesives. Here, the effect of different ions in low concentrations on enzymatic polymerization of LS was investigated, including salt solutions consisting of mono‐ and dicarboxylic acids, sulfate, phosphate and chloride with sodium as counter ion. LS polymerization was followed by viscometry and size exclusion (SEC) chromatography. Interestingly, there was only a small effect of ions on the activity of the laccase on standard substrate ABTS, while the effect on polymerization of LS was substantially different. The presence of acetate led to a 39 % higher degree of polymerization (DP) for LS. Small angle X‐ray scattering (SAXS) revealed that the structure of the enzyme was largely unaffected by the ions, while the determination of the zeta potential showed that those ions conveying higher negative surface charges onto LS particles showed lower DPs, than those not affecting the surface charge. Further, electron paramagnetic resonance (EPR) spectroscopy showed 5‐times higher intensity in phenoxyl radicals for the monovalent ions compared to the divalent ones. It was concluded that the DPs of LS could be tuned in the presence of certain ions, by facilitating the interaction between the laccase substrate‐binding site and the LS molecules. [ABSTRACT FROM AUTHOR]

Background: Laccase (LAC) gene family plays a pivotal role in plant lignin biosynthesis and adaptation to various stresses. Limited research has been conducted on laccase genes in common beans. Results: 29 LAC gene family members were identified within the common bean genome, distributed unevenly in 9 chromosomes. These members were divided into 6 distinct subclades by phylogenetic analysis. Further phylogenetic analyses and synteny analyses indicated that considerable gene duplication and loss presented throughout the evolution of the laccase gene family. Purified selection was shown to be the major evolutionary force through Ka / Ks. Transcriptional changes of PvLAC genes under low temperature and salt stress were observed, emphasizing the regulatory function of these genes in such conditions. Regulation by abscisic acid and gibberellins appears to be the case for PvLAC3, PvLAC4, PvLAC7, PvLAC13, PvLAC14, PvLAC18, PvLAC23, and PvLAC26, as indicated by hormone induction experiments. Additionally, the regulation of PvLAC3, PvLAC4, PvLAC7, and PvLAC14 in response to nicosulfuron and low-temperature stress were identified by virus-induced gene silence, which demonstrated inhibition on growth and development in common beans. Conclusions: The research provides valuable genetic resources for improving the resistance of common beans to abiotic stresses and enhance the understanding of the functional roles of the LAC gene family. [ABSTRACT FROM AUTHOR]

In this work, we present a comprehensive investigation of the entrapment of laccase, a biotechnologically relevant enzyme, into levan-based nanoparticles (LNPs). The entrapment of laccase was achieved concomitantly with the synthesis of LNP, catalyzed by a truncated version of a levansucrase from Leuconostoc mesenteroides. The study aimed to obtain a biocompatible nanomaterial, able to entrap functional laccase, and characterize its physicochemical, kinetic and thermal stability properties. The experimental findings demonstrated that a colloidal stable solution of spherically shaped LNP, with an average diameter of 68 nm, was obtained. An uniform particle size distribution was observed, according to the polydispersity index determined by DLS. When the LNPs synthesis was performed in the presence of laccase, biocatalytically active nanoparticles with a 1.25-fold larger diameter (85 nm) were obtained, and a maximum load of 243 μg laccase per g of nanoparticle was achieved. The catalytic efficiency was 972 and 103 (μM·min)-1, respectively, for free and entrapped laccase. A decrease in kcat values (from 7050 min-1 to 1823 min-1) and an increase in apparent Km (from 7.25 μM to 17.73 μM) was observed for entrapped laccase, compared to the free enzyme. The entrapped laccase exhibited improved thermal stability, retaining 40% activity after 1 h-incubation at 70°C, compared to complete inactivation of free laccase under the same conditions, thereby highlighting the potential of LNPs in preserving enzyme activity under elevated temperatures. The outcomes of this investigation significantly contribute to the field of nanobiotechnology by expanding the applications of laccase and presenting an innovative strategy for enhancing enzyme stability through the utilization of fructan-based nanoparticle entrapments. [ABSTRACT FROM AUTHOR]

Incorporation of aminated starch (AS)-polyphenol conjugates to improve corn starch (CS) film properties were explored. AS-polyphenols (gallic acid (GA), syringic acid (SA) and vanillic acid (VA)) conjugates were synthesized by laccase and the grafting rates of GA, SA and VA were 36.59, 69.71 and 68.85%, respectively. Incorporation of the three conjugates (AS-GA, AS-SA and AS-VA) into the corn starch film improved a variety of properties of the CS film. The CS film without the conjugates was smooth and flat, while surface of the CS film with the conjugates became relatively rough and wrinkled and there were some small particles on the surface. Incorporation of the conjugates improved the thermal stability of the CS film during the polymer depolymerization phase. The thickness of the CS film with AS-VA conjugates was significantly larger than that of the CS film and the incorporation of the conjugates improved the hydrophobicity of the CS film. The tensile strength (TS) of both the CS films with AS-GA conjugates and AS-VA conjugates was significantly enhanced, while the TS of the CS film with AS-SA conjugates was significantly reduced. The incorporation of AS-polyphenol conjugates could improve the antioxidant ability and antibacterial ability of the CS film, especially the CS films containing AS-GA conjugates. [ABSTRACT FROM AUTHOR]

This article discusses the issue of negative interference from calcium dobesilate in the Roche sarcosine oxidase creatinine assay and proposes a solution using CuO. The study found that calcium dobesilate can cause false decreased creatinine values in patients, which poses challenges to clinical diagnosis and treatment. The researchers conducted experiments using CuO as an oxidant to correct the interference of calcium dobesilate on serum creatinine measurements. The results showed that CuO effectively corrected the interference, making it a cost-effective and easily implementable solution compared to other methods. However, further testing is needed to determine its application in other detecting systems. [Extracted from the article]

This work investigated non-polar solvent hexane and polar solvents methanol and ethanol as inducers besides a well-known inducer, copper, for laccase production with and without mesoporous silica-covered plastic packing under sterilised and unsterilised conditions. The potential of waste-hexane water, which is generated during the mesoporous silica production process, was also investigated as a laccase inducer. During the study, the free and immobilised laccase activity on the packing was measured. The results showed that the highest total laccase activity, approximately 10,000 Units, was obtained under sterilised conditions with 0.5 mM copper concentration. However, no immobilised laccase activity was detected except in the copper and ethanol sets under unsterilised conditions. The maximum immobilised laccase activity of the sets that used waste hexane as an inducer was 1.25 U/mg packing. According to its significant performance, waste hexane can be an alternative inducer under sterilised conditions. Concomitant immobilised packing showed satisfactory laccase activities and could be a promising method to reduce operation costs and improve the cost-efficiency of enzymatic processes in wastewater treatment plants. [ABSTRACT FROM AUTHOR]

Bisphenol A diglycidyl ether (BADGE), a derivative of the well-known endocrine disruptor Bisphenol A (BPA), is a potential threat to long-term environmental health due to its prevalence as a micropollutant. This study addresses the previously unexplored area of BADGE toxicity and removal. We investigated, for the first time, the biodegradation potential of laccase isolated from Geobacillus thermophilic bacteria against BADGE. The laccase-mediated degradation process was optimized using a combination of response surface methodology (RSM) and machine learning models. Degradation of BADGE was analyzed by various techniques, including UV-Vis spectrophotometry, high-performance liquid chromatography (HPLC), Fourier transform infrared (FTIR) spectroscopy, and gas chromatography-mass spectrometry (GC-MS). Laccase from Geobacillus stearothermophilus strain MB600 achieved a degradation rate of 93.28% within 30 min, while laccase from Geobacillus thermoparafinivorans strain MB606 reached 94% degradation within 90 min. RSM analysis predicted the optimal degradation conditions to be 60 min reaction time, 80°C temperature, and pH 4.5. Furthermore, CB-Dock simulations revealed good binding interactions between laccase enzymes and BADGE, with an initial binding mode selected for a cavity size of 263 and a Vina score of -5.5, which confirmed the observed biodegradation potential of laccase. These findings highlight the biocatalytic potential of laccases derived from thermophilic Geobacillus strains, notably MB600, for enzymatic decontamination of BADGE-contaminated environments. [ABSTRACT FROM AUTHOR]

Laccase (LAC) is a potent multicopper oxidase that relies on O2 for its catalytic activity. LAC has been affirmed as an environmentally friendly biocatalyst that often catalyzes a wide array of phenolic substrates. Bacterial‐derived LACs have been less investigated for non‐phenolic substrates in contrast to fungi‐derived LAC. To comprehend the substrates (3,4‐Dimethoxybenzyl alcohol, and Dimer (Guaiacyl 4‐O‐5 guaiacyl) binding interactions of LAC (Thermus thermophilus HB27) was carried out and contrasted with fungal‐derived Lignin peroxidase (LiP) (Trametes cervina) exploiting computational methods, including physicochemical properties, Sequence Annotated by Structure (SAS), Extra precision docking (Glide), and DESMOND‐directed MD‐ simulation. Protein structures exhibited by LAC, and LiP have diverse dissimilar component architects. The XP docking suggested LiP‐Dimer seems to have a comparatively lowest binding affinity (−8.413 kcal/mol), with an MMGBSA score of −33.249 kcal/mol. Further, docked complexes were validated leveraging 50 ns NPT system‐based MD‐simulation for structural and functional stability. The system achieved equilibrium and stability at the end of the simulation, with only the LiP‐3,4‐Dimethoxybenzyl alcohol complex maintained stability. The results of this study offer a framework for improving the binding ability of substrates by way of the use of in‐silico protein engineering, which might eventually result in more effective catalytic applications. [ABSTRACT FROM AUTHOR]

Cell wall is involved in plant growth and plays pivotal roles in plant adaptation to environmental stresses. Cell wall remodelling may be crucial to salt adaptation in the euhalophyte Salicornia europaea. However, the mechanism underlying this process is still unclear. Here, full‐length transcriptome indicated cell wall‐related genes were comprehensively regulated under salinity. The morphology and cell wall components in S. europaea shoot were largely modified under salinity. Through the weighted gene co‐expression network analysis, SeXTH2 encoding xyloglucan endotransglucosylase/hydrolases, and two SeLACs encoding laccases were focused. Meanwhile, SeEXPB was focused according to expansin activity and the expression profiling. Function analysis in Arabidopsis validated the functions of these genes in enhancing salt tolerance. SeXTH2 and SeEXPB overexpression led to larger cells and leaves with hemicellulose and pectin content alteration. SeLAC1 and SeLAC2 overexpression led to more xylem vessels, increased secondary cell wall thickness and lignin content. Notably, SeXTH2 transgenic rice exhibited enhanced salt tolerance and higher grain yield. Altogether, these genes may function in the succulence and lignification process in S. europaea. This work throws light on the regulatory mechanism of cell wall remodelling in S. europaea under salinity and provides potential strategies for improving crop salt tolerance and yields. Summary Statement: SeXTH2 and SeEXPB enhance cell size, two SeLACs promote xylem differentiation and increase the secondary cell wall thickness, which contribute to plant salt tolerance. [ABSTRACT FROM AUTHOR]