Effect of cultivation conditions on ligninolytic enzyme production by Ganoderma carnosum

Ganoderma carnosum has been found in Europe only, at coniferous trees and it is difficult to distinguish it morphologically from G. lucidum. Since G. carnosum has not been studied biochemically yet, the aim of this study was to analyse its ability to produce Mn-dependent peroxidase (MnP), versatile peroxidase (VP) and laccase (Lac) under conditions of solid-state fermentation of selected plant raw materials as carbon sources, in the presence of different nitrogen concentrations in the medium. Wheat straw, corn stem, oak and grapevine sawdust were the analysed plant raw materials. Nitrogen source in synthetic medium was NH4NO3 and its concentrations were: 10mM N and 20 mM N. Enzyme activity was determined spectrophotometrically, using ABTS and phenol red, as the substrates for Lac and Mn-oxidizing peroxidases, respectively. Maximum level of MnP activity (56.82 U/l) was obtained in the medium with wheat straw and nitrogen concentration of 10 mM. Best carbon source for VP production was grapevine sawdust at nitrogen concentration of 10 mM (80.80 U/l). The obtained Lac activity was very low in the medium with wheat straw (1.80 U/l), while it was not detected in the presence of other three analyzed plant raw materials. Maximum of total protein content (0.06 mgml-1) was noted in the medium where oak sawdust was carbon source and nitrogen concentration was 20 mM.Vrste roda Ganoderma su producenti mnogih biološki aktivnih supstanci pa su objekti proučavanja mnogih medicinskih i farmaceutskih studija. U novije vreme se sve više pažnje poklanja upoznavanju njihovog ligninolitičkog enzimskog sistema, u cilju uspešne primene u različitim biotehnološkim procesima. Za razliku od G. lucidum, čiji je ligninolitički sistem intenzivno proučavan, ostale vrste ovog roda su retko bile objekti istraživanja. G. carnosum se teško morfološki razlikuje od G. lucidum. i još uvek nije biohemijski proučena, pa je cilj našeg istraživanja bio analiza produkcije Mn-oksidujućih peroksidaza i lakaza u uslovima čvrste kultivacije, na različitim biljnim ostacima (pšenična slama, stabljike kukuruza, piljevina hrasta i vinove loze) u prisustvu azota u obliku NH4NO3 i u koncentracijama od 10 mM, odnosno 20 mM. Maksimum produkcije Mn-zavisne peroksidaze dobijen je na pšeničnoj slami pri koncentraciji azota od 10 mM (56.82 U/l). Piljevina vinove loze i koncentracija azota od 10 mM bili su optimalni za sintezu verzatil peroksidaze (80.80 U/l). Dobijena aktivnost lakaza je bila izuzetno niska u medijumu sa pšeničnom slamom (1.80 U/l), dok na ostalim biljnim ostacima nije zabeležena. Maksimalni sadržaj ukupnih proteina je bio najveći u medijumu sa piljevinom hrasta kao izvorom ugljenika i koncentracijom azota od 10 mM.Projekat ministarstva br. 14304

WHEAT STRAW CONVERSION BY ENZYMATIC SYSTEM OF GANODERMA LUCIDUM

The purpose of this study was to resolve the question of whether various nitrogen sources and concentrations affect characteristics of selected G. lucidum ligninolytic enzymes participating in wheat straw fermentation. This is the first study reporting the presence of versatile peroxidase activity in crude extract of G. lucidum culture, as well as isoforms profile of Mn-oxidizing peroxidases. NH4NO3 was the optimum nitrogen source for laccase and Mn-dependent peroxidase activity, while peptone was the optimum one for versatile peroxidase activity. Four bands with laccase activity were obtained by native PAGE and IEF separations from medium enriched with inorganic nitrogen source, and only two bands from medium containing organic source. Medium composition was not shown to affect isoenzyme patterns of Mn-oxidizing peroxidases. Four isoforms of Mn-dependent peroxidase and three of versatile peroxidase were obtained on native PAGE. By IEF separation, five isoforms of Mn-dependent peroxidase and only two of versatile peroxidase were observed. The results demonstrated that G. lucidum has potential for mineralization and transformation of various agricultural residues and should take more significant participation in large-scale biotechnological processes

Effect of cultivation conditions on ligninolytic enzyme production by Ganoderma carnosum

Ganoderma carnosum has been found in Europe only, at coniferous trees and it is difficult to distinguish it morphologically from G. lucidum. Since G. carnosum has not been studied biochemically yet, the aim of this study was to analyse its ability to produce Mn-dependent peroxidase (MnP), versatile peroxidase (VP) and laccase (Lac) under conditions of solid-state fermentation of selected plant raw materials as carbon sources, in the presence of different nitrogen concentrations in the medium. Wheat straw, corn stem, oak and grapevine sawdust were the analysed plant raw materials. Nitrogen source in synthetic medium was NH4NO3 and its concentrations were: 10mM N and 20 mM N. Enzyme activity was determined spectrophotometrically, using ABTS and phenol red, as the substrates for Lac and Mn-oxidizing peroxidases, respectively. Maximum level of MnP activity (56.82 U/l) was obtained in the medium with wheat straw and nitrogen concentration of 10 mM. Best carbon source for VP production was grapevine sawdust at nitrogen concentration of 10 mM (80.80 U/l). The obtained Lac activity was very low in the medium with wheat straw (1.80 U/l), while it was not detected in the presence of other three analyzed plant raw materials. Maximum of total protein content (0.06 mgml-1) was noted in the medium where oak sawdust was carbon source and nitrogen concentration was 20 mM.Vrste roda Ganoderma su producenti mnogih biološki aktivnih supstanci pa su objekti proučavanja mnogih medicinskih i farmaceutskih studija. U novije vreme se sve više pažnje poklanja upoznavanju njihovog ligninolitičkog enzimskog sistema, u cilju uspešne primene u različitim biotehnološkim procesima. Za razliku od G. lucidum, čiji je ligninolitički sistem intenzivno proučavan, ostale vrste ovog roda su retko bile objekti istraživanja. G. carnosum se teško morfološki razlikuje od G. lucidum. i još uvek nije biohemijski proučena, pa je cilj našeg istraživanja bio analiza produkcije Mn-oksidujućih peroksidaza i lakaza u uslovima čvrste kultivacije, na različitim biljnim ostacima (pšenična slama, stabljike kukuruza, piljevina hrasta i vinove loze) u prisustvu azota u obliku NH4NO3 i u koncentracijama od 10 mM, odnosno 20 mM. Maksimum produkcije Mn-zavisne peroksidaze dobijen je na pšeničnoj slami pri koncentraciji azota od 10 mM (56.82 U/l). Piljevina vinove loze i koncentracija azota od 10 mM bili su optimalni za sintezu verzatil peroksidaze (80.80 U/l). Dobijena aktivnost lakaza je bila izuzetno niska u medijumu sa pšeničnom slamom (1.80 U/l), dok na ostalim biljnim ostacima nije zabeležena. Maksimalni sadržaj ukupnih proteina je bio najveći u medijumu sa piljevinom hrasta kao izvorom ugljenika i koncentracijom azota od 10 mM.Projekat ministarstva br. 14304

Antioxidative and antimicrobial potentials of Parmelia saxatilis and Pseudoevernia furfuracea

Even though numerous lichen species possess significant medical potentials they are still unexplored, and particularly species and strains originating from Serbia. Therefore, the aim of this study was to evaluate the antioxidative and antimicrobial potential of ethanol extracts of Parmelia saxatilis and Pseudoevernia furfuracea collected in Serbia. The tested extracts were good scavengers of DPPH radicals, with capacities ranging from 14.76% to 79.76% in P. saxatilis and from 21.39% to 90.04% in P. furfuracea. In P. saxatilis level of DPPH• neutralisation was highly correlated with phenol content (r2 = 0.9981) and in P. furfuracea with amount of total flavonoides (r2 = 0.9641). The extract of P. furfuracea inhibited the growth of all tested microorganisms with exception of Aspergillus flavus, while P. saxatilis extract affected only growth of bacterial species. Among tested microorganisms, Staphylococcus aureus and Klebsiella pneumoniae were the most sensitive, while Enterococcus faecalis, Pseudomonas aeruginosa as well as micromycetes were the least sensitive to tested extracts. Because of these potentials and the fact that their long term usage does not have any negative side effects on organism and development of microbial resistance, the extracts could be included in conventional therapy. [Projekat Ministarstva nauke Republike Srbije, br. 173032

Ganoderma lucidum - from tradition to modern medicine

Ganoderma lucidum has a long tradition of use in folk medicine of the Far East, which is documented in the oldest Chinese pharmacopoeia, written in the first century B.C, declaring it a superior medicine. The healing properties of G. lucidum reflected on folk names such as: Reishi, Mannentake, Ling Zhi etc., which mean “herb of spiritual power”, “mushroom of immortality” or “10,000-year mushroom”, respectively. It has been known, for thousands of years, that this species extends life span, increases youthful vigour and vitality and it was used in the treatments of hepatitis, kidneys’ disease, hypertension, arthritis, asthma, bronchitis, arteriosclerosis, ulcers and various types of cancer. However, Western civilisation did not discover its healing properties until the 20th century. Modern scientific researches and numerous clinical trails, conducted in recent decades, have confirmed the ancient knowledge of Eastern nations and given them a scientific basis. These studies have demonstrated many biological activities of G. lucidum extracts and compounds, including: immunomodulating, antioxidative, cytotoxic, hypoglycaemic, anti-inflammatory, antiallergic, antimicrobial, etc. It has been reported that its extracts play important role in detoxification of the body and protecton of the liver, as well as in reducing cardiovascular problems, stress and anxiety. However, its most important effect is undoubtedly immunostimulating one as it is the basis of many other positive effects. The Japanese government introduced G. lucidum on the official list of auxiliary agents for the treatments of various cancers, Alzheimer’s disease, diabetes and chronic bronchitis. Many chemical components have been isolated from G. lucidum, but polysaccharides and terpenoids are the main carriers of its bioactivities

Ligninolytic enzyme production in Pleurotus eryngii depending on the medium composition and cultivation conditions

Pleurotus eryngii produced laccase (Lac) both under conditions of submerged fermentation (SF) and solid-state fermentation (SSF) using all of the investigated carbon and nitrogen sources, while significant peroxidases production occurred only under SSF conditions. The highest levels of Lac activity were found under SF conditions of dry ground mandarine peels (999.5 U/l). After purification of extracellular crude enzyme mixture of P. eryngii which was grown under SF conditions with dry ground mandarine peels it was revealed two peaks of Lac activity and one peak of activity against phenol red in absence of external Mn2+ which was very low (1.4 U/l). Results obtained by purification also showed that the levels of phenol red oxidation in absence of external Mn2+ were higher than phenol red oxidation levels in presence of external Mn2+. In the medium with the best carbon source for Lac production (dry ground mandarine peels), (NH4)2SO4, with a nitrogen concentration of 20 mM, was the most optimum nitrogen source among 8 investigated sources

Intraspecific Diversity within Ganoderma lucidum in the Production of Laccase and Mn-Oxidizing Peroxidases During Plant Residues Fermentation

Comparison of the potential for laccase and Mn-oxidizing peroxidases synthesis by ten strains of Ganoderma lucidum, originating from different worldwide areas, during solid-state fermentation of selected plant raw materials was the aim of this study. The great intraspecific variability in the production of analyzed enzymes as well as the dependence of the enzyme activity on plant raw materials were reported. The strain HAI 957 was the best laccase producer in the presence of corn stem, as a unique carbon source (129.46 U/L). The highest level of Mn-dependent peroxidase activity was noted after wheat straw fermentation by G. lucidum HAI 246 (78.64 U/L), while the maximal versatile peroxidase production (59.72 U/L) was observed in strain HAI 957 in the medium with oak sawdust.Ministry of Science and Technological Development of Serbia [143041

Intraspecific Diversity within Ganoderma lucidum in the Production of Laccase and Mn-Oxidizing Peroxidases During Plant Residues Fermentation

Comparison of the potential for laccase and Mn-oxidizing peroxidases synthesis by ten strains of Ganoderma lucidum, originating from different worldwide areas, during solid-state fermentation of selected plant raw materials was the aim of this study. The great intraspecific variability in the production of analyzed enzymes as well as the dependence of the enzyme activity on plant raw materials were reported. The strain HAI 957 was the best laccase producer in the presence of corn stem, as a unique carbon source (129.46 U/L). The highest level of Mn-dependent peroxidase activity was noted after wheat straw fermentation by G. lucidum HAI 246 (78.64 U/L), while the maximal versatile peroxidase production (59.72 U/L) was observed in strain HAI 957 in the medium with oak sawdust.Ministry of Science and Technological Development of Serbia [143041

A Comparative Assessment of the Potential of Polysaccharide Production and Intracellular Sugar Composition within Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (W.Curt.:Fr.)P. Karst. (Aphyllophoromycetideae)

Ganoderma lucidum is a well-known medicinal mushroom species in which polysaccharides are one of the major sources of biological activity. The species was considered as a species-complex due to significant variations in morphological, biochemical, and genetic features among populations with a worldwide distribution. This fact was the basis for setting the aim of this research: to study intraspecific diversity in polysaccharide production and intracellular sugar composition among selected G. lucidum strains. The presence of intraspecific diversity among 10 G. lucidum strains, from different areas worldwide, was noted. Values of produced mycelia biomass and intracellular polysaccharides were found in wide ranges (3.1-28.2 g L-1 and 20.0-53.3 mg g(-1), respectively), while differences in extracellular polysaccharide amounts were minor (0.2-1.5 mg mL(-1)). The significant quantitative and qualitative differences in intracellular sugar composition were noted. Glucose was the predominant sugar in almost all strains except one (HAI 447), where sucrose was dominant. The potential of polysaccharide production and intracellular sugar composition could be one more taxonomic criterion for strain characterization within G. lucidum. The differences in intracellular sugar composition and proportions could be reflected in features of produced polysaccharides and also in their biological activities.Ministry for Science and Technological Development of Serbia [173032, 173040

Antioxidative potential of daedaleopsis tricolor basidiocarps and mycelium

Although some members of the genus Daedaleopsis have been prized for their medicinal and spiritual powers since Neolithic times, modern science has not yet sufficiently dealt with their bioactivities. This study aims at defining the antioxidative ac­tivities of extracts of Daedaleopsis tricolor wild and cultivated basidiocarps and mycelium and assessing their dependance on substrate type. Ethanol extracts (at a concentrations from 0.25 mg/mL to 16.00 mg/mL) of mycelium and wild fruiting bodies showed a considerable antioxidative potential (88.65% and 81.57%, respectively), which was almost the same as the commercial antioxidant BHA (88.91%). These radical scavenging abilities were reflected in EC50 values, which were 12.45 mg/mL for the extract of cultivated basidiocarps, 8.29 mg/mL for the extract of wild basidiocarps, 7.93 mg/mL for mycelium one, and 0.10 mg/mL for commercial antioxidant. Despite the fact that phenol proportion in the extracts was no neg­ligible (between 20.41 μg GAE/mg of the extract of dry wild basidiocarps and 146.37 μg GAE/mg of the extract of dry cultivated basidiocarps), its correlation with antioxidative activity was moderate. Flavonoids, in significant concentration, were detected only in the extract of cultivated fruiting bodies (28.64 μg QE/mg of dry extract), but no correlation with radical scavenging capacity was noted. A remarkable antioxidant potential, especially of the submerged cultivated mycelium, put D. tricolor high on the list of promising new natural antioxidants. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 173032