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Effect of selected plant raw materials and nitrogen concentrations on Lac activity. Gray bar 10 mM nitrogen concentration, white bar 20 mM nitrogen concentration, circle specific activity (data represent mean value of activities of three different samples. Variations are given as standard errors)
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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...
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To get a better insight into the ligninolytic system of Lenzites betulinus, the effect of wheat straw and oak sawdust, as carbon sources, on production of Mn-oxidizing peroxidases and laccase, under solid-state and submerged fermentation, was studied. Obtained results revealed considerable differences related to the both factors affecting enzyme ac...
Citations
... As G.resinaceum mushrooms rare in nature and traditional cultivation on logs is not an ecologically and economically justified method, current study is finding of alternative media for production. Whether wheat straw, paper wastes, poplar trees and rice hulls, the most abundant materials in Iraq could be form (with CaCO3, CaSO4 and sucrose) a novel media for G. resinaceum cultivation, so not all components suitable for G. resinaceum cultivation i.e. oak sawdust, was less favorable for ligninolytic enzymes activities that are responsible for efficient medium degradation and mushroom biomass production [21]. ...
Ganoderma was isolated from an apple tree in the Tikrit region (Iraq), according to phenotypic characters and phylogenetic analyses based on internal transcribed spacer ITS 1 - 5.8S - ITS4 rDNA sequences, this isolate identified as G. resinaceum and registered with accession number MN448375 in the NCBI Database. We described a novel media for artificial cultivation of G. resinaceum, composed of wheat straw as basic substrate amended with premix (paper wastes, poplar sawdust and rice hulls at 1:1:1 (W:W:W) the most suitable medium was wheat straw amended with 20 % of premix, in this medium the highest yield (biological efficiency) of G. resinaceum basidiocarps was recorded. The cultural G. resinaceum characteristics grown in all tested media were also describted. HIGHLIGHTS Isolation and identification of the fungus Ganoderma resinaceum was very important for the purpose of adding this fungus to the mycoflora The production of the fruit bodies of Ganoderma resinaceum is one of the important economic factors, as these fruits are the raw source for the production of many medicinal materials The components of the nutrient medium that are uniquely mentioned in this study are the main key to the growth of this mushroom and the production of its fruiting bodies The production of mushroom Ganoderma resinaceum using agricultural waste, such as wheat straw and paper waste, contributes to the environmental cleaning of these wastes and enters the recycling of resources to produce useful products GRAPHICAL ABSTRACT
... Differences in the levels of ligninolytic enzyme activity on the same substrate could be explained by the genetic basis, since significant differences were previously observed in different species/strains of the genus Ganoderma. 14,25,30,32 The genetic predisposition for laccase synthesis was also confirmed by isoelectric focusing of active isoforms of this enzyme in G. lucidum strains grown on the same substrate. 19,32 In addition to the substrate composition and the species/strain selection, the type of cultivation also has a significant effect on the level of ligninolytic activity. ...
... 14,25,30,32 The genetic predisposition for laccase synthesis was also confirmed by isoelectric focusing of active isoforms of this enzyme in G. lucidum strains grown on the same substrate. 19,32 In addition to the substrate composition and the species/strain selection, the type of cultivation also has a significant effect on the level of ligninolytic activity. Thus, Songulashvili et al. 14 recorded extremely high laccase activity in G. ...
"The present study aimed to determine the capacity of Ganoderma lucidum and G. tsugae for pretreatment of common agroforestry residues. G. lucidum dominated in Mn-dependent peroxidase activity (1625.00 U L-1) after corn stalks fermentation, while the peak of Mn-independent peroxidase activity (2458.33 U L-1) was detected on raspberry sawdust fermented by G. tsugae. Laccases were dominant in the ligninolytic cocktail, with the maximal activity of 42480.09 U L-1 synthetized by G. lucidum cultivated on plum sawdust. The reduction of the lignocellulosic dry matter ranged between 11.00% and 33.00% in oak sawdust and wheat straw, respectively, fermented by G. tsugae, while the highest extent of lignin, cellulose and hemicellulose consumption (54.78%, 40.30% and 54.91%, respectively) was obtained on corn stalks. On the other hand, the smallest cellulose removal (6.26%) was detected after raspberry sawdust fermentation by G. lucidum, which, besides moderate ligninolysis, induced the highest selectivity index (2.87). The obtained differences in enzyme activities and lignocellulosics degradation extent could be the consequence of the different composition of the studied residues, especially macro- and microelements content. The obtained results clearly indicated the huge potential of G. lucidum and G. tsugae enzyme cocktails for the transformation of lignocellulosic residues that present abundant sources of numerous value-added products."
... U L -1 after only 15 days of oak sawdust solid-state fermentation. This could be explained either by the fact that laccase activity decreases with cultivation period which was already shown in previous reports (Ćilerdžić et al. 2014 or by the influence of medium composition. ...
Grifola frondosa HAI 1232 was tested for ligninolytic enzyme activities and for lignin, cellulose and hemicellulose degradation during cultivation on eight common agro-forestry residues in Serbia. Wheat straw was favorable lignocellulosic for the production of Mn-dependent and Mn-independent peroxidases (2513.89 and 354.17 U L-1, respectively), while selected residues inhibited the synthesis of laccases. The highest lignin removal was observed during fermentation of blackberry sawdust (36.75%), while the highest selectivity index was recorded on oak sawdust (4.34). The dry matter loss varied between 8.17% in corn stalks and 14.16% in apple sawdust. According to the presented results, it can be concluded that G. frondosa HAI 1232 could be an important participant in various biotechnological processesdue to its high capacity to selectively degrade different agro-forestry residues.
... 1,8 The most efficient participants in pretreatment, the first phase in the process of bioethanol production, are white-rot fungi, due to their remarkable capacity to produce ligninolytic enzymes, which oxidize lignin and facilitate access to holocellulose. 9 However, the efficiency of these enzymes varies from species to species, even from strain to strain, and depends on cultivation conditions. [10][11][12] Therefore, the most effective and selective lignin remover and optimized conditions for the production of its most active enzyme isoforms should be found. ...
Hypsizygus marmoreus is an edible and medicinal commercially cultivated mushroom, whose ligninolytic potential is still insufficiently known. Therefore, the goals of this study were to profile its Mn-oxidizing peroxidases and laccases activity and its potential to depolymerize common agro-forestry residues. Raspberry and blackberry sawdust induced the production of highly active laccases (26006.83 U L-1 and 17576.79 U L-1, respectively), while the activity peaks of Mn-dependent peroxidase (134.47 U L-1) and Mn-independent peroxidase (184.40 U L-1) were recorded on plum sawdust and wheat straw, respectively. The pretreatment of corn stalks resulted in the highest lignin removal (57.15%), while the highest lignin/cellulose removal ratio (selectivity) was noted on oak sawdust (17.69). The maximal (20%) and the minimal (8.30%) dry matter loss were obtained for corn stalks and oak sawdust, respectively. The presented results showed the exceptional potential of the ligninolytic system of H. marmoreus HAI 605 and the possibility of its application in numerous biotechnological processes.
... So far, a large number of fungi have been confirmed as producers of laccases, especially Basidiomycetes and Ascomycetes fungi. Four acidic laccase isoforms have been detected in Ganoderma lucidum BEOFB 431 [3], and recently, an extracellular laccase (LccH) from the newlyisolated Basidiomycetous fungus Hexagonia hirta MSF2 was also reported [4]. ...
Laccases are multi-copper oxidases with important industrial value. In the study, a novel laccase gene (mco) in a Staphylococcus haemolyticus isolate is identified and heterologously expressed in Escherichia coli. Mco shares less than 40% of amino acid sequence identities with the other characterized laccases, exhibiting the maximal activity at pH 4.0 and 60 °C with 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) as a substrate. Additionally, the Mco is tolerant to a wide range of pH, heavy metal ions and many organic solvents, and it has a high decolorization capability toward textile dyes in the absence of redox mediators. The characteristics of the Mco make this laccase potentially useful for industrial applications such as textile finishing. Based on a Blastn results, mco is found to be widely distributed in both the bacterial genome and bacterial plasmids. Its potential role in oxidative defense ability of staphylococci may contribute to the bacterial colonization and survival.
... Since laccases are glycoproteins, another source of variability comes from different posttranslational glycosylation levels [16]. Purified and electrophoretic homogeneous Lac 3.2 presents a relative molecular mass of 60 kDa, and an isoelectric point of 4.6, which are within the values reported for fungal laccases [17,18]. The effects of pH and temperature on its catalytic activity reveal the same optimum pH of 3.0 for ABTS or DMP, while, higher than previously reported [16,19,20] for other Ganoderma laccases, 70°C is its optimum reaction temperature (Fig. 1). ...
Using laccase as a case study, an alternative methodology for discrimination of bisubstrate mechanism types, useful for Bi-Bi reactions when water is one of the products, is presented. A purified enzyme from fungal cultures of Ganoderma resinaceum and two reducing substrates: 2,2´-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) and 2,6-dimethoxyphenol (DMP) are used. Kinetic assays of ABTS or DMP oxidation using variable initial concentrations of dioxygen are performed in a closed system and depletion rates of dioxygen are measured using polarography. Seven bisubstrate models based on Michaelis-Menten Bi-Bi mechanisms are applied to the experimental data. Akaike information criterion, a penalized likelihood analysis, and Akaike weights are used for model selection. Results show that the more likely bisubstrate models for ABTS and DMP oxidation are ping-pong and Theorell-Chance, respectively. However, the catalytic efficiency of oxygen reduction to water exhibits similar values, and is compatible with a process relatively independent of reducing substrates and the bisubstrate mechanism type. Finally, Michaelis-Menten constants towards dioxygen, 233 μM with ABTS and 700 μM with DMP, indicate that the soluble electron acceptor available for laccase-catalyzed reactions, is far removed from saturation.
... A similar effect was described by D'Souza et al. [27], which showed that although the presence of Pinus strobus resulted in laccase production, the addition of syringic acid as an inducer tripled laccase activity. On the other hand, Ćilerdžić et al. [28] showed that both wheat-straw and oak sawdust were able to induce laccase activity on G. lucidum in submerged and solid-state fermentation, without the addition of other inducers. ...
Laccases are multicopper oxidases with high potential for industrial applications. Several basidiomycete fungi are natural producers of this enzyme; however, the optimization of production and selection of inducers for increased productivity coupled with low costs is necessary. Lignocellulosic residues are important lignin sources and potential inducers for laccase production. Pinus taeda, a dominant source of wood‐based products, has not been investigated for this purpose yet. The aim of this study was to evaluate the production of laccase by the basidiomycete fungus Ganoderma lucidum in the presence of different inducers in submerged and solid‐state fermentation. The results of submerged fermentation in presence of 5 μM CuSO 4, 2 mM ferulic acid, 0.1 g/L P. taeda sawdust, or 0.05 g/L Kraft lignin indicated that although all the tested inducers promoted increase in laccase activity in specific periods of time, the presence of 2 mM ferulic acid resulted in the highest value of laccase activity (49 U/L). Considering the submerged fermentation, experimental design following the Plackett–Burman method showed that the concentrations of ferulic acid and P. taeda sawdust had a significant influence on the laccase activity. The highest value of 785 U/L of laccase activity on submerged fermentation was obtained on the seventh day of cultivation. Finally, solid‐state fermentation cultures in P. taeda using ferulic acid or CuSO 4 as inducers resulted in enzymatic activities of 144.62 and 149.89 U/g, respectively, confirming the potential of this approach for laccase production by G. lucidum.
... So far, a large number of fungi have been confirmed as producers of laccases, especially Basidiomycetes and Ascomycetes fungi. Four acidic laccase isoforms have been detected in Ganoderma lucidum BEOFB 431 [3], and recently, an extracellular laccase (LccH) from the newlyisolated Basidiomycetous fungus Hexagonia hirta MSF2 was also reported [4]. ...
Fusarium oxysporum trypsin (FOT) is a fungal serine protease similar to mammal trypsin. The FOT could be successfully expressed in Pichiapastoris by engineering the natural propeptide APQEIPN. In this study, we constructed two recombinant enzymes with engineered amino acid sequences added to the N-terminus of FOT and expressed in P. pastoris. The N-terminal residues had various effects on the structural and functional properties of trypsin. The FOT, and the recombinants TE (with peptide YVEF) and TS (with peptide YV) displayed the same optimum temperature (40°C) and pH (8.0). However, the combinants TE and TS showed significantly increased thermal stability at 40°C and 50°C. Moreover, the combinants TE and TS also showed enhanced tolerance of alkaline pH conditions. Compared with those of wild-type FOT, the intramolecular hydrogen bonds and the cation π-interactions of the recombinants TE and TS were significantly increased. The recombinants TE and TS also had significantly increased catalytic efficiencies (referring to the specificity constant, kcat/Km), 1.75-fold and 1.23-fold than wild-type FOT. In silico modeling analysis uncovered that the introduction of the peptides YVEF and YV resulted in shorter distances between the substrate binding pocket (D174, G198, and G208) and catalytic triad (His42, Asp102, and Ser180), which would improve the electron transfer rate and catalytic efficiency. In addition, N-terminal residues modification described here may be a useful approach for improving the catalytic efficiencies and characteristics of other target enzymes.
... pulmonarius, P. floridanus, P. ostreatus and P. sajor-caju). Silva et al. [11], Camarero et al. [19], Stajić et al. [20], Manavalan et al. [22] and Ćilerdžić et al. [23] also reported remarkable level of intraspecific diversity in laccase and MnP activities into Cerrena maxima, G. lucidum, and numerous Pleurotus species. Thus, Silva et al. [11] noted considerable diversity in activity of laccase produced during submerged wheat bran fermentation with four G. lucidum strains, i.e. activities were ranged from 0.58 U L −1 to 49,519 U L −1 . ...
... Thus, Silva et al. [11] noted considerable diversity in activity of laccase produced during submerged wheat bran fermentation with four G. lucidum strains, i.e. activities were ranged from 0.58 U L −1 to 49,519 U L −1 . After 14 days of wheat straw submerged fermentation with four G. lucidum strains studied by Ćilerdžić et al. [23] laccase activity was in the range from 153.5 U L −1 in strain BEOFB 432 to 5921.5 U L −1 in strain BEOFB 434. Likewise, activity of MnP produced during 21-old day solid-state fermentation of sugarcane bagasse by G. lucidum strain studied by Manavalan et al. [22] was significantly lower in comparison with value reported for G. lucidum BEOFB 435 (70 U g −1 substrate vs.~900 U 2 g −1 of substrate). ...
... Contrary to only two laccase isoforms produced during cultivation of G. lucidum in malt medium [28] and three isoenzymes in Korean strain grown in liquid glucose/peptone/yeast extract medium [29], Manavalan et al. [22] visualized even five isoforms in strain cultivated on solid sugarcane bagasse substrate. However, Stajić et al. [10] and Ćilerdžić et al. [23] demonstrated higher effect of genetic on isoenzyme profile. Namely, two G. lucidum strains of different origin, HAI 447 and BEOFB 431, synthetized four and three isoforms, respectively, in cultivation under the same conditions, in solid-state wheat straw/NH 4 NO 3 medium. ...
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.
... (D'Souza et al. 1996;1999;Ko et al. 2001;Silva et al. 2005;Stajić et al. 2010). In previous studies, laccases were the main ligninolytic enzymes studied because they are the dominant, and sometimes the solely produced, enzymes (D'Souza et al. 1999;Ko et al. 2001;Ćilerdžić et al. 2014b). On the contrary, Mn-oxidizing peroxidases of the same species have not been sufficiently researched, despite their high potential in applications such as biopulping, biobleaching, and bioremediation (Janusz et al. 2013). ...
Trunks and stumps of various deciduous species act as natural habitats for Ganoderma lucidum. The chemical composition of their cell wall affects the development of fungal ligninolytic enzyme system as well as its ability to degrade lignin from the plant cell wall. Additionally, numerous compounds structurally similar to lignin can be degraded by the G. lucidum enzyme system which could take important roles in various biotechnological processes. The laccases, which are the dominant enzymes synthesized by G. lucidum, have been studied more extensively than the Mn-oxidizing peroxidases. Therefore, this study aimed to create the dynamics profile of Mn-oxidizing peroxidases activities in four G. lucidum strains, classifying and determining their properties depending on the cultivation type and plant residue as a carbon source in the medium, as well as to establish whether intraspecific variety exists. The findings suggest that submerged cultivation appeared to be a more appropriate cultivation type for enzyme activities compared with solid-state cultivation, and oak sawdust was a better carbon source than wheat straw. Under the optimum conditions, on day 14, G. lucidum BEOFB 431 was characterized by the highest levels of both Mn-dependent and Mn-independent peroxidase activities (4795.5 and 5170.5 U/L, respectively). Strain, cultivation type, and carbon source were factors that affected the profiles of Mn-oxidizing peroxidases isoenzymes.
