Copper increases laccase gene transcription and extracellular laccase activity in Pleurotus eryngii KS004.

The white-rot fungus Pleurotus eryngii secretes various laccases involved in the degradation of a wide range of chemical compounds. Since the laccase production is relatively low in fungi, many efforts have been focused on finding ways to increase it, so in this study, we investigated the effect of copper on the transcription of the pel3 laccase gene and extracellular laccase activity. The results indicate that adding 0.5 to 2 mM copper to liquid cultures of P. eryngii KS004 increased both pel3 gene transcription and extracellular laccase activity in a concentration-dependent manner. The most significant increase in enzyme activity occurred at 1 mM Cu2+, where the peak activity was 4.6 times higher than in control flasks. Copper also induced the transcription of the laccase gene pel3. The addition of 1.5 and 2 mM Cu2+ to fungal culture media elevated pel3 transcript levels to more than 13-fold, although the rate of induction slowed down at Cu2+ concentrations higher than 1.5 mM. Our findings suggest that copper acts as an inducer in the regulation of laccase gene expression in P. eryngii KS004. Despite its inhibitory effect on fungal growth, supplementing cultures with copper can lead to an increased extracellular laccase production in P. eryngii.

Extracellular ligninases production and lignin degradation by Paenibacillus polymyxa.

Bacteria represent an attractive source for the isolation and identification of potentially useful microorganisms for lignin depolymerization, a process required for the use of agricultural waste. In this work, ten autochthonous bacteria isolated from straw, cow manure, and composts were characterized for potential use in the biodelignification of the waste. A comparison of the ability to degrade lignin and the efficiency of ligninolytic enzymes was performed in bacteria grown in media with lignin as a sole carbon source (LLM, 3.5g/L lignin-alkali) and in complex media supplemented with All-Ban fiber (FLM, 1.5g/L). Bacterial isolates showed different abilities to degrade lignin, they decreased the lignin concentration from 7.6 to 18.6% in LLM and from 11.1 to 44.8% in FLM. They also presented the activity of manganese peroxidase, lignin peroxidases, and laccases with different specific activities. However, strain 26 identified as Paenibacillus polymyxa by sequencing the 16S rRNA showed the highest activity of lignin peroxidase and the ability to degrade efficiently lignocellulose. In addition, P. polymyxa showed the highest potential (desirability ≥ 0.795) related to the best combination of properties to depolymerize lignin from biomass. The results suggest that P. polymyxa has a coordinated lignin degradation system constituted of lignin peroxidase, manganese peroxidase, and laccase enzymes.

Laccase-Catalyzed Synthesis of Added-Value Polymers from Cork and Grape Extracts.

The development of products from natural plant sources, including agriculture and food wastes, contributes significantly to the circular economy and global sustainability. Cork and grape wastes were employed as the primary sources in this study to obtain compounds of interest under mild extraction conditions. Laccase was applied to oxidize the cork and grape extracts, with the aim of producing value-added molecules with improved properties. Ultraviolet-visible (UV-vis) spectroscopy was assessed to monitor the oxidation process, and characterization of the end products was performed by matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) spectroscopy. The antioxidant and antiaging properties were evaluated by means of ABTS, DPPH, FRAP, and SPF testing. Overall, as compared to their monomeric counterparts, the polymeric compounds displayed remarkable antioxidant and antiaging characteristics after laccase oxidation, showing tremendous potential for applications in the food, pharmaceutical, cosmetic, and textile industries.

Characterization of white rot fungi from wood decayed for lignin degradation.

The present study was conducted to isolate and identify white rot fungi (WRF) from wood decayed and to determine their ability to produce lignin-modifying enzymes (LMEs), specifically laccase (Lac), lignin peroxidase (LiP), and manganese peroxidase (MnP), on solid and liquid media supplemented with synthetic dyes namely 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), azure B, and phenol red. A total of 23 isolates of WRF were isolated from decayed wood and identified as eight different species namely Phanerochaete australis, Perenniporia tephropora, Lentinus squarrosulus, Ganoderma australe, Trametes polyzona, Lentinus sajor-caju, Gymnopilus dilepis, and Fomitopsis palustris based on morphological characteristics, DNA sequences of the internal transcribed spacer (ITS) region, and phylogenetic inference. The fungal isolates can be divided into four groups based on the type of LMEs produced, namely A (Lac-LiP-MnP) with 16 isolates, B (Lac-MnP) (three isolates), C (Lac) (three isolates), and D (MnP) (one isolate). This study highlights P. australis (BJ38) as the best producer of Lac and LiP, while L. squarrosulus (IPS72) is the best producer of MnP. The present study is the first reported P. australis as an efficient lignin degrader by demonstrating the highest activity of two important LMEs.

The Involvement of the Laccase Gene Cglac13 in Mycelial Growth, Germ Tube Development, and the Pathogenicity of Colletotrichum gloeosporioides from Mangoes.

Colletotrichum gloeosporioides is one of the most serious diseases that causes damage to mangoes. Laccase, a copper-containing polyphenol oxidase, has been reported in many species with different functions and activities, and fungal laccase could be closely related to mycelial growth, melanin and appressorium formation, pathogenicity, and so on. Therefore, what is the relationship between laccase and pathogenicity? Do laccase genes have different functions? In this experiment, the knockout mutant and complementary strain of Cglac13 were obtained through polyethylene glycol (PEG)-mediated protoplast transformation, which then determined the related phenotypes. The results showed that the knockout of Cglac13 significantly increased the germ tube formation, and the formation rates of appressoria significantly decreased, delaying the mycelial growth and lignin degradation and, ultimately, leading to a significant reduction in the pathogenicity in mango fruit. Furthermore, we observed that Cglac13 was involved in regulating the formation of germ tubes and appressoria, mycelial growth, lignin degradation, and pathogenicity of C. gloeosporioides. This study is the first to report that the function of laccase is related to the formation of germ tubes, and this provides new insights into the pathogenesis of laccase in C. gloeosporioides.

Comprehensive analysis of the laccase gene family in tea plant highlights its roles in development and stress responses.

BACKGROUND: Laccase (LAC) is the pivotal enzyme responsible for the polymerization of monolignols and stress responses in plants. However, the roles of LAC genes in plant development and tolerance to diverse stresses are still largely unknown, especially in tea plant (Camellia sinensis), one of the most economically important crops worldwide. RESULTS: In total, 51 CsLAC genes were identified, they were unevenly distributed on different chromosomes and classified into six groups based on phylogenetic analysis. The CsLAC gene family had diverse intron-exon patterns and a highly conserved motif distribution. Cis-acting elements in the promoter demonstrated that promoter regions of CsLACs encode various elements associated with light, phytohormones, development and stresses. Collinearity analysis identified some orthologous gene pairs in C. sinensis and many paralogous gene pairs among C. sinensis, Arabidopsis and Populus. Tissue-specific expression profiles revealed that the majority of CsLACs had high expression in roots and stems and some members had specific expression patterns in other tissues, and the expression patterns of six genes by qRT‒PCR were highly consistent with the transcriptome data. Most CsLACs showed significant variation in their expression level under abiotic (cold and drought) and biotic (insect and fungus) stresses via transcriptome data. Among them, CsLAC3 was localized in the plasma membrane and its expression level increased significantly at 13 d under gray blight treatment. We found that 12 CsLACs were predicted to be targets of cs-miR397a, and most CsLACs showed opposite expression patterns compared to cs-miR397a under gray blight infection. Additionally, 18 highly polymorphic SSR markers were developed, these markers can be widely used for diverse genetic studies of tea plants. CONCLUSIONS: This study provides a comprehensive understanding of the classification, evolution, structure, tissue-specific profiles, and (a)biotic stress responses of CsLAC genes. It also provides valuable genetic resources for functional characterization towards enhancing tea plant tolerance to multiple (a)biotic stresses.

Copper-Guanosine Nanorods (Cu-Guo NRs) as a Laccase Mimicking Nanozyme for Colorimetric Detection of Rutin.

Inspired by laccase activity, herein, Cu-guanosine nanorods (Cu-Guo NRs) have been synthesized for the first time through a simple procedure. The activity of the Cu-Guo NR as the laccase mimicking nanozyme has been examined in the colorimetric sensing of rutin (Rtn) by a novel and simple spectrophotometric method. The distinct changes in the absorbance signal intensity of Rtn and a distinguished red shift under the optimum condition based on pH and ionic strength values confirmed the formation of the oxidized form of Rtn (o-quinone) via laccase-like nanozyme activity of Cu-Guo NRs. A vivid and concentration-dependent color variation from green to dark yellow led to the visual detection of Rtn in a broad concentration range from 770 nM to 54.46 µM with a limit of detection (LOD) of 114 nM. The proposed methodology was successfully applied for the fast tracing of Rtn in the presence of certain common interfering species and various complex samples such as propolis dry extract, human biofluids, and dietary supplement tablets, with satisfactory precision. The sensitivity and selectivity of the developed sensor, which are bonuses in addition to rapid, on-site, cost-effective, and naked-eye determination of Rtn, hold great promise to provide technical support for routine analysis in the real world.

Oil body extraction from high-fat and high-protein soybeans by laccase cross-linked beet pectin: physicochemical and oxidation properties.

BACKGROUND: Soybean oil bodies (SOB) are droplets of natural emulsified oil. Soybean oil emulsifies well but it is easily oxidized during storage. Beet pectin is a complex anionic polysaccharide, which can be adsorbed on the surface of liposomes to improve their resistance to flocculation. Laccase can covalently cross-link ferulic acid in beet pectin, and its structure is irreversible, which can improve the stability of polysaccharides. RESULTS: At pH 2.5, laccase cross-linked beet pectin high-oil soybean oil body (HOSOB) and high-protein soybean oil body (HPSOB) emulsions showed obvious aggregation and severe stratification, and the oxidation of the emulsions was also high. The flocculation of emulsions decreased with an increase in the pH. The effect of pH on the flocculation of emulsion was confirmed by confocal laser electron microscopy. The ζ potential, emulsification, and rheological shear force increased with increasing pH whereas the particle size and surface hydrophobicity decreased with increasing pH. CONCLUSION: This experiment indicates that the physicochemical stability of the two composite emulsions was strongly affected under acidic conditions but stable under neutral and weakly alkaline conditions. Under the same acid-base conditions, the degree of oxidation of HPSOB composite emulsion changes substantially. The results of this study can provide a basis for the design of very stable emulsions to meet the demand for natural products. © 2023 Society of Chemical Industry.

Characterization of the biosurfactant production and enzymatic potential of bacteria isolated from an oil-contaminated saline soil.

Biosurfactants are amphiphilic compounds with extensive applications in oily contaminated environments to remove hydrocarbons. Moreover, enzymes such as laccase and manganese peroxidase are responsible for the oxidation of a variety of phenolic compounds and aromatic amines. Therefore, in the present study, bacteria with the potential to produce biosurfactants and enzymes (namely, laccase, manganese peroxidase, and endoglucanase carboxymethyl cellulose (CMCase)) were isolated from petroleum oil-contaminated soil. From 15 isolated bacteria, three isolates were selected as the best producers of biosurfactants according to the related tests, such as tests for surface tension reduction. These three bacteria indicated tolerance to a salinity test and were classified as resistant and very resistant. The isolates 3, 12, 13, and 14 showed positive results for the degradation of guaiacol, phenol red, and carboxymethylcellulose, as well as the decoloration of methylene blue by the creation of a clear halo around the bacterial colony. Upon the quantitation of the laccase and manganese peroxidase activities, 22.58 U/L and 21.81 U/L, respectively, were measured by isolate 13. Furthermore, CMCase activity was recorded with 0.057436 U/ml belonging to isolate 14. Bacterial strains with appreciable laccase, peroxidase, CMCase activity, and biosurfactant production potentials were identified through 16S rDNA sequence analysis as Bacillus sp. (isolate 3), Bacillus toyonensis (isolate 12), Bacillus cereus (isolate 13), and Bacillus tropicus (isolate 14), and their nucleotide sequences were deposited in the GenBank. The potentials for the industrial applicability of the biosurfactants and enzymes abound, and production needs to be optimized by the selected bacterial strains.

The preparation of soy glycinin/sugar beet pectin complex network gels catalyzed by laccase under weakly acidic conditions.

BACKGROUND: Traditional soy protein gel products such as tofu, formed from calcium sulfate or magnesium chloride, have poor textural properties and water retention capacity. Soy glycinin (SG) is the main component affecting the gelation of soy protein and can be cross-linked with polysaccharides, such as sugar beet pectin (SBP), and can be modified by changing system factors (e.g., pH) to improve the gel's properties. Soy glycinin/sugar beet pectin (SG/SBP) complex double network gels were prepared under weakly acidic conditions using laccase cross-linking and heat treatment. The structural changes in SG and the properties of complex gels were investigated. RESULTS: Soy glycinin exposed more hydrophobic groups and free sulfhydryl groups at pH 5.0. Under the action of laccase cross-linking, SBP could promote the unfolding of SG tertiary structures. The SG/SBP complex gels contained 46.77% ß-fold content and had good gelling properties in terms of hardness 290.86 g, adhesiveness 26.87, and springiness 96.70 mm at pH 5.0. The T22 relaxation time had the highest peak, and magnetic resonance imaging (MRI) showed that the gel had even water distribution. Scanning electron microscopy (SEM) and confocal scanning laser microscopy (CLSM) indicated that the SG/SBP complex network structure was uniform, and the pore walls were thicker and contained filamentous structures. CONCLUSION: Soy glycinin/ sugar beet pectin complex network gels have good water-holding, rheological, and textural properties at pH 5.0. The properties of soy protein gels can be improved by binding to polysaccharides, with laccase cross-linked, and adjusting the pH of the solution. © 2022 Society of Chemical Industry.

Transcriptome Analysis of Podoscypha petalodes Strain GGF6 Reveals the Diversity of Proteins Involved in Lignocellulose Degradation and Ligninolytic Function.

The present study reports transcriptomic profiling of a Basidiomycota fungus, Podoscypha petalodes strain GGF6 belonging to the family Podoscyphaceae, isolated from the North-Western Himalayan ranges in Himachal Pradesh, India. Podoscypha petalodes strain GGF6 possesses significant biotechnological potential as it has been reported for endocellulase, laccase, and other lignocellulolytic enzymes under submerged fermentation conditions. The present study attempts to enhance our knowledge of its lignocellulolytic potential as no previous omics-based analysis is available for this white-rot fungus. The transcriptomic analysis of P. petalodes GGF6 reveals the presence of 280 CAZy proteins. Furthermore, bioprospecting transcriptome signatures in the fungi revealed a diverse array of proteins associated with cellulose, hemicellulose, pectin, and lignin degradation. Interestingly, two copper-dependent lytic polysaccharide monooxygenases (AA14) and one pyrroloquinolinequinone-dependent oxidoreductase (AA12) were also identified, which are known to help in the lignocellulosic plant biomass degradation. Overall, this transcriptome profiling-based study provides deeper molecular-level insights into this Basidiomycota fungi, P. petalodes, for its potential application in diverse biotechnological applications, not only in the biofuel industry but also in the environmental biodegradation of recalcitrant molecules. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-022-01037-6.

Improved laccase production from Pleurotus floridanus using deoiled microalgal biomass: statistical and hybrid swarm-based neural networks modeling approach.

Fungal laccases are versatile biocatalyst and occupy a prominent place in various industrial applications due to its broad substrate specificity. The simplest method to enhance the laccase production is by usage of cheap substrates in the fermentation processes incorporating modeling approaches for optimization. Integrated biorefinery concept is receiving wide popularity by making use of various products from microalgal biomass. The research aimed to identify the potential of deoiled microalgal biomass (DMB), a waste product from algal biorefinery as a nutrient supplement to enhance laccase production in Pleurotus floridanus by submerged fermentation. The maximum production was obtained in the presence of DMB as an additional nutrient supplement and copper sulfate as an inducer. The predictive capabilities of the two methodologies Response Surface Methodology (RSM) and hybrid Particle swarm optimization (PSO)-based Artificial Neural Network (ANN) were compared and validated. The results showed that ANN coupled with PSO predicted with more accuracy with an R 2 value of 0.99 than the RSM model with an R 2 value of 0.97. The optimized condition as predicted by superior model hybrid PSO-based ANN was glucose (3.51%), DMB (0.545%), pH (4.9), temperature (24.68 â„ƒ) and CuSO4 (1.35 mM). The experimental laccase activity was 80.45 ± 0.132 U/mL which was 1.3 fold higher than unoptimized condition. This study promotes the usage of DMB as a novel supplement for the improved production of Pleurotus floridanus laccase. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03404-y.

Overexpression of SmLAC25 promotes lignin accumulation and decreases salvianolic acid content in Salvia miltiorrhiza.

Laccase (LAC) is a blue multicopper oxidase that contains four copper ions, which is involved in lignin polymerization and flavonoid biosynthesis in plants. Although dozens of LAC genes have been identified in Salvia miltiorrhiza Bunge (a model medicinal plant), most have not been functionally characterized. Here, we explored the expression patterns and the functionality of SmLAC25 in S. miltiorrhiza. SmLAC25 has a higher expression level in roots and responds to methyl jasmonate, auxin, abscisic acid, and gibberellin stimuli. The SmLAC25 protein is localized in the cytoplasm and chloroplasts. Recombinant SmLAC25 protein could oxidize coniferyl alcohol and sinapyl alcohol, two monomers of G-lignin and S-lignin. To investigate its function, we generated SmLAC25-overexpressed S. miltiorrhiza plantlets and hairy roots. The lignin content increased significantly in all SmLAC25-overexpressed plantlets and hairy roots, compared with the controls. However, the concentrations of rosmarinic acid and salvianolic acid B decreased significantly in all the SmLAC25-overexpressed lines. Further studies revealed that the transcription levels of some key enzyme genes in the lignin synthesis pathway (e.g., SmCCR and SmCOMT) were significantly improved in the SmLAC25-overexpressed lines, while the expression levels of multiple enzyme genes in the salvianolic acid biosynthesis pathway were inhibited. We speculated that the overexpression of SmLAC25 promoted the metabolic flux of lignin synthesis, which resulted in a decreased metabolic flux to the salvianolic acid biosynthesis pathway.

Pectin-based microgels for rheological modification in the dilute to concentrated regimes.

HYPOTHESIS: A novel range of microgel particles of different internal cross-linking densities can be created by covalently cross-linking sugar beet pectin (SBP) with the enzyme laccase and mechanically breaking down the subsequent parent hydrogels to sugar beet pectin microgels (SBPMG) via shearing. The bulk rheological properties of suspensions of the different SBPMG are expected to depend on the microgel morphology, elasticity (crosslinking density) and volume fraction respectively. EXPERIMENTS: The rheology of both dilute and concentrated dispersions of SBPMG were studied in detail via capillary viscometry and shear rheometry, supplemented by information on particle size and shape from static light scattering, confocal microscopy and electron microscopy. FINDINGS: For dilute suspensions of SBPMG, data for viscosity versus effective volume fraction (ɸeff) falls on a 'master' curve for all 3 types of SBPMG. In the more concentrated regime, the softer microgels allow greater packing and interpenetration and give lower viscosities at the same ɸeff, but all 3 types of microgel give much higher viscosities than the equivalent concentration of 'non-microgelled' pectin. The firmer microgels can be concentrated to achieve elasticities equivalent to the original parent hydrogel. All SBPMG suspensions were extremely shear thinning but showed virtually no time-dependence.

Microbial degradation and transformation of benzo[a]pyrene by using a white-rot fungus Pleurotus eryngii F032.

Polycyclic aromatic hydrocarbons (PAHs) are environmentally recalcitrant contaminants formed from naturally or incomplete combustion of organic materials and some of them are difficult to degrade due to their hydrophobicity and persistency. Benzo [a]pyrene (BaP), is one of PAHs that having five fused benzene and reported as mutagenic, carcinogenic and teratogenic compounds. Biodegradation is one of promising techniques due to its relatively low economic cost and microorganism is a natural capacity to consume hydrocarbons. In this investigation, Pleurotus eryngii F032 was grown in 20 mL of modified mineral salt broth (MSB) supplemented with BaP under static and agitated culture. Within 20 days, static culture removed 59% of BaP, whereas agitated culture removed the highest amount (73%). To expedite BaP elimination, the mechanism and behavior of BaP biosorption and biotransformation by Pleurotus eryngii F032 were additionally examined by gas chromatography-mass spectrometer (GC-MS). The optimal conditions for P. eryngii F032 to eliminate BaP were 25 °C, a C/N ratio of 8, pH 3 and 0.2% inoculum concentration. At an initial BaP content of 10 mg/L, more than 50% was effectively eliminated within 20 days under these conditions. Salinity, glucose, and rhamnolipids were the most important factors impacting BaP biodegradation. GC-MS found degradation products such as BaP-3,6-quinone, indicating plausible metabolic routes. Finally, it may be assumed that the primary mechanism by which white-rot fungi eliminate BaP is by the utilization of biotransformation enzymes such as laccase to mineralize the PAHs. Hence, Pleurotus eryngii F032 could be an ideal candidate to treat PAHs contaminated soils.

Assessment of the Antioxidant Activity of Catechin in Nutraceuticals: Comparison between a Newly Developed Electrochemical Method and Spectrophotometric Methods.

The analysis of antioxidants in different foodstuffs has become an active area of research, which has led to many recently developed antioxidant assays. Many antioxidants exhibit inherent electroactivity, and, therefore, the use of electrochemical methods could be a viable approach for evaluating the overall antioxidant activity of a matrix of nutraceuticals without the need for adding reactive species. Green tea is believed to be a healthy beverage due to a number of therapeutic benefits. Catechin, one of its constituents, is an important antioxidant and possesses free radical scavenging abilities. The present paper describes the electrochemical properties of three screen-printed electrodes (SPEs), the first one based on carbon nanotubes (CNTs), the second one based on gold nanoparticles (GNPs) and the third one based on carbon nanotubes and gold nanoparticles (CNTs-GNPs). All three electrodes were modified with the laccase (Lac) enzyme, using glutaraldehyde as a cross-linking agent between the amino groups on the laccase and aldehyde groups of the reticulation agent. As this enzyme is a thermostable catalyst, the performance of the biosensors has been greatly improved. Electro-oxidative properties of catechin were investigated using cyclic voltammetry (CV) and differential pulse voltammetry (DPV), and these demonstrated that the association of CNTs with GNPs significantly improved the sensitivity and selectivity of the biosensor. The corresponding limit of detection (LOD) was estimated to be 5.6 × 10-8 M catechin at the CNT-Lac/SPE, 1.3 × 10-7 M at the GNP-Lac/SPE and 4.9 × 10-8 M at the CNT-GNP-Lac/SPE. The biosensors were subjected to nutraceutical formulations containing green tea in order to study their catechin content, using CNT-GNP-Lac/SPE, through DPV. Using a paired t-test, the catechin content estimated was in agreement with the manufacturer's specification. In addition, the relationship between the CNT-GNP-Lac/SPE response at a specific potential and the antioxidant activity of nutraceuticals, as determined by conventional spectrophotometric methods (DPPH, galvinoxyl and ABTS), is discussed in the context of developing a fast biosensor for the relative antioxidant activity quantification.

Co-cultivation of filamentous microorganisms in the presence of aluminum oxide microparticles.

In the present work, the approaches of submerged co-cultivation and microparticle-enhanced cultivation (MPEC) were combined and evaluated over the course of three case studies. The filamentous fungus Aspergillus terreus was co-cultivated with Penicillium rubens, Streptomyces rimosus, or Cerrena unicolor in shake flasks with or without the addition of aluminum oxide microparticles. The influence of microparticles on the production of lovastatin, penicillin G, oxytetracycline, and laccase in co-cultures was compared with the effects recorded for the corresponding monocultures. In addition, the quantitative analyses of morphological parameters, sugars consumption, and by-products formation were performed. The study demonstrated that the influence of microparticles on the production of a given molecule in mono- and co-culture may differ considerably, e.g., the biosynthesis of oxytetracycline was shown to be inhibited due to the presence of aluminum oxide in "A. terreus vs. S. rimosus" co-cultivation variants but not in S. rimosus monocultures. The differences were also observed regarding the morphological characteristics, e.g., the microparticles-induced changes of projected area in the co-cultures and the corresponding monocultures were not always comparable. In addition, the study showed the importance of medium composition on the outcomes of MPEC, as exemplified by lovastatin production in A. terreus monocultures. Finally, the co-cultures of A. terreus with a white-rot fungus C. unicolor were described here for the first time. KEY POINTS: • Aluminum oxide affects secondary metabolites production in submerged co-cultures. • Mono- and co-cultures are differently impacted by the addition of aluminum oxide. • Effect of aluminum oxide on metabolites production depends on medium composition.

Laccase production by Pleurotus ostreatus using cassava waste and its application in remediation of phenolic and polycyclic aromatic hydrocarbon-contaminated lignocellulosic biorefinery wastewater.

The treatment of contaminants from lignocellulosic biorefinery effluent has recently been identified as a unique challenge. This study focuses on removing phenolic contaminants and polycyclic aromatic hydrocarbons (PAHs) from lignocellulosic biorefinery wastewater (BRW) applying a laccase-assisted approach. Cassava waste was used as a substrate to produce the maximum yield of laccase enzyme (3.9 U/g) from Pleurotus ostreatus. Among the different inducers supplemented, CuSO4 (0.5 mM) showed an eight-fold increase in enzyme production (30.8 U/g) after 240 h of incubation. The catalytic efficiency of laccase was observed as 128.7 ± 8.47 S-1mM-1 for syringaldazine oxidation at optimum pH 4.0 and 40 °C. Laccase activity was completely inhibited by lead (II) ion, mercury (II) ion, sodium dodecyl sulphate, sodium azide and 1,4 dithiothretiol and induced significantly by manganese (II) ion and rhamnolipid. After treating BRW with laccase, the concentrations of PAHs and phenolic contaminants of 1144 µg/L and 46160 µg/L were reduced to 96 µg/L and 16100 µg/L, respectively. The ability of laccase to effectively degrade PAHs in the presence of different phenolic compounds implies that phenolic contaminants may play a role in PAHs degradation. After 240 h, organic contaminants were removed from BRW in the following order: phenol >2,4-dinitrophenol > 2-methyl-4,6-dinitrophenol > 2,3,4,6-tetrachlorophenol > acenaphthene > fluorine > phenanthrene > fluoranthene > pyrene > anthracene > chrysene > naphthalene > benzo(a)anthracene > benzo(a)pyrene > benzo(b)fluoranthene > pentachlorophenol > indeno(1,2,3-cd)pyrene > benzo(j) fluoranthene > benzo[k]fluoranthène. The multiple contaminant remediation from the BRW by enzymatic method, clearly suggests that the laccase can be used as a bioremediation tool for the treatment of wastewater from various industries.

Immobilization of Laccase Through Inorganic-Protein Hybrids Using Various Metal Ions.

In this study, the inorganic-protein hybrid strategy was employed for immobilization of laccase from Rhus vernicifera (Rvlac) using various metals calcium, cobalt, copper, and zinc (Zn). The efficient synthesis of hybrids for Rvlac immobilization was noted at 4 °C for incubation of 24 h. Among these hybrids, the maximum encapsulation yields (EY) of 90.1% and relative activity (RA) of 225% to free enzyme were recorded for Zn and Rvlac based inorganic-protein hybrids as Zn3(PO4)2-Rvlac. The upper optimum pH, and temperature values were observed of 4.0, and 45 °C after immobilization as compared to 3.5, and 40 °C for the free enzyme, respectively. After encapsulation, Rvlac showed a significant improvement up to 11.4-fold in pH and 5.7-fold in temperature the activity profiles. Free enzyme completely lost its activity at 60 °C after 2 h of incubation, whereas Zn3(PO4)2-Rvlac retained its residual activity of 56.7% under similar conditions. After ten cycles of reusability, Zn3(PO4)2-Rvlac possessed high residual activity of 90.8%. This study showed that the variation in the metal ions for immobilization of Rvlac as inorganic-protein hybrids significantly altered EY and RA. Also, Zn3(PO4)2-Rvlac proved more efficient as compared to free laccase that can be beneficially employed for biotechnological applications. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-022-01000-5.

Degradation of sulfonamides in aquaculture wastewater by laccase-syringaldehyde mediator system: Response surface optimization, degradation kinetics, and degradation pathway.

As a new type of environmental pollutant, environmental antibiotic residues have attracted widespread attention, and the degradation and removal of antibiotics has become an engaging topic for scholars. In this paper, Novozym 51003 industrialized laccase and syringaldehyde were combined to degrade sulfonamides in aquaculture wastewater. Design Expert10 software was used for multiple regression analysis, and a response surface regression model was established to obtain the optimal degradation parameters. In the actual application, the degradation system could maintain a stable performance within 9 h, and timely supplement of the mediator could achieve a better continuous degradation effect. Low concentrations of heavy metals and organic matter would not significantly affect the degradation performance of the laccase-mediator system, making the degradation system suitable for a wide range of water quality. Enzymatic reaction kinetics demonstrated a strong affinity of sulfadiazine to the substrate. Ten degradation products were speculated using high-resolution mass spectrum based on the mass/charge ratios and the publication results. Four types of possible degradation pathways of sulfadiazine were deduced. This work provides a practical method for the degradation and removal of sulfonamide antibiotics in actual sewage.