The chemical profiling and assessment of antioxidative, antidiabetic and antineurodegenerative potential of Kombucha fermented Camellia sinensis, Coffea arabica and Ganoderma lucidum extracts.

The scientific interest in the medicinal properties of Kombucha beverages, a carbonated drink with live microorganisms, has increased recently. Hence, the aim of this study was to determine the chemical profile and to examine the antioxidant, antidiabetic and antineurodegenerative potential of unfermented and also Kombucha fermented Camellia sinensis (green tea), Coffea arabica (coffee), and Ganoderma lucidum (Reishi) extracts. The extracts were prepared as follows: the first (unfermented) set contained 1 L of water, 50 g of sucrose and 20 g of dried and ground green tea, coffee, or Reishi basidiocarp, while the second (fermented) set contained all of the aforementioned ingredients individually inoculated with Kombucha and fermented for 21 days. The chemical analysis was conducted using liquid chromatography-mass spectrometry (LC-MS). The antioxidant activity was assessed by DPPH, total reducing power (TRP), and ß-carotene bleaching assays. The inhibition of α-amylase and α-glucosidase activity was used to estimate the antidiabetic potential, while the level of inhibition of acetylcholinesterase (AChE) and tyrosinase (TYR) was used to evaluate the antineurodegenerative activity. The results suggested that the fermented extracts of green tea, coffee, and Reishi exert significant antioxidant effects, although they were lower compared to the unfermented extracts. The unfermented green tea extract exhibited the highest DPPH-scavenging activity (87.46%) and the highest preservation of ß-carotene (92.41%), while the fermented coffee extract showed the highest TRP (120.14 mg AAE per g) at 10 mg mL-1. Although the extracts did not inhibit the activity of α-amylase, they were quite effective at inhibiting α-glucosidase, especially the unfermented Reishi extract, inhibiting 95.16% (at a concentration of 10 mg mL-1) of α-glucosidase activity, which was slightly higher than the positive control at the same concentration. The most effective AChE inhibitor was unfermented green tea extract (68.51%), while the fermented coffee extract inhibited 34.66% of TYR activity at 10 mg mL-1. Altogether, these results are in accordance with the differences found in the extracts' chemical composition. Finally, this is the first report that highlights the differences in the chemical profile between the unfermented and Kombucha fermented green tea, coffee and Reishi extracts, while it also reveals, for the first time, the antineurodegenerative potential of Kombucha fermented Reishi extract. The examined extracts represent potent functional foods, while their more detailed mechanisms of action are expected to be revealed in future research.

Immunomodulatory Effects of Extract of Lingzhi or Reishi Medicinal Mushroom Ganoderma lucidum (Agaricomycetes) Basidiocarps Cultivated on Alternative Substrate.

Ganoderma lucidum is a medicinal mushroom exhibiting numerous health benefits primarily based on strong immunostimulatory effects. The study aimed to investigate if there were differences in effects of extracts of commercially (GC) and alternatively (wheat straw) (GA) cultivated G. lucidum basidiocarps on properties of peritoneal macrophages (PM) and monocyte-derived dendritic cells (MoDCs). Differences in immunomodulatory effects of GC/GA extracts were studied. The viability of treated PMs, their adhesive and phagocytic capability, and their capacity to produce reactive oxygen species (ROS) and NO were tested. Immature MoDCs generated from human monocytes were treated with poly I:C (10.0 µg/ml) and loxoribine (34.0 µg/ml), a selective TLR3 and TLR7 agonists, respectively, and with/without GC/GA extract (100.0 µg/ml). The effect of each combination on phenotypic properties, cytokines production by MoDCs, and their proliferation and Th polarizing capacity was studied. GA extract stimulated the metabolic and phagocytic activity of PMs, their adhesion capability, and ability to produce ROS and NO more strongly compared to GC. Both tested extracts significantly increased allostimulatory and Th1 polarization capacity of simultaneous TLR3 and TLR7-activated MoDCs, but GA extract was more effective. The extract of alternatively cultivated G. lucidum basidiocarps increased production of ROS and NO by TLR4 stimulated PMs and upregulated production of certain cytokines as well as allostimulatory and Th1 polarization capacity of MoDCs. GA extract could be a potent immunostimulatory agent for activation of MoDCs with the simultaneous engagement of TLRs, which seems to be a promising strategy for the preparation of DC-based anti-tumor vaccines.

From pomiculture waste to biotechnological raw material: efficient transformation using ligninosomes and cellulosomes from Pleurotus spp.

The goal of this study was to determine the capacity of Pleurotus spp. lignocellulosome to transform frequent pomiculture residues (grapevine-, plum-, and raspberry sawdust) into raw materials for biotechnological processes. All three lignocellulosics induced the synthesis of ligninolytic and cellulolytic enzymes in the tested species. Laccase was dominant in the ligninolytic cocktail, with a maximum activity of 40,494.88 U L-1 observed after the cultivation of P. pulmonarius on grapevine sawdust. Grapevine sawdust also proved to be the optimal substrate for the synthesis of versatile peroxidases especially in P. eryngii (1010.10 U L-1), while raspberry sawdust favored the production of Mn-dependent peroxidase in P. pulmonarius (479.17 U L-1). P. pulmonarius was the dominant cellulolytic agent and raspberry sawdust was optimal for the synthesis of xylanases, and endo- and exo-cellulases (15,746.35 U L-1, 9741.56 U L-1, and 836.62 U L-1), while grapevine sawdust mostly induced ß-glucosidase activity (166.11 U L-1). The degree of residues delignification was more substrate- than species-dependent, ranging between 6.44 and 23.72% after the fermentation of grapevine and raspberry sawdust with P. pulmonarius. On the other hand, the lowest level of cellulose consumption was also observed on raspberry sawdust after the cultivation of P. eryngii, which together with high delignification also induced the highest selectivity index (1.27). The obtained results show the exceptional lignocellulolytic potential of Pleurotus spp. enzyme cocktails which opens up many possibilities for their application in numerous biotechnological processes.

Obtaining Cellulose-Available Raw Materials by Pretreatment of Common Agro-Forestry Residues With Pleurotus spp.

The goals of the present study were to characterize the profile of ligninolytic enzymes in five Pleurotus species and determine their ability to delignify eight common agro-forestry residues. Generally, corn stalks were the optimal inducer of Mn-dependent peroxidase activity, but the activity peak was noted after wheat straw fermentation by P. eryngii (3066.92 U/L). P. florida was the best producer of versatile peroxidase, especially on wheat straw (3028.41 U/L), while apple sawdust induced the highest level of laccase activity in P. ostreatus (49601.82 U/L). Efficiency of the studied enzymes was expressed in terms of substrate dry matter loss, which was more substrate-than species-dependent. Reduction of substrate dry mass ranged between 24.83% in wheat straw and 8.83% in plum sawdust as a result of fermentation with P. florida and P. pulmonarius, respectively. The extent of delignification of the studied substrates was different, ranging from 51.97% after wheat straw fermentation by P. pulmonarius to 4.18% in grapevine sawdust fermented by P. ostreatus. P. pulmonarius was also characterized by the highest cellulose enrichment (6.54) and P. ostreatus by very low one (1.55). The tested biomass is a highly abundant but underutilized source of numerous value-added products, and a cocktail of ligninolytic enzymes of Pleurotus spp. could be useful for its environmentally and economically friendly transformation.

Pleurotus ostreatus and Laetiporus sulphureus (Agaricomycetes): Possible Agents against Alzheimer and Parkinson Diseases.

Previous studies showed that some mushrooms are highly efficient in inhibiting acetylcholinesterase and tyrosinase, the increased activity of which can trigger the development of Alzheimer and Parkinson diseases. Starting from the fact that free radicals at high concentrations could cause neurodegenerative disorders as well as great interest in new, natural antineurodegenerative drugs, the goal of this study was to determine the in vitro antioxidative and neuroprotective potentials of various Pleurotus ostreatus and Laetiporus sulphureus extracts. L. sulphureus was a better antioxidative agent; it showed higher reducing power, was a more efficient scavenger of DPPH and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radicals, and was an Fe3+ reducer. The most efficient acetylcholinesterase inhibitor was hot water extract of P. ostreatus fruiting body, which was slightly weaker than the commercial preparation, galantamine. However, in comparison with α-kojic acid, tested extracts were weaker tyrosinase inhibitors. Considering that tested extracts were rich in phenols and that their amounts were in positive correlation with the extent of radical neutralization and acetylcholinesterase and tyrosinase inhibition, it is assumed that these compounds are the potential carriers of the neuroprotective activities. Owing to the significant antioxidative and antineurodegenerative capacity of these species, they can be suggested as novel nutraceuticals and pharmaceuticals.

Stimulation of Wood Degradation by Daedaleopsis confragosa and D. tricolor.

Biological pretreatment of the lignocellulosic residues, in which white-rot fungi have a crucial role, has many advantages compared to the chemical, physical, and physico-chemical methods of delignification and therefore attracts increasing scientific attention. Regarding the fact that properties and capacities of the ligninolytic enzymes of Daedaleopsis spp. are still unknown, the aim of this study was to research how nitrogen sources and inducers affect the potential of Daedaleopsis confragosa and Daedaleopsis tricolor to degrade cherry sawdust. NH4NO3, (NH4)2SO4, and peptone were tested as nitrogen sources, while veratryl alcohol, p-anisidine, vanillic acid, and phenylmethylsulfonyl fluoride were the studied inducers. As Mn-dependent peroxidase and laccase were the leader enzymes and cherry sawdust/peptone medium the best stimulator of their activities, the effect of inducers on delignification potential of these species was studied during fermentation of that substrate. Veratryl alcohol was the best stimulator of laccase and phenylmethylsulfonyl fluoride of Mn-dependent peroxidase activity (27,610.0 and 1338.4 U/L, respectively). These inducers also increased cherry sawdust delignification selectivity, particularly in D. tricolor in the presence of phenylmethylsulfonyl fluoride (lignin:hemicellulose:cellulose = 32.1%:0.9%:11.7%). Owing to the presented results, studied species could have an important role in the phase of lignocellulose pretreatment in various biotechnological processes.

Neuroprotective Potential and Chemical Profile of Alternatively Cultivated Ganoderma lucidum Basidiocarps.

Various neurodegenerative diseases are the main challenges to the modern medicine and there is a great need for novel, natural, neuroprotective agents. Ganoderma lucidum is a well-known medicinal mushroom, which health benefits have been confirmed by numerous studies. As demand for its basidiocarps is increased and traditional cultivation on hardwoods is not environmentally friendly and economically justified, finding of alternative substrates is necessary. The aim of the study was to assess the effect of alternative cultivation substrates on the chemical profile of G. lucidum basidiocarps and their capacity to inhibit acetylcholinesterase and tyrosinase, which higher activity is directly associated with neurodegenerative processes. Extracts of basidiocarps cultivated on alternative substrates, especially on clear wheat straw, showed significantly higher inhibition capacities than extracts of commercially-grown ones. These extracts were considerably different chemically from commercial basidiocarps extracts and even nine new compounds were isolated from them. Our results suggest that cultivation substrate greatly affect the chemical profile and neuroprotective capacity of obtained basidiocarps and wheat straw is a promising cultivation substrate.

Mushrooms as Potent Sources of New Biofungicides.

BACKGROUND: Microfungi are causal agents of numerous diseases and disorders of agricultural plants, farm mushrooms and animals as well as human, which results are serious global reduction of the food amount, decrease of life quality, the severe life-threatening diseases and enormous economic losses. METHODS: In spite of organism innate ability to combat against pathogens, in invasions of some pathogens, support of additional antimycotic agents to defence system is required. Nowadays, common "fighters" against the microfungi are numerous synthetic fungicides that, besides benefits, have also side effects on host and environment and can cause the development of fungicide resistance in the pathogens. Therefore, the creation of new natural fungicides with different modes of action, strengthening the defense system and increase of organism resistance to pathogens are the main requirements of modern society. RESULTS: Numerous mushrooms produce chemically various intra- and extracellular metabolites with antifungal potential, among which the most potent ones are polysaccharides, proteins, and phenolic compounds. They act as immunostimulators, inhibitors of pathogen development and virulence and/or activators of pathogens` autolytic system. CONCLUSION: Therefore, mushroom-based antimycotic agents could be successfully applied in the diseases treatments as accessories or alternatives to commercial therapies and in such a way contribute to environmentally friendly combat against pathogens, i.e. decrease or complete substitution of commercial synthetic fungicides with natural ones.

Potential of selected fungal species to degrade wheat straw, the most abundant plant raw material in Europe.

BACKGROUND: Structural component of plant biomass, lignocellulose, is the most abundant renewable resource in nature. Lignin is the most recalcitrant natural aromatic polymer and its degradation presents great challenge. Nowadays, the special attention is given to biological delignification, the process where white-rot fungi take the crucial place owing to strong ligninolytic enzyme system. However, fungal species, even strains, differ in potential to produce high active ligninolytic enzymes and consequently to delignify plant biomass. Therefore, the goals of the study were characterization of Mn-oxidizing peroxidases and laccases of numerous mushrooms as well as determination of their potential to delignify wheat straw, the plant raw material that, according to annual yield, takes the first place in Europe and the second one in the world. RESULTS: During wheat straw fermentation, Lentinus edodes HAI 858 produced the most active Mn-dependent and Mn-independent peroxidases (1443.2 U L-1 and 1045.5 U L-1, respectively), while Pleurotus eryngii HAI 711 was the best laccase producer (7804.3 U L-1). Visualized bends on zymogram confirmed these activities and demonstrated that laccases were the dominant ligninolytic enzymes in the studied species. Ganoderma lucidum BEOFB 435 showed considerable ability to degrade lignin (58.5%) and especially hemicellulose (74.8%), while the cellulose remained almost intact (0.7%). Remarkable selectivity in lignocellulose degradation was also noted in Pleurotus pulmonarius HAI 573 where degraded amounts of lignin, hemicellulose and cellulose were in ratio of 50.4%:15.3%:3.8%. CONCLUSIONS: According to the presented results, it can be concluded that white-rot fungi, due to ligninolytic enzymes features and degradation potential, could be important participants in various biotechnological processes including biotransformation of lignocellulose residues/wastes in food, feed, paper and biofuels.

Species of Genus Ganoderma (Agaricomycetes) Fermentation Broth: A Novel Antioxidant and Antimicrobial Agent.

The bioactivity of Ganoderma lucidum basidiocarps has been well documented, but there are no data on the medicinal properties of its submerged cultivation broth nor on the other species of the genus Ganoderma. Thus the aim of this study was to test the potential antimicrobial and antioxidant activity of fermentation broth obtained after submerged cultivation of G. applanatum, G. carnosum, and G. lucidum. DPPH· scavenging ability, total phenols, and flavonoid contents were measured to determine the antioxidative potential of Ganoderma spp. fermentation filtrates, whereas their antimicrobial potential was studied using the microdilution method. DPPH· scavenging activity of G. lucidum fermentation filtrates was significantly higher than that of G. applanatum and G. carnosum, with the maximum (39.67%) obtained from strain BEOFB 432. This filtrate also contained the highest concentrations of phenols (134.89 µg gallic acid equivalents/mL) and flavonoids (42.20 µg quercetin equivalent/mL). High correlations between the activity and phenol content in the extracts showed that these compounds were active components of the antioxidative activity. G. lucidum strain BEOFB 432 was the most effective antibacterial agent, whereas strain BEOFB 434 has proven to be the most effective antifungal agent. The study showed that Ganoderma spp. fermentation filtrates are novel potent antioxidative and antimicrobial agents that could be obtained more quickly and cheaper than basidiocarps.

Induction of wheat straw delignification by Trametes species.

Wheat straw is the major crop residue in European countries which makes it the most promising material for bioconversion into biofuels. However, cellulose and hemicellulose are protected with lignin, so delignification is an inevitable phase in lignocellulose processing. The organisms predominantly responsible for its degradation are white-rot fungi and among them Trametes species represent promising degraders due to a well-developed ligninolytic enzyme system. Although numerous studies have confirmed that low molecular weight compounds can induce the production and activity of ligninolytic enzymes it is not clear how this reflects on the extent of delignification. The aim of the study was to assess the capacity of p-anisidine and veratryl alcohol to induce the production and activity of Mn-oxidizing peroxidases and laccases, and wheat straw delignification by six Trametes species. Significant inter- and intraspecific variations in activity and features of these enzymes were found, as well as differences in the potential of lignocellulose degradation in the presence or absence of inducers. Differences in the catalytic properties of synthesized enzyme isoforms strongly affected lignin degradation. Apart from enhanced lignin degradation, the addition of p-anisidine could significantly improve the selectivity of wheat straw ligninolysis, which was especially evident for T. hirsuta strains.

Genoprotective Capacity of Alternatively Cultivated Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), Basidiocarps.

Ganoderma lucidum is traditionally used in Eastern medicine to preserve vitality, promote longevity, and treat disease. It possesses immunomodulatory, antitumor, antimicrobial, and antiaging activities, among others, but one of the most important is its antioxidant property, which is the basis for other effects, because free radicals trigger many diseases. The substrate commonly used for commercial cultivation of G. lucidum is not environmentally friendly nor economically justified, so there is a need to find new alternative substrates. The aim of this study was to analyze the effect of substrate composition on the bioactivity of G. lucidum basidiocarps. G. lucidum was cultivated on 2 different substrates: (1) a mixture of wheat straw, grapevine branches, and wheat bran, and (2) wheat straw. Commercial fruiting bodies, cultivated on oak sawdust, were used as the control. 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability, total phenols, and flavonoid content were determined spectrophotometrically to define the antioxidative potential of basidiocarp extracts. The comet test was performed to detect the degree of DNA damage in the cells that were exposed to G. lucidum extracts before and after the effect of oxidants. Higher antioxidative potential was observed for the extract of G. lucidum basidiocarps cultivated on wheat straw compared with that from the mixed substrate and especially with commercial ones. The alternatively cultivated basidiocarps also showed stronger antigenotoxic potential compared with commercial ones. The study showed that fruiting bodies produced on wheat straw, one of the most accessible and cheapest crop residues, are more potent antioxidant and antigenotoxic agents than commercially cultivated ones.

Potential of Submergedly Cultivated Mycelia of Ganoderma spp. as Antioxidant and Antimicrobial Agents.

The study aimed to evaluate the antiradical and antimicrobial (antibacterial and antifungal) potentials of ethanol mycelial extracts of selected Ganoderma species and strains and to define interand intraspecies diversity among Ganoderma species and strains. Ganoderma lucidum strains were good DPPH• scavengers (neutralizing up to 57.12% radicals), contrary to G. applanatum (20.35%) and G. carnosum (17.04%). High correlations between the activities and contents of total phenols in the extracts showed that these compounds were carriers of the activity. Results obtained by both discdiffusion and microdilution methods indicated that the extract of G. lucidum BEOFB 433 was the most potent antibacterial agent that inhibited growth of almost all bacterial species at a concentration of 1.0 mg/mL. Salmonella typhimurium was the most sensitive species to the mycelium extracts. Extracts of G. lucidum BEOFB 431 and BEOFB 434 showed the best antifungal activity since in concentration of 0.5 mg/mL inhibited the growth of Aspergillus glaucus (BEOFB 431) and the growth of A. glaucus and Trichoderma viride (BEOFB 434). Extracts of G. applanatum and G. lucidum BEOFB 431 had the strongest fungicidal effects, with lethal outcomes for A. glaucus and T. viride, respectively, being noted at a concentration of 1.17 mg/mL. Aspergillus niger was proved as the most resistant species.

Effect of Selenium Enrichment of Lenzites betulinus and Trametes hirsuta Mycelia on Antioxidant, Antifungal and Cytostatics Potential.

The aim of the study was an assessment of the antioxidant, antifungal and cytotoxic potentials of L. betulinus and T. hirsuta mycelia extracts and the effect of selenium on these activities. Extracts of L. betulinus were twice as efficient in DPPH• scavenging as those of T. hirsuta. The phenol content in Se-enriched L. betulinus extracts was higher than in non-enriched extracts, in contrast to the effect of Se-enrichment on T. hirsuta extracts, and a direct correlation between the amount and DPPH• scavenging effect was observed. Ethanol extracts exhibited fungistatic but not fungicidal activity against a range of micromycetes, and mycelium enrichment with selenium inhibited this effect. Although the extracts showed low cytotoxic activity against HeLa and LS174 cells, T. hirsuta extracts, especially those enriched with selenium, had better potential. L. betulinuss extracts showed better antioxidant and antifungal activity than T. hirsuta extracts which were more active cytotoxic agents. The presence of selenium stimulated antioxidant and cytotoxic, and inhibited antifungal activity in L. betulinus, while in T. hirsuta its effect was slight.

Antioxidant and antifungal potential of Pleurotus ostreatus and Agrocybe cylindracea basidiocarps and mycelia.

Basidiocarps of Pleurotus ostreatus and Agrocybe cylindracea are characterized by high nutritional value and numerous medicinal activities, though bioactivities of their mycelia have not been sufficiently studied. The aim of the study was to evaluate antioxidant and antifungal potentials of P. ostreatus and A. cylindracea basidiocarp extracts, as well as those obtained from mycelia cultivated in common synthetic medium and media enriched with various agro-industrial residues. The free radical scavenging activity of the extracts was determined spectrophotometrically, based on DPPH• reduction while antifungal potential was studied by a microdilution method. The highest level of DPPH• scavenging ability was obtained by an extract of P. ostreatus mycelium cultivated in wheat bran-enriched medium, while control medium favoured the antioxidant potential of A. cylindracea mycelium. Phenol compounds were the main carriers of antioxidant activity that was demonstrated by high coefficients for correlations between total phenol contents in extracts and level of DPPH• scavenging (0.94 for P. ostreatus extracts and 0.91 for A. cylindracea extracts). Only the extracts of A. cylindracea basidiocarp and control mycelium, at a concentration of 1.33 mg/mL, inhibited the growth of Microsporum gypseum and Aspergillus flavus, while basidiocarp extract at 1.67 mg/mL also had fungicidal effect against M. gypseum. These results revealed a considerable antioxidant potential of submergedly-cultivated mycelium which showed higher antioxidant activity than basidiocarp extracts. This is very important because significant amounts of mycelium biomass could be obtained more easily, cheaper and in a more controllable way than basidiocarps cultivation.

Biological activity of Ganoderma lucidum basidiocarps cultivated on alternative and commercial substrate.

ETHNOPHARMACOLOGICAL RELEVANCE: Ganoderma lucidum (W. Curt.:Fr.) P. Karst. has been used for several thousand years in traditional medicine of the Far East for the treatment of a various diseases including: cancer, immunological disorders, hepatitis, arthritis, nephritis, bronchitis, asthma etc. Ganoderma species are natural sources of various antimicrobial, antioxidant and antitumor active compounds which can be alternative to commercial antibiotics, antimycotics, antioxidants and cytostatics. As the amount of wild Ganoderma lucidum basidiocarps is not sufficient, the aim of this study was to compare the bioactivity of basidiaocarps obtained on alternative substrate (wheat straw) with commercially cultivated ones. MATERIALS AND METHODS: Antimicrobial potential of the tested Ganoderma lucidum extracts was studied by disc-diffusion and microdilution method. Cytotoxic effect of tested extracts on two cancer cell lines was determined by microculture tetrazolium test. DPPH scavenging ability, total phenols and flavonoids content were measured in order to determine the antioxidative potential of Ganoderma lucidum extracts. RESULTS: Ganoderma lucidum BEOFB 434 extract was the most effective antibacterial agent, while commercial strain was the weakest. DPPH· scavenging activity of extracts of basidiocarps from alternative substrate was also significantly higher than that of commercially obtained basidiocarps. The extracts of Ganoderma lucidum BEOFB 431 exhibited the strongest cytotoxic activity significantly higher than those in commercial strain. CONCLUSION: The study showed that fruiting bodies produced on one of the most accessible and cheapest crop residue, wheat straw, are better antimicrobial, antioxidant and cytotoxic agents than those obtained on conventional substrate, oak sawdust.

Lignin degradation by selected fungal species.

As biological decomposition of plant biomass represents a popular alternative environmental-friendly and economically justified process, screening of ligninolytic enzyme systems of various fungal species is a topical study area. The goal of the study was to obtain clear insight into the dynamics of laccase, Mn-dependent peroxidase, and Mn-independent peroxidase activity and levels of wheat straw lignin degradation in seven wood-rotting fungi. The best laccase producers were Pleurotus ostreatus and Pleurotus eryngii. Lenzites betulinus and Fomitopsis pinicola were the best Mn-dependent peroxidase producers, and P. ostreatus the weakest one. The peak of Mn-independent peroxidase was noted in Dichomytus squalens, and the minimum value in P. ostreatus. The profiles of the three enzymes, obtained by isoelectric focusing, were variable depending on the species and cultivation period. D. squalens was the best lignin degrader (34.1% of total lignin amount), and P. ostreatus and P. eryngii the weakest ones (7.1% and 14.5%, respectively).

Oxidative stress and species of genus Ganoderma (higher Basidiomycetes).

Oxidative stress, which is a factor in the aging process and in a series of serious disorders, arises when the reactive oxygen or nitrogen species are produced in excess and the capacity of cellular antioxidant defense is insufficient to detoxify and remove them. An internal antioxidant system is not always active enough to protect the human body from oxidative stress and, therefore, it needs the help of either synthetic or natural antioxidants. Nowadays, there is a growing interest in the substitution of synthetic antioxidants, which could have toxic and mutagen effects, with natural antioxidants. Recent studies revealed that besides their high nutritional value, mushrooms have great potential as antioxidant agents. Species of the genus Ganoderma, especially G. lucidum, are well-known medicinal mushrooms that traditionally are used in the prevention and treatment of many diseases and possess appreciable antioxidant potential.

Morpho-physiological diversity between Lingzhi or Reishi medicinal mushroom Ganoderma lucidium (W. Curt.:Fr.) P. Karst. and G. carnosum pat.

Two weakly differentiated taxa, Ganoderma lucidum and G. carnosum, were compared in their sufficient morphological and physiological features. The obtained results showed that dimensions of basidiospores and pileocystidia were insignificantly different, while pore shape and dimensions have shown greater diversity with average diameter of 138.46 µm in G. carnosum and 238.34 µm in G. lucidum. Mycelial growth rate was higher in G. lucidum (8.39 mm day-1) than in G. carnosum (6.02 mm day-1). G. lucidum was also a slightly better producer of biomass and extracellular polysaccharides (28.16 g L-1 and 1.42 mg mL-1, respectively) than G. carnosum (23.68 g L-1 and 0.35 mg mL-1, respectively). However, a higher amount of synthesized intracellular polysaccharides was noted in G. carnosum than in G. lucidum (40.00 mg g-1 and 30.00 mg g-1 of dry biomass, respectively). Higher activity levels of Mn-oxidizing peroxidases were obtained in G. carnosum, while G. lucidum was a better laccase producer. In G. carnosum, corn stem/NH4NO3 medium with nitrogen concentration of 20 mM was the optimum for Mn-dependent peroxidase production (88.00 U L-1), while the highest versatile peroxidase activity was detected in the medium with grapevine sawdust and 10 mM of nitrogen (80.80 U L-1). Wheat straw was the best carbon source for laccase synthesis in G. lucidum (55.75 U L-1).