Isoelectric Point Of pH Research Articles

Heme-peroxidase from medicinal plant Artocarpus lakoocha: Purification, characterization and wound healing studies

Artocarpus lakoocha is a medicinal plant used in cosmetics and traditional remedies of many diseases. We have purified a protein from its latex using anion exchange chromatography and gel filtration, which resulted in 28.82% yield of protein, with specific activity of 10.8 units/mg. FPLC spectroscopy and isoelectrofocusing demonstrated that purified protein has molecular mass of 53 kDa with isoelectric point of pH 7.4. Isolated peroxidase consists of 17 tryptophan, 14 tyrosine, and 8 cysteine residues per molecule with extinction coefficient of 16.3 M−1 cm−1. The pH and temperature optima for this peroxidase are 6.0 and 55 °C respectively. MALDI-TOF/TOF was done and MASCOT analysis specified that isolated protein is a peroxidase. Isolated peroxidase shows oxidizing activity with various phenolic substrates like guaiacol, o-phenylenediamine, aminoantipyrine and pyrogallol with Kcat 1.90, 0.74, 3.22, 1.49 s−1 respectively. Stability experiments with chemical denaturants, metal ions and organic solvents indicate that isolated protein is very stable under studied conditions. Spectroscopic studies, like absorbance, fluorescence, and circular dichroism reveal that the peroxidase has α/β type secondary structure with approximately 42.6% α-helix, 9.7% β-sheet. In vitro scratch wound healing assay and oxidative damage protection experiments via MTT assay demonstrate that purified peroxidase have wound healing properties. In conclusion, this novel peroxidase is stable at various conditions and helps in healing which hints it׳s potential to be used in inflammation prevention, wound healing, and other biotechnological and industrial applications.

Purification strategies and properties of a low-molecular weight xylanase and its application in agricultural waste biomass hydrolysis

Xylanases are hydrolytic enzymes responsible for the depolymerization of xylan, and they are of great interest because of their wide industrial applications. Extracellular low molecular weight xylanase was purified from a submerged culture of a thermophilic fungus, Scytalidium thermophilum ATCC No. 16454 grown on corn cobs, and the enzyme was characterized. Among the investigated low-cost purification techniques, the highest recovery (79%) with 2.7-fold purification, was obtained using an aqueous two phase system. The maximal purity (4.3-fold) was achieved by ultrafiltration and generated a yield of 25%. A two-step chromatography procedure consisting of gel filtration and anion exchange without any additional pre-purification steps yielded ca. 21.8-fold xylanase purification to apparent homogeneity with 9.6% recovery as demonstrated by SDS-PAGE. This study is the first to report a complement purification procedure and the corresponding results for S. thermophilum low molecular weight xylanase isolation from culture medium. The enzyme has a molecular weight of 21kDa and an isoelectric point of pH 8.6, indicating that it belongs to GH family 11. The low molecular weight of the enzyme provides the important advantage of making it easy to access in the lignocellulosic network, and it efficiently degrades hemicellulose. The optimum temperature and pH values of purified xylanase were 65°C and 6.5, respectively. Xylanolytic activity was most stable at pH 7.0 and 40°C. Km and Vmax values of xylanase on beechwood xylan were predicted as 2.4mg xylan/ml and 168.6IU/ml, respectively. The potential of xylanase in biomass hydrolysis was also investigated, as lignocellulosic biomass has attracted increasing attention as an important raw material for second generation biofuel production. Among the investigated commercial and lignocellulosic substrates, purified xylanase showed the maximum specificity for beechwood xylan and wheat bran. The hydrolysis of an inedible and waste lignocellulosic substrate, corn cob, by xylanase was studied. A release of reducing sugars and apparent morphological changes in the corn cob structures were observed by SEM analysis after enzymatic treatment.

Determination of the second virial coefficient of bovine serum albumin under varying pH and ionic strength by composition-gradient multi-angle static light scattering.

Composition-gradient multi-angle static light scattering (CG-MALS) is an emerging technique for the determination of intermolecular interactions via the second virial coefficient B22. With CG-MALS, detailed studies of the second virial coefficient can be carried out more accurately and effectively than with traditional methods. In addition, automated mixing, delivery and measurement enable high speed, continuous, fluctuation-free sample delivery and accurate results. Using CG-MALS we measure the second virial coefficient of bovine serum albumin (BSA) in aqueous solutions at various values of pH and ionic strength of a univalent salt (NaCl). The systematic variation of the second virial coefficient as a function of pH and NaCl strength reveals the net charge change and the isoelectric point of BSA under different solution conditions. The magnitude of the second virial coefficient decreases to 1.13 x 10(-5) ml*mol/g(2) near the isoelectric point of pH 4.6 and 25 mM NaCl. These results illuminate the role of fundamental long-range electrostatic and van der Waals forces in protein-protein interactions, specifically their dependence on pH and ionic strength.

The leucine-rich amelogenin protein (LRAP) is primarily monomeric and unstructured in physiological solution.

Amelogenin proteins are critical to the formation of enamel in teeth and may have roles in controlling growth and regulating microstructures of the intricately woven hydroxyapatite (HAP). Leucine-rich amelogenin protein (LRAP) is a 59-residue splice variant of amelogenin and contains the N- and C-terminal charged regions of the full-length protein thought to control crystal growth. Although the quaternary structure of full-length amelogenin in solution has been well studied and can consist of self-assemblies of monomers called nanospheres, there is limited information on the quaternary structure of LRAP. Here, sedimentation velocity analytical ultracentrifugation (SV) and small angle neutron scattering (SANS) were used to study the tertiary and quaternary structure of LRAP at various pH values, ionic strengths, and concentrations. We found that the monomer is the dominant species of phosphorylated LRAP (LRAP(+P)) over a range of solution conditions (pH 2.7-4.1, pH 4.5-8, 50 mmol/L(mM) to 200 mM NaCl, 0.065-2 mg/mL). The monomer is also the dominant species for unphosphorylated LRAP (LRAP(-P)) at pH 7.4 and for LRAP(+P) in the presence of 2.5 mM calcium at pH 7.4. LRAP aggregates in a narrow pH range near the isoelectric point of pH 4.1. SV and SANS show that the LRAP monomer has a radius of ∼2.0 nm and an asymmetric structure, and solution NMR studies indicate that the monomer is largely unstructured. This work provides new insights into the secondary, tertiary, and quaternary structure of LRAP in solution and provides evidence that the monomeric species may be an important functional form of some amelogenins.

Gliadin-based nanoparticles: Fabrication and stability of food-grade colloidal delivery systems

There is great interest in converting gluten, a by-product from wheat starch isolation, into a value added functional food ingredient. The insolubility of monomeric gluten, i.e. gliadin, in water makes these proteins interesting materials to produce nanoparticles using liquid antisolvent precipitation. The effect of different production parameters (such as mixing speed, time, sonication parameters, gliadin concentration) on the properties of gliadin particles formed by liquid antisolvent precipitation was investigated. The produced particles were also hardened using glutaraldehyde (0–0.25%). The stability of the produced gliadin particles was investigated in different pH and salt conditions relevant for food processing and upon short term temperature treatments and isothermal long-term storage. Gliadin nanoparticles (diameter < 200 nm) could be produced, however, they had poor stability in different conditions relevant to food processing: they redissolved below pH 4.0 and flocculated near their isoelectric point (pH ≈ 6.5), at elevated salt levels, and after heat treatment (T > 40 °C for 30 min). A slight improvement in particle stability was obtained by chemical hardening (0.25% glutaraldehyde). Gliadin nanoparticles may be useful in the development of delivery systems to encapsulate, protect, target and release active ingredients during food processing or after ingestion. However, additional strategies to stabilize the particles should be explored.

Modification of Nickel Ferrite with Cationic Surfactant: Dye Removal from Textile Wastewater Using Magnetic Separation

AbstractIn this paper, the modification of nickel ferrite nanoparticles with cationic surfactant [cetyltrimethyl ammonium bromide (CTAB)] and its capability as a magnetic adsorbent for the removal of direct dyes (Direct Red 31, Direct Red 81, and Direct Red 80) from textile wastewater have been investigated. The Fourier-transformm infrared (FTIR) analysis, X-ray diffraction (XRD), and scanning electron microscopy (SEM) images, and the determination of isoelectric point (pH at point of zero charge), were used to analyze the morphology to characterize the synthesized magnetic adsorbent. The effect of operating parameters including pH, initial dye concentration, adsorbent dosage, and inorganic salts on the dye removal efficiency was studied. The results showed that varying the pH of solution did not affect the dye removal efficiency. It was also concluded that by increasing the adsorbent dosage the available adsorption sites and the dye removal efficiency would increase. Adsorption isotherms such as Langmuir...

Self-assembled pH-responsive films prepared from mussel anchoring threads.

The byssus is a series of collagen-rich fibers securing mussels to surfaces. The complex but elegant heterogeneous assembly of the various proteins in the threads is responsible for their remarkable mechanical properties combining strength and extensibility. Along with the well-known biocompatibility and biodegradability attributed to collagen-based materials, these mechanical properties are highly desirable to produce biomaterials for soft tissue engineering and drug delivery applications. In order to replicate the byssus natural features and properties, we prepared a soluble byssus protein hydrolyzate (BPH) that can generate water-insoluble self-standing films. Atomic force and scanning electron microscopy revealed the presence of self-assembled collagen-like fibrils at the surface of the films. Infrared spectroscopy analysis of the film formation showed that insolubility is caused by the self-assembly of polypeptides from the hydrolyzate into antiparallel β-sheets, aggregated β-strands and collagen triple-helix structures. The mechanical properties and water swelling measurements on the films can be reversibly pH-modulated by modifying the electrostatic interactions between the ∼30 mol% of charged amino acids. Optimal mechanical properties and minimum swelling are obtained at the isoelectric point (pH 4.5). Higher or lower pH treatment reversibly decreases their stiffness and strength and increases their swelling ratio. Altogether, our results show that byssus proteins are an interesting sustainable feedstock for preparing new solid-state pH-tuneable biomaterials.

Effect of nitrogen and temperature on the transcription of an ACAT gene in Isochrysis galbana.

Thiolases are functionally divided into two groups: 3-ketoacyl-CoA thiolase and acetoacetyl-CoA thiolase (ACAT). Acetoacetyl-CoA thiolase plays a key role in the mevalonate pathway. In this study, a novel gene, IgACAT, which encodes ACAT was cloned from Isochrysis galbana and characterized. The cDNA of IgACAT was 1551bp in length, consisting of an open reading frame of 1173bp, a 5' untranslated region of 69bp and a 3' untranslated region of 309bp. The deduced amino acid sequence of IgACAT was 390 amino acid residues in length with a predicted molecular weight of 53.59kDa and an isoelectric point of pH 9.04. The triterpenes content and the expression of IgACAT under nitrogen and temperature stress were analyzed. When I. galbana was treated with excessive nitrogen and at 35°C, respectively, both the triterpenes content and the abundance of IgACAT gene transcript increased. Our findings will facilitate the regulation of gene expression and genetic modification of the triterpenes synthesis pathway of I. galbana.

A phenylalanine ammonia-lyase ortholog (PkPAL1) from Picrorhiza kurrooa Royle ex. Benth: molecular cloning, promoter analysis and response to biotic and abiotic elicitors.

Picrorhiza kurrooa Royle ex Benth. is a highly reputed medicinal herb utilised in the preparation of a number of herbal drug formulations, principally due to the presence of novel monoterpene iridoid glycosides kenned as picrosides. Phenylalanine ammonia-lyase catalyses an important rate-limiting step in phenylpropanoid pathway and supplies precursors like cinnamic acid, vanillic acid, ferulic acid, etc., to a variety of secondary metabolites including picrosides. The imperilled status of P. kurrooa coupled with lack of information regarding biogenesis of picrosides necessitates deciphering the biosynthetic pathway for picrosides. In the present study, a PAL gene, designated PkPAL1 was isolated from P. kurrooa. The cDNA is 2312 bp in length, consisting of an ORF of 2142 bp encoding for a 713 amino acid protein having a predicted molecular weight of 77.66 kDa and an isoelectric point of pH 6.82. qRT-PCR analysis of various tissues of P. kurrooa showed that PkPAL1 transcript levels were highest in the leaves, consistent with picroside accumulation pattern. Using Genome walking, a 718 bp promoter region was also isolated resulting in identification of distinct cis-regulatory elements including TGA-element, TGACG-motif, CGTCA-motif, etc. qRT-PCR indicated up-regulation of PkPAL1 by methyl jasmonate, salicylic acid, 2,4-dicholorophenoxy acetic acid and UV-B elicitations that corroborated positively with the identified cis-elements within the promoter region. Moreover, altitude was found to have a positive effect on the PkPAL1 transcript levels, driving the expression of PkPAL1 abundantly. Based on docking analysis, we identified eight residues as potentially essential for substrate binding in PkPAL1.

Thermo-mechanical and hydrophilic properties of polysaccharide/gluten-based bioplastics

The influence of adding different polysaccharides (locust bean gum, LBG; methyl cellulose, MC; and carboxymethyl cellulose, CMC) to gluten-based biodegradable polymeric materials was assessed in this work. Gluten/polysaccharide/plasticiser bioplastics were prepared at different polysaccharide concentrations (0–4.5%) and pH values by mixing in a two-blade counter-rotating batch mixer (at 25°C under adiabatic conditions) and thermomoulding at 9MPa and 130°C. Bioplastic probes were evaluated through dynamic mechanical thermal analysis, tensile strength and water absorption capacity tests. Results pointed out that a moderate enhancement of the network structure may be achieved by adding polysaccharide at a pH close to the protein isoelectric point (pH 6), which also conferred a further thermosetting capacity to the system. Moreover, the addition of MC and CMC was found to significantly enhance material elongation properties. However, the presence of charges induced by pH leaded to a higher incompatibility between the polysaccharide and protein domains forming the composite. The pH value played a relevant role in the material water absorption, which significantly increased under acidic or basic conditions (particularly at pH 3).

Cloning and Expression Analysis of Sugarcane Alkaline/Neutral Invertase Gene SoNIN1

Neutral/alkaline invertase irreversibly catalyzes sucrose into glucose and fructose, and plays an important regulating role in plant growth and development. In the present study, the whole length of neutral/alkaline invertase gene cDNA was cloned and sequenced from leaf of sugarcane variety GT28 using RT-PCR(reverse transcription-polymerase chain reaction) and RACE(rapid amplification of cDNA ends), and named as SoNIN1. The gene consists of 2289 bp with an ORF(open reading frame) of 1812 bp encoding a polypeptide of 603 amino acids with calculated protein molecular weight of 67.79 kD, and an isoelectric point of pH 6.41. SoNIN1 gene encodes a protein that is close to that of Zea mays, Oryza sativa, and Lolium perenne, belonging to the same evolutionary branch. The putative protein contains the conserved domain of alkaline/neutral invertase, and no membrane-spanning domain and signal peptide at N-end. The gene promoter sequence was cloned by using genome walking, which contains the basic components of CAAT-boxes and TATA-boxes, as well as specific acting elements such as cis-regulatory element involved in meristem activation, methyl jasmonate response and circadian rhythm, plus drought-induced MYB binding sites, abscisic acid response element, gibberellin response element, etc. Real-time PCR results showed that expression of SoNIN1 could be detected in the roots, stalks, leaves, flowers and buds, among which the highest was found in leaves, the medium in buds, and the lowest in internode +1(enrolled by the top visible dewlap leaf). The expression of SoNIN1 gene in roots and leaves of sugarcane seedling was induced by PEG and NaCl.

Hybrid Nanomaterial Complexes for Advanced Phage-guided Gene Delivery.

Developing nanomaterials that are effective, safe, and selective for gene transfer applications is challenging. Bacteriophages (phage), viruses that infect bacteria only, have shown promise for targeted gene transfer applications. Unfortunately, limited progress has been achieved in improving their potential to overcome mammalian cellular barriers. We hypothesized that chemical modification of the bacteriophage capsid could be applied to improve targeted gene delivery by phage vectors into mammalian cells. Here, we introduce a novel hybrid system consisting of two classes of nanomaterial systems, cationic polymers and M13 bacteriophage virus particles genetically engineered to display a tumor-targeting ligand and carry a transgene cassette. We demonstrate that the phage complex with cationic polymers generates positively charged phage and large aggregates that show enhanced cell surface attachment, buffering capacity, and improved transgene expression while retaining cell type specificity. Moreover, phage/polymer complexes carrying a therapeutic gene achieve greater cancer cell killing than phage alone. This new class of hybrid nanomaterial platform can advance targeted gene delivery applications by bacteriophage.

Identification and Characterization of TEX101 in Bovine Epididymal Spermatozoa.

Several studies exhibit the presence of Ricinus Communis Agglutinin I (RCA) binding glycocalyx in mammalian spermatozoa. However, the molecular characterization of RCA binding glycocalyx in sperm membranes and its mechanism of action are poorly understood. The objective of the study was to identify and to characterize RCA binding glycoprotein of the bovine sperm plasma membranes (PM). Lectin blots of caput and cauda sperm PM revealed a 38 kDa polypeptide exhibiting the highest affinity to RCA among the several major RCA binding polypeptides. The 38 kDa RCA binding polypeptide of cauda sperm PM was purified and exhibited a charge train of three distinct spots with isoelectric points (pH 5.3 and 5.8). Proteomic identification yielded ten peptides that matched the sequence of Testis Expressed 101 protein (TEX101). Western blots data revealed that bovine sperm TEX101 is present in both testicular and epididymal sperm PM fractions. The native TEX101 polypeptide contains ~17 kDa N-linked oligosaccharides and the polypeptide is anchored to sperm membrane via a glycosylphosphatidylinositol lipid linkage. Immunofluorescence staining of sperm with anti-TEX101 demonstrated that the polypeptide is localized at the head of cauda sperm. Our biochemical results provide evidence on the presence of TEX101 in bovine epididymal sperm plasma membranes and may have a potential role in sperm-egg interaction.

Composition and properties of the surface of oil bodies recovered from Echium plantagineum

Neutral-lipids within oilseeds are most commonly stored in oil bodies, small spherical organelles with oleosin proteins inserted through a phospholipid monolayer. Oil bodies extracted from Echium plantagineum are highly enriched in polyunsaturated fatty acids and are stable to coalescence and oxidation. This stability has been attributed to the strong association between the phospholipid monolayer and oleosin proteins. To better understand this association the phospholipid fatty acyl groups of E. Plantagineum oil bodies were determined for the first time; a large proportion (≈70%) of saturated fatty acids were present, and this may aid in oleosin anchorage and thus contributes to oil body stability. The effect of oil body washing on surface charge was also observed (using turbidity, zeta and streaming potentials), and dependent on the washing protocol, E. Plantagineum oil bodies had an isoelectric point of pH 4–5. This is significantly different to pI values for oil bodies from a range of other seeds reported in the literature using isoelectric focusing; a possible explanation for this discrepancy is discussed.

Stability and cell uptake of calcium carbonate templated insulin microparticles

Therapeutic proteins are an integral part of today’s pharmaceutical practice, but they still present challenges from the drug delivery point of view. In this work, a new approach is studied based on hard templating for fabrication of microparticles composed of pure insulin, which may enable effective delivery, for instance pulmonary delivery. The approach is both simple and versatile: the protein particles are prepared by selective precipitation into porous CaCO3 microtemplates, followed by full decomposition of the template at the isoelectric point of the protein (pH 5.2). Control over the main material parameters (mechanical properties, porosity, morphology and stability at physiological conditions) are critical for the envisioned application in drug delivery. It is demonstrated that these critical parameters can be significantly tuned by a slight final pH variation around the isoelectric point (pH range 4–6) and by the denaturation degree of insulin. Electrostatic interactions and inter-protein crosslinking in the protein particles as well as their internal structure are considered, to explain the variation in the particle properties. The particle property parameters are explored using atomic force microscopy, optical microscopy and circular dichroism spectra. Finally, phagocytic clearance of the protein particles in vitro was studied to explore possible enhancements in particle fabrication to improve the efficiency of insulin delivery by inhalation.

Modification of montmorillonite with sodium sulfide for prebiotic chemistry studies: Characterization using spectroscopy methods and X-ray diffractometry

Changes in the mineral characteristics of montmorillonite (Mt) resulting from exposure to sodium sulfide (Na2S) or artificial seawater revealed the capacity for clays to absorb and protect amino acids or nucleic acids from hydrolysis or UV radiation on the prebiotic Earth. The pH of isoelectric point (pHiep) of the modified montmorillonite (MMt) increased probably due to the adsorption of Na+ from the solution of Na2S, and would likely enhance the adsorption of biomolecules that are not protonated at higher pH. However, the exchange of cations with Na+ from artificial seawater or Na2S solutions likely contributed to the observed decrease in the interlayer distance of all treated samples. The associated release of Fe3+ from treated Mt likely decreased the capacity for the clay to adsorb biomolecules. The amount of Fe2+ in solution decreased by almost 50% when Mt was mixed with artificial seawater, but this decrease was not observed when Mt exposed to Na2S and artificial seawater. Given the capacity for Fe2+ to react with nucleic acids, this protection could be important for establishing chemical environments on the prebiotic Earth.

Isolation and characterization of protein isolated from defatted cashew nut shell: Influence of pH and NaCl on solubility and functional properties

This work reports the isolation of protein from defatted cashew nut shell (CNS), with the crude protein product containing 91.07% protein. Under its natural conditions, the solubility of this protein isolate is comparable (74.02%) to that of mustard green meal protein. The solubility of the protein isolate decreases with decreasing pH, with the minimum solubility observed at its isoelectric point (pH 3). The water holding capacity, oil holding capacity, foaming capacity, foam stability, emulsifying capacity and emulsion stability were found to be 2.56 cm3 H2O/g protein, 4.28 cm3 oil/g protein, 76.88%, 70.98%, 62.0% and 79.0%, respectively. The profiles of these functional properties were determined with varying pH values and NaCl concentrations, and improved properties were observed in the alkaline pH range and in the presence of NaCl. Electrophoretic analysis showed that the high molecular weight protein globulin was the major protein in the protein isolate.

Cloning, expression and stress-respondent transcription of long-chain acyl-coenzyme A synthetase cDNA gene of Nannochloropsis gaditana and its involvement in the biosynthesis of eicosapentaenoic and decosahexaenoic acids

A novel cDNA gene, NgLACS, that encodes a long-chain acyl-CoA sythetase (LACS), was cloned from Nannochloropsis gaditana and characterized. The cDNA was 2,360bp in length, consisting of an ORF of 1,950bp, a 5'-untranslated region of 88bp and a 3'-untranslated region of 322bp. The deduced amino acid sequence of LACS was 649 amino acid residues in length with a predicted molecular weight of 71kDa and an isoelectric point of pH 7.8. When the alga was treated with excessive nitrogen and iron, and at 15°C, the proportion of long-chain polyunsaturated acyl-CoAs in the total acyl-CoAs and the abundance of NgLACS cDNA gene transcript were up-regulated. Over-expression of NgLACS in Saccharomyces cerevisiae caused the accumulation of eicosapentaenoic acid and docosahexaenoic acid.

Stabilization of Fish Oil‐in‐Water Emulsions with Oleosin Extracted from Canola Meal

International dietary guidelines advocate replacement of saturated and trans fat in food with unsaturated oils. Also, there is growing interest in incorporating highly unsaturated omega-3 oils in to food products due to beneficial health effects. A major obstacle to incorporating highly unsaturated oils in to food products is the extreme susceptibility to oxidative deterioration. Oil bodies were prepared from tuna oil, oleosin, and phospholipid mimicking natural oil bodies within oilseed. Oleosin was extracted from canola (Brassica napus) meal by solubilization in aqueous sodium hydroxide (pH 12) and subsequent precipitation at its isoelectric point of pH 6.5. The tuna oil artificial oil bodies (AOBs) readily dispersed in water to produce oil-in-water (o/w) emulsions, which did not coalesce on storage and were amenable to pasteurization using standard conditions. Accelerated oxidation studies showed that these AOB emulsions were substantially more resistant to lipid oxidation than o/w emulsions prepared from tuna oil using Tween40, sodium caseinate, and commercial canola protein isolate, respectively. There is potential to use commercial canola meal, which is cheap and abundant, as a natural source of oleosin for the preparation of physically and oxidatively stable food emulsions containing highly unsaturated oils.

The secretome of Trametes versicolor grown on tomato juice medium and purification of the secreted oxidoreductases including a versatile peroxidase

The present work was carried out with the aim to analyze the secretome of Trametes versicolor BAFC 2234 grown on tomato juice medium supplemented with copper and manganese. T. versicolor BAFC 2234 was selected among diverse wood dwelling agaricomycetes from Argentina by its ability to cause a strong white rot on hardwood and in addition to show high tolerance toward phenolic compounds. A considerable number of the identified proteins were related to the degradation/modification of lignocelluloses. Hydrolases, peroxidases and phenoloxidases were the most abundant enzymes produced under the above-mentioned culture conditions. The lignin-modifying oxidoreductases laccase, manganese peroxidase (MnP) and versatile peroxidase (VP) were successfully purified – the latter for the first time from T. versicolor. The native VP protein has a molecular mass of 45kDa and an isoelectric point of pH 3.7. The study clearly shows that complex plant-based media being rich in phenolics, such as tomato juice, can stimulate the secretion of a broad set of extracellular lignocellulolytic enzymes. Using such natural products as fungal culture media may give the opportunity to investigate plant biomass decomposition as well as the biodegradation of organic pollutants in an environment close to nature.